schedule1 nmd dbase programme

December 2012 1 Schedule 1 National Microbiological Database Programme

Prelims

Schedule 1 National Microbiological Database

Programme

Prelims

December 2012

Table of Contents

Prelims ...................................................................................................................... 1

1. Administration and Participation ................................................................. 6

1.1 Species to which the National Microbiological Database

Programme Applies ............................................................................... 6

1.2 Participation........................................................................................... 7

1.3 NMD Administration .............................................................................. 7

1.4 Health of Personnel ............................................................................... 8

1.5 Training of Samplers ............................................................................. 8

1.6 NMD analyses ....................................................................................... 8

2. NMD Sampling Requirements - Red Meat and Poultry .............................. 9

2.1 Products sampled ................................................................................. 9

2.2 Number of samples per week ............................................................... 9

2.3 Summary Table ...................................................................................13

2.4 Post Chill Carcass Sampling Programme ...........................................14

2.5 Salmonella Sampling Programme ......................................................14

2.6 Poultry Campylobacter Sampling Programme ....................................17

2.7 Sampling Location within the Process ................................................17

2.8 Carcass Sample Sites .........................................................................23

2.9 Analysis Required ...............................................................................37

2.10 Summary .............................................................................................38

2.11 Weekly Sampling Plan for Red Meat – Random Rotational Sampling40

2.12 Poultry Broiler Sampling Requirements ..............................................46

3. Sampling ......................................................................................................50

3.1 Manufacture of Diluents for Sampling and Analysis ...........................50

3.2 Templates Required for Swab Sampling ............................................52

3.3 Collection of Samples .........................................................................54

3.4 Sampling Method ................................................................................55


Prelims

3.5 Transportation of Samples to the Laboratory for All Species .............61

4. Laboratory Analysis ....................................................................................68

4.1 Dilutions Required ...............................................................................68

4.2 Duplicate Plating/Analytical Precision .................................................69

4.3 Preparation of Dilutions .......................................................................70

4.4 Preparation of Swab Samples for APC and E. coli Analysis ..............70

4.5 Preparation of Red Meat Separate Carcass Site Samples

Swabbed for Salmonella Analysis .......................................................71

4.6 Preparation of Bulk Meat (Whole Tissue) Samples for APC, E. coli ...72

4.7 Aerobic Plate Count, APC30 ................................................................72

4.8 Escherichia coli PetrifilmTM ..................................................................76

4.9 Salmonella ...........................................................................................77

4.10 Poultry Carcass Campylobacter Direct Plate Enumeration Method ...81

5. Results and Formulas .................................................................................86

5.1 Result Calculation (Manual) Red Meat ...............................................86

5.2 Result Calculation (Manual) Poultry ....................................................88

5.3 Result Calculation (Automated) ..........................................................89

5.4 Limits of Detection ...............................................................................89

5.5 Reporting of Results ............................................................................91

6. Targets – Premises Analysis and Interpretation, Independent

Verification .............................................................................................................93

6.1 Premises Level Targets ......................................................................93

6.2 Bovine Species NMT: Escherichia coli Moving Window ....................93

6.3 Bobby Calf NMT: Escherichia coli Moving Window ...........................94

6.4 Ovine NMT 95th Percentile m Limits: APC ..........................................96

6.5 NMT: Documentation and Record Keeping .......................................97

6.6 Ranked List .........................................................................................98

6.7 Salmonella Performance Standards (SPS) .......................................100

6.8 Poultry Campylobacter Performance Target (CPT) ..........................102

6.9 Verification Requirements .................................................................108

7. References .................................................................................................109


Prelims

Interpretation In this Schedule, unless the context otherwise requires,-

“alert” response means an immediate review of the process by an operator to identify and

document factors that may have compromised hygienic processing, and if appropriate, take

corrective and preventative action.

batch of samples means all the samples collected in one sampling period.

biosecurity (Campylobacter) means measures undertaken to prevent the introduction and

spread of Campylobacter into grower flocks.

birds means poultry broilers.

bobby calf means bovine calf that is at least 4 days old, generally unweaned and less than

45 kilograms dressed carcass weight.

bovine means beef, with 3 classes; bull, cow and prime.

broiler chicken means a male or female chicken kept primarily for meat production, but

does not include poussins.

bulk meat means bulk packed meat, not including IW cuts.

caprine means red meat species, goat.

carcass post slaughter and dressing means a red meat carcass after completion of

slaughter and dressing, after post-mortem inspection, prior to chiller entry at cold/warm

boning premises or just prior to quartering at hot-boning premises. For the purposes of NMD

post mortem inspection is considered to include removal of navel in bobby calves.

cervine means deer, with 2 classes defined for NMD; fallow and other (wapiti, red and

hybrid).

domestic premises means premises that supply the domestic market only.

dsBPW means double strength buffered peptone water.

equine means red meat species which includes horse, mules and donkeys.

extremes of temperature means, in relation to samples during collection and transport, less

than 0oC and greater than 25oC.

EU and US listed premises means premises that are listed for export to the EU and/or US

markets or premises supplying red meat to markets specifying EU and/or US.

farm means the location at which the poultry sheds are situated

HACCP means Hazard Analysis Critical Control Points.

initial item means is the first carcass, cut or bulk carton within the run to be sampled.


Prelims

initiation of analysis means

a. In relation to APC and E.coli analyses, from the time the sample is suspended and ready

for dilution; and

b. In relation to Salmonella analysis, the commencement of incubation of the BP

pre-enrichment.

ILCP means Inter-Laboratory Comparison Programme.

insulated container means a sample collection or transport container that is insulated.

IW cut means an individually wrapped cut.

MEGAREG means slang for US Pathogen Reduction HACCP Final Rule.

MIMM 2005 means Meat Industry Microbiological Methods, Edition Four, March 2005.

moving window means addition of 5 new samples to the bottom of the window to displace 5

samples from the top of the window.

NMT means National Microbiological Target.

non EU and non US premises means premises that do not export to EU and/or US, or any

markets specifying EU and/or US.

off-site laboratory means an independent consulting laboratory to which the operator of a

premises has appointed responsibility to enter results directly to NMD.

on-site laboratory means a laboratory facility located within the physical boundaries of a

premises.

ovine means species with 2 classes; sheep and lamb.

porcine means red meat species pig.

processing plant means poultry broiler slaughter house.

processing season means bovine, caprine, cervine, ovine, ostrich and emu, poultry, 1

October to 30 September or from the commencement of a short season such as bobby

calves, which may be before 1 October.

primal cuts means a primal cut from red meat carcass.

post chill carcass means carcass after chilling.

post slaughter and dressing carcass means a carcass after post mortem inspection of

carcasses. In the case of bobby calves after inspection, after the umbilicus/navel is removed

and prior to any further trimming.

PSW means a primary sampling window re Salmonella sampling programme.

RMP means a Risk Management Programme.

ssBPW means single strength buffered peptone water.


Prelims

shed means building purpose built for sustenance of a flock of poultry broilers.

SSW means secondary sampling window in relation to the Salmonella sampling programme.

sub-contracted Laboratory means where a premises has a LAS approved laboratory but

sub-contracts an off-site laboratory for specific analyses, the on-site laboratory may be sub-

contracted or seconded to train personnel to collect samples for the off-site laboratory.

TNTC means too numerous to count.

To means the time a poultry carcass takes to move from the point of selection on the chain to

the first drop point.

verifier means “official assurance verifier” which means a person accredited under section

103 of the Animal Products Act 1999 to undertake official assurance verification and includes

animal product officers employed by MPI.

VLT means very low throughput.


Administration and Participation

1. Administration and Participation

1.1 Species to which the National Microbiological Database Programme Applies

The species to which the NMD applies are bovine, ovine, bobby calf, caprine, cervine,

ostrich, emu, poultry and porcine. The NMD operates on the same principles for each

species with slight variations. A summary of the requirements for each species is listed

below in Table 1.

Note: If your premises is EU and/or US listed, whether or not you are exporting, you must

comply with the NMD programme for all species processed that are covered by the NMD

programme. If you are currently EU listed or US listed it is irrelevant whether the product is

going to an EU or US market that week, NMD sampling/analysis must be conducted each

processing week.

Domestic and export premises; even if not processing, a report to NMD is required to confirm

that there was no processing that week.

Table 1: Domestic and export application of NMD programme.

Species Domestic, non EU and non US Listed Premises

EU Listed Premises US Listed Premises

Bovine (excluding bobby calves)

Every processing week Every processing week Every processing week

Ovine Every processing week Every processing week Every processing week

Bobby Calves Every processing week Every processing week Every processing week

Caprine Every processing week Every processing week Every processing week

Cervine (farmed)

Every processing week Every processing week Currently no requirements for US. Domestic, non EU and non US or EU apply

Ostrich and emu Every processing week Every processing week Every processing week

Porcine Every processing week Exporting to the EU is prohibited unless prior notification is obtained from the Director General

Comply with the US Pathogen Reduction HACCP Final Rule. Must notify Director General of intentions

Poultry (broiler chickens)

Every processing week Exporting to the EU is prohibited unless prior notification is obtained from the Director General

Exporting to the US is prohibited

Ducks, geese and guineas

Not required Not required Exporting to the US is prohibited

Horses, mules, and other equines

Not required Not required Comply with the US Pathogen Reduction HACCP Final Rule. Must notify Director General of intentions.



Table 1: Domestic and export application of NMD programme (continued).

Species Domestic, non EU and non US Listed Premises

EU Listed Premises US Listed Premises

Wild game Not required Not required Wild games, excluding feral goats and pigs, can be exported to US. Not required

Game estate Not required Not required No negotiated entry requirements. Not required

1.2 Participation

All premises referred to in Table 1 must participate in the NMD programme as part of the

New Zealand standard. Once a premises is participating it is mandatory to comply with all

requirements of the NMD programme.

Premises may voluntarily participate in additional NMD programmes but if submitting results

to MPI must follow all requirements of the NMD.

The NMD sampling plan applies to all premises irrespective of throughput. Allowances are

made for premises classified as “VLT”; (see section 2.3 Table 2).

1.3 NMD Administration

On commencing participation in the NMD an operator must submit completed demographic

questionnaires to the NMD Administrator (these can be found on the MPI website).

If any of the following details change, the operator must inform the NMD Administrator within

7 working days of the change occurring.

a. name of premises;

b. name of plant manager and contact details: phone, cell phone and email;

c. address of premises, physical and postal;

d. name of NMD controller and contact details: phone, cell phone and email;

e. name of LAS approved laboratory conducting NMD analysis and coordinating the NMD

sampling programme;

f. name of the data submitter and whether this is a person employed by the premises or

the LAS approved laboratory conducting analysis;

g. species required to be sampled for NMD;

h. if species processed is VLT or standard throughput;

i. any process details as outlined on demographics questionnaire;



j. location and reference number for each farm and reference number for each shed for

which the operator processes poultry.

1.4 Health of Personnel

Personnel handling product for the NMD programme must comply with the specifications and

market access requirements for health of personnel, refer to Part 3 of the Animal Products

(Specifications for Products Intended for Human Consumption) Notice 2004.

1.5 Training of Samplers

All NMD sampling must be undertaken by persons trained and deemed competent for the

species being sampled. Persons who have attended an LAS approved sampling course

related to the species they will be sampling are collectively called “Certified Trainers”.

Certified trainers are qualified to select personnel who are technically competent and are

sufficiently familiar with the process associated with the species to be sampled, to be trained

as “Associate Trainers”. Both Certified and Associated Trainers may train others to sample.

Persons trained by Associate Trainers cannot train others. Samplers trained by Certified or

Associate Trainers who are not qualified to train others are referred to as “restricted

samplers”.

1.6 NMD analyses

All sample analysis must be performed in LAS approved laboratories. Modification or

substitution of methods is not permitted unless approved by LAS, and documented in LAS.

Each NMD analysis must be performed according to the method listed in section 4 of the this

Schedule. These analyses are based on those described in Meat Industry Microbiology

Methods Edition Four (MIMM 2005) or later edition, general procedures. NMD analyses

have been modified from those published in MIMM where appropriate, to reflect the specific

requirements of the NMD programme. Methods as written in NMD procedures for

application to the NMD programme take precedence over the MIMM 2005 or later edition.

As required under LAS all quality assurance functions and quality control procedures for

methods and media as per MIMM 2005 or later edition, Chapter 2 must be followed.


NMD Sampling Requirements - Red Meat and Poultry

2. NMD Sampling Requirements - Red Meat

and Poultry

2.1 Products sampled

Non EU and non US listed premises, including domestic premises must sample the

following:

• Bobby Calves and Caprine: all product types (fresh carcasses, primal cuts and bulk

meat) processed under their RMP

• Bovine (excluding bobby calves), Ovine, Ostrich and Emu: fresh carcasses

• Poultry broiler: fresh carcasses

• Cervine: fresh carcasses and primal cuts

• Porcine: fresh carcasses.

EU and US listed premises must sample the following:

• Bovine (excluding bobby calves), Bobby Calves and Caprine: all products types

(carcasses, primal cuts and bulk meat) processed under their RMP

• Ovine, Ostrich and Emu: fresh carcasses only

• Cervine: fresh carcasses and primal cuts

2.2 Number of samples per week

The number of samples is dependant on:

• whether the product is for the domestic market, non EU and non US markets or to

be exported to EU and/or US; and

• the species processed; and

• whether the species to be sampled is classified as standard throughput or VLT; and

• the type of analyses required.

(Refer also to section 2.3 Summary, Table 2: Number of samples required per week).

2.2.1 VLT provisions for domestic only and non EU and/or non US Iisted premises

These VLT provisions

• do not apply to ovine and porcine species; and



• apply to premises that process product destined only for the local market and/or

markets other than EU and US or any markets specifying EU and US; and

• apply to bovine (excluding bobby calf) premises which process product destined only

for local markets and/or markets other than EU and US or any other markets

specifying EU and US, regardless of throughput.

Domestic only and non EU and/or non US listed red meat premises processing bovine as

above must sample:

• one fresh carcass each week of processing, starting from the first week of operation

and continuing for at least 16 processing weeks:

• where a VLT processing season is less than 16 weeks, the number of samples

collected per product per week must be increased to ensure that 16 samples are

collected within a season (for example an 8 week season would require collection of

2 samples per week for 8 weeks).

VLT premises for bobby calf and/or caprine are premises which process product destined

only for local markets and/or markets other than EU and US or any other markets

specifying EU and US and have a throughput of less than 400 bobby calf/caprine animals

in each processing week throughout the season.

Domestic only and non EU and/or non US listed premises processing bobby calf and/or

caprine qualifying as VLT premises must sample as follows:

• from the first week or part week of operation, including short seasonal operations or

where processing is intermittent, sampling of one of each product type (fresh

carcasses, primal cuts and bulk) must be conducted each processing week or part

week for at least 16 weeks or, if the number of processing weeks in the season is

less than 16 weeks, conducted each processing week or part week; or

• if at any time during the current season the weekly bobby calf and/or caprine

throughput exceeds 400 animals in a processing week, sampling of five samples of

each product type (fresh carcasses, primal cuts and bulk) each processing week is

required.

VLT premises for cervine species are defined as those that slaughter or cut/bone product

from 100 cervine animals or less per week (this is based on throughput over a whole

season). If at any time during the current season the weekly cervine throughput exceeds

100 cervine animals, cervine sampling must revert to standard throughput requirements.

Premises qualifying as VLT for cervine species must sample as follows:

• one of each of the following product types; fresh carcasses and primal cuts each

week of processing, starting at the first full week of operation within a season, and

continuing for at least 16 processing weeks:



• where a VLT processing season is less that 16 weeks, the number of samples

collected per product per week must be increased to ensure that 16 sampled are

collected within a season (for example an 8 week season would require collecting on


2.2.2 VLT for EU and US listed premises processing bobby calf, bovine, cervine and caprine

VLT premises are defined as those that slaughter or cut/bone product from 100

bovine/cervine animals or less, and/or 400 bobby calf/caprine animals or less per week (this

is based on throughput over a whole season).

If at any time during the season the weekly throughput exceeds the VLT limit, samples must

be collected at the standard rate for the remainder of the season.

Premises qualifying as VLT for a given bobby calf, bovine, caprine or cervine species or

product type of one of those species must sample as follows:

• at least one item of each product each week of processing, starting at the first full

week of operation within a season, and continuing for at least 16 processing weeks:

• where a VLT processing season is less than 16 weeks, the number of samples

collected per product per week shall be increased to ensure that 16 samples are

collected within a season (for example an 8 week season would require collection of


2.2.3 Discontinuing VLT sampling

These VLT provisions do not apply to porcine and ovine. The following are the only

circumstances when VLT sampling may be discontinued:

• Bovine/bobby calf: These species have National Microbiological Targets (NMTs),

see sections 6.2.1 NMT (bovine species): Escherichia coli moving window (m) and

6.2.4 Bobby calf NMT 80th percentile m limits: Escherichia coli re limits specified for

E. coli. So, in addition to the above 16 week requirement, NMD sampling at the

required rate may only be discontinued after E. coli moving window of 15

consecutive samples that does not elicit an “m alert”, is completed:

• VLT premises processing cervine and caprine may discontinue weekly testing of a

product type after 16 samples of that product type have been collected.

VLT premises processing bobby calf, bovine, caprine and cervine may continue to sample as

per NMD requirements throughout the season if they wish. The NMD Administrator will

continue to accept results.



2.2.4 Recommencing VLT sampling

The VLT sampling requirements above/in section 2 are to be recommenced each season

(which usually commences on 1st October) or whenever the process is changed such that

new process conditions could affect microbiological outcomes.

2.2.5 VLT for porcine premises

VLT porcine premises are defined as those that slaughter product from 10,000 pigs or fewer

per annum.

Porcine premises qualifying as VLT must sample at least one carcass each week or part

week of processing for analyses required.

2.2.6 VLT for poultry premises

VLT poultry premises are defined as those that slaughter product from one million

(1,000,000) birds or fewer per annum.

Poultry premises qualifying as VLT for carcasses must sample at least three carcasses each

week or part week of processing for analyses required.



2.3 Summary Table

Table 2: Number of samples required per week

Species Product type

Technique Domestic and non EU and non US listed

EU and US listed standard throughput

EU and US listed VLT

Bovine Carcass Multiple Swab Technique

1 per week 5 per week 1 per week

Primal Cuts Multiple Swab Technique

Not required 5 per week 1 per week

Bulk Meat Product

Whole Tissue Composite Sampling

Not required 5 per week 1 per week

Post Chill Carcass

Multiple Swab Technique

Not required 5 per week for 6 weeks

1 per week for 6 weeks

Ovine Carcass Multiple Swab Technique

5 per week 5 per week Not applicable

Porcine Carcass Multiple Swab Technique

5 per week; VLT 1 per week

Bobby Calf and Caprine

Carcass Multiple Swab Technique

5 per week VLT: 1 per week for 16 weeks

5 per week 1 per week




Bulk Meat product

Whole Tissue Composite Sampling



Post Chill Carcass

Multiple Swab Technique

Not required 5 per week for 6 weeks

1 per week for 6 weeks

Cervine Carcass Multiple Swab Technique

3 per week VLT: 1 per week



2 per week VLT: 1 per week


Ostrich/Emu Carcass Multiple Swab Technique

1 per week 2 per week Not applicable



Table 2 continued: Number of samples required per week

Species Product type

Technique Domestic and non EU and non US listed

EU and US listed standard throughput

EU and US listed VLT

Poultry Carcass Whole Carcass Rinse Method

3 per day VLT: 3 per week

Not applicable Not applicable

2.4 Post Chill Carcass Sampling Programme

Post chill carcass sampling is only required for bovine, bobby calf, and caprine species for

EU and US listed premises.

• Post chill carcass sampling must be commenced on starting the NMD programme

and then be undertaken annually at the start of the season.

• Post chill carcass sampling is required when the chiller operating conditions changes

significantly (calibration). The season is classified as beginning at 1st October or the

commencement of a short season such as bobby calves, which may be before 1st

October.

• Post chill carcass sampling provides a microbiological profile of chillers at normal

operating capacity for the particular species being tested.

• Post chill carcass sampling is required until 30 carcasses (at 5 carcasses per week

for 6 consecutive processing weeks) have been monitored. If sampling takes more

than 6 weeks it must be justified (for example, chiller capacity being lower than the

normal for the species, multiple species and concurrent Salmonella sampling

obligations exceeding laboratory capability for the sample collection and/or analysis).

• Post chill carcass sampling must be rotated on a weekly basis around the chillers, to

a maximum of 6 chillers (5 samples per chiller) during each annual period.

• Premises that qualify as VLT for bovine, bobby calf and/or caprine species are

required to have one post chill carcass sample per week for 6 weeks for each

season.

2.5 Salmonella Sampling Programme

There is a performance standard for Salmonella sampling for all products (excluding chilled

carcasses) from the following species:

• bovine



• bobby calf

• caprine

• cervine

• ostrich/emu

• porcine (see section 2.5.2)

• poultry

(Note: Salmonella testing is not required on ovine species).

Salmonella sampling and performance standards are split into three groups:

Table 3: Salmonella groups

Group 1 – Salmonella Group 2 – Salmonella Group 3 – Salmonella

Bovine Bobby calf Caprine Cervine

Ostrich/Emu Porcine

Poultry

2.5.1 Group 1 (bovine, bobby calf, caprine, cervine) – seasonal Salmonella

Seasonal Salmonella sampling is required for each product type processed. The processing

season for bovine, caprine, and cervine is 1st October – 30th September. For bobby calves

the processing season is taken from the commencement of bobby calf processing during the

calendar year 1st January– 31st December.

An initial “primary sampling window” (PSW) of 16 consecutive weeks, within a processing

season must be conducted. Where a premises processing season is shorter than 16 weeks

the PSW carries over to the subsequent processing season.

A premises that has not detected Salmonella in a given product type of a given species in

the preceding PSW must test product for the initial six consecutive processing weeks of the

next season. (See Figure 1). This reduced sampling frequency is defined as a “secondary

sampling window” (SSW).

Where a premises doesn’t complete either a PSW or SSW in their processing season, this

then carries over to the new processing season. The carried over PSW or SSW does not

displace the new processing season requirement. The new seasonal Salmonella sampling

must commence on the completion of the carried over one. Examples:

• A bobby calf processing season, usually starting between March and August, might

commence with PSW15; indicating a carry over from the previous season. In the first

week of the new season the PSW of the previous season would be complete at

PSW16 (providing no Salmonella detection occurs). The processor is then required

to start a secondary sampling window (SSW1) in the following week.



• Where a SSW reads SSW2, SSW3, SSW4, SSW5 or SSW6 at the beginning of a

new processing season this indicates an incomplete SSW from the previous season.

The full SSW of the previous season must be completed and the new season SSW1

must be started in the next processing week (providing no Salmonella detection

occurs).

A premises that detects Salmonella in a given product type of a given species during a PSW

or SSW must immediately begin another 16 week PSW for that species/product type. A

“sampling window” terminates on detection of Salmonella and week one of a new PSW

commences for that particular species/product type in the following processing week.

Figure 1: Salmonella sampling windows succession

Salmonella: detected (+) not detected (-)

• An operator who demonstrates to a verifier that Salmonella was not detected in

either a PSW or a SSW may, with approval of MPI VA, cease testing of that product

for the remainder of the processing season. Unless the PSW or SSW was carried

over from last season, where a SSW needs to be commenced for the current

processing season following completion of the carried over seasonal window.

Closures (seasonal or maintenance): An incomplete PSW or SSW carries over to the next

processing week following completion of maintenance.

Changes to process: A premises that is substantially modified in such a manner that it could

affect expected microbiological outcomes of the process must immediately begin a 16 week

PSW when processing recommences. Substantial modification includes:

• new premises under the same licence/registration number at the same or different

location; and

• major changes in processing methods (for example, change from traditional to

inverted dressing, installation of robotics, etc.) where changes in microbiological

performance are possible or probable.



2.5.2 Group 2 (Ostrich/Emu and Porcine) - Salmonella

An operator must conduct ostrich/emu and porcine Salmonella sampling each processing

week.

Porcine Salmonella sampling is a 27 month programme which commenced week beginning

5 October 2009 and will cease on the week beginning 2 January 2012.

2.5.3 Group 3 (Poultry) – Salmonella

Poultry Salmonella sampling must be conducted by operators of;

• Standard throughput premises; each processing day

• Very Low Throughput premises; on one processing day of each processing week.

2.6 Poultry Campylobacter Sampling Programme

The Campylobacter sampling programme for poultry carcasses must be conducted by

operators of;

• Standard throughput premises; each processing day

• Very Low Throughput premises; on one processing day of each processing week.

2.7 Sampling Location within the Process

2.7.1 Red meat product selection

Refer to section 2.3 for products (carcasses post slaughter and dressing, post chill

carcasses, primal cuts and bulk meat) required to be sampled for each particular red meat

species NMD programme.

2.7.1.1 Carcasses post slaughter and dressing

NMD sampling for post slaughter and dressing aims is conducted to gain a representative

picture of microbiological contamination during the slaughter and dressing process.

Procedures that result in a significant transfer of microbes to the carcass are completed

before the post mortem inspection station. “Post slaughter and dressing” is defined, for the

purposes of NMD, as after post mortem inspection and after final dressing procedures

specified below.

The NMD sampling for post-slaughter and dressing is as follows:

• NMD sampling must commence as soon as is practical, but no later than 30 minutes

after post mortem inspection of carcasses.

• Carcasses detained by AsureQuality must not be sampled.



• Carcasses must be sampled BEFORE any procedure that may impact on the NMD

carcass sampling sites. Processes following the post mortem inspection station may

redistribute contamination over a carcass, add some contamination by handling from

workers, or remove contamination due to particular process interventions. Such

procedures include carcass stacking (contact), gross hot trimming, carcass washing,

rail stimulation and worker contact during marshalling.

• In the case of bobby calves, samples are to be collected immediately after

inspection, after the navel is removed and prior to any further trimming.

• Positioning of carcass for sampling:

(i) Ovine, porcine, caprine and bobby calf carcasses can be moved to a dead rail for

sampling, or another suitable position that does not mask the microbiological

consequences of slaughter and dressing.

(ii) Bovine and cervine carcass sampling must be conducted on a moving rail (as

the carcass cannot be manually moved from the rail). This requires a greater level

of training to ensure sample collection is consistently from the correct sites, the

correct carcass side and with the correct sampling technique.

• The specified carcass sampling sites require wet/dry swab sampling. The sampling

sites will vary in location according to the species. The sites have been selected to

represent the points where microbes are most likely to be transferred to the carcass

during dressing procedures.

• Record the time of sampling, run, shift, species, class and process type; inverted,

traditional, gas de-pelted.

• Rotation of chains on a weekly basis in premises with multiple slaughter chains is

required.

2.7.1.2 Post chill carcasses

Post chill carcasses of cold and warm boning premises require wet/dry swab sampling. Refer

section 2.8.8. Post chill carcass sampling is not required for hot boning premises.

Cold and warm boning premises must sample the post chill carcass:

• no later than 24 hours after the onset of chilling; or

• when the chilling cycle is completed in less than 24 hours, at the completion of the

chilling cycle; and in either case

• in the chilling area itself prior to wrapping, transportation, freezing, boning or loadout.

Records must be taken of the:

• location of the chilled carcass at time of sampling; identifying the cooling floor,

chiller, side chiller, quarter chiller or other,



• boning process (cold or warm), and

• number of hours chilled (timed from the onset of chilling).

2.7.1.3 Primal cuts and bulk meat product

Primal cuts and bulk meat product the microflora comprises organisms from two sources;

• those which remain attached to the carcass after the initial transfer during slaughter

and dressing; and

• those acquired by contact of the cut surfaces with other surfaces or materials (for

example condensate, aerosols) in the processing environment (cross

contamination).

Primal cuts and bulk meat manufactured at a premise, but from carcasses supplied by

another premises, shall be regarded as product of the boning premises.

Boning premises must record the premises of a carcass origin and enter that information in

the boning premises NMD report under the product description column of the NMD

database.

Only fresh product may be sampled for NMD (secondary processes and product that has

either been chilled in the carton, frozen or thawed from frozen is not required to be sampled

for NMD).

Primal cuts (cold, warm or hot boned, bone-in or bone-out).

Wet/dry swab samples must be taken from an outside (fell) surface of the cut, not an

exposed, freshly cut internal surface. The sampling site selected on the primal cut should

correspond to the equivalent carcass sampling site for the species concerned. Primal cut

samples are to be taken immediately prior to vacuum packaging, wrapping or bulk meat

packing into cartons.

• Bovine, bobby calf, caprine: hindquarter cuts or hindleg cuts

(refer Table 4).

• Venison: hindquarter cut (rump/topside) and forequarter cut (shank/shoulder). Removal of the silver skin (de-sinewing) of cuts is more commonly carried out in

venison further processing. Samplers must record whether the venison cuts

sampled are de-sinewed or intact.



Table 4: Primal cut descriptors

Species Primal Cut Descriptor

Bobby Calf Hindleg Outside hindleg

Bovine ATS Bottom Round Eye of Round Flat Inside Outside Less Eye of Round Outside Round Outside Smalls SAT Silverside Topside

Caprine Hindleg

Cervine Sinew Intact – Hindquarter, rump Sinew Intact – Hindquarter, topside Sinew Intact – Hindquarter, outside Sinew Intact – Forequarter, shank Sinew Intact – Forequarter, shoulder Sinew Intact – Forequarter, shoulder shank on Sinew Intact – Forequarter, foreleg Sinew Intact – Forequarter, knuckle Desinewed – Hindquarter, rump Desinewed – Hindquarter, topside Desinewed – Hindquarter, outside Desinewed – Forequarter, shank Desinewed – Forequarter, shoulder Desinewed – Forequarter, shoulder shank on Desinewed – Forequarter, foreleg Desinewed – Forequarter, knuckle

Ovine No primal cut NMD required

Porcine No primal cut NMD required

Poultry No primal cut NMD required

Ratite No primal cut NMD required



Figure 2: Beef carcass, location of cuts

The bovine primal cut descriptors are highlighted in yellow:



Bulk meat and trim.

The micro organisms on the surface of bulk meat or trimmings will be distributed throughout

the carton. Bulk meat will eventually be ground into a homogenous comminuted product and

the original micro organisms will become evenly distributed throughout. Whole tissue

samples from the original bulk meat carton are expected to give a good representation of the

microflora. Boneless bulk meat product of 85-95 VL, not including individually wrapped (IW)

product, is to be selected for sampling. Where a premises does not produce boneless bulk

packed meat, cartons of trim in the 60-85VL range will be acceptable. Whole tissue samples

are to be taken from bulk meat product immediately prior to closing and strapping the full

carton before equilibration or freezing.

Table 5: Bulk meat pack descriptors

Species Bulk Product Descriptor

Bobby Calf Trim Trunk Veal Inside

Bovine Fores and Hinds Trim Shanks

Caprine Trim Trunks

Cervine No bulk NMD required

Ovine No bulk NMD required

Poultry No bulk NMD required

Porcine No bulk NMD required

Ratite No bulk NMD required

The time of sampling must be recorded (those conducting only further processing will enter

this time on the NMD data entry sheet), species, class, description of boning process (cold,

warm, hot) primal cut type, VL/CL grade percentage or estimate of and product description of

bulk.

Premises with multiple boning rooms must rotate sampling between boning rooms on a

weekly basis.



2.7.2 Poultry carcasses selection

• Whole poultry carcasses are to be selected in order for the whole carcass rinse

method to be applied

• For premises where carcasses are mixed in bins at manual grading, immediately

prior to grading (for example off the drain-table, after draining)

• For premises where carcasses are re-hung on the chain after immersion chilling, or

are air chilled, the carcass must be selected immediately prior to the first drop point,

or similar, at which the carcasses enter secondary processing or are bagged for

retail.

Should the position at which the carcass is released from the chain not be readily accessible,

the carcasses for sampling may be selected:

• At the last readily accessible position on the chain or from the drain-rack. In order to

drain excess water, the carcasses must be hung prior to being sampled or “bagged

for shipment to the laboratory” for the same time (T0) as the carcass would take to

move from the point of selection to the first drop point.

Note: This time (T0) will vary at each premises, but is representative of “normal” processing

at that premises.

Carcasses for sampling must be handled aseptically for removal from the drain table or main

chain and for transfer into the sampling bag.

Figure 3: Selection of poultry carcass

2.8 Carcass Sample Sites

Individual samples are to be collected (ICMSF, 1986; MIRINZ, 1971) from the species

specific sites described below and illustrated in Figure 4 (bovine), Figure 5 (ovine and

caprine), Figure 6 (bobby calf), Figure 7 (cervine – traditional), Figure 8 (cervine – inverted),

Grading

T0

Immersion chill

Drain table or

re-hanging

Air chill

Selection of carcass

Pre-wash

Drip-line

Bagging or

further processing



Figure 9 (porcine) and Figure 10 (ostrich/emu). Figure 11 illustrates sampling from the

opposite sides of the carcass for APC/E. coli and Salmonella samples.



2.8.1 Bovine

Three sites need to be swabbed on bovine. The three sites are as follows:

Rump: (situated on the hindleg, refer Figure 4)1: The 100cm2 site is centred on the fascia

overlying the semitendinosis muscle. The muscle is cut, packed and sold as the ‘eye round’.

The centre of the sampling site is halfway along the muscle on its superficial lateral margin.

Lateral to semimembranosis is the gluteobiceps muscle. This muscle is cut, packed, and

sold as the ‘outside flat’. The 100cm2 template may overlap both muscles.

Flank: The 100cm2 site is centred on the fascia overlying the cutaneous trunci muscle. The

centre of the site is 10cm lateral to the umbilicus on a line drawn along the ninth rib from the

spine and continued on to cut edge of the abdominal wall.

Brisket: The 100cm2 site is centred on the fascia overlying the cranial ventral part of the

cutaneous trunci muscle. The centre of the sampling area is 10cm off the central midline.

The margins of the sampling site may overlap onto the fascia overlying the caudal margin of

the pectoral profundis muscle. The lower edge of the sampling site is an imaginary line

drawn transversely along the thoracic wall at the level of the point of the elbow.

1The naming of the bovine hindleg sample site as rump is required under our equivalence agreement with the US.



Figure 4: Sampling sites for bovine carcass, 100cm2

Rump site situated on the

hindleg

Flank site

Brisket site

Photographs courtesy of David Walkinshaw, Taylor Preston Limited



2.8.2 Ovine and Caprine

2.8.2.1 Ovine: traditional and inverted

A single ovine carcass site needs to be swabbed. The site is as follows:

Forequarter opening Y-cut: The Y-cut site is defined as the anterior aspect of the humero-

radial joint close to the brachial vein, which is clearly visible at the site but a little erratic in its

course.

2.8.2.2 Caprine

Three carcass sites need to be swabbed for caprine. The three sites are as follows:

Outside hindleg: The outside hindleg site is defined as an area one third up on a vertical

line originating at the midpoint of a line between the ischial crest and stifle, and extending to

a line horizontal to the cut end of the hock.

Flap: The flap site is defined as an area ~50mm from the flap edge, midway between the

flap joint and the xiphoid cartilage.

Forequarter opening Y-cut: The Y-site is defined as the anterior aspect of the humero-

radial joint close to the brachial vein, which is clearly visible at the site but a little erratic in its

course.



Figure 5: Ovine sampling site and caprine sampling sites, 5cm2 sampling area.

Ovine carcasses (both traditional and inverted dressing) require sampling of the opening Y

cut site only.

Caprine carcasses require sampling of all three sites shown below.

Photographs courtesy of Sandy Moorhead (AgResearch) and Monique Biss (MAF VA)

Outside leg

Opening Y-cut

Flap



2.8.3 Bobby calf

Three sites need to be swabbed on bobby calves. The three sites are: Fore rump: An area adjacent to the rectal canal, no greater than 50mm horizontally from the

edge of the rectal canal, vertically with midpoint on a horizontal line dissecting the forward

(lower) edge of the rectal canal.

Flank: The site is defined as an area ~50mm from the flank edge, midway between the flank

joint and the xiphoid cartilage.

Foreleg: An area on the outside surface

of the lateral head of the triceps brachii.

The side (left or right of the animal) of the carcass APC/E. coli swabs are collected from

must be recorded for bobby calf samples.



Figure 6: Sampling sites for bobby calf carcasses, 25 cm2 sampling area



2.8.4 Cervine

2.8.4.1 Cervine – traditional

Three sites need to be swabbed on traditional cervine. The three sites are as follows:

Posterior hindleg: The hindleg sample site is centred longitudinally and laterally on an

imaginary line drawn between the posterior aspect of the aitch bone and the archilles

tendon.

Sternum: The sternum site is immediately adjacent to the intersect of the abdominal cavity

opening and brisket, but outside the area of “standard brisket trim” (by <10mm at its closest

edge). Note: Tissue exposed by the standard trim must not be sampled.

Inside foreleg: The inside foreleg site is defined as the anterior aspect of the humero-radial

joint close to the brachial vein, which is clearly visible at the site but a little erratic in its

course.

2.8.4.2 Cervine – inverted

Three sites need to be swabbed on inverted cervine. The three sites are as follows:

Inside hindleg: The sample site is on the medial side and proximal end of the hindleg

immediately adjacent to the pelvic symphysis (midline).

Brisket: The brisket sample site is centred longitudinally on the brisket, but immediately

outside the area of “standard brisket trim” (by <10mm at its closest edge). Note: Tissue

exposed by the standard trim must not be sampled.

Inside foreleg: The inside foreleg site is defined as the anterior aspect of the humero-radial

joint close to the brachial vein, which is clearly visible at the site but a little erratic in its

course.



Figure 7: Sampling sites for cervine carcasses (traditional), 25cm2 sampling area



Figure 8: Sampling sites for cervine carcasses (inverted), 25cm2 sampling area



2.8.5 Porcine

Three sites need to be swabbed on porcine carcasses. The three sites are as follows:

Outside hindleg: Midway between the stifle and hip joint.

Lower flap: 50mm from the abdominal incision midway between the two lower nipples.

Outside shoulder: Above the forward top end of the shoulder blade.

Figure 9: Sampling sites for porcine carcasses, 25cm2 sampling area Photos courtesy Land Meats

Porcine NMD site 1: Outside Hindleg Porcine NMD site 2: lower flap between the

lower two nipples

Porcine NMD site 3: outside shoulder site

Whole carcass view of NMD sites



2.8.6 Ostrich and emu

LEG HUNG

A single ostrich and emu carcass site needs to be swabbed. The site is as follows:

Inside leg: Located longitudinally on the opening cut line and laterally where the edge of the

remaining flap contacts the leg. It is recommended that this site be alternated between left

right sides.

Figure 10: Sampling sites for leg hung ostrich and emu carcasses, 25cm2 sampling area

Photo courtesy of Venison Packers Fielding Limited

WING HUNG

No site identified at present as no NZ premises are using wing hung slaughter and dressing

practice.

Inside hindleg



2.8.7 Salmonella sampling

Where Salmonella sampling is being undertaken the same sites on the opposite side of the

carcass from APC/E. coli samples are to be selected and sampled.

In the case of adult cattle the carcass is physically split. Substitute a side from another

carcass for sampling if the opposite side to that sampled for APC/E. coli is not available due

to condemnation.

Figure 11: Illustrates sampling from opposite sides of the carcass as required for APC/E.

coli and Salmonella samples.

2.8.8 Post chill carcass sampling

• Where post chill sampling is being undertaken, select the same sites on the opposite

side of the carcass from APC/E. coli sampling. For example, the carcasses post

slaughter and dressing can be tagged and re-sampled on the opposite side of the

carcass after chilling.

• Where Salmonella sampling of carcasses post slaughter and dressing coincides with

post chill sampling choose another carcass for post chill sampling that has



undergone the same chilling conditions adjacent to the carcass originally selected

for carcass post slaughter and dressing sampling.

• Where matching is impractical (for example, pre-chill carcasses spread over several

chillers) post-chill carcasses may be selected randomly from a single chiller, rotating

weekly through all available chillers.

• NEVER sample a site that has been previously swabbed because the microbial load

will have been removed/modified during the first swab sampling procedure.

• Note: post-chill carcass sampling can be conducted on the same day as fresh

carcasses are sampled.

2.8.9 Poultry carcass sampling

The whole carcass is sampled for poultry.

2.9 Analysis Required

The following analyses are required:

Table 6: NMD analyses

Product Analyses required

Bovine, bobby calf, caprine, cervine, ostrich and emu, and porcine carcasses post slaughter and dressing

Aerobic plate count E. coli Salmonella

Ovine carcasses post slaughter and dressing

Aerobic plate count

Post chill carcasses Aerobic plate count E. coli

Primal cuts Aerobic plate count E. coli Salmonella

Bulk meat Aerobic plate count E. coli Salmonella

Poultry whole carcass Salmonella Campylobacter



2.10 Summary

Table 7: Summary of NMD sampling requirements

Product to be sampled

Frequency Sites to be tested Analyses

Carcasses Post Slaughter and Dressing Bovine, bobby calf, caprine, cervine and porcine.

Weekly • 3 sites, one side of

carcass, alternating

between the leading

and trailing sides

• 3 sites, other side of

carcass

• APC/E. coli

• Salmonella (each

seasonal sampling

window, except porcine-

refer 2.5.2)

Carcasses post Slaughter and Dressing Ovine

Weekly • 1 site, one side of


between the leading

and trailing sides.

• APC only.

• Salmonella is not

required

Carcasses Post Slaughter and Dressing Ostrich and emu

Weekly • 1 site, one side of


between the leading

and trailing sides

• 1 site, other side of

carcass

• APC/ E. coli

• Salmonella

Carcasses Post Chill Only required for bovine, bobby calf and caprine species Not required for hot boning premises

Weekly, for a 6 consecutive week annual sampling window, at the start of the season

• 3 sites, one side of


between the leading

and trailing sides

• APC/ E. coli

• Salmonella is not

required



Table 7: Summary of NMD sampling requirements (continued)

Product to be sampled

Frequency Sites to be tested Analyses

Primal Cuts

Bovine, bobby calf,

caprine, and cervine

Weekly • 1 site on required

number of separate

cuts (same site as

on carcass)

• APC/ E. coli

• Diluent remaining after

APC/E.coli test is

composited for

Salmonella


seasonal sampling

window)

Bulk Meat

Bovine, bobby calf,

and caprine.

Weekly • 10g from 5 locations

within each carton

composited, from

which a 25g

analysis sample is

taken

• APC/ E. coli

• Diluent remaining after

APC/E. coli tests is

composited for

Salmonella


seasonal sampling

window)

Poultry Whole Carcass Rinse

Every

processing

day

One

processing

day a week for

VLT

• Whole carcass is

rinsed with ssBPW

• Campylobacter for each

carcass.

• Additionally - Salmonella

for one carcass.



2.11 Weekly Sampling Plan for Red Meat – Random Rotational Sampling

To ensure that a complete evaluation of the process is made, samples must be selected

randomly such that every animal has an equal chance of selection.

The sampling plan for red meat must include a randomly selected time each week to sample

all products types (carcasses, cuts and bulk as per premises scope and as required for a

particular species) for each species.

When there are multiple red meat species processed at a premises, a single selected time

period to sample all species can be used (same day, shift and run for all species) if you have

enough samplers and laboratory personnel to process the samples. Alternatively, a different

day of the week or time in the day may be randomly selected for the other species from the

remaining days or part days.

Red meat VLT premises must base their sampling plan on the farmed animal/cervine plan as

above and randomly select a day, shift and run for each week or part week of processing.

2.11.1 Sample selection

First priority: to sample the required carcasses, cuts and bulk for each species each week.

MPI can provide a random sample selection macro available to assist (contact the NMD

Administrator).

The order of selection; day, class, shift, run, time can be varied. The selection can be made

in any order as long as all the parameters required by NMD random rotational sampling

programme are met.

2.11.1.1 Day: random/rotational selection of day of week

Ascertain the days in the week processing for each particular species is taking place.

All processing or part processing days (including statutory holidays and weekends) must be

eligible for selection.

Randomly select an order of days from days available by randomly selecting one and

rotation through the remainder (in any order).

Random sampling only, without rotation through all available days is not permitted. All

available processing days must be sampled from.

For example if there are 5 days in the week available for processing over 5 weeks all days

will be selected for sampling.



Table 8: Random rotational of weekdays

Week number

1 2 3 4 5

Day Wednesday Monday Friday Tuesday Thursday

Ovine species: Random/rotational sampling of sampling days is required for ovine species.

However, where reasonable practical constraints exist, a specified day may be excluded

from sampling without the scientific proof required under section 2.11.3 and on agreement

with MPI VA. Where not all days in a processing week are available for sampling,

random/rotational sampling must occur through the remaining days.

2.11.1.2 Class: random/rotational sampling of class

Where there is more than one class random rotational sampling of classes is required. This

is not dependent on throughput.

The proportion of livestock classes processed at premises is irrelevant under the NMD

programme and must not be used to deliberately bias the selection. Throughput only affects

the number of samples collected if the premises is classified as Very Low Throughput.

The class selected should be used for all product types (carcasses, cut, bulk) of that species

that week.

Ovine – 1 class, lamb, is required to be sampled except on processing weeks where only

sheep is being processed, in which case sheep must be sampled.

Ostrich and emu – 2 classes if both ostrich and emu are being processed.

Cervine – 2 classes; fallow and ‘other’ - wapiti, red, hybrid and sika grouped as one class.

Similar to tossing a coin for heads or tails randomly select the first class of fallow or ‘other’ to

be sampled every 2 weeks.

Bovine – 3 classes; bull, cow, prime. Randomly select one of the 3 classes for week 1, then

randomly selected week 2 from the remaining two classes. The class remaining will be the

one for week 3.

Porcine – 4 classes: suckers, porkers, baconers, choppers. Randomly select one of the 4

classes for week 1, randomly select week 2 from the remaining three classes, and week 3

from the remaining two classes and the class remaining will be the one for week 4. Variation

to this may be required to ensure choppers are selected for one week of each month if

choppers are being processed that month.



Table 9: Random rotation of classes

Week number

1 2 3 4 5 6

Ostrich/Emu Ostrich Emu ostrich Emu Emu ostrich

Cervine Fallow ‘other’ ‘other’ Fallow ‘other’ Fallow

Bovine Cow Prime Bull Bull Cow Prime

Porcine Suckers Porkers Choppers Baconers Choppers Suckers

The italicised class indicates where the rotation starts for each species.

2.11.1.3 Shift: random/rotational sampling of shift

Samples must be collected from all processing shifts. When multiple shifts are running for a

species or a product type of a particular species rotate through all shifts.

Ovine species: Rotational sampling through all processing shifts is required for ovine

species. However, where reasonable practical constraints exist, a specified shift may be

excluded from sampling without the scientific proof required under section 2.11.3 and on

agreement with MPI VA. Sampling must be carried out on the other shift on the selected day

of that week or from the same shift on another randomly selected day of that week.

Bovine, bobby calf, caprine cervine, ostrich, emu and porcine where there are two shifts:

alternate between day and night shift such that over two weeks both shifts are sampled.

Table 10: Alternating shifts

Week number

1 2 3 4 5 6

Shift Day Night Day Night Night Day

2.11.1.4 Random selection of run

It is important that the choice of run from which samples are to be collected is randomly

selected. Every carcass, primal cut or bulk carton has to have an equal chance of being

selected during the chosen run.

2.11.1.5 Random selection of time

• Select the initial item by randomly selecting the time in the run. No element of

routine timetable practises is acceptable.

• Only the initial item needs to be randomly selected.

• Subsequent, but not consecutive, items can be selected in sequence after the

previous item has been sampled and returned to the rail or the conveyor. Selection

of items will be at time intervals equivalent to the time required to sample. Selecting

5 primal cuts at once from the conveyor belt and placing them on a table to swab at



the same time is NOT ACCEPTABLE, except where there is insufficient product type

remaining in the week to make up the samples.

• Another acceptable method is selecting the first carcass randomly from carcass

ticket numbers in that run.

• The time may be chosen by the 24 hour clock system or minutes into the run with a

sampling error of 10 minutes either side. For example, 15:33 ± 10 minutes (the

range is between 15:23 – 15:43).

• If time segments are used the time segments are to be no greater than 20 minutes.

No time either side of the time segments is permitted. It is NOT 20 minutes plus or

minus 10 minutes at each end, allowing 40 minutes altogether.

• If the carcass, cut or bulk sample available at that time is unsuitable (e.g. carcass

goes on to detain rail), then select the next available appropriate carcass, cut or

bulk.

• The time recorded at time of sampling (am/pm, 24 hour clock, carcass ticket number

or minutes into a run) must be in a format that can be interpreted/translated to 24

hour clock units in the final results submitted to NMD.

Table 11: Random/rotational selection of day, random selection of run and time

Week number

1 2 3 4 5 6

Day Wednesday Thursday Friday Monday Tuesday Tuesday

Run 4 3 1 4 2 2

Time (using ticket number)

4045 3125 0215 3701 2245 1523

Time (24 hour clock)

15:33 13:01 08:22 14:45 11:55 10:37

Time (minutes into run)

15 78 61 23 42 36

2.11.2 Departure from the originally selected sampling period

Any changes from the originally selected sampling period must be justified.

If cuts and bulk cannot be sampled in the same run or day selected for carcasses, randomly

reselect for cuts and bulk.

If the production schedule cause days available to sample for a species to alter from what

was expected, randomly reselect from remaining times available for that week.

The most important feature is to complete the required sampling and analyses for each

species each week.



Regarding random run and time selection, it is vital there is adequate communication

between the samplers and the process personnel on the day. The slaughter floor should

notify the sampler if they suspect processing may finish early or that there may be other

interruptions that may have an impact on sampling. The sampler must then randomly

reselect from remaining time available that day or reselect the day and time from remaining

days available that week.

Where the random time selection is near the end of the run sampling may commence 10

minutes prior to the randomly selected time. Any remaining samples must be collected at

the beginning of the next run unless not available (refer below for further options to complete

sampling).

The key task is to collect the carcass, cut and bulk samples required for each species each

week. Where there is insufficient product in the selected sampling period the following

remedial actions can be taken to complete the sampling requirements.

In order of priority, collect samples from:

• incomplete cartons;

• across multiple runs within a shift;

• multiple product types: bulk 95CL, 60CL, 80CL within a class;

• different classes;

• across multiple shifts;

• across multiple days.

Any of the changes described above (for example, alterations to shift, day, class run and or

time) must be documented along with reasons for the departure from the original randomly

selected parameter.

2.11.3 Modifications to the random sampling plan

Modifications to the random sampling plan at individual premises for all species, except

ovine species, may be approved by MPI.

Examples of modifications include:

1. Use of dayshift sampling only instead of day and night shift sampling.

2. Choosing only some particular days of the week for sampling instead of all available

processing days.

Sound statistical analysis will be required to show that microbiological outcomes will not be

compromised by any modifications. It is recommended that where there are shifts that data

is collected for shift comparisons and that verification of no significant differences between

shifts is completed before progressing to statistical comparison between days of the week.



It must also be shown that by electing to sample from only one shift and/or particular day of

the week that this is not bias the coverage of all possible processing scenarios.

The statistical analysis and conclusions must be fully documented by the premises and

approved by MPI VA. MPI VA may choose to confirm the statistical conclusions drawn with

MPI before granting approval. Once approved the premises must notify the NMD

Administrator of the modified sampling plan and provide a copy of the approved justification.

Ovine species: Modification of random sampling plans is prohibited except on occasion

where practical constraints prevent the required sampling. Practical constraints such as

courier schedules must be identified and documented by the premises and agreed with MPI

VA.

Premises are required to document and maintain records of the departure from sampling a

particular shift (day or night shift) and of the particular days of the week nominated for ovine

sampling. These records must be available for MPI VA to view at any time. The NMD

Administrator must be notified of the modified ovine sampling plan.

2.11.4 Practical constraints and technical failures

Due to the premises production schedules, breakdowns, sample transport problems, or

other, meeting the random sampling plan and number of weekly samples required may not

always be possible.

Should a technical problem occur – randomly reselect from the remaining available times

that week (This applies to red meat standard throughput and VLT premises).

If no other times are available in that sampling week, two sets of samples are not required in

the following week.

When expected sample results are non-existent due to technical failure or production

schedule constraint, the factors involved must be recorded by the premises and supplied to

the NMD Administrator promptly. The NMD Administrator will note this in the sample ledger

for future reference.

If the practical constraint is routine, for example no sampling on Friday afternoons because

of courier problems, a detailed justification must be documented. This must show that all

alternatives have been researched and are impossible to implement. This must be approved

by MPI VA who may choose to discuss this with MPI before granting approval. Once

approved the premises must notify the NMD Administrator of the approved derogation and

supply a copy of the supporting documents.



2.12 Poultry Broiler Sampling Requirements

2.12.1 Number of samples

A set number of poultry broiler samples must be taken:

Standard throughput: select 3 carcasses each processing day:

• all 3 carcasses must be analysed for Campylobacter,

• randomly select 1 of the 3 carcasses to be analysed for Salmonella in addition to

Campylobacter,

VLT; 3 carcasses must be selected on one* processing day per week;

• all 3 carcasses must be analysed for Campylobacter,

• randomly select one of the 3 carcasses to be analysed for Salmonella.

The operator must ensure 3 carcasses are collected each processing day for standard

throughput and 3 carcasses are collected on one processing day each processing week for

VLT. The operator must communicate production schedules and any variation to production

schedules to the laboratory at the earliest opportunity.

* Operators of a VLT premises may apply to the NMD Administrator to modify the collection

of 3 carcasses in one week to be conducted over more than one processing day (where

there is more than one processing day available in a particular processing week). The

alternative sample selection programme must be fully documented by the operator and

approved by MPI VA prior to application.

2.12.2 Selection of sampling days

Standard throughput: Sampling must be conducted every processing day.

VLT: 3 samples from one single randomly selected day of the processing days available that

week.

The table below gives two examples of how this requirement can be met.



Table 12: VLT sampling days

Monday Tuesday Wednesday Thursday Friday Saturday Sunday Total Samples

Randomly select 1 sampling day from processing days available

5 processing days, Monday – Friday:

Exclude Exclude 3 Exclude Exclude Np Np 3

1 processing day

3 Np Np Np Np Np Np 3

2.12.3 Shift; random/rotational sampling of shift

If shifts are undertaken then take samples from alternate shifts.

Table 13: Standard throughput shifts

Processing day

1 2 3 4 5 6

Shift day Night day night day Night

Table 14: VLT Shifts

Week number

1 2 3 4 5 6

Shift day Night day night day Night

2.12.4 Random selection of time within a processing day

• All processing times during that processing day must be available for selection.

• You must not alter the randomly selected time to allow for convenient times to

sample or according to flocks, sheds or cuts or any other preference.

Random selection of time

• From unique carcass identification numbers across the entire days processing, or

• From all times processing is underway that day (in minutes)

• The randomly selected time (in minutes) must be recorded prior to sampling.

• If it is not practicable to conduct the test on the specified minute a window of up to

10 minutes either side of the originally selected random time is permitted in which to

conduct the sampling. The reason for departure from the originally selected time

must be recorded by the sampler.



• The actual sample time must be recorded in a format (am/pm, 24 hour clock, unique

carcass number) that can be interpreted/translated to 24 hour clock units in the final

results submitted to NMD.

Standard throughput: Each of the three carcasses needs to be independently randomly

selected from all the times available in each processing day. Where premises are operating

day and night shifts they must collect all three samples from one shift each day as per Table

13 above.

VLT: The initial carcass must be randomly selected. Collect the remaining carcasses at time

intervals equivalent to the time required to bag or rinse each carcass. An alternative is to

independently randomly select all 3 carcasses across the processing day as per standard

throughput. Carcasses for the day must not be collected all at once from the line and then

bagged or rinsed, unless there are no other opportunities to complete collection of the 3

carcasses.

2.12.5 Changes to processing schedules

If processing schedules change in a manner that means that the original random sampling

plan can not be followed any remaining samples that day (standard throughput) or week

(VLT) must be randomly selected from the remaining available processing time.

For VLT premises if there is doubt on the number of days (or which single day that week)

processing is to be undertaken, ensure that sampling is undertaken on the first processing

day. This variation to random selection of sampling day must be recorded.

For standard throughput and VLT; if less than the required samples are taken each missed

sample will be recorded on the database as non-compliant for the poultry Campylobacter

performance target (CPT).

2.12.6 Poultry Campylobacter Performance Target (CPT) programme sampling

requirements

The required number of samples must be taken, as specified in 2.12.1. There are no

practical constraints allowed for within the CPT Programme. If the required numbers of

samples are not taken, the default will be to register the missed sample result/s as a result

greater than the CPT moving window limit.

Where there are technical failures such as samples not delivered to the laboratory within the

correct time, or a laboratory error during analysis, these will be recorded on the database as

technical failures.

Where premises processing schedules change suddenly, there are breakdowns, sample

transport problems or laboratory errors, variations to the above random sample selection

protocols are permitted to allow sampling to be undertaken from the remaining processing

that day or week to make up the required numbers. Such variations must be recorded.



The operator may wish to undertake extra sampling routinely to allow for practical constraints

and technical failures. Where extra sampling is undertaken, the operator must identify the

original NMD selection and the extra sample/s selected. The Campylobacter enumeration

results entered in the NMD database will commence using the original NMD samples in the

first instance followed with the extra sample/s if necessary, to give a total of 3 results over 1

processing day for standard throughput and 3 results over one processing day per week for

VLT.


Sampling

3. Sampling

3.1 Manufacture of Diluents for Sampling and Analysis

A component of the equipment required for sampling will be sterile diluents for moistening

swabs. Diluent aliquots may already be added to vials that swabs will be directly placed into

at time of sampling.

Sterile diluents are used for:

1. Moistening APC/E. coli carcass and primal cut swabs.

2. Moistening APC/E. coli/Salmonella primal cut swabs.

3. Suspension of APC/E. coli bulk meat samples.

4. In dilution series for APC and E. coli analyses.

5. Rinsing whole poultry carcasses.

Whether sterile diluents have been manufactured by the laboratory or an approved supplier

the expiry dates must be recorded and no sterile diluent outside of expiry date used for

sampling.

Peptone diluent

0.1% peptone with 0.85% NaCI.

Note: for bobby calves where the chemical intervention, acidified sodium chlorite (ASC), has

been applied prior to sampling, the addition of Sodium thiosulphate to dechlorinate the

sampled is required.

Table 15: Peptone diluent manufacture

0.1% Peptone Ingredients Gram/litre

Peptone 1.0

Sodium chloride (NaCI) 8.5

Distilled water 1000 ml

Reference: MIMM 2005 or later edition, 5.7.2 (0.1% peptone manufacture) which applies to

all species and 11.1.5 (dechlorination of water sample using sodium thiosulphate) which

applies to bobby calf samples post ASC intervention only.


Sampling

Table 16: The volume of peptone diluent required

Species Sterile one diluent volume peptone

Zero (swab, whole tissue immersion) dilution

Ovine, caprine 10ml

Bobby calf, bovine, cervine, ostrich, emu, porcine

15ml

Bulk meat sample suspension 225ml Buffered Peptone Water (BPW)

Single strength buffered peptone water (ssBPW)

Sterile ssBPW is used for

1. Moistening swabs for collection of carcass samples solely for Salmonella analysis.

2. Carcass composite Salmonella analysis making up the volume required for the sample

pre-enrichment broth.

3. Poultry whole carcass rinse diluent for poultry broiler carcasses sampled for

Campylobacter and Salmonella testing. 400ml of ssBPW per carcass is required for all

poultry rinses.

For poultry whole carcass rinse samples and bobby calf Salmonella samples where

chlorinated antibacterial agents are used the addition of sodium thiosulphate is required.

The proportion of sodium thiosulphate to add to ssBPW is listed in Table 17. Where

antibacterial agents other than chlorine based ones are used suitable non-antimicrobial

neutralising additives must be determined.

Table 17: ssBPW manufacture

ssBPW Ingredients gram/litre

Peptone 10.0

Sodium chloride (NaCl) 5.0

Disodium phosphate (Na2HPO4) 3.5

Potassium phosphate (KH2PO4) 1.5

Sodium thiosulphate (Na2S2O3) where chlorinated antibacterial agents are used to decontaminate carcasses (poultry and bobby calves only).

1.0ml of a 3% (Na2S2O3) solution

Distilled water 1000ml

Final pH 7.2+/-0.2 at 25ºC

Reference: MIMM 2005 or later edition, Chapter 7.7.4


Sampling

Double strength buffered peptone water (dsBPW).

Sterile dsBPW is used for:

1. For primal cut swab sample composite Salmonella analysis; addition of equal volume of

dsBPW with remaining peptone diluent to constitute as ssBPW pre-enrichment broth.

2. For bulk meat sample composite Salmonella analysis addition of equal volume of

dsBPW with remaining peptone diluent to constitute an ssBPW pre-enrichment broth.

Table 18: dsBPW manufacture

dsBPW Ingredients Gram/litre

Peptone 20.0

Sodium chloride (NaCl) 10.0

Disodium phosphate (Na2HPO4) 7.0

Potassium phosphate (KH2PO4) 3.0


Final pH 7.2+/-0.2 at 25ºC

Note: this is a formulation specific to NMD requirements.

3.2 Templates Required for Swab Sampling

Sample collection for carcasses and primal cuts requires the use of templates to delineate

the site to be swabbed.

3.2.1 Template materials

Materials from which templates can be constructed:

1. Hoops (circular or square) made of 3mm stainless steel rod with handle that can be

hooked over a finger.

2. Flexible material (plastic or cardboard).

3. Flat rigid sheet metal. Unsuitable for 100cm2 templates due to the large area and

curved/irregular nature of the surface to be sampled.

The size of all templates used for sampling must be verified in laboratory calibration records.


Sampling

Examples of template designs:

Table 19: Suitable template designs

Area to be sampled 5cm2 Ovine, caprine

25cm2 Bobby calf, cervine, ostrich and emu, and

porcine

100cm2 Bovine

Area measurements

Circular – diameter/mm

25.2mm 56.4mm 112.8mm

Square – Side/mm

22.4mm x 22.4mm 50.0mm x 50.0mm 100.0mm x 100.0mm

Hoops (circular or square) made of 3mm stainless steel

rod with handle

Suitable Suitable Suitable

Flexible material – plastic or cardboard

Suitable Suitable Suitable

Flat rigid sheet metal Suitable Suitable not suitable

3.2.2 Sterilisation of templates

It is preferable to use pre-sterilised (autoclaved) multiple templates.

Otherwise for metal hoops and templates after each sample sterilise these for the next

sample by:

• 70% ethanol for flame sterilisation; or

• Ethanol/iso-propanol based disinfectant wipes.


Sampling

3.2.3 Sterile cotton tipped swabs

Table 20: Swabs required for sampling

SPECIES Ovine, caprine Bobby calf, cervine,

ostrich/emu, porcine

Bovine Poultry

Number of swabs required per

carcass site or cut site

2 1 wet, 1 dry

4 2 wet, 2 dry

6 3 wet, 3 dry

No swabs Whole carcass rinse method

Swabs must be sterilised before use. A pack of about 30 for use with each sampling batch is

a practical number. Any remaining swabs must be re-sterilised before re-use in another

sampling batch.

Note: Commercial swabs supplied in individual plastic tubes and returned after sampling to

those individual plastic tubes can be used however certain conditions apply. All tubes must

be labelled. The suspension buffer required for each site sampled must be used to rinse out

the inside of all the tubes of swabs associated with that site in order to remove any bacteria

that may have been smeared onto the inner walls by the swabs during sampling and

transport.

3.3 Collection of Samples

Trained sample collectors must be used for sample collection.

Refer to Part 1 clause 4 and Schedule 1 section 1.5 of this Notice.

3.3.1 Equipment required for sampling

For swabbed sites the following requirement is required:

1. Templates – size required for species.

2. Sterile vials containing 4 – 9 plastic or glass beads with or without diluent. If the vials do

have diluent ensure the diluent volume is correct for the species to be sampled.

3. Diluent for swab moistening as required for the type of sampling.

4. Sterile cotton tipped swabs.

5. Flaming alcohol or wipes if required for sterilising templates.


Sampling

For bulk meat:

For bulk meat the following equipment is required:

1. Knives (sharpened with appropriate equipment or by experts on site) or scalpels and

blades.

2. Tweezers if required.

3. Plastic sample bags with twist ties or equivalent for example, whirl pac bags.

4. Gloves (vinyl or latex as required) or plastic bags (these must be clean. Sterilisation not

required) to cover hands.

5. Flaming 70% ethanol or ethanol/iso-propanol containing disinfectant wipes for sterilising

the knife, or scalpel blade, and tweezers.

For whole rinse poultry carcasses:

For whole rinse poultry carcasses plastic sample bags with twist ties or equivalent for

example, poultry rinse bags with a tear tie are required.

General:

In general the following equipment is required:

1. Appropriate headwear, overalls/coats and trousers, boots, aprons for entering an edible

area.

2. Pens and paper to record sampling details and indelible pen to label vials as required.

3. Spare sterile templates, vials, swabs and/or gloves/plastic bags.

4. Insulated containers with ice packs (slicker pads or bags of shaved ice).

5. Ladders or stools for access to sampling sites where necessary.

3.4 Sampling Method

3.4.1 Wet/dry swabbing technique

The following is the wet/dry sampling technique:

1. Fill out the laboratory sample log and label all vials (with indelible pen or coded

adhesive labels) to correspond with date, time, sample sites, product type, class and

species.

2. Loosen the caps of vials, but do not open.

3. Document the time that sampling begins.

4. Unwrap a sterile template or alcohol flame/wipe a hoop or metal template. Ensure that

the template does not become contaminated. Hold the template in one hand.

5. Carefully extract the required number of swabs from their sterile wrap.


Sampling

6. Hold the shaft/s of half the number of swabs required per site between the fingers and

thumbs of one hand and moisten in the diluent for 5 seconds. Leave the other half of the

sterile swabs dry and hold either separately like chopsticks in the same hand or the hand

in which the template is being gripped.

7. Press the sterile template onto the surface to be sampled. Hold firmly. Care is required

not to smear the template over the sampling area.

8. Take the wet swab/s and rub them over the area defined by the template. Rub the wet

swab/s vertically, then horizontally, then diagonally across the entire surface. The

sampler is required to swab in all three directions to cover the surface area adequately

but not necessarily in the order stated. Rub in all three directions, one way following the

other. Rub for at least the specified times in Table 21.

Table 21: Time required to rub wet swabs

Ovine, caprine 5 cm2

Bobby calf, cervine, ostrich/emu, porcine

25 cm2

Bovine 100 cm2

10 seconds swabbing time 20 seconds swabbing time 20 seconds swabbing time

It is important that the swab/s are rotated slowly and continually, by rolling the shafts

between the fingers and the thumb (take care not to drop or contaminate the dry swab in the

process). Use as much pressure as possible, but avoid breaking the shaft. Do not hold a

swab on such an oblique angle that the shaft contacts the sample site or template. Only the

bud should make contact with the sample, not the shaft.

If a swab becomes contaminated by the contact with foreign non-target surfaces discard the

contaminated swabs and resample another carcass or primal cut.

9. Hold the wet swabs to one side, and repeat swabbing with dry swab/s until the area

being sampled is dry.

10. Insert the wet and dry swabs into a labelled transport vial. The vial may or may not

contain the diluent zero dilution volume required for number of swabs/species. Break

the swabs by pressing the buds against the inside of the vial near the rim and press

down on the lid to aid in snapping the shafts from the buds. Care is required with

aseptic technique and dexterity to prevent fingers contaminating the swabs, inside of lid

or inside of vial. Discard and resample from a new carcass or cut if any contamination

event occurs at this point.

11. Close the vials and place in an insulated container avoiding direct contact with the ice

pack.

12. Disinfect templates between sample sites and carcasses/cuts, or use separate pre-

sterilised templates for each sample.


Sampling

3.4.2 Whole tissue composite sampling technique for bulk meat

The following is the whole tissue composite sampling technique for bulk meat:

1. Fill out the laboratory sample log and label all plastic sample bags with twist ties or

equivalent for example, whirl pac bags (with indelible pen or coded adhesive labels) to

correspond with date, time, sample sites, product type class and species.

2. Document the time that sampling begins.

3. Select a carton after packing and before strapping.

4. Open the carton to be tested and carefully peel back the plastic liner.

5. Select 5 whole tissue samples, each of approximately 10g in volume, from the 4

diametrically opposite corners of the full carton and 1 from the centre of the full carton.

No extra whole tissue samples are needed when Salmonella testing is required.

Figure 12: The position of the 5 sites to take whole tissues samples from in a bulk meat

carton

6. Using a glove or a plastic bag inverted over your hand or sterile equipment such as meat

hooks, tweezers or knives, reach into one of the 5 sites in the carton identified in figure

12. Cut out a whole tissue sample by trimming a thin layer from an original external

carcass surface if possible. Each whole tissue sample weighing approximately 10 gram

to result in at least 50 gram collected per carton.

7. Deposit the sample aseptically into a sterile sample bag.

8. Repeat at the 4 remaining sites in the carton depositing samples into the same sample

bag.

9. Seal the sample bag with tape or twist tie.

10. Place the sample bag into an insulated container avoiding direct contact with ice packs.


Sampling

11. Re-sterilise knife with alcohol flame/wipes (and tweezers if you are using these) or

change scalpel blade between cartons.

12. Make sure all implements used (knives, tweezers, gloves or plastic bags, ties) are not

left inside the carton after completion of sampling.

3.4.3 Poultry: whole carcass rinse

Each poultry whole carcass rinse sample will be analysed for Campylobacter. In addition one

whole carcass rinse sample of the set taken on the day of sampling will be analysed for

Salmonella as specified in section 2.12.1.

1. Fill out the laboratory sample log and label all plastic sample bags (with indelible pen or

coded adhesive labels) to correspond with date and time of sampling.

2. Sampling periods:

a. Standard throughput; 3 samples per processing day are required. Each of these

samples will be taken at separate randomly selected times. There must be

three discrete sampling times across that days processing. For each of the

three sampling periods the date and time must be recorded.

b. VLT: Select the first carcass of 3 for sampling (or bagging) from the chain.

Record the time of selection of the first carcass. Collect the remaining

carcasses in this single sampling period, ensuring that carcasses are not taken

consecutively from the chain.

3. If the carcass is collected prior to the first drop point suspend the carcass for Tº to drain

excess water from the carcass. This procedure will mimic the usual handling and

draining of carcasses at the premises.

4. Invert the bag over the hand to form a glove. Take care not to contaminate the exposed

internal surface of the bag.

5. Open the sample bag aseptically.

6. Pick up the “drained” bird/carcass by the legs (or air-chilled bird from the chain) using the

“bagged” hand and reverse the bag over the carcass.

7. Bottles (400ml) of sterile ssBPW for rinsing the carcass for Campylobacter, E coli and

Salmonella analyses will have been prepared as per section 3.1 Table 17 and must be

pre-cooled to 4ºC for use at 4ºC at time of sampling.

8. The carcass can be rinsed on selection or transported to the laboratory for rinsing in

which case the carcass must be transported to the laboratory in an insulated container

with ice packs, but not in direct contact with ice packs, and not frozen; arriving at the

laboratory within 15-30 minutes of carcass selection. The temperature of the carcass on

arrival at the laboratory does not need to be measured.


Sampling

3.4.3.1 Rinse procedure

1. Rinsing involves pouring 400ml of ssBPW diluent over the carcass in the bag, expelling

most of the air from the bag then securing the bag tightly by hand or with a twist tie.

2. While securely holding the bag, rinse the outside of the carcass, including the wings,

legs, and the inside cavity by gently rocking the carcass in an arching motion,

transferring the weight of the carcass from one hand to the other. The hands can

simultaneously be used to massage the carcass surface, particularly around the wings

and the legs.

3. Time require for the poultry whole carcass rinse procedure is two minutes (120

seconds). Commence by rocking the carcass permitting the rinsate to pass across the

external surfaces. The rinsate will appear yellowy in colour with suspended fats.

Periodically position the bird and clear any fat or skin covering the vent and tilt such that

the rinsate can enter the cavity. Swirl to rinse the entire internal cavity. The rinsate on

exit from the cavity should be a pink to red colour. Continue the rocking motion to pass

the rinsate over external surfaces and complete an internal cavity rinse at least once

more during the two minutes. After the two minutes a rinsate with a reddish tint and

thicker with suspended fats and cells should be visible.

Table 22: Time required

Poultry carcass whole rinse

120 seconds (2 minutes) of rocking for 400ml rinsate

4. To transport the rinse sample to a laboratory place the sample bag/vial into a second

bag, secure the opening and place in an insulated container with ice packs for transport

to the laboratory. Avoid direct contact of the rinse sample with the ice packs.


Sampling

Figure 13: Poultry whole carcass rinsing

Arrival of poultry broiler sample;

400ml of ssBPW to be added.

Rocking the bird

Yellowish rinsate after the rocking

procedure, tipped into a corner

Opening the vent entry into the

cavity.

Rinsate after passage through cavity Final rinsate view after 2 minutes.


Sampling

5. Isolating the carcass rinse for analysis

• Reset the bag with the carcass on a flat surface. Open the bag and remove the

carcass aseptically using a gloved hand. Expel the air (to ease packing) and secure

the top of the stomacher bag to prevent contamination and/or spillage of the rinse

sample;

OR

• Prior to removal of the carcass aseptically transfer at least 70ml of the rinse sample

to a sterile vial for transportation of the laboratory. Cutting a corner of the bag off is

one practical means to achieve this. 70ml will provide enough rinse sample for

analysis plus back up.

6. Disposition of carcasses

• Carcasses rinsed in the processing room may be returned to the immersion chiller.

• Carcasses subjected to air chilling shall be rinsed with potable water prior to return

to the chain.

• Carcasses rinsed in the laboratory must not be returned to the processing room.

3.5 Transportation of Samples to the Laboratory for All Species

In order to protect samples from extremes of temperature and minimise changes to the

microflora during collection and transport, all samples must be stored and transported in an

insulated container. For NMD purposes extremes of temperature are defined as <0˚C and

>25˚C.

All samples must be subjected to sufficient “frozen coolant” during sample collection and

transportation to keep the samples cold, but not frozen, regardless of whether the sample is

to be immediately analysed, transported off-site or held for a period of time.


Sampling

3.5.1 Red meat species samples

Premises may choose to have samples transported to an on-site lab for some analyses and

off-site for others according to schedules, laboratory scope of testing and other factors.

Figure 14: Red meat species sampling timetable

ssss

3.5.2 Transportation temperature and times for red meat samples

• Sample collection temperatures – The temperature within the insulated container

used at the time of sampling should be maintained between 0˚C and 5˚C (not

frozen), and must not exceed 10ºC.2

• Storage temperatures – A refrigerator operating in the 0ºC – 5ºC range must be

used to store samples and diluents.

• Transportation temperatures – The temperature of samples within the insulated

container used for transport should be maintained between 0˚C and 5˚C (not frozen),

and must not exceed 10ºC in transit.

• Controls - Transport sample temperature shall be measured/monitored (using a

control vial/sample to prevent contamination) at a site within the transportation

container that will most likely reflect the temperature of the samples;

2 Four Small Studies for NMD, AgResearch 2001


Sampling

o for swab samples stored transported dry, immediately adjacent to the

swabs;

o for swab samples stored/transported in liquid medium, within the medium;

o for whole tissue samples, immediately adjacent to tissue samples.

• Frozen - Samples that are received frozen shall be rejected by the laboratory and

replacement samples collected.

• High Temperature - Samples received that exceed 10ºC must be rejected and

replacement samples sought.

• Sample receipt - The laboratory receiving the samples must:

o record in the sample register the sample temperature on receipt.

o verify that analysis can be initiated within 24h, or 30h under exceptional

circumstances, from time of collection of the first sample. Reference: ISO

17604:2003. Extensions to a maximum of 30 hours are permitted where

practical constraints dictate, but not routinely. Extensions must be justified

with reasons recorded.

o verify that the sampler recorded as collecting the samples was a currently

listed Certified, Associate or restricted sampler.

• Verification of temperatures – the laboratory must use data loggers to verify

temperatures of insulated containers/samples from time of sampling to receipt by

laboratory. Verification of temperatures is required every 6 months.

Premises must implement procedures to ensure that requirements for all analyses of samples,

including Salmonella analysis, are complied with within the time periods specified.

Laboratories, including off-site laboratories, are responsible for training on-plant personnel

seconded to collect samples.

Off-site laboratories are responsible for ensuring that transport companies, for example, on-site

personnel, couriers are aware of the transport and storage temperature and time requirements.

Off-site and on-site laboratories are responsible to ensure night shifts are informed of storage

temperatures, transport to laboratory and time requirements.

3.5.3 Collection and transport of whole tissue samples

All bulk meat whole tissue samples; cold boned, hot or warm boned must be placed in an

insulated container with sufficient frozen coolant at time of sampling to maintain a temperature

between 0˚C and 5˚C (not frozen), and not exceeding 10ºC .

Hot/warm boned whole tissue samples must be rapidly cooled to <5ºC but not frozen. Trials

must be conducted to demonstrate;


Sampling

• temperature falls rapidly to <5ºC,

• samples do not freeze,

• that transport arrangements can still be met, and

• that E. coli NZRM 916 when inoculated into the whole tissue samples can be

recovered.

All bulk meat whole tissue samples requiring storage before transport and/or analysis must be

stored in a refrigerator calibrated to 0 - 5ºC.

All bulk meat whole tissue samples must be transported in an insulated container maintaining

between 0˚C and 5˚C (not frozen), and not exceeding 10ºC temperature in transit.

The laboratory must conduct six monthly verification of temperature ranges by datalogging the

temperature of bulk meat samples from time of sampling until commencement of analysis.

3.5.4 Transport of red meat Salmonella samples

Salmonella samples include: carcass Salmonella swabs, APC primal cut swabs, the bulk meat

sample itself, primal cut and bulk original peptone APC/E. coli suspensions, and the final BPW

suspensions.

All Salmonella samples must be in an insulated container maintained between 0˚C and 5˚C (not

frozen), and not exceeding 10ºC in transit, and/or stored in a refrigerator operating between 0˚C

and 5ºC until commencement of analysis.

Following receipt of samples by the laboratory initiation of Salmonella analysis is defined as the

commencement of incubation of the BPW pre-enrichment broth. This may mean cooling of these

original suspensions during APC/E. coli analysis and use of pre-chilled dsBPW to <5ºC to make

up the final Salmonella sample suspension.


Sampling

3.5.5 Poultry samples

Figure 15: Poultry sampling timetable

Select whole carcass in process room day shift or

night shift. Record time of collection of

first carcass.*

Initiate analysis within 24 hours after first carcass

selected.

Rinse carcass immediately in process room.

Transport carcass in an insulated container to the laboratory within 15 – 30 minutes of

sampling.

Rinse carcass immediately.

Commence analysis immediately or store at <5ºC.

Transport sample to on-site or off-site laboratory.

Insulated, arriving at or below 10ºC and stored at

<5ºC.

The maximum period may be extended to 30 hours, not routinely, with reasons recorded

*First carcass in each discrete consignment of samples dispatched to the laboratory for that processing day.


Sampling

3.5.6 Preparation of poultry samples for transportation

See also 3.4.3

The process for the preparation of poultry samples for transportation is as follows:

1. Tightly seal the sample bag, transportation vial, or bag containing the carcass to be

sampled; and

2. All samples must be immediately placed into an insulated container with frozen coolant

ensuring the sample bags or transport vials are not in direct contact with the frozen

coolant.

3.5.7 Transport and temperature requirements for poultry whole carcass samples

The carcass rinse sample or the carcass itself must be transported to the laboratory in an

insulated container with ice packs, but not in direct contact with ice packs, and not frozen.

1. Whole carcasses not rinsed immediately in the process room, but transported to the

laboratory for rinsing must be transported in insulated containers with sufficient frozen

coolant to cool the sample. The temperature of the carcass on arrival at the laboratory

does not need to be measured.

2. Whole carcass for rinsing in the laboratory must be preferably delivered to the laboratory

within 15 minutes (no later than 30 minutes) of selection of the whole carcass from the

processing chain and immediately rinsed.

3. Rinse samples transported to the laboratory must be transported in insulated containers

with frozen coolant. The temperature of rinse samples within the insulated container

used for transport should be maintained between 0˚C and 5ºC (not frozen), and must not exceed 10ºC. The temperature of the diluent, not just the airspace in the container

must be maintained in this temperature range.

4. All samples are to be delivered to the laboratory as soon as possible after collection.

5. The time of collection of the whole carcass sample must be recorded.

6. The temperature of the rinse sample on arrival at the laboratory must be recorded.

7. Samples that are frozen shall be rejected by the laboratory and replacement samples

collected from the same days processing if possible.

8. Premises must be able to verify that analysis can be initiated within 24h from time of

collection for the first sample. This may be extended to 30h under exceptional

circumstances, not routinely, with reasons recorded.

9. Laboratories, including off-site laboratories, are responsible for training on-plant

personnel seconded to collect samples. Off-site laboratories are responsible for


Sampling

ensuring that transport companies, e.g. on-site personnel, couriers are aware of the

transport and storage temperature and time requirements.

10. Off-site and on-site laboratories are responsible to ensure night shifts are informed of

storage temperatures, transport to laboratory and time requirements.


Laboratory Analysis

4. Laboratory Analysis

4.1 Dilutions Required

Table 23: Dilutions required

Analyses Red Meat Poultry

APC30 zero to 10-4 (dilution series of 5)

E. coli zero to 10-3 (dilution series of 4) 100 to 10-2 (dilution series of 3)

Campylobacter 100 and 10-1 plus higher dilutions if required.

Note: zero is the undiluted sample.

4.1.1 High end dilutions

All dilutions must be plated in duplicate until the premises data indicates at what dilution a

colony count of greater than the maximum allowable is never, or rarely exceeded.

The maximum allowable counts are:

300 CFU for spread plates;

150 CFU for spread plate half plates;

250 CFU for PetrifilmTM APC;

150 CFU for PetrifilmTM E. coli.;

150 CFU for mCCDA direct plate Campylobacter

When a valid count exceeds (>300, >250 or >150 on the highest dilution) the dilution series

shall be immediately extended until such time as the premises demonstrates that the higher

counts are not a consistent trend.

It is very important to have a valid actual count of higher results for statistical purposes. A

TNTC count can be any value greater than the valid count value of 300, 250 or 150 on the

highest dilution tested and is thus more variable and unknown than a not detected count. A

TNTC result can range from as low as a 2 log10 value up to any higher value (infinite). It

could be 5 log10 or 10 log10. If a dilution series is not great enough this will lead to TNTC

results and such high counts cannot be included in the database, which, in turn, leads to a

poor representation of the true range of results.

When insufficient dilutions are prepared and counts are above the maximum allowable count

on the lowest dilution the laboratory must report to NMD a too numerous to count result

which will be classed as a technical failure.

A TNTC must be regarded as “detection” and/or “higher than expected count” when

assessing compliance with premises performance criteria under HACCP and RMPs.


Laboratory Analysis

Note that the area sampled will have an effect on the number of dilutions required. More

bacteria are expected to be recovered from a larger sample area therefore more dilutions will

be required.

As regulatory limits will be applied to poultry broiler carcass Campylobacter enumeration

results laboratories need to ensure that a suitable series of dilutions is plated to minimise the

chance of TNTC results. If a TNTC result occurs it will be recorded as exceeding the CPT

limit. Refer sections 4.10 and 6.9.

When a TNTC count occurs the test does not need to be repeated.

Upon receipt of a TNTC result the laboratory must extend the dilution series, in anticipation

of higher counts, for at least 3 subsequent poultry processing days (standard throughput) or

3 subsequent processing weeks (VLT) following the TNTC result notification.

4.1.2 Low end dilutions

All cases – Undiluted (zero dilution) plates or PetrifilmTM plates that contain less than the

lowest counting range of 30, 25, or 15 are to be counted and reported.

Table 24: Summary of acceptable counting ranges

Dilutions Spreadplate APC

Spreadplate APC ½ plate

mCCDA direct plate

PetrifilmTM APC PetrifilmTM

E. coli

100 0-300 0-150 0-250 0-150

10-1, 10-2,…..10-n 30-300 15-150 25-250 15-150

4.2 Duplicate Plating/Analytical Precision

Plating must be performed in duplicate, i.e. two agar plate, both sides of a ½ plate or two

PetrifilmTM inoculated from each dilution until laboratory precision data (for all analysts

performing testing) indicates otherwise. Duplicate plating (APC30 and E. coli) must be

undertaken to give statistical confidence to the results obtained. Use the Analytical Precision

calculation (for a single operator) in chapter 5 Appendix 1.7 of MIMM to determine a Z score

which is then compared to a limit of 1.96.

Record all duplicate plate counts that fall within the acceptable counting ranges as specified

in section 4.1.

If a laboratory demonstrates that it meets the Analytical Precision criteria outlined in MIMM

2005 or later edition, it may use singlet plating for all dilutions other than the undiluted

(zero) dilution sample.

For ILCP analyses related to NMD APC and E. coli tests use the NMD procedures described

in this publication. Although successful precision analysis may have eliminated the need for

duplicates on all dilutions for routine NMD analyses, duplicates are required for all dilutions

for the corresponding ILCP tests.


Laboratory Analysis

4.3 Preparation of Dilutions

4.3.1 Introduction

When conducting a quantative bacterial analysis, sample suspensions must be diluted to an

appropriate level that will result in 30 – 300 colony forming units (CFU) on the plate, 25 – 250

CFU if using APC PetrifilmTM or 15 – 150 CFU if using half plates, E. coli PetrifilmTM or

mCCDA direct plate.

Dilutions for viable bacterial counts are aseptically prepared in ten fold steps.

All dilutions are performed using 0.1% peptone + 0.85% NaCI diluent. Reference: ISO

17604:2003.

The dilutions volume used must be sufficient to inoculate duplicate plate count agar plates,

duplicate PetrifilmTM, enough volume for the composite Salmonella pre-enrichment if

required and leaving enough volume in case of error.

Two dilution volumes are recommended:

a. 9ml in 25ml universal bottles or vials, with 1ml transfer volume.

b. 4.5ml in microcentrifuge or microdilution tubes, with 0.5ml transfer volume.

The use of pre-poured dilution volume is allowed only if a laboratory can provide evidence

that they do not lose volume during autoclaving (as defined in MIRINZ Bulletin 35 or MIMM

2005 or later edition, Section 2.5.6), and that they have an appropriate QA programme in

place for ongoing verification.

Always use a fresh pipette or pipette tip for each transfer of diluted sample.

Ensure that dilutions are as homogenous as possible at all stages by:

• Carefully following the instructions for the use of micropipettors;

• Mixing each dilution thoroughly prior to the next dilution step to ensure that the

bacteria are randomly distributed;

4.3.2 Definition of zero dilutions for calculation purposes

The swab suspension (swabs immersed in the initial volume of either 10ml or 15ml) is an

undiluted sample referred to as the “zero dilution”. The swab suspension is entered into the

calculations as the 100 dilution.

The whole tissue suspension is dilution but in order to maintain consistency with plate

labelling, it is entered into the calculation as the 100 dilution (undiluted) and accounted for in

the final numeric multiplier.

4.4 Preparation of Swab Samples for APC and E. coli Analysis

Add to each vial with swabs.


Laboratory Analysis

Table 25: Swab suspension volumes required

Bovine, bobby calf, cervine, ostrich/emu, porcine

Ovine and caprine

15ml sterile 0.1% peptone diluent to each vial

10ml sterile 0.1% peptone diluent to each vial

Make certain there are 4 – 9 sterile glass or plastic beads in each vial.

Close the vial and shake vigorously to loosen the cotton bud of the swab for 2 minutes

(ensure consistency). Shaking can be performed by hand or by using a combination of initial

shaking by hand and a vortex mixer in 3 x 10 second bursts. Note that shaking by hand is

usually sufficient and enables several vials to be processed at once.

The cotton buds should be well broken up by this treatment. The cotton swabs must be left

in the diluent until all the dilution and plate inoculation procedures are completed. Do not

remove the swabs from the vials at any stage.

4.5 Preparation of Red Meat Separate Carcass Site Samples Swabbed for Salmonella

Analysis

Note: Salmonella testing to support official assurances for beef exported to Sweden and

Finland or to any other country with the same requirements, eg Iceland, must use the

required sampling and method specified for routine official test 2.4.2.

Table 26: Preparing Salmonella sample suspensions

Species Bovine 5 carcasses

Bobby calf, Porcine 5 carcasses

Cervine 3 carcasses

Ostrich/Emu 2 carcasses

Caprine 5 carcasses

Total number of swabs

6 swabs x 3 sites x 5 carcasses =

90 swabs


60 swabs


36 swabs

4 swabs x 1 site x 2 carcasses = 8

swabs


30 swabs

Volume of ssBPW

250 ml 150 ml 150 ml 30 ml 75 ml

VLT (Very low throughput) premises with only one carcass sample (no composite)

Species Bovine VLT Bobby calf, Porcine VLT

Cervine VLT Ostrich/Emu VLT

Caprine VLT

Total number of swabs

6 swabs x 3 sites x 1 carcass =

18 swabs


12 swabs


12 swabs

4 swabs x 1 site x 1 carcass =

4 swabs


6 swabs

Volume of ssBPW

50 ml 30 ml 30 ml 15 ml 15 ml

Add an appropriate number of glass beads to the suspension, close the bottle and shake

vigorously for 2 minutes to remove the bacteria from the swabs into the diluent.


Laboratory Analysis

4.6 Preparation of Bulk Meat (Whole Tissue) Samples for APC, E. coli


Finland or to any other country with the same requirements, for example, Iceland, must use

the required sampling and method specified for routine official test 2.4.2

• Place the whole tissue samples (5 X ~10g) onto a sterile chopping board and finely

dice (size reduce) with a sterile knife or other suitable aseptic means.

• Re-sterilise or use separate chopping boards and knives etc for each sample.

• Aseptically weigh 25g of diced tissue directly into a stomacher bag.

• To each bag add 225ml of sterile diluent (0.1% peptone + 0.85% NaCl).

• Stomach for 2 minutes.

4.7 Aerobic Plate Count, APC30

Applicable to red meat NMD programmes.

Not applicable to poultry NMD programmes.

The following APC methods are approved for the red meat NMD programmes:

1. Spread Plate method (Routine Official test 2.1.2)

2. PetrifilmTM Aerobic Plate Count method (Routine official test 2.1.3)

3. Spiral Plater method (Routine official test 2.1.4)

4.7.1 General considerations for plating

The following are general considerations for plating:

1. Pre-poured plate count agar plates must be dried before use. Plates may be dried

using one of the following methods:

i. incubation for 24h at 30-37ºC inverted with lids in place;

ii. inverting with lids ajar in a laminar flow cabinet for 20-30 minutes;

iii. inverting with lids ajar in a still air incubator at 30-37ºC for 1 ½ to 2h;

iv. inverting with lids ajar in a forced air incubator at 30-37ºC for 30 minutes.

2. The swab samples in diluent with beads must be shaken for 2 minutes, bulk meat

samples 25g/225ml peptone diluent must be stomached for 2 minutes before

commencing analysis.

3. The same pipette/tip may be used when working up a dilution series from the most dilute

level to the most concentrated level.

4. The required incubation temperature for the APC is 30ºC +/- 1ºC (ISO 17604:2003).


Laboratory Analysis

5. Initiation of analysis for APC is defined as the time when the sample is suspended and

ready for serial dilution.

6. The incubation time is 48 hours. Where plates cannot be counted at 48h plates may be:

I. incubated for up to but not longer than 72h

II. removed from the incubator and stored for no longer than 48h in a laboratory

refrigerator set at not greater than 5ºC.

The extra storage time of the plates must be recorded on the result sheet.

7. Count only plates with between 30 to 300 colonies, 25 to 250 colonies for PetrifilmTM

APC, 15 to 150 colonies for ½ plates and PetrifilmTM E. coli plates (ISO 17604:2003).

See table 22: Acceptable counting ranges.

8. If colonies are indeterminate at 48h estimate a count, re-incubate for a further 18-24h

and recount. The total incubation time must be recorded on the result sheet.

9. Express the result as the number of colony forming units (CFU)/cm2 or /g of sample.

When there are nil counts on both of the duplicate zero dilution plates, express the result

as “not detected”.

4.7.2 APC30 by spread plate method

(Reference MIMM 2005, Chapter 6 or later edition). Carry out all quality control procedures

for the media and methods using Pseudomonas aeruginosa NZRM 981 or Lactobacillus

viridescens NZRM 3313 as the positive control.

4.7.2.1 Method

The following is the APC30 by spread plate method:

1. Dispense 0.1ml (100µl) volumes of the appropriate dilutions (100 non-diluted, 10-1, 10-2,

10-3, 10-4) onto duplicate, dried plate count agar plates, or a single ½ plate.

2. Start with the highest (most dilute) level of dilution. Mix the dilutions thoroughly before

dispensing. The same pipette/tip may be used for all inoculations.

3. Spread the inoculum over the surface of the agar as evenly and as quickly as possible

using a sterile spreader (bent glass rods or disposable plastic spreaders). In order to

prevent cross-contamination, sterilise the spreader between plates both within and

between dilutions.

4. Allow the inoculum to soak into the agar surface. This should occur within 15 minutes. If

liquid does not soak in (i.e. the plates were not dried sufficiently), the viable bacteria

present in the inoculum may start to replicate and spread, possibly resulting in an

inaccurate count.


Laboratory Analysis

5. Invert the plates before incubation so that the agar is in the upper half of the petri dish.

Inversion prevents condensation dropping onto the surface of the agar reducing

contamination and preventing the spread of motile micro-organisms. Stack if necessary,

and incubate at 30±1ºC for not less than 48h.

6. Count all colonies. Record all counts on the undiluted plates and only those between

30-300 on full plates or 15-150 on half plates.

7. Express the results as the number of colony forming units CFU/cm2 or /g of sample.

8. If colonies are not detected on both of the zero dilution plates express the results as “not

detected”.

9. Where counts are greater than 300 on full plates or 150 on half plates on the highest

dilution plate or half plate record the result as to numerous to count (TNTC). The TNTC

must be regarded as a “detection” and /or “higher than expected count” when assessing

compliance with premises performance criteria under HACCP and RMPs.

4.7.3 APC30: PetrifilmTM Aerobic Count Plate Method

(Reference MIMM 2005, Chapter 6 or later edition).

4.7.3.1 Method

The following is the APC30 PetrifilmTM Aerobic Count Plate Method:

1. Carry out all quality control procedures for the media and methods using Pseudomonas

aeruginosa NZRM 981 or Lactobacillus viridescens NZRM 3313 as the positive control.

2. Place the PetrifilmTM Aerobic Count plate onto a flat surface, label and lift the top film.

Do not allow the plate to curl.

3. Inoculate and process each PetrifilmTM plate individually. Under no circumstances

inoculate several at the same time.

4. Lift the top film, and carefully dispense 1ml of the appropriate dilution onto the agar.

5. Slowly roll the top clear plastic film down onto the inoculum to prevent entrapment of air

bubbles.

6. Distribute the sample evenly using the spreader provided (ridge side down). PetrifilmTM

Aerobic Count plates do not have a foam dam that’s why the spreader is used ridge side

down. Apply a gentle even pressure, but do not twist or slide the spreader. Carefully lift

the spreader clear or the PetrifilmTM plate.

7. Move the completed PetrifilmTM plate to one side and allow one (1) minute for the agar to

set.


Laboratory Analysis

8. In the meantime inoculate and process the next PetrifilmTM plate.

9. Incubate all PetrifilmTM plates for not less than 48h at 30ºC +/- 1ºC clear side up and in

stacks of no more than 20.

10. Count only plates with 25-250 colonies, the exception being the undiluted (zero dilution)

plate if counts are less than 25.

11. Express the results as the number of colony forming units CFU/cm2 or /g of sample.

12. If colonies are not detected on both of the zero dilution plates express the result as “not

detected”.

13. Where counts are greater than 250 on the highest dilution plate record the result as too

numerous to count (TNTC). A TNTC must be regarded as a detection “and/or “higher

than expected count” when assessing compliance with premises performance criteria

under HACCP and RMPs.

4.7.4 APC30: Sprial Plater Method

(Reference MIMM 2005, Chapter 6 or later edition).

4.7.4.1 Considerations

The quality of pre-poured agar is paramount to the successful use of the Spiral Plater.

Plates must be prepared according to the manufacturer’s guidelines supplied with the Spiral

Plater.

The plate surface must be smooth, the depth uniform and the surface uniformly dry but not

dehydrated.

Pre-poured agar plates must be dried before use.

4.7.4.2 Method

The following is the Spiral Plater Method:

1. Carry out all quality control procedures for the media and methods using Pseudomonas

aeruginosa NZRM 981 or Lactobacillus viridescens NZRM 3313 as a positive control.

2. Follow all instructions for use in the manual supplies with the spiral plater.

3. After inoculation, leave the plates on the bench right-way-up for 10 minutes to allow the

inoculum to soak into the agar.

4. Invert the plates, stack if necessary, and incubate at 30ºC +/- 1ºC for no less than 48h.

5. Obtain a count either manually according to the manual supplied with the spiral plater or

automatically using a computerised plate scanner.


Laboratory Analysis

6. Do not count spiral plates with an irregular distribution of colonies caused by dispensing

errors.

7. Express the result as the number of colony forming units; CFU/cm2 or /g of sample.

8. If colonies are not detected on the plate express the result as “not detected”.

9. Where counts out of range on the highest dilution plate record the result as too

numerous to count (TNTC). A TNTC must be regarded as a “detection” and/or “higher



4.8 Escherichia coli PetrifilmTM

Note that red meat samples will be in a peptone diluent suspension.

The PetrifilmTM E. coli method is the only method of E. coli analysis approved for the red

meat NMD Programme.

Reference MIMM 2005, chapter 8.4 or later edition.

4.8.1 Method

1. Carry out all quality control procedures for the media and methods using Escherichia coli

NZRM 916 as the positive control and Klebsiella pneumoniae NZRM 482 or Enterobacter

aerogenes NZRM 798, as a negative control.

2. Initiation of analysis is defined as when the sample has been suspended as is ready for

serial dilution.

3. Place a PetrifilmTM E. coli plate on a flat surface and hold the plate flat. Do not allow the

plate to curl.

4. Mix the dilutions thoroughly before dispensing. The same pipette tip may be used for all

inoculations if you commence with the highest (most dilute) dilution.

5. Lift the top film and carefully dispense 1ml of the appropriate dilution into the circular well

containing the agar. Note: Do not inoculate several at the same time. It is imperative to

inoculate and lower the top film before proceeding to the next.

6. Slowly roll the top clear plastic film down on to the inoculum to prevent entrapment of air

bubbles.

7. If necessary use the spreader provided flat side down, as PetrifilmTM E. coli plates have

a rim around the well, to distribute the sample evenly over the well. Apply a gentle even

pressure to the spreader, but do not twist or slide the spreader. Carefully lift the

spreader clear of the plate.

8. Move the completed plate to one side and allow one (1) minute for the agar to set.

9. In the meantime inoculate and process the next plate.


Laboratory Analysis

10. Incubate all plates for 18-24h at 35 +/- 1ºC or 37 +/- 1ºC (36 +/- 2ºC is not acceptable)

clear side up and in stacks of no more than 20.

11. Where plates cannot be counted at 24h the plates may be removed from the incubator

and stored for no longer the 72h in a laboratory refrigerator set at not greater than 5ºC,

or freezer. Storage time/temperature must be recorded on the result sheet.

12. If colonies are indeterminate at 24h, count as best as possible, re-incubate for a further

18-24h and recount. Record the incubation time on the result sheet.

13. Count as E. coli all blue colonies with or without gas bubbles (3M approved).

i. Do not count colonies that have grown on the foam dam as the may not have been

subjected to the selective influence of the medium thus might not be E. coli.

ii. Count only plates with between 15 and 150 counts, the exception being if the plate

from the non-diluted (zero) dilution sample has less than 15 colonies.

iii. High numbers of E. coli will turn the medium blue and high numbers of

Enterobacteriaceae will turn the medium red. Both situations make accurate

enumeration improbable. Should this occur the highest dilution plated the samples

must be recorded as (TNTC) and the operator immediately notified.

14. Express the results as the number of E. coli CFU/cm2 or /g or /ml of sample. If blue

colonies with or without gas are NOT detected express the results as “not detected”.

15. Where counts are greater than 150 on the higher dilution plate record the result as too

numerous to count (TNTC). A TNTC must be regarded as a “detection” and/or “higher



4.9 Salmonella

This method is applicable to all NMD Programmes; red meat and poultry.


Finland or to any other country with the same requirements, for example, Iceland, must use

the required sampling and method specified for routine official test 2.4.2

4.9.1 Sample preparation

Reference MIMM 2005, Chapter 7.7.

1. Carry out all quality control procedures for the media and methods using Salmonella

menston NZRM 383 as the positive control and E. coli NZRM 916 as the negative

control.

2. Sample preparation. Samples are derived either from separate carcass swabs taken

from a specific site sampled for Salmonella analysis only or from the primal cut swab,

bulk meat or whole carcass (poultry) suspension.


Laboratory Analysis

4.9.1.1 Red meat carcasses with a separate site swabbed for Salmonella analysis

See section 4.5 Table 26.

4.9.1.2 Poultry carcass rinse samples and red meat primal cut and bulk meat swab suspensions for Salmonella analysis

Poultry carcass rinse samples.

On receipt of the poultry carcass rinse sample transfer 30ml to a separate sterile vial for use

as the BPW suspension described in section 4.9.2.

Red meat.

See also Section 2.10 Table 7 for red meat species requiring Salmonella sampling/analysis

for cuts and or bulk.

For each of the primal cut initial swab suspensions take a specified volume of the diluent and

composite into one vial then add an equal volume of dsBPW to the composite sample.

Note: For VLT premises there will be only one cut and bulk sample each week. Thus there

will be no composite sample.

Table 27: Primal cut Salmonella pre-enrichment composite sample from swab suspensions

Bovine 100cm2 Bobby calf 25cm2 Cervine 25cm2 Caprine 5cm2

5 samples of 8ml = 40ml + 40ml dsBPW

= 80ml in total


= 80ml in total


= 32ml in total


= 10ml in total

VLT (very low throughput) premises with only one sample (no composite)

1 sample of 8ml = 8ml + 8ml dsBPW

= 16ml in total


= 16ml in total


= 16ml in total


= 2ml in total

Bulk meat – bovine, bobby calf, caprine species Salmonella enrichment composite

sample from five 25g bulk meat stomached suspensions:

• 5 x 1ml suspension diluent = 5ml + 5ml dsBPW = 10ml in total.

• For samples of 60-80VL trim add 0.05ml (50µl) of Tween 80 or 0.03ml (30µl) of

Terigol 7 to the enrichment volume.

For VLT premises where there is only one bulk meat sample:

• To 1ml of suspension dilution add 1ml dsBPW = 2ml in total.

• For samples of 60-80VL trim add 0.01ml (10µl) of Tween 80 or 0.006ml (6µl) of

Terigol 7 to the enrichment volume.

Bulk meat samples are not required for cervine, ostrich and emu, ovine or poultry.


Laboratory Analysis

4.9.1.3 Transport or retention of Salmonella samples for later analysis

For red meat carcass sample where the Salmonella sample sites/swabs are separate:

• Whether or not the swabs are suspended in ssBPW they must be transported to the

laboratory under normal transport requirements; maintaining temperatures between

0 ºC and 5ºC (not frozen), and not exceeding 10ºC.

For primal cut swabs and bulk meat ex APC and E. coli testing:

• These samples suspensions are then composited and mixed with dsBPW to form

the composite BPW suspension for Salmonella analysis. These BPW suspensions

must be transported or stored maintaining temperatures between 0 ºC and 5ºC (not

frozen), and not exceeding 10ºC. The original suspensions also need to be kept

below 10ºC during APC/E. coli analysis.

All Salmonella analysis must commence within 24 hours of the original sample collection.

Under special circumstances this may be extended to 30 hours, with reasons recorded.

4.9.2 Method

1. BPW suspensions must not be stored or left on the bench for long periods (such as over

the weekend) before incubation commences. Incubation must commence as soon as

possible after the BPW suspension is prepared.

Initiation of Salmonella analysis is defined as from the time the BPW suspension is

placed in the incubator, the start of the BPW pre-enrichment step.

2. Incubate the pre-enrichment samples at 35+/-1ºC or 37+/-1ºC (36+/-2ºC is not

acceptable) for 18-24 hours. Reference: Mills and Barea, (2003) AgResearch Client

Report (CR) 907.

After initiation of analysis, refrigeration of this pre-enrichment broth and/or the RVS

selective enrichment broth is not permitted at any time. All steps in the analysis must

be completed within the time temperature bounds described in this method.

3. Transfer 0.1ml of the BPW enrichment culture to 10ml of RVS selective enrichment broth

pre-warmed to 42ºC.

4. Incubate the RVS selective enrichment broth for 24 hours +/- 2 hours at 42+/-0.2ºC in a

water bath (preferable) or incubator certified as capable of control to this specific

temperature range. The temperature is critical for maximum recovery of Salmonella.

5. Transfer a loop-full (10µl) of the RVS culture to one plate of BGM agar and one plate of

XLD agar selective plating media. Plates must be labelled with agar type (BGM or XLD)

as the agar colour can change during incubation making the two types of media

indistinguishable. Streak to obtain single, well isolated colonies.


Laboratory Analysis

6. Incubate both plates for 18-24 hours at 35+/-1ºC or 37+/-1ºC (36+/-2ºC is not

acceptable).

7. Examine the plates for typical Salmonella colonies (refer to MIMM 2005 or later edition,

chapter 7.7, Table 1).

BGM agar: Pink surrounded by bright red medium.

XLD agar: Red with a black centre. H2S negative serotypes have red colonies without a black

centre.

If swarming bacterial growth covers the plates refer to MIMM 2005 or later edition, Chapter 7.7.5

for further instruction.

4.9.3 Confirmation of presumptive positive Salmonella

Select five or more single, well isolated, typical colonies from the selective plating media.

Use biochemical tests and/or commercial kits as listed in MIMM 2005 or later edition,

Chapter 7. Perform the agglutination reaction as per manufacture’s instructions. The five

colonies may be tested sequentially and if any one of these colonies is positive the sample is

deemed positive. The remaining colonies need not be tested.

4.9.3.1 Obtaining pure cultures of Salmonella for serotypical analysis

Streak colony from either selective media onto MacConkey agar. Incubate for 24h at 35+/-

1ºC or 37+/-1ºC (36+/-2ºC is not acceptable). Subculture typical yellow colonies to plate

count agar and incubate overnight at 35+/-1ºC or 37+/-1ºC (36+/-2ºC is not acceptable).

Refer to MIMM 2005 or later edition, Chapter 7.7.7 and Table 4 or 7.7.7.

4.9.3.2 Final confirmation and serotyping

Colonies may be confirmed in-house using Poly O and Poly H anti-sera. If either is positive,

submit purified colonies to ESR.

Where presumptive Salmonella has been confirmed as in 4.9.3 by latex agglutination purified

colonies must be submitted directly to ESR Kenepuru Science Centre, Porirua for final

confirmation and serotyping.

4.9.3.3 Reporting results

Where samples are composited for Salmonella testing express the result as Salmonella

“detected in a composite sample” or “not detected in a composite sample”. For poultry whole

carcass rinse samples report the results as “not detected/carcass”.


Laboratory Analysis

4.10 Poultry Carcass Campylobacter Direct Plate Enumeration Method

This method is applicable to the NMD poultry broiler programme for carcasses selected for

Campylobacter testing rinsed with 400ml of ssBPW.

If chemical decontamination washes were used during processing, ensure that non-

antimicrobial neutralising additives tailored to the decontamination concerned were added to

the ssBPW used for rinsing. Refer section 3.1.

4.10.1 Plating Media

Selective enrichment medium:

Modified charcoal cefoperazone desoxycholate agar (mCCDA).

Basal medium: (commercially available)

Lab-lemco powder 10.0g/l

Peptone 10.0g/l

Sodium chloride 5.0g/l

Bacteriological charcoal 4.0g/l

Casien hydrolysate 3.0g/l

Sodium desoxycholate 1.0g/l

Ferrous sulphate 0.25g/l

Sodium pyruvate 0.25g/l

Agar 12.0g/l


pH 7.4+/-0.2

Suspend 22.75g of dehydrated mCCDA base in 500ml or distilled water, mix well and boil to

dissolve the agar. Sterilise by autoclaving at 121ºC for 15 minutes.

mCCDA Selective Supplement (commercially available)

Vial quantities suitable for 500ml basal medium are:

Cefoperazone 16mg

Amphotericin B 5mg

Complete mCCDA medium

Cool autoclaved basal medium to about 50ºC. Aseptically add 1 vial of each selective

supplement to 500ml of mCCDA base, mix thoroughly and pour into Petri plates.

Plates can be stored for up to two weeks in sealed containers at 4ºC.

Prior to use, air dry plates (do NOT use an laminar flow cabinet), either by leaving unopened

on the bench overnight, or when plates are used on the day of preparation, in an incubator at

42ºC.


Laboratory Analysis

4.10.2 Direct Plating

Taking 2ml of rinsate from the rinse bag or sample container apply over six labelled,

appropriately dried, mCCDA plates. All of the 2ml must be dispensed onto the six plates.

The exact amount per plate of the 2ml does not need to be measured as all six plates will be

counted. Spread each aliquot on the agar surface with a sterile spreader. Allow the 6 plates

to remain upright at room temperature to permit the sample to soak in before inverting.

Plate out an additional 0.1ml of the rinsate onto each of an additional two mCCDA plates.

If higher numbers are expected set up a further 10 fold dilution series in ssBPW and plate

0.1ml of each dilution (in duplicate) onto mCCDA plates as described above. Note that a

suitable dilution series must be set up to minimise the occurrence of TNTC results.

Incubate in a microaerobic atmosphere (5% O2, 10% CO2, 85% N2). Ensure an appropriate

number of sachets for the jar/container are used. If too many sachets are used a moist

environment and condensate on plates will occur, which could influence colony growth and

cause colony spreading. Too many, or too few sachets, will not create the correct gaseous

mixture in the container.

Incubate for 48 hours +/- 2 hours at a single temperature of 42+/- 0.5ºC. The microaerobic

atmosphere must be maintained at all stages of incubation.

4.10.3 Presumptive Count

Select plates from dilutions of the 2ml set of 6 plates, or 0.1ml duplicate plates with not more

than 150 colonies on any plate. Counts of over 150 on any plate are considered TNTC as

Campylobacter colonies are so variable in size.

Examine these plates for characteristic thermotolerant Campylobacter spp colonies: flat grey

and moistened, variable size for 1mm – 5mm in diameter, from pinpoint colonies to roundish

2ml rinsate over 6 plates

0.1ml

0.1ml

0.1ml

0.1ml

0.1ml 0.1ml

Zero dilution 2ml rinsate plated over all 6 plates

0.1ml rinsate plated onto each plate

- 1 dilution 1:10 ssBPW dilution of rinsate, 0.1 plated onto each plate

- 2 dilution 1:10 ssBPW dilution of above, 0.1 plated onto each plate

DILUTION SERIES


Laboratory Analysis

or flattened out colonies with irregular spreading margins. Count and record the numbers of

characteristic thermotolerant Campylobacter spp. colonies on this set of plates.

Other organisms that may be present on mCCDA plates include creamy coloured yeast

colonies, which can easily be distinguished by 10x magnification, Arcobacter and

Pseudomonads.

Photograph of Campylobacter jejuni colonies and some creamy white yeast colonies on a

mCCDA plate, courtesy of Tegel Foods Limited, Christchurch.

4.10.4 Confirmation

Two methods of confirmation are permitted. Laboratories may elect which they will use

routinely, but must use their elected method on a regular basis to comply with their

accreditation requirements.

Confirmatory testing should be conducted on fresh cultures from mCCDA agar plates

immediately following incubation and presumptive count.

4.10.4.1 Oxidase/Latex confirmation

Select five characteristic colonies ensuring that any variation in colony size and shape is

included in the selection (or all colonies if there are less than 5 colonies).

a. Screen all 5 colonies for oxidase activity. Record result.

b. On the first colony that is oxidase positive carry out a latex agglutination test for

Campylobacter. The latex agglutination test confirms three Campylobacter species: C.


Laboratory Analysis

jejuni, C. coli, and C. lari. If the latex test is positive assume all 5 are Campylobacter

positive. If not conduct the latex test on the remaining oxidase positive colonies.

c. PCR may be used as an alternative to the latex agglutination assay confirmatory test.

d. If none of the five characteristic colonies selected confirm as Campylobacter, select a

further five atypical colonies and repeat steps (a-c). If there are no atypical colonies

present no further confirmation is required.

e. Small colonies may not contain enough cells for the oxidase test and latex test. It is

recommended that small colonies are subcultured to blood agar prior to confirmation.

Validation: Once a week choose a set of 5 characteristic Campylobacter colonies from

mCCDA agar plates that are oxidase positive and test all 5 with latex to validate the

presumption that an oxidase positive test will be latex positive.

Table 28: Example of confirmatory results by Oxidase/Latex

Typical colonies selected

Oxidase Latex Result per colony

1 positive Positive Campylobacter

2 positive Presume remaining oxidase positive are latex positive

Campylobacter

3 positive Campylobacter



Where the first oxidase positive colony selected of 5 records a latex negative result:

Typical colonies selected

Oxidase Latex Result per colony

1 negative not conducted Negative

2 positive Negative Negative



5 negative not conducted Negative

4.10.4.2 Confirmation and species identification as per MIMM sections 7.3.7 and 7.3.8

Select five characteristic colonies as per 4.10.4.1 (or all colonies if there are less than five

colonies).

a. For each colony carry out gram stain, motility, oxidase, hippurate hydrolysis and

antibiotic sensitivity using Nalidixic acid and Cephalothin as per MIMM sections 7.3.7 and

7.3.8.

b. Record as positive colonies that are gram stain negative with small “gull” shaped rods,

exhibiting wet mount motility, are oxidase positive and where the hippurate hydrolysis

result and antibiotic sensitivity confirm positive as C. jejuni, C. coli or C. lari.


Laboratory Analysis

c. If none of the five characteristic colonies selected confirm as Campylobacter, select a

further five atypical colonies and repeat steps (a-b). If there are no atypical colonies

present no further confirmation is required.

Table 29: Example of confirmatory results by MIMM

Colonies Selected MIMM 7.3.7 and 7.3.8 confirmation and species identification to C. jejuni, C. coli or C. lari

Result per colony

1 C. jejuni Campylobacter

2 Negative Negative

3 C. coli Campylobacter



4.10.5 Reporting results

Final count:

• If no thermotolerant Campylobacter spp colonies were confirmed report as “not

detected”. Refer to section 5.4, Table 30.

• Where thermotolerant Campylobacter spp colonies were confirmed the counts

obtained must be calculated according to section 5.2.2.

Report result as per the other NMD poultry analysis (Salmonella).


Results and Formulas

5. Results and Formulas

5.1 Result Calculation (Manual) Red Meat

Calculate the number of colony forming units (CFU) per cm2 or per g using the following

formulae.

Note:

1. That the swab suspension is not a dilution and is entered into calculations as the 100

dilution or zero dilution (non-diluted).

2. Although the whole tissue suspension is a 1:10 dilution, in order to maintain consistency

in plate labelling it is entered into calculations as the 100 dilution or zero dilution.

3. The dilution series values for the calculations below are to be mathematically expressed

as 100, 10-1, 10-2, 10-3, 10-4… Refer to section 4.1.

5.1.1 Aerobic plate count (spread plate)

Swab samples – 100cm2 area and 15ml suspension volume



Whole tissue samples – 25g sample + 225ml diluent = 250ml total volume

2 100cm

15ml x

0.1ml

1 x

Dilution

1 x

2

2 Count 1 Count = cm sq. / CFU

+

2 25cm

15ml x

0.1ml

1 x

Dilution

1 x

2


+

2 cm 5

10ml x

0.1ml

1 x

Dilution

1 x

2


+

25g

250ml x

0.1ml

1 x

Dilution

1 x

2

2 Count 1 Count = g / CFU

+



5.1.2 Aerobic plate count (PetrifilmTM)




Whole tissue samples – 25g sample + 225ml diluent + 250ml total volume

5.1.3 Aerobic plate count (spiral plater)

Calculate the number colony forming units per cm2 or per g using the procedures outlined in

the spiral platers operating instructions. The typical volume for the inoculum is 50µl = 0.05ml

or 100µl = 0.1ml.




2 5cm

10ml x

ml Inoculum

1 x

Dilution

1 x

2


+

2 100cm

15ml x

1ml

1 x

Dilution

1 x

2


+

2 25cm

15ml x

1ml

1 x

Dilution

1 x

2


+

2 5cm

10ml x

1ml

1 x

Dilution

1 x

2


+

2 100cm

15ml x

ml Inoculum

1 x

Dilution

1 x

2


+

2 25cm

15ml x

ml Inoculum

1 x

Dilution

1 x

2


+

25g x

1ml

1

Dilution

1

2


+ x x

250ml




5.1.4 Escherichia coli (PetrifilmTM)





5.2 Result Calculation (Manual) Poultry

Calculate the number of colony forming units (CFU) per carcass of rinse sample using the

following formulae.

5.2.1 Campylobacter enumeration

For counts obtained from 2ml spread over six (6) plates:

CFU/carcass = *(number colonies confirmed as Campylobacter/n) x count characteristic

Campylobacter morphology colonies (plate 1 + plate 2 + plate 3 + plate 4 + plate 5 + plate 6)

x 400ml/2ml

= number of Campylobacter organisms/poultry carcass sample.

For duplicate plates of higher dilutions:

CFU/carcass = *(number colonies confirmed as Campylobacter/n) x count characteristic

Campylobacter morphology colonies (plate 1 + plate 2)/2 x 400ml/0.1ml x 1/dilution

= number of Campylobacter organisms/poultry carcass sample.

n = number or characteristic colonies examined, usually 5 unless there are less than 5

characteristic colonies altogether.

25g

250ml x

ml Inoculum

1 x

Dilution

1 x

2


+

2 100cm

15ml x

ml 1

1 x

Dilution

1 x

2


+

2 25cm

15ml x

ml 1

1 x

Dilution

1 x

2


+

2 5cm

10ml x

ml 1

1 x

Dilution

1 x

2


+

25g

250ml x

ml 1

1 x

Dilution

1 x

2


+



* Usually five/five if the first colony of five is confirmed as positive. Will be a proportion of

five if the remaining colonies are required to be confirmed e.g. three/five.

5.3 Result Calculation (Automated)

Laboratories and operators must use web based data entry or other that would generate

NMD data for submission to NMD in an equivalent manner. The web based data entry takes

raw counts and dilution data and automatically performs all calculations, provides descriptive

statistics, and graphical profiles, and provides an ongoing database for storage of

information.

5.3.1 Procedure

Enter premises demographics and sample descriptors such as shift, day, time of day, etc., by

selecting check boxes, etc., within the spreadsheet.

Enter data into web based data entry or Microsoft Excel spreadsheet. Follow instructions

supplied with the software.

Authorise results:

a. Laboratory – Reconcile data entered into the spreadsheet against the laboratory

workbook, and sign off by the LAS signatory.

b. Premises – Where premises personnel independent of the laboratory enter authorised

laboratory results into the spreadsheet, the results for submission to the NMD must be

signed off by authorised, technically competent premises personnel.

5.4 Limits of Detection

Not detected results are considered to be results less than the lowest limit of detection

(LLD), which is Count plate 1 = 0, Count plate 2 = 1 (or vice versa) on a zero dilution pair of

duplicates.

Note: Because CFU/cm2 results (arithmetic) are transformed to logarithmic Log CFU/cm2

results for NMD reporting all results above the LLD, even though they may be negative

logarithmic values, are to be considered as detections/counts. For example the LLD value of

0.075 CFU/ cm2 for a PetrifilmTM E. coli 100 cm2 sample site converts to – 1.12 Log CFU/ cm2

which represents a count, a detection, even though it is a negative log value.

The values used to represent LLD and “not detected” results in the NMD database are listed

in the table below.



Table 30: Limits of detection

Lowest Limit of Detection Value “Not Detected”

APC CFU/cm2 or /g

APC Log10 value

E. coli CFU/cm2 or /g or /ml

E. coli Log10 value

Log10 value

Spreadplate 0.1ml

5cm2

25cm2

100cm2

Bulk Meat (25g)

10

3

0.75

50

1.00

0.48

-0.12

1.70

Not applicable

Not applicable

-0.31

-0.53

-1.13

-0.31

PetrifilmTM 1ml

5cm2

25cm2

100cm2

Bulk Meat (25g)

1

0.3

0.075

5

0.00

-0.52

-1.12

0.70

1

0.3

0.075

5

0.00

-0.52

-1.12

0.70

-0.31

-0.53

-1.13

-0.31

Spiral plater 50µl

5cm2

25cm2

100cm2

Bulk Meat (25g)

20

6

1.5

100

1.30

0.78

0.18

2.00

Not applicable

Not applicable

-0.31

-0.53

-1.13

-0.31

Campylobacter direct plating

CFU/carcass

Log10 value per carcass

“Not detected” Log10 value per carcass

2ml 200 2.30 2.00

If the number of colonies on the highest dilution plate exceeds the maximum allowable

count; for example>300 APC spreadplate,>150 PetrifilmTM E. coli enter that result as TNTC.

Such a result is classified as a “too numerous to count” (TNTC) technical failure.

Notwithstanding omission from the database, the result must be regarded as a “detection”

and/or “higher than expected count” when assessing compliance with premises performance

criteria under HACCP and RMPs.

The TNTC result must still be used at premises level for assessing compliance with “in-

house” targets and HACCP performance criteria.

For mCCDA Campylobacter where the colonies on the highest dilution plate exceed the

maximum allowable count the result must be classified as TNTC, reported and will default to

a greater than 3.78 Log10 CFU/carcass result of 3.79 Log10 CFU/carcass on the database.

See section 6.8.1.



5.5 Reporting of Results

5.5.1 General requirements

The following are general requirements for process and sample descriptors to be used for

reporting results:

• All reports to the NMD must clearly indicate the premises from which samples were

taken and any required process descriptor.

• All reports to the NMD must have complete sample description information

according to the product.

• Red Meat: species, APC method used, sampling week, class, shift, time, boning

process, cut product description, cervine cuts (skinned or not skinned), bulk meat

VL/CL grade, bulk product description, chiller identification, chilling period of time.

• Poultry: sample date, shift, sample time, method of drainage, carcass rinse location,

1% Tween 80 addition, rinse diluent temperature, carcass to laboratory time (when

carcass is transported from the premises to be rinsed in the laboratory), time

analysis initiated, cut number, average age of birds, farm reference number, shed

number.

• Results are to be submitted to NMD in the required format, in Log10cfu/cm2 or /g, /ml

or carcass units, or “detected”/”not detected” for pathogen presence/absence tests.

• If colonies are not detected express the results as the “not detected” assigned value

e.g. -1.13, -0.53, -0.31.

• Results must be entered directly by web based data entry to MPI.

• Data provided manually will not be accepted unless agreed by the NMD

Administrator.

• Entering a subset of results from more substantial sampling programme is allowed.

However, in order to avoid biasing the NMD, the samples from which results would

be entered must be selected prior to collection of the samples, as per random

sample selection, not after the results had been obtained.

5.5.2 Reporting for the database to premises and use of data

The NMD administration must:

• Post on the MPI web site, summary reports to the NMD National Microbiological

performance profiles on a quarterly basis.

• Use the RMP identifier when reporting the ranked list to the operator.

Quarters for reporting are defined as follows:

1st Quarter: January 1 – March 31



2nd Quarter: April 1 – June 30

3rd Quarter: July 1 – September 30

4th Quarter: October 1 – December 31

• Provide reports as soon as possible after completion of a quarter, preferably within

one (1) month.

• hold results from individual premises in the strictest confidence from other

contributors. Individual premises data will only be available to the operator who

provided it, and any LAS approved laboratory supplying data on behalf of the

operator.


Targets – Premises Analysis and Interpretation, Independent Verification

6. Targets – Premises Analysis and

Interpretation, Independent Verification

6.1 Premises Level Targets

All premises shall set premises level microbiological targets (all species) and Salmonella

performance standards (excluding ovine species), and present NMD data in a manner that

facilitates early identification and visualization of loss of process control.

6.2 Bovine Species NMT: Escherichia coli Moving Window

The NMT consist of two levels of target administered on an ongoing basis by the operator:

Moving window (m) based on the percentage of samples that exceed the 80th percentile. For

bovine E. coli the 80th percentile approaches not detected. Thus E. coli detected/not detected

(presence/absence) is used as the limit. For m-alerts a carcass is considered a single entity. A

detection of E. coli on any carcass site renders the carcass as a whole as detected.

An upper limit (M) based on the 98th performance percentile (M) of E. coli numbers, as specified

in the US Pathogen Reduction/HACCP Final Rule (“MegaReg”), which is 100cfu/cm2. This is

converted to logarithmic notation for NMT purposes.

100cfu/cm2 = 2.00 log10cfu/cm2. Results from each carcass site are considered separately. The

3 carcass site results are not composited.

6.2.1 NMT (bovine species): Escherichia coli moving window (m)

A five (5) sample increment “moving window”. Addition of 5 new samples to the bottom of the

window displaces 5 samples from the top.

An overall window size of 15 samples (3 weeks). NMT shall be implemented for each

product: carcasses (post-slaughter and dressing), primal cuts, and bulk meats.

In this case, the carcass is considered a single entity. If any of the three sites of a carcass

are positive, the carcass as a whole is considered positive.

A premises may detect E. coli in four (4) samples of product type within the 15 sample (3 week)

window, without response.

Detection of E. coli in five (5) samples or more shall elicit an “Alert” response.

An “Alert” response shall consist of immediate review of the process by the operator to

identify and document factors, that may have compromised hygienic processing, and if

appropriate, take corrective and preventative action.



MPI VA need not be notified. However, the company response to an “Alert” shall be verified

during routine MPI VA audits.

6.2.2 NMT (bovine species): Escherichia coli upper limit (M)

A NMT must be implemented for each bovine product: carcass post slaughter and dressing,

primal cuts, and bulk meats. This NMT must be implemented for each site on the bovine

carcass; thus there are three (3) sets of five (5) results for carcasses each week, five (5)

rump (hindleg), five (5) flank and five (5) brisket sets of results to consider.

• For M-alerts results from each carcass site sample are considered separately. The

3 carcass site results are not composited.

• Premises may detect E. coli in numbers greater than 2.00 log10cfu/cm2 in one (1)

carcass site, cut or bulk sample for bovine product type in any given sampling week.

• Detection of E. coli in numbers greater than 2.00 log10cfu/cm2 in two (2) carcass

sites, cut or bulk sample, in a sampling week must elicit an “Alert” response.

Table 31: Carcass example (units 2.00 log10cfu/cm2)

Carcass Site Rump (hindleg) Flank Brisket

Carcass 1 >2.00 <2.00 <2.00

Carcass 2 >2.00 <2.00 <2.00

Carcass 3 <2.00 <2.00 <2.00

Carcass 4 <2.00 <2.00 >2.00

Carcass 5 <2.00 <2.00 <2.00

M-Alert Yes No No

Bovine carcasses must be listed with M-alert status if one or more carcass sites generate a

M-alert.

An “Alert” response must include immediate review of the process by the premises to identify

and document factors that may have compromised hygienic processing, and, if appropriate,

take corrective and preventative action.

The premises MPI VA verifier must immediately be notified, and must prior to company

review of the process, assess and approve the review procedures according to the principles

outlined in Section 5.10 (Processing Conformance) or IS-8 (ref. 5.1 & 5.2).

6.3 Bobby Calf NMT: Escherichia coli Moving Window

6.3.1 Bobby calf product types

For m-alerts a carcass is considered a single entity. The arithmetic (cfu/cm2) results from the

three separate carcass sites samples are composited by averaging and the expressing this

arithmetic average as a single log10cfu/cm2 results per carcass.



For M-alerts results from each carcass site sample are considered separately. The 3

carcass site results are not composited.

Primal cut and bulk meat results are taken as individual sample results in log10cfu/cm2 or /g

units.

6.3.2 Bobby calf NMT 80th percentile m limits: Escherichia coli

Analysis of data from a 15 sample moving window, (15 carcass, 15 cuts and 15 bulk results)

canvassing 3 weeks is required. The addition of the 5 latest samples to the window

displaces the oldest dated 5 samples.

The m-alert limit is bases on the 80th percentile E. coli values of NZ industry 25cm2 bobby

calf data from July 2002 to December 2002.

Carcasses post slaughter and dressing 1.23 log10cfu/cm2 (whole carcass average)

Primal Cuts 0.32 log10cfu/cm2

Bulk meat 2.08 log10cfu/g A premises may detect E. coli >80th percentile limit in four (4) samples of a product type

within a 15 (3 week) sample window without response.

Detection of E. coli counts exceeding the 80th percentile in five (5) or more samples within a

15 sample (3 week) window for any product type shall elicit a m-alert response.

The m-alert response requires an immediate review of the process by the company to

identify and document factors that may have compromised hygienic dressing. Corrective

and preventative actions shall be taken if required.

MPI VA need not be notified. The company must record its responses to the m-alert, as this

will be reviewed during routine MPI VA audits.

6.3.3 Bobby calf NMT M limits: Escherichia coli

M-alerts apply to each carcass site of five (5) carcass samples, and five (5) cut and bulk

sample results (one week’s results). There are three (3) sets of five (5) results for carcasses

each week, five (5) fore rump, five (5) flank and five (5) fore leg sets of results to consider.

The bobby calf M-alert is based on the 98th percentile of E coli results from NZ industry

25cm2 bobby calf data from July 2002 to December 2002. The M value is set at not less

than the FSIS “M” of 100 cfu/cm2 = 2.00 log10cfu/cm2.

Carcasses post slaughter and dressing 2.11 log10cfu/cm2

Primal Cuts 2.00 log10cfu/cm2

Bulk meat 2.98 log10cfu/g Premises may detect E. coli in numbers greater than “M” in not greater than one (1)

sample for a bobby calf product type in any given sampling week (5 samples).



Detection of E. coli in numbers greater than “M: in two (2) or more samples for any bobby

calf product type in sampling week (5 samples) shall elicit an M-alert response.

Table 32: Carcass example (units log10 cfu/cm2)

Carcass Site Fore Rump Flank Fore Leg

Carcass 1 >2.11 >2.11 >2.11

Carcass 2 <2.11 <2.11 <2.11

Carcass 3 <2.11 >2.11 <2.11

Carcass 4 <2.11 >2.11 <2.11

Carcass 5 >2.11 <2.11 <2.11

M-Alert Yes Yes No

Bobby calf carcasses must be listed with M-alert status if one or more carcass sites

generate an M-alert.

An M-alert response must include immediate notification to the MPI VA and immediate

review of the process by the premises to identify and document factors that may have

compromised hygienic processing. When factors are identified corrective and preventative

action is to be implemented.

MPI VA must, prior to company review of the process, assess and approve the review of

procedures according to the principles outlined in Section 5.10 (Processing Conformance)

of IS-8 (ref, 5.1 & 5.2).

6.4 Ovine NMT 95th Percentile m Limits: APC

The ovine species APC NMT requires analysis of APC data from a fifteen (15) sample

moving window; single Y-cut site, five (5) carcasses, three (3) week period. Addition of the 5

most recent samples to the window displaces the 5 least recent samples.

The ovine APC m-alert is based on the 95th percentile of APC results from NZ industry ovine

fresh lamb carcass Y-cut site 5cm2 APC data over two years of NMD data from the

beginning of January 2002 to the end of December 2003. The 95th percentile ovine APC m-

alert value over this range of results is 4.65 log10 CFU/cm2.

Detection of an APC value greater than “m” (4.65 log10 CFU/cm2) in three (3) or more

individual samples in a 15 sample (3 week) moving window is classified as an “m-alert”.

In contrast to the NMT for other species, the moving window “resets” on eliciting an “m-alert”.

Non-conforming samples (3 or more APC values greater than “m”) do not carry over into

subsequent sampling windows, i.e. they contribute towards just one “m-alert”.

Responses to ovine “m-alerts”:

1. First “m-alert”. Premises shall:



• review the process,

• identity contributing factors,

• propose preventative measures, and

• reset window.

2. Second “m-alert” in the moving window immediately after the first “m-alert” Premises

must:

• inform MPI VA (premises level), who will review the process and the effectiveness

of the implementation of the preventative measures or changes proposed in

response to the first “m-alert”, and

• reset window.

3. Third “m-alert” in the moving window immediately after the second “m-alert” Premises

must:

• inform MPI, who will carry out an independent regulatory review,

• carry out a full review of HACCP plans,

• implement process changes as required,

• reset window.

Actions taken, including any sanctions, will be subject to ongoing verification of compliance.

6.5 NMT: Documentation and Record Keeping

All operators must document the system by which they monitor performance against the

NMT, and perform any required process reviews, according to the requirements of section

5.10 (Process Conformance) of IS-8.

Documentation must include:

• authorities;

• review procedures on notification of an “Alert”;

• corrective actions;

• preventative actions.

The verifier must validate documentation as per the requirements for implementation of IS-8,

and shall assess and approve the documented process review procedures prior to

implementation.

All operators shall record the results of review procedures and consequent corrective

and preventative actions implemented when exceeding a premises level target or on

notification of an “Alert”.



All documents shall be reviewed during routine MPI VA verification audits.

6.6 Ranked List

6.6.1 Bovine and bobby calf

For bovine and bobby calf the percentage of APC results greater than the published national

80th percentile (Table 33) will be calculated for each premises each report quarter. Bovine

and bobby calf E. coli results will be compared against the values specified by the bovine

and bobby calf Escherichia coli NMT targets (sections 6.2 and 6.4) each quarter. Note that

for bovine E. coli the 80th percentile is taken as “not detected” and the percentage detected

is calculated for ranking purposes.

Table 33: Bovine and bobby calf APC national 80th percentile NMT limits

BOVINE3 BOBBY CALF4

Bovine Sites/Cuts/Bulk

Bovine APC NMT Bobby Calf Sites/Cuts/Bulk

Bobby Calf APC NMT

Carcass (rump hindleg) Carcass (flank) Carcass (brisket) Primal cuts Bulk meat

1.77 log10 CFU/cm2

1.69 log10 CFU/cm2

0.95 log10 CFU/cm2 2.01 log10 CFU/cm2 3.53 log10 CFU/g

Carcass (fore-rump) Carcass (flank) Carcass (foreleg) Primal cuts Bulk meat

2.03 log10 CFU/cm2 2.85 log10 CFU/cm2 2.13 log10 CFU/cm2 2.82 log10 CFU/cm2 4.07 log10 CFU/g

6.6.2 Ovine

Each quarter ranked lists will be published based on fresh carcass APC results for the

opening Y cut site. Premises will be ranked in descending order of the percentage of results

greater than the national profile ovine carcass opening Y cut APC 80th percentile all data to

date value.

Premises with percentage APC exceeding the 80th percentile that fall into the following

category

• greater than 2 x the average, and prevalence statistically (Chi-square) of P <0.05

will be bolded for microbiological performance information purposes only. Alerts as

described in section 6.6.4 will not be elicited.

3Derived from 100cm2 NMD data for the year to 31 December 2007.

4All bobby calf 25cm2 data from July 2002 to December 2002.



6.6.3 Caprine, cervine, ostrich and emu and porcine

For cervine, caprine, ostrich and emu, and porcine the national profiles 80th percentile values

for APC and E. coli all data to date will be used to compare premises data against to compile

the quarterly ranked lists.

6.6.4 Ranking and alerts

Premises will be ranked in descending order of APC/E. coli prevalence based on the 80th

percentiles/NMT for the particular species. The average prevalence will be calculated.

Premises with percentage APC/E. coli exceeding the 80th percentile that fall into the

following two categories shall elicit an “Alert”:

• greater than 2 x the average, and prevalence statistically (Chi-square) of P <0.05

• less than 0.5 x the average, and prevalence statistically (Chi-square) of P<0.05

Premises that elicit an upper or lower “Alert” shall be highlighted in bold on a ranked list

issued by MPI on a quarterly basis.

6.6.4.1 Upper alert

Notified premises must review the process to identify and document factors that resulted in a

prevalence significantly higher than the industry average. The review is likely to result in

modification of process control programmes programmes (pre-requisite and HACCP).

6.6.4.2 Lower alert

A lower alert highlights a very low percentage prevalence. It is accepted that this result may

be a reflection of consistent application of good hygienic dressing practices by the premises

rather than any issue with implementation of the NMD sampling programme.

Notified premises must review the NMD sampling programme; in particular sample

collection, transportation (time/temperature), and laboratory procedures, to identify and

correct any deficiencies that may impact on microbiological results.

Where there are successive recurring lower alerts and the following points are met, the MPI

verifier may decide that further reviews are no longer required:

• There is a history of compliance with NMD microbiological sampling requirements;

and

• Review of both past approved laboratory audits (refer NMD Notice clause 10, sub

clause (5)), and NMD verifications completed by the MPI verifier are found to have

been acceptable;

If after a period time outside the lower alert category the premises ranked list includes a

lower alert then the review process must be reinstated.



6.6.5 Poultry quarterly report ranked list

Poultry premises are ranked in descending order of their Campylobacter averages over the

last completed quarter.

Poultry premises are bolded high and low where the premises average is greater or less

than +/- 0.5 log10 CFU/carcass of the average of all poultry results for that quarter.

6.7 Salmonella Performance Standards (SPS)

6.7.1 Group 1 and porcine Group 2– Salmonella performance standard

Absence of Salmonella is the desired objective, and the presence of these organisms must

not be overlooked, if and when detected. The contributing factors and source of Salmonella

must be investigated.

Upon detection of Salmonella the operator must undertake the following actions addressing

product, livestock and personnel:

• Immediately inform the MPI VA verifier of the detection.

• Ensure the laboratory has submitted purified cultures of isolates detected to ESR

Enteric Reference Laboratory at National Centre for Biosecurity and Infectious

Disease (NCBID), Wallaceville for serotypic confirmation.

• Ensure records from the original product sampled are traced back to the catchment

area of the stock being processed.

• Consult the laboratory(s) providing an animal health service in the stock catchment

area, and inquire as to any increase in reported cases of animal salmonellosis.

• Determine if symptoms of salmonellosis have been reported by any personnel.

In addition to the above actions related to product, livestock and personnel the operator must

undertake the following process responses which escalate according to the frequency of

Salmonella detections in the season.

Monitoring and control of this pathogen is also of interest to overseas authorities. The

appropriateness of country listings of an operator will be reconsidered by MPI VA when there

are ongoing detections of Salmonella at the premises.

6.7.1.1 First detection

On the first detection for the season in a PSW or SSW the operator must:

• Review the entire process.

• Identify contributing factors.

• Implement corrective and preventative measures.



6.7.1.2 Second detection

On the second detection in a subsequent PSW or first PSW following a first detection in a

SSW the operator must:

• Under take a further review as per the first detection.

• Review the effectiveness of the corrective and preventative actions implemented in

the first detection.

• Ensure any new or existing preventative and corrective actions are implemented.

• Inform the MPI VA primary verifier, who will review the process and the effectiveness

of the operator’s actions in response to the first and second detections.

6.7.1.3 Third detection

On the third detection in a subsequent third PSW or second PSW if the first detection was a

SSW the operator must:

• Undertake a further review as per the first detection.

• Carry out a full review of HACCP plans.

• Reassess the effectiveness of corrective and preventative actions implemented in

the first and second detections.

• Implement further corrective and preventative actions to address previous or newly

identified contributing factors.

• The premises shall submit a Salmonella Management Plan, describing

process/HACCP reviews and the measures implemented to reduce the prevalence

of pathogens, to MPI VA.

The MPI VA verifier must:

• Review operator corrective and preventative actions; and

• Review the operators Salmonella Management Plan and submit to MPI VA

Specialist Advisor.

Actions taken, including any sanctions, will be subject to ongoing verification of compliance.

6.7.2 Group 3 – Salmonella performance standard

The Poultry Salmonella performance standard is based on the US Performance Standard.

Operators are recommended to monitor the prevalence of Salmonella according to the US

Performance Standard, PR/HACCP Salmonella Performance Standards described in Table

2 Federal Register/Volume 61, No. 144 page 38867.



The US Standard states that Salmonella may be detected in no more than 12 of 51

consecutive poultry samples.

It is recommended that on breaching the US Performance Standard operators immediately

review the process and livestock Salmonella status to identify and document factors that

resulted in breach of the performance standard. This may lead to modification of process

control programmes (pre-requisite and HACCP).

6.8 Poultry Campylobacter Performance Target (CPT)

The poultry Campylobacter performance target (CPT) consists of two regulatory limits

requiring Campylobacter analysis of poultry broiler carcass rinse samples over set

processing periods as follows:

• Standard processing premises, a total of three samples must be taken per

processing day. Each of the three samples must be collected at a separate

randomly selected sampling time per processing day. A processing period is five

days processing equalling a total of 15 samples.

• Very low throughput (VLT) premises, a total of three samples on a single

randomly selected day of one processing week must be randomly selected over

available processing times. A processing period is one processing week equalling a

total of three samples.

Table 34: Table of CPT sampling requirements

Sampling period Standard throughput Very low throughput (VLT)

A moving window of

three processing

periods.

45 samples over 15

processing days.

9 samples over 3 weeks.

The moving window is defined as three processing periods. The addition of the samples of

the latest processing period displaces the samples of the oldest processing period.

Two CPT regulatory limits are used to determine compliance over each moving window:

• Number of samples with a result of greater than 6000 CFU/carcass, 3.78

log10CFU/carcass and

• Number of positive samples; those samples with a result of 2.30 log10CFU/carcass

or higher representing Campylobacter detection.5

5 A ‘not detected’ Campylobacter result will be recorded as 2.00 log10CFU/carcass on the

NMD database.



Transitional arrangements:

The new requirements in this Schedule will be implemented from Monday 7 January 2012.

The first processing period under the new system will start on the first day of processing on

or after that date. All premises will be reset to “compliant” on that date.

6.8.1 Recording of sample descriptors and default results

Each sample must have its sample descriptors recorded on the NMD database; sample time,

farm reference number, shed number, cut number and average age of birds in that cut.

Failure to sample

If less than the required number of samples have been collected during a processing period

the missed samples will each default to a greater than 3.78 log10 CFU/carcass result

recorded as 3.79 Log10 CFU/carcass on the database.

Technical Failures

Samples which have been collected, but where a technical failure (TF) has not permitted a

result the sample descriptors must be entered as proof of sampling with TF recorded in the

result field. Entering the sampling descriptors of samples taken ensures that a 3.79 default

result is not generated.

Too numerous to count results

Too numerous to count (TNTC) results must be reported. Each TNTC result will default to a

greater than the 3.78 log10 CFU/carcass result; recorded as 3.79 Log10 CFU/carcass on the

database.

6.8.2 CPT non-compliance

There are two classes of CPT non-compliance

(1) Enumeration Failure (EF):

An EF will be generated upon detection of a value greater than 6000 CFU per carcass

(3.78 log10CFU/carcass) in:

a. Standard throughput: premises: seven (7) or more out of 45 individual

carcass samples taken from a 3 successive processing period moving

window; OR

b. VLT premises: two (2) or more out of 9 individual carcass samples taken

from a 3 successive processing period moving window.

(2) Detection Failure (DF)



A DFwill be generated upon a result of 2.30 log10CFU/carcass or greater in:

a. Standard throughput: premises: 30 or more out of 45 individual carcass

samples taken from a 3 successive processing period moving window; OR

b. VLT premises: Six (6) or more out of 9 individual carcass samples taken

from a 3 successive processing period moving window.

If the premises has an EF, a DF or both for a moving window it is counted as one non-compliant

window. Responses to CPT escalate according to the number of consecutive non-compliant

moving windows. To clear the non-compliance a moving window without an EF and without a DF

is required. The database then resets to zero to show that the premises is compliant.

Note that a noncompliance will be recorded in the database as soon as the EF or DF becomes

evident (which may be before the results from all samples for that moving window have been

entered). This enables corrective actions to be initiated at the earliest opportunity.

6.8.3 Required responses to CPT non-compliance

The premises NMD controller must check the NMD results at least once every processing period to

determine whether or not the premises is CPT compliant.

The NMD Controller must notify the operator and the MPI-assigned verifier within 24 hours of

determining each non-compliant moving window.

Responses escalate with each consecutive non-compliant moving window. With each non-

compliant moving window the investigations, corrective actions undertaken and further actions

planned to restore control must be recorded by the NMD Controller in the NMD ledger.

The following responses must be undertaken.

Consecutive non-

compliant moving windows

Response Required

1 Within 1 week of a noncompliant window being reported in NMD:

• the NMD Controller must:

o notify the Operator and the , MPI-assigned premises

verifier, and

o indicate that this has been done in the NMD ledger, and

• the Operator must initiate corrective actions to restore control.

2 As soon as a 2nd consecutive noncompliant window is reported in NMD:

• the NMD Controller must notify the Operator, and

• the Operator must document the investigations done, corrective

actions taken to date and further actions planned to restore



control, and

• the Operator must copy this information to the MPI-assigned

premises verifier, and

• the NMD Controller must indicate that this has been done in the

NMD ledger.

As soon as possible the MPI- assigned premises verifier must:

• review the actions and if necessary visit the premises or ask for

additional information to ensure that the actions are appropriate,

and

• indicate that this has been done in the NMD alert screen, and

• report any concerns to nominated MPI managers/technical

people.

If the MPI-assigned premises verifier or an MPI manager/technical person

is not satisfied that the actions are appropriate they may notify this to an

appropriate MPI Director who may require an immediate response as per

7 consecutive non-compliant windows.

3 As for non-compliance 2 with information updated on the NMD ledger by

both the NMD controller and the MPI-assigned premises verifier.



5 As for non-compliance 4 with information updated with information

updated on the NMD ledger by both the NMD controller and the MPI-

assigned premises verifier.



The operator must document any product disposition options they could

implement in order to minimise the risk of contaminated product reaching

the consumer. The product disposition options must be provided to the

MPI-assigned premises verifier.



Responses to be undertaken continued:

Non-compliant

moving window

Expected response



The MPI-assigned premises verifier must:

• provide all information received to date to a nominated MPI

Campylobacter expert.

The MPI Campylobacter expert must:

• review the actions taken and the results to date then recommend

to an MPI Director whether or not to initiate a Campylobacter

Response Team (CRT) visit to the non-compliant premises and

which experts should be in the team.

The MPI Director must:

• sign-off the decision to initiate the CRT visit, and nominate a CRT

Leader and any other relevant experts to form the team, or

• sign a statement declining the recommendation with associated

justification.

If authorised by the MPI Director, the CRT must visit the premises at the

first available opportunity to:

• review all actions to date and recommend to the Operator other

corrective actions likely to bring the premises into compliance,

and

• where necessary, require corrective actions;

• where necessary, recommend the application of sanctions as per

non-compliance level 8 under Section 89 of the Animal Products

Act 1999 to protect the consumer.

The Response Team Leader must:

• provide a report to the Operator summarising the visit findings, a

required action plan and recommendations.

• copy the report to the MPI-assigned premises verifier, and the

MPI Director who approved the visit.

The Operator must:



• comply with the required action plan unless an alternative is

agreed and signed off by the Response Team Leader and copied

to the premises verifier, and

• consider the recommendations.

The MPI-assigned premises verifier must:

• monitor the actions taken and

• report any concerns to the CRT Leader.

If actions are not taken as agreed then non-compliance 8 response is

required.

8 MPI must apply sanctions under Section 89 of the Animal Products Act to

protect the consumer. The sanctions may include, but are not limited to

one or more of the following:

• Revisit(s) by the CRT and further required actions /

recommendations by the CRT

• Increased verification

• Full-time supervision of processing

• Introduction of further interventions

• Product disposition

• Further sampling and research initiatives and

• Premises closure.

When any of the above sanctions are applied they must remain in place

until revoked by an Animal Products Officer

• After the premises has a compliant moving window or

• At the direction of an MPI Director.



6.9 Verification Requirements

MPI VA/the verifier must verify that premises comply with the NMD Programme according to

the Performance-Based Verification procedures to be issued by MPI. In addition, on two

occasions in each processing season (Oct 1 to Sept 30), NZFSA VA/the verifier shall verify

the collection, packaging and reporting of a complete NMD sample set against the NMD

specification, by:

• observing the rotational and/or random selection of sampling day, shift, run, time and

carcass/cut/carton to be sampled.

• observing the collection of samples, including documentation of sample descriptors

and chronological information.

• apply a MPI carton seal to a set of NMD samples to permit a check with the

receiving laboratory to verify samples are received in the correct

condition/temperature and that analyses are commenced in the required time from

time of sampling.

• observing the storage of samples prior to packaging, and subsequent packaging of

samples for transport.

• requesting and observing the laboratory records and reports for samples transported

under the MPI carton seal.

• observe and verify chiller exit sampling.

• confirm submission of current demographics to NMD Administrator.

• confirm commencement of appropriate Salmonella sampling programme (16 week

PSW, 6 week SSW, or continuation of PSW requirements for the season).

• confirm reporting of Salmonella sampling programme results and completion of PSW

or SSW for the season.

• confirm implementation of Campylobacter sampling programme where required.


References

7. References

Animal Welfare (Broiler Chickens: Fully Housed) Code of Welfare, Biosecurity NZ, 25 July

2003.

AOAC (1990) Official Methods of Analysis of the Association of Official Analytical Chemists,

15th edition, Vol. 1, (ed. K. Helrich), Association of Official Analytical Chemists Inc., Arlington,

Virginia, USA.

Bell, R.G., J.C.L. Harrison and A.R. Rogers (1994) Distribution of microbial contamination on

beef and lamb carcasses. Proc. 28th Meat Industry Research Conference, Meat Ind. Res.

Inst. N.Z. Publ. No. 942.

Bell, R.G and S.C. Hathaway (1996) The hygienic efficiency of conventional and inverted

lamb dressing systems. J. App;. Bact. 81: 225-234.

Busta, F.F., E.H. Peterson, D.M. Adams and M.G. Johnson (1984) Colony count methods.

In: Compendium of Method for the Microbiological Examination of Foods, 2nd edition (ed.

M.L. Speck), pp.62-83, American Public Health Association, Washington DC.

Cook, R.L. (1991) Microbiological Methods of the Meat Industry, 2nd Edition. Meat Ind. Res.

Inst. N.Z., Publ. No. 873.

Donnison Procedures for Sampling, Handling & Storage of Campylobacter Isolates PDF 545

kb) July 2003.

Donnison Isolation of Thermotolerant Campylobacter – Review & Methods for New Zealand

Laboratories (PDF 1,809 kb) Revised May 2003.

FSIS, National Advisory Committee on Microbiological Criteria for Foods, Analytical Utility of

Campylobacter Methodologies, adopted September 28, 2005.

Hathaway, S.C. & R.L. Cook and P. van der Logt New Zealand National Microbiological

Database for “Minor Species” Summary of Findings and Recommendations for Sheep.

ICMSF (1978) Indicator microorganisms. In: Microorganisms in Food 1. Their Significance

and Methods of Enumeration. 2nd ed. Pp. 3-13. University of Toronto Press, Toronto.

ICMSF (1986) Microoganisms in Foods 2. Sampling for Microbiological Analysis: Principles

and Specific Applications. 2nd ed. International Commission on Microbiological

Specifications for Foods. University of Toronto Press, Toronto.

ISO (1990) General Requirements for the Competence of Calibration and Testing

Laboratories. ISO/IEC Guide 25:1990.

ISO 17604:2003 Microbiology of food and animal feeding stuffs – Carcass sampling for

microbiological analysis


References

Meat Industry Microbiological Methods Edition Four, March 2005, AgResearch, Hamilton,

New Zealand. Abbreviated as MIMM 2005.

Mills J. & H. Barea (2003), Effect of pre-enrichment incubation time on recovery of

Salmonella spp. – A Review, AgResearch Client Report (CR) 907.

MIRINZ (1971) MIRINZ Specification for conditioning and aging of beef. Meat Ind. Res. Inst.

N.Z. Publ. No. 227.

MIRINZ (1973) Specifications for lamb conditioning and aging. Meat Ind. Res. Inst. N.Z.

Publ. No. 303.

NZFSA December 2006, Proposal for Regulatory Monitoring for Campylobacter in Chickens

and on Poultry Meat.

Roberts, T.A., Hudson, W.R., Whelehan, O.P., Simonsen, B., Olgaard, K., Labots, H.,

Snijders, J.M.A., Van Hoof, J., Debevere, J., Dempster, J.F., Devereux, J., Leistner, L.,

Gehra, H., Gledel, J. & Fournard J. (1984) Number and distribution of bacteria on some beef

carcasses at selected abattoirs in some member states of the European Communities. Meat

Sci. 11, 191-205.

Stern, N.J., Line, J.E., Comparison of Three Methods for Recovery of Campylobacter spp.

From Broiler Carcasses, 1991.

Issued under section 167 of the Animal Products Act 1999.

Date of notification in Gazette:

This notice is administered in Ministry for Primary Industries.

schedule1 nmd dbase programme

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