second workshop of national reference …old.iss.it/binary/crlp/cont/cyprus.1190119812.pdf ·...
TRANSCRIPT
SECOND WORKSHOP OF NATIONAL REFERENCE LABORATORIES FOR
PARASITES14-15 JUNE, 2007
ISTITUTO SUPERIORE DI SANITURoom Daniel Bovet, Viale Regina Elena, 299
00161 Rome Italy
Dr Andreas PhylactouVeterinarian
CYPRUS
Information
• The third-largest island in the Mediterranean Cyprus lies off the southern coast of Turkey and the western shore of Syria. The highest peak is Mount Olympus at 6,406 ft (1,953 m).
• Land area: 3,568 sq mi (9,241 sq km);
• Total area: 3,571 sq mi • Population (2006 est.): 784,301
(growth rate: 0.5%); life expectancy: 77.8; density per sq mi: 220
Veterinary Services of Cyprus
Veterinary Services Organogram
DIRECTOR
REGISTRY ACCOUNTS
STORE
EUROPEAN UNION AND
INTERNATIONAL ORGANIZATIONS SECTION INTERNAL AUDITING SECTION
ANIMAL HEALTH AND WELFARE
DIVISION
VETERINARY PUBLIC HEATLH
DIVISION
VETERINARY LABORATORIES SECTION
DRUGS AND SUPPLIES SECTION
D.V.O.
NICOSIA
D.V.O. LIMASSOL
D.V.O. LARNACA
D.V.O. AMMOCHOSTOS
D.V.O. PAPHOS
R.V.S. AVDEMOU
R.V.S. SKARINOU
R.V.S. P. CHRYSOCHOUS
R.V.S. NISOU R.V.S. AKAKI
Laboratory for the Control of Food of Animal Origin
VETERINARY LABORATORIES
QUALITY MANAGERDEPUTY SAFETY MANAGER
A. PHYLACTOU
SAFETY MANAGERDEPUTY QUALITY MANAGER
G. CHRISTOFI
HEAD OF VETERINARYLABORATORIES
KYRIAKOS GERGIOU
VIROLOGY
DIRECTOR OF VETERINARY SERVICES GIORGOS NEOPHYTOU
SCIENTIFIC STAFFK. GEORGIOU
(TECH. MANAGER)
P. STYLIANOUA.MAZERIS
(QUALITY MANAGER)
TECHNICAL STAFF
A. SHIAKALLISS. PAPANEOPH.M. NIKOLAOUM HATJIYANNI
THEOPHANOUSS. MINA
D. THEMISTOKL.
SCIENTIFIC STAFFI. IOANNOU)
TECHNICAL STAFF
N. CHRISTOFIC. ZINIERIS
SCIENTIFIC STAFFI. IOANNOU
(TECH. MANAGER)
TECHNICAL STAFFM. SAVVIDES
Z.POLYCARPOUT. DEMETRIOUS. DAMIANOU
SCIENTIFIC STAFFSOTIRIA GEORGIADOU
(TECH. MANAGER)
TECHNICAL STAFF
N. THEOPHILOU
SCIENTIFIC STAFFM. LIAPI
(TECH. MANAGER)
TECHNICAL STAFFM. AVRAAME. TSINTIDOUM. ANDREOUP. IOANNOU
N .PAPAEFSTATHIOU
LCFAOBACTERIOLOGY/
SEROLOGY HISTOPATHOLOGY
POST MORTEM-MICROBIOLOGY
PARASITOLOGY
TECHNICAL STAFFK. ACHILLEOS
F. TOUMBAM. SHIAKALLISA.TOFALLIDOU
S. CONSTANTINOUA. DIMOSTHENOUS
M. OMIROUAFROYLA SKOTTI
SCIENTIFIC STAFFANDREAS PHYLACTOU
C. ECONOMIDESMARIA EMMANUEL
CLEANERSA. NIKOLAOUK. KYRIACOU
A. PAVLOUC. MONOGIOU
BIOLOGICALPRODUCTS
VETERINARY LABORATORIES
Veterinary Services of Cyprus • Laboratory for the Control of Foods of Animal Origin
(LCFAO) is the Cyprus National Reference Laboratory for Trichinella spiralis
• Dr Kyriakos GeorgiouHead of Veterinary Laboratories• Dr Andreas PhylactouHead of LCFAO• Dr Ioannis IoannouOfficer in Charge for the Laboratory of Parasites
Mission of Veterinary Services
• The Cyprus Veterinary Services are the Competent Authority for the control of foods of animal origin.
• LCFAO provides services through microbiological, parasitological, presence of residues and physicochemical examinations on foods of animal origin including water.
• Control of exported and imported foods of animal origin for their suitability.
• LCFAO• Is based in Nicosia in the same block with the Headquarters of the
Veterinary Services.• Is responsible for the control of foods of animal origin for the whole
island.
PersonnelHead of LCFAO (Vet. Off.)Quality Manager (Vet. Off.)
Technical Manager (Vet. Off.)Technical Manager Assistant (Vet. Off.)
8 Laboratory Technicians1 Secretariat
1 Glassware cleaning & Sterilization1 Cleaning Staff
Structure of LCFAO• Sample Registration, Management and Dispatching
Unit
• Media Preparation Unit
• Raw Milk Unit• Total bacterial count at 30 οC (ISO 6610/1992)• Somatic cells (ISO 13366-1, Microscopic method)
(ISO 13366-3, Fluoro-opto-electronic method)• Freezing point (Dir. 91/180 EEC)• Presence of antibiotics (DELVO-Test)
Pasteurized Milk and Milk Products Unit• Total count at 30 οC (ISO 6610/1992)• Staphylococcus aureus (ISO 6888-1/1999)• Clostridium perfringens (Iron sulfit agar)• Bacillus cereus (ISO 7932/1998)• Coliforms (ISO 5541-1/1986)• Escherichia coli (ISO 16649-2/2001)• Yeast and Molds (ISO 7954/1987)• Psychrotrophic microorganisms (IDF 132A)• Phosphatase (Lactognost®)• Peroxydase (Dir. 91/180 EEC)
• Meat and Meat Products, Fish, Eggs, Water Unit• Total bacterial count at 30 οC (ISO 6610/1992)• Staphylococcus aureus (ISO 6888-2/1999)• Clostridium perfringens (Iron sulfit agar)• Coliforms (Meat: ISO 4832/2001)
Water: ISO 9308-2MPN/1990)• Escherichia coli 0157(Meat:16654/2001
Water: ISO 9308-2 MPN/1990)• Trichinella spiralis (Trichinoscope, Digestion
method, Commission regulation 2075/2005/EC)
• Food borne Pathogens, Mastitis Unit• Salmonella spp. (ISO 6579/2002)• Salmonella Susceptibility testing (NCCLS
Guidelines)• Listeria (ISO 11290-1/1996, 11290-2/1998)• Campylobacter (direct plating on Camby-agar)• E. coli 0157 (ISO 16654/2001)• Mastitis (antibiogramme)
• Veterinary Drug Residues and Staphylococcal Enterotoxin Unit:
• FPT (Dir. 86/469/EEC,96/23/EEC)• Sulfonamides (ELISA, TLC)• Staphylococcal Enterotoxin (Transia -
Reference method of the CRL, V.2, September 2005)
• Histamine detection (TLC)
Monitoring Programs of LCFAO
• Somatic Cell Count in raw milk• Total bacterial count of raw milk• Antibiotics in raw milk• Veterinary drug residues in meat• Salmonella, Listeria, Campylobacter in
poultry carcasses• Trichinella spiralis
Accreditation - ISO 17025/2005
8 July 2003 accredited for ISO 17025/1999by the Greek Accreditation Body ESYD• Total Bacterial Count in Raw and Pasteurized milk.• Somatic Cell Count-Microscopic method• Somatic Cell Count-Fluoro-opto-electronic method• Listeria Detection• Salmonella Detection• Four Plate Test (FPT)• Staphylococcal Enterotoxin
Trichinella spiralis
• CLASS: ADENOPHOREA • SUBCLASS: ENOPLIA • ORDER: STICHOSOMIDA
(TRICHURATA) • SUPERFAMILY:
TRICHOCEPHALOIDEA • FAMILY: TRICHINELLIDAE
Scientific name - Trichinella spiralis
• Disease - trichinosis
Life Cycle
• This nematode has no stages outside a host. The adult worms are found attached to or buried in the mucosa of the duodenum. Aftermating the males die. Females (ovoviviparous) produce living young (approximately 1,500 per female over a period of 4 to 16 weeks) and then die. Juveniles enter the lymphatics and mesenteric veins and are found throughout the arterial circulation between the 7th and 25th day after infection. They travel in the hepatoportal system through the liver, then to the heart, lungs, and the arterial system, which distributes them throughout the body. They are transported to striated muscles, penetrate individual fibers, and cysts are formed around the juveniles. Within cysts, juveniles remain viable for many years, up to 25 years in man and 11 years in pigs. There is agreement that there are four juvenile stages but disagreement in the literature as to whether or not nematode development occurs within the cyst. When viable encysted juveniles are ingested, they are digested from the cysts and pass to the duodenum where they mature. Humans are essentially a dead end for the parasites.
Symptoms-Pathogenicity
• Nausea, dysentery, colic due to invasion by adult worms. • Migrating juveniles cause pain as they invade muscle
tissue; there may also be edema (swelling), delirium, cardiac and pulmonary difficulty, pneumonia, nervous disorders, deafness and delayed or lost reflexes.
• Many cases are never diagnosed because of the vaugeness of the symptoms. Muscle biopsy can be conducted and involves pressing muscle between glass plates to look for cysts. Xenodiagnosis involves feeding suspected muscle to laboratory rats.
Management - Prevention
• Cook pork thoroughly (also flesh of bear, walrus, wild pigs). • Cook all garbage fed to hogs. • Proper meat handling, ordinary curing and salting of pork products will not kill
encysted juveniles. • Freezing is effective if carried out properly. The freezing requirements differ with the
size of the meat. Pieces not exceeding 6 inches in thickness require 20 days at 5F, 10 days at -10F, 6 days at -20F. Larger pieces require longer periods. Quick freezing and storage for 2 days is effective.
• Although there have been marked reductions in the percentage of hogs carrying encysted juveniles in the United States, there is considerable variation from one part of the United States to another. It is therefore a wise precaution to avoid uncooked pork products. In the United States Trichinella occurred in 0.4 to 2.5% of the prepared pork products examined in some areas (1961); however some areas had as much as 10% infection. In Chile 0.22% of 500,000 hogs were infected (1964). In Germany only 0.00026% of 14,000,000 hogs were infected (1965). It has been estimated that 3/4 of the persons infected with Trichinella are from North America.
• Treatment of Host: No cure. Relieve symptoms with analgesics and corticosteroids.
Characteristics
• The males measure 1.4 to 1.6 mm long and are more slender at the anterior than the posterior end. The anus is nearly terminal and has a large papilla on each side of it. A copulatory spicule is absent. Stichocytes are arranged in a row following a short muscular esophagus.
• Females are about twice the size of males, also tapering toward the anterior end. The anus is nearly terminal. The vulva is located near the middle of the esophagus, which is about a third the length of the body. The single uterus is filled with developing eggs in its posterior portion, whereas the anterior portion contains fully developed, hatching juveniles.
• Adult female male infective muscle larvae
Historical Data of Trichinella in Cyprus
• Trichinellosis is a notifiable parasitic disease in Cyprus. • However it has never been observed in the island. • During 1969 and annually afterwards, pig carcasses were periodically
tested for the presence of Τrichinella spiralis from slaughterhouses all over Cyprus.
• Since 1999 a more systematic surveillance program for Trichinella testing has been established. Samples were collected from the pillarsof the diaphragm of pig carcasses including sows and boars, from all the pig farms all over the island. The last seven years (1999-2005) animals from the wild life (foxes, stray dogs and cats, rodents) werealso included.
• From 1969 until January 2004, the Trichinella spiralis testing was performed centrally in the Laboratory for the Control of Foods of Animal Origin of the Cyprus Veterinary Services.
• Originally meat samples were examined with the Trichinoscope whilethe last 10 years the method of choice is the digestion method with the use of Pepsin and Hydrochloric acid.
• Since January 2004, the examination of fresh meat derived of domestic swine is performed at the slaughterhouses laboratories.
• During 2004 a number of 357638 swine meat samples were examined. These animals originated from all 121 piggeries of Cyprus and covered80% of the target population.
• During 2005 and 2006 all pigs slaughtered in Cyprus were tested for Trichinella.
• The testing of samples from the wild life and from other domestic animals is performed only at the LCFA. (2006-108 samples)
• There are no piggeries in the non controlled by the Government of Cyprus area. The reason for this is that Muslim population (Turkish Cypriots, Settlers and soldiers from Turkey) living in this part of the island, are not consuming pork meat of religious reasons. They are Muslims.
• There are no wild pigs in Cyprus and the fox population is very low.
Wild life
• During the last seven years 1288 samples from the wild life were tested.
• All samples were found negative for Trichinella.• The last few years a great number of stray dogs and foxes were
examined for Trichinella, after were captured and killed by United Nations in the buffer zone, separating the controlled and the non controlled area by the Government of Cyprus, based on the antiechinococcosis campaign programme supported by UN (UNOPS).
• None of these animals was found positive for Trichinella.• The same was the case with the testing of rats for Trichinella where
many rats were also captured from areas close to the buffer zone.
Wild life
1288355104052487210692Total
1----1--Goats
8---62--Hares
12--13332Mouflons
1-----1-Horses
361235-718Foxes
938354-386196-11Rats
4----2-2Mice
5---41--Stray cats
283-81534639469Stray dogs
Total
2005 (-May)200420032002200120001999Animal
species
Trichinella testing in Cyprus after 1974
NIL19241994
NIL41741993
NIL3181992
NIL2471991
NIL101990
NIL4721989
NIL2141988
NIL1071987
NIL1881986
NIL4241985
NIL7221984
NIL1341983
Positive samplesNo of samples examinedYear
Trichinella testing in Cyprus after 1974
NIL6477292006
NIL6658782005
NIL3576432004
NIL23992003
NIL12902002
NIL13392001
NIL14122000
NIL8281999
NIL4171998
NIL3021997
NIL1811996
NIL1871995
Positive samplesNo of samples examinedYear
Trichinella spiralis has never existed in CyprusSurveillance program is in place according to Directive 77/96
4209
386 (weasels)
3
15
3805
2003
-208 (weasels)
910Other Animals
2004200220012000
255292
131113421412Total
3537Foxes
8376394Dogs
255281
106112671301Swine
4209
386 (weasels)
3
15
3805
2003
-208 (weasels)
910Other Animals
2004200220012000
255292131113421412Total
3537Foxes
8376394Dogs
255281106112671301Swine
Toxoplasmosis
• Serological examinations (indirect haemagglutination test) in sheep and goats are showing positive results and the disease is considered to be one cause of abortions.
• In humans although there are no official reports for the disease, it is considered to be a problem in the female population.
• The problem needs further investigation.
Cryptosporidiosis
• Only feaces sample from diarrheic calves, lambs and kids examined
Method usedRapid testModified Zichl Nielsen coloring
Cryptosporidiosis
124
3121Total
3382006
242005
592004
CalvesLamb and kidsYear
E. Echinococcosis
• 1971-1985: Eradication program total eradication
• 1994: New eradication program
Surveillance program in place monitors:
• Registration and identification of all dogs with microchip (obligatory) • Dog treatment:
– preventive treatment with praziquantel (required also for imported dogs)
• Stray dog control – praziquantel baits to the buffer zone
• Dog testing: – faeces samples tested with Coproantigen ELISA test
Surveillance program in place monitors:
• Slaughtering inspections: – inspection of all kinds of carcasses
• Dead animals:– Proper disposal of carcasses
• Public enlightening: – lectures at clubs, schools, army camps etc.
Echinococcus
ANIMALS (cows, sheep & goats) INFECTED WITH CYSTS
13
19
13
2
9
1
8
10
0
2
4
6
8
10
12
14
16
18
20
1997 1998 1999 2000 2001 2002 2003 2004 2005YEAR
NO
. OF
ANIM
ALS
NO. OF ANIMALS INFECTED
Farm animals found infected in slaughterhouse inspectionsDogs: Since 1995 no infected dog has been found
00000≈ 2500002005
100109812602004
8023310098952003
100109781862002
900909392822001
201109274322000
13011028994461999
1915948962261998
1310938954711997
TOTALMoufflonsGoatsSheepBovine
INFECTED ANIMALSAnimals slaughtered
Year
Μόλυνση µε Κύστεις Εχινόκοκκου –Αίγες
____14,362____130,5651985
0,0119,348____103,4141984
0,01110,084____99,5701983
0,01110,220____100,4371982
0,03412,391____101,8381981
0,131612,715____117,0121980
0,224118,250____119,7121979
0,326620,570____131,8871978
0,679013,476____111,6091977
1,831699,243____101,5381976
3,913178,1010,007689,4571975
4,932655,3790,0181583,6201974
8,841,45116,4050,0896122,9341973
12,511,32110,5560,966575,6251972
15,013682,4511,460041,5331971
ΠοσοστόΜόλυνσης
Μολυσµένα ΖώαΑίγες άνω των 2 ετών
ΠοσοστάΜόλυνσης
Μολυσµένα ΖώαΑίγες κάτω των 2 ετών
Έτος
Μόλυνση µε Κύστεις Εχινόκοκκου – Πρόβατα
0,112421,779__198,1231985
0,498216,576__158,1321984
0,7913416,884__154,7491983
1,0819017,512__164,4381982
3,0457018,754__158,1001981
4,1578018,809__176,7311980
7,271,67723,080__180,2581979
12,763,09724,262__187,3971978
14,862,63717,746__140,7071977
24,093,20013,2830,00091113,7321976
30,443,77812,4100,0252391,4461975
43,413,4968,0530,0898124,4551974
10,731,70215,8610,21304146,7321973
53,856,17511,4661,291,21893,9921972
49,631,8193,6652,281,36859,8761971
ΠοσοστόΜόλυνσης
Μολυσµένα ΖώαΠρόβατα άνω των 2 ετών
ΠοσοστόΜόλυνσης
ΜολυσµέναΖώα
Πρόβατα κάτω των2 ετών
Έτος
Μόλυνση µε Κύστεις Εχινόκοκκου – Βοοειδή
0,0412,321____6,0131985
0,6291,457____6,0401984
1,38191,376____6,3431983
3,41401,174____5,0641982
5,85871,487____5,2201981
2,63351,333____5,5471980
5,42721,329____5,1301979
5,65921,627____5,8501978
4,08591,446____4,9181977
8,051151,4290,0734,5001976
10,201541,5120,0734,1211975
21,03671,7430,2094,4261974
35,101,1593,3060,26228,4051973
30,68612,8120,9485,0741972
38,901,0322,6521,8915,2751971
ΠοσοστόΜόλυνσης
Μολυσµένα ΖώαΒοοειδή άνω των 2 ετών
ΠοσοστόΜόλυνσης
Μολυσµένα ΖώαΒοοειδή κάτωτων 2 ετών
Έτος
Μόλυνση µε Κύστεις Εχινόκοκκου – Χοίροι
____5,565____294,2171985
____5,269____298,8861984
____4,431____269,1431983
____3,514____246,9241982
____2,691____221,4021981
____3,695____181,9441980
____4,164____195,9501979
____6,281____186,1251978
0,0425,584____157,8971977
0,2751,8500,0012136,5641976
4,71282,7200,0023123,4971975
10,23803,7340,02738142,8221974
7,44365,8840,0799134,9061973
10,04404,3980,2219788,7511972
18,607644,1160,8257570,0011971
ΠοσοστόΜόλυνσης
Μολυσµένα ΖώαΧοίροι άνω των 2 ετών
ΠοσοστόΜόλυνσης
ΜολυσµέναΖώα
Χοίροι κάτω των 2 ετών
Έτος
Αριθµός δολωµάτων Praziquantel πουχρησιµοποιήθηκαν
88741163711241Σύνολο
100023501367Πάφος
176611641400Αµµόχωστος
30101188976Λεµεσός
180036435248Λάρνακα
129632922250Λευκωσία
200620052004Επαρχία
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