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A World of Food Solutions

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Importance of Microbial Analysis in Food Industry

Key Note Address

A World of Food Solutions 2

Introduction:

Food as excellent source of nutrients for the consumers & microbial growth.

Spoilages and nightmares to Consumers

Importance of proactive detection and control of pathogens and spoilage organisms to balance the food nutritionally & make it microbiologically safe.

Outbreaks of food borne illness occur daily in all the countries, from the most to the least developed. Most of the cases are not reported, the true dimension of the problem is unknown with the absence of reliable data & hence effective solutions often fail.

In developed countries the public awareness on the food safety risk is high with clear commitments of the government & in developing countries there are competing priorities in health agenda.

Innovative strategies and methods are needed for laboratory based surveillance system with international networking on food borne disease and contaminations.

The evaluation procedures should be based on internationally agreed principles.

Methods shall ensure that product is safe before consumption.


Why?

Food Safety is global

Food Safety Concern

Disease burden on

Various countries

Food-related outbreak

Product recall Vs

economic loss

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Why?

Legal fees

Medical claims

Lost wages

Cleaning and sanitizing cost

Food that must be discard

Bad publicity

Loss of confidence in facility

Embarrassment

Closing of facility 4


Food Safety Hazards: Contribution of pathogens

5

Hazard

Physical Chemical Biological

Glass, plastic, heavy

metal, wood, rubber,

stone, bone, dust etc.

Dioxins and

polychlorinated

biphenyls

Listeria, Salmonella, Staph,

E.Coli, Campylobacter etc


Contribution of pathogens

6

About 70-72% of food safety hazards are attributed to

biological contamination.

Source: http://www.nbafoodadvocate.com/2010/08/22/cdc-foodborne-illness-statistics/


Pathogens-Contribution

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Source: http://www.defendingfoodsafety.com/articles/food-safety-news/


Food Vs Food borne diseases

According to the Centers For Disease Control (CDC) and Prevention, also known as the

CDC, every year 48 million people in the U.S. become ill from food borne diseases, and

3,000 will die. The prevention of a single case of E. Coli 0157 could save an estimated $7

million.

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Source: http://www.royalcrownconsultinggroup.com/?page_id=362


In India

No formal surveillance system for food borne

disease reporting and statistical analysis.

Even educated people keep food borne

illness to them

Quantum of disease outbreaks not known

Investigations limits to major disease

outbreaks such as some functions like

marriages

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Where to attack in Analysis?

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Source: http://www.huffingtonpost.com/2012/10/11/fda-food-regulation_n_1955074.html


Food spoilages

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Food spoilages

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Factors promoting microbial growth and food Spoilage:

Intrinsic Factors Extrinsic factors Implicit factors Processing

factors

Nutrient content Temperature Synergism Irradiation

pH Relative humidity Antagonism Washing

Redox potential Gaseous

atmosphere

Commensalism Slicing

Water activity Growth rate Pasteurization

Antimicrobial

constitutes &

barriers

Packaging

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A variety of intrinsic and extrinsic factors determine whether microbial growth will preserve or spoil foods


Microbial Spoilage of Food :

It is apparent that practically all kinds of microorganisms are potential contaminants. Given a chance to grow, the microorganisms will cause changes in appearance( Color changes, pockets of gas/swelling)’ Texture(Soft or mushy);Odour; Flavor or Slime formation.

Putrefaction:

Protein foods + proteolytic microorganism amino acids + amines + ammonia + H2S

Fermentation:

Carbohydrate foods + saccharolytic microorganism organic acids + alcohol + gases

Rancidity :

Fatty foods + lipolytic microorganisms fatty acid + glycerol

The various sources through which microorganisms gain entry into the foods

- Microflora present in soil, water, air, plant & plant products, animal feeds, animal hides, Intestinal tracts of humans & animals, food utensils and equipments.

To manage microbial contamination and growth from the farm to consumer, HACCP approach has become mandatory for the food processing industries to ensure safe foods to the public.

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Food spoilage and microorganisms in different category of Food:

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Types of food spoilage with some examples of casual organisms Food Types of Spoilage Microorganisms involved

Bread Moldy Rhizopus nigricans, Penicillium

Ropy Bacillus subtilis

Pickles, Sauerkraut Film/ Pink Yeasts Rhodotorula

Fresh Meats Putrefaction Alcaligenes, Clostridium, Proteus vulgaris

Pseudomonas fluorescens

Cured Meats Moldy Rhizopus, Aspergillus, Penicillium

Souring Pseudomonas, Micrococcus

Greening,Slime Lactobacillus, Leuconostoc

Fish Discolouration Alcaligens,Pseudomonas, Flavobacterium

Poultry Slime, Odor Alcaligens,Pseudomonas,

Eggs

Colorless & Green Rots Alcaligenes, Proteus, Pseudomonas, P. fluorescens

Concentrated Juices Off flavor' Acetobacter,lactobacillus,Leuconostoc

Fresh Fruits and Vegetables Soft rots Rhizopus, Erwinia

Milk / Cream Gray & black mold Botrytis,Aspergillus niger

Ropiness Alcaligenes viscolactis,Micrococcus, Enterobacter, Lactobacillus,Streptococcus, Bacillus

Decomposition of fats Yeast,molds,Alcaligenes, Proteus, Pseudomonas, Micrococcus, Bacillus, Clostridium

Alkali formers Alcaligenes visolactis, P.fluorescens

Flavor changes Lactobacillus, Sreptococcus, Leuconostoc


Food Spoilage and microorganisms in Meat:

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Types of Spoilage in Meat Aerobic or Anaerobic Spoilage Defect Microorganisms involved

Aerobic ( Bacterial) Surface Slimes Alcaligenes,Pseudomonas,Lactobacillus

Leuconostoc,Micrococcus, Bacillus

Bloom of Discolorations Lactobacillus, Leuconostoc

Rancidity Pseudomonas,Achromobacter

Phosphorescence Photobacterium spp

Red spot Serratia marcescens

Yellow discolorations Micrococcus,Flavobacterium

Off-odors off tastes (Taints) Lactic acid bacteria,Actinomycetes

Aerobic ( Yeasts ) Discolorations/Taints/Sliminess/lipolysis Rhodotorula

Aerobic ( Molds) Stickiness Rhizopus,Aspergillus,Penicillium

Whiskers Rhizopus,Thamnidium,Mucor

Black spot Cladosporium herbarium

White Spot Sporotrichum carnis

Green patches Peniclllium expansum,P.oxalicum

Decomposition of fats Lipolytic molds

Taints Penicillium, Thamnidium

Anaerobic ( Bacterial) Souring Clostridium Spp

Putrefication Alcaligenes,Pseudomonas, Clostridium spp

Taints Alcaligenes,Pseudomonas


Food Borne Diseases:

Any illness resulting from the consumption of contaminated food with one or more disease producing agents such as bacteria, parasites, viruses, fungi and their products as well as toxic substances not of microbial origin.

WHO estimates that 1.5 billion cases of food borne illness cause about 3 million deaths each year costing up to $ 40 billion in health care & job related absenteeism. More than 250 different food borne diseases have been described with different symptoms.

The microbes or toxins enter through gastrointestinal tract often causes first common symptoms of nausea, vomiting, abdominal cramps & diarrhea in many food borne diseases.

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Types of Food borne diseases:

TYPES OF FOOD BORNE DISEASES:

a. Food infection:

Occurs by consumption of contaminated food & water with pathogenic microorganisms.

e.g. Salmonella in meat

b. Food intoxication:

Occurs due to the consumption of a preformed toxin in food elaborated by microorganisms

e.g. Staphylococcal enterotoxins

c. Food toxi-infection:

Condition wherein bacteria are consumed in contaminated food & water but the bacterial cells either sporulate or die and release toxin in the gut to produce symptoms

e.g. Bacillus cereus spp. Gastroenteritis

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FOOD BORNE DISEASES

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FOOD BORNE DISEASES

Poisonings Infectious

Bacterial Chemical Poisoning Intoxication E.Coli

Salmonellosis

Poisonous Plant tissues Poisonous Animal tissue Microbial Intoxications Bacillus cereus

C.perfringens

Listeriosis

Algal toxins Mycotoxins Bacterial toxins Shigellosis

Yersiniosis

Neurotoxins Entero toxins Enteric virus

Clostridium botulinum Staphylococcus aureus Protozoan

Helminithic


Type of Microbial Food Borne Diseases:

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Type of Disease Causative bacteria Kind & nature of the bacteria

Major Symptoms

Intoxication

Staph poisoning Staphylococcus aureus strains Gram-positive cocci, present in pairs, short chans or bunched grape like clusters.

GIT

Botulism Clostridium botulinum strains Anaerobic, Gram-positive spore forming rod.

Non-gastric neurological signs of paralysis of different involuntary muscles.

Infection

Salmonellosis Over 2000 Salmonella spp.(except S.typhi and S.paratphi)

Rod shaped, motile, non spore forming Gram-negative bacteria

GIT

Campylobacter enteritis Camphlobacter jejuni & Cam.coli strains

Gram-negative,slender, curved, motile, microaerophillic rod

GIT

Yersiniosis Yersinia enterocolitica Small rod shaped, Gram-negative bacterium

GIT

Enter hemorrhagic E.coli colitis

E.coli 0157:H7 Gram-negative,motile, non-sporulating, rod shaped facultative anaerobic bacterium

GIT and non-gastric

Non-hemorrhagic E.coli strains

Shiga-like toxin producing E.coli strains like E.coli 026:H11

Gram-negative, motile,non-sporulating, rod shaped facultative anaerobic bacterium.

GIT

Listeriosis Listeria monocytogenes Motile,Gram-positive bacteria GIT and non-gastric

Shigellosis Four shigellaspps.( Sh. dysenteriae)

Gram negative, non-motile, non spore forming rods

GIT

Vibrio parahemolyticus gastroenteritis

Vibrio parahemolyticus Gram negative,curved bacteria GIT

Vibrio vulnificus infection Vibrio vulnificus Lactose-fermenting, halophilic, gram-negative, opportunistic pathogen

GIT and non-gastric

Brucellosis Brucella abortus Gram-negative bacteria GIT and non-gastric


Type of Microbial Food Borne Diseases: Continued…

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Type of Disease Causative bacteria Kind & nature of the bacteria

Major Symptoms

Toxicoinfection

Clostridium perfringens gastroenteritis

Clostridium perfringens Anaerobic, Gram-positive spore forming rod.

GIT

Bacillus cereus gastroenteritis

Bacillius cereus Gram-positive,facultative areobic spore former rods

GIT

E.coli gastroenteritis Enteropathogenic and enterotoxigenic E.coli

Gram-negative,motile, non-sporulating, rod shaped facultative anaerobic bacterium

Cholera Vibrio cholera Gram-negative,curved bacteria

GIT

Gastroenteritis by opportunististic pathogens

Aeromonas hydrophila gastroenteritis

Aeromonas hydrophila Gram-negative bacteria GIT

Plesiomonas shigelloides gastroenteritis

Plesiomonas shigelloides Gram-negative,rod shaped bacterium

GIT


PATHOGENS OF FOOD BORNE ILLNESS IN MEAT:

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PATHOGENS OF FOOD BORNE ILLNESS FOOD PRODUCTS HAZARD

Beef Lamb Pork Poultry Carcasses, carcass parts, and variety Meats

Salmonella, Escherichia coli O157:H7

Salmonella Salmonella Salmonella, Campylobacter

Raw (ground and not ground) products



Thermally processed commercially sterile products

Clostridium botulinum Clostridium botulinum

Clostridium botulinum

Clostridium botulinum

Not heat treated shelf stable products

Listeria monocytogenes, Staphylococcus aureus

Listeria monocytogenes, Staphylococcus

aureus


aureus


aureus

Fully cooked not shelf stable products

Salmonella, Escherichia coli O157:H7, Listeria monocytogenes, Cl.

perfringens, Cl. botulinum

Salmonella, Listeria

monocytogenes, Cl. perfringens, Cl. botulinum

Salmonella , Listeria

monocytogenes, Cl. perfringens, Cl. botulinum

Salmonella, Campylobacter,

Listeria monocytogenes, Cl. perfringens, Cl.botulinum

Heat-treated not fully-cooked and not shelf stable



Product with secondary inhibitors not shelf stable

Salmonella, Escherichia coli O157:H7,

Staphylococcus aureus

Salmonella, Staphylococcus

aureus

Salmonella, Staphylococcus

aureus

Salmonella, Campylobacter, Staphylococcus

aureus


Why it happens?

Food from unsafe source

Improper holding of temperature during processing

Inadequate cooling during storage

Contaminated equipments and utensils

Usage of contaminated water during processing

Poor personal hygiene

Adding contaminated ingredients to cooked foods

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Microbial Control Measures in Poultry Integration :

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Microbial Control Measures in Poultry Supply Chain: Supply Chain Control CONTROL MEASURES VERIFICATION

Breeder Farms 1.Control measures and Monitoring Plan for vertical transmission of Salmonella and Mycoplasma. 2. Suitable Vaccination Schedule, Preventive Medications and control through other Competetive Exclusion Products ( Feed Acidifiers & Probiotics) 3. Sera Monitoring schedule to evaluate the field Challenges 4. Ensure Pest and Rodent control. 5. Biosecurity Measures

BIOSECURITY & ANIMAL WELFARE AUDIT

Hatchery 1.Ensuring incoming HE for proper Sanitation. 2.Unidirectional flow for the Hatchery to avoid cross contamination. 3.Effective Sanitation & Hygiene Programme for the Hatchery operations. 4.Monitoring the chick pullouts for DIS and chick deformities. 5. Biosecurity Measures


Feed Mill 1.Monitoring Incoming Ingredient Analysis for Nutrients & Microbes. 2.Proper handling and Storage of RM & FG. 3.Pelletization of feed with heat treatment to reduce the contaminations & improve NV. 4.Feed without Animal Protein source 5. Biosecurity Measures


Commercial Broiler Farms 1.All in and All out System 2.Monitoring Health Status through sera Monitoring 3.Disease Prevention through Vaccination Schedule 4.Biosecurity measures at Farm 5.Health certificate of the flocks for Zoonotic Diseases, Feed and Drugs withdrawls as specified before sending the live birds to Plant.



Microbial Control Measure in Poultry Processing Plant:

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Microbial Control Measures in Poultry Processing Plant:

Supply Chain Control CONTROL MEASURES VERIFICATION

Chicken Processing Plant

1. All birds arriving at Plant are tested and certified for there Health. 2. Chicken Processing & deboning as per HACCP plan (ISO 22000). 3. Dressed Chicken is chilled on line to < 4° C and the boneless meat temp is < 7.2° C when packed & thereafter immediately stored in Chiller < 4°C. 4.Every batch of fresh boneless undergoes microbial test. External lab testing Twice a year Residue Monitoring for Antibiotics, Pesticides, Insecticides and Heavy Metals and the microbial Test. 5. Meat is transported and delivered to customer at < 4°C.

HACCP & ANIMAL WELFARE AUDIT

Major Challenge in the Fresh meat procurement is the conventional Microbial

Test which takes three days for the report.

Conventional Method is followed due to Prohibitive cost for Rapid Test.

We have to wait for report and consume the meat within 5 days of its shelf life.


Case Study on Eggs to understanding Food Safety Challenges in Market :

FARM SELECTION

PROJECTS TO UNIVERSITY ON

EVALUATIONS OF MARKET EGGS Vs SELECTED FARM FRESH EGGS

1. PHYSIOCHEMICAL PARAMTERS

2. MICROBIAL & CHEMICAL ANALYSIS

3. SANITISERS & THEIR VARIOUS CONCENTRATIONS Vs SHELF LIFE OF EGGS AT REFRIGERATED TEMPERATURE

Evaluating & Selecting layer farm on the basis of resources such as location, incoming chicks, water & water sanitation, feed ingredient & formulations, Flock monitoring, vaccination & preventive medication, GMP and Biosecurities.

1.Egg Weight, Shape, Shell thickness, Yolk

Index & Haugh Unit.

2. Microbial – E.coli, Coliforms, Staph, Salmonella ,Campylobacter & yeast & Mould Residue Monitoring - Antibiotics & Pesticide residue

3. Trials on Sanitizers with different levels of concentrations to Evaluate the efficacy against concern microbial for their counts for determining the Shelf life

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Fresh Chilled washed & sanitized Egg

Process Implementation and Monitoring:

IMPLEMENTING EGG WASHING PROCESS WITH HACCP APPROACH

LAYER FLOCKS & EGG MONITORING PRACTICES

1. FARM

2. EGG WASHING PROCESS

Eggs are graded, washed, sanitized, candled, dried, oil sprayed, packed, stored and transported in refrigerated temperature to the QSR’s Store.

1.Flocks are monitored frequently for Salmonella & other poultry diseases.

2. Farm Biosecurity and Animal Welfare Audit is conducted.

1. Washed & Sanitised Eggs are regularly monitored for the microbial contaminations

2. Egg washing process is Audited for the GMP and HACCP Audit

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Conventional Methods of Microbial Testing:

Conventional Methods I II III IV

Cultural and Microscopic methods

Physical Method Chemical Method Immunological Methods

Standard Plate Count ( SPC )

Impedance Thermostable nuclease ( Staph aureus )

Fluorescent antibody

Most Probable Number (MPN )

Microcalorimetery Limulus lysate for endotoxins Enrichment serology

Dye reduction Method ( MBRT )

Flow cytometry ATP measurement (luciferase emits Light)

Radioimmunoassay

Direct Microscopic Count ( DMC)

Radiometry

Glucuronidase assay for E.coli

DISADVANTGE

Technique is cumbersome

Material Intensive

Time Consuming


RAPID DETECTION OF PATHOGENS IN

FOODS :

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RAPID DETECTION OF PATHOGENS IN FOODS DNA probe based

detection PCR based detection

Real Time PCR

Recent advances in biotechnology have considerably simplified microbiological

procedures for detecting pathogens and have reduced the assay time. These

assay procedures include use of DNA probes and PCR based test.

With increasing regulation of food products, in response to consumer concern, food

industry needs efficient quality assurance programme to monitor the safety &

quality of foods before reaching the consumer.

The sooner the food Industry adopts and implements these biotechnological

methods, the better for it to make its presence felt both in National and

International market.


Thank you

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