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  NATIONAL INSTITUTE OF VETERINARY RESEARCH ENTERIC DISEASES DIAGNOSTIC REQUEST FORM NIVR – Attn: Department of Bacteriology (Dr Cu Huu Phu) Postal Address 86 Truong Chinh Rd Dong Da-Hanoi Phone 84 48693923 Fax 84 48694082 Please complete ALL sections Complete ONE form for each sow Multiple piglets from the same sow can be entered on a SINGLE form Farm Owner: NIVR Use Only: Number of sows: TEST FARM NUMBER: Address: LABORATORY SUBMISSION NUMBER: Phone: Fax: Email: Mobile: Contact details of person collecting specimen: Name: Occupation: Address: Province: Postcode: Telephone: Fax: Email: Date of Collection: Sow ID Par-ity Breed: Date of birth Litter size Foster pigs Number of piglets with diarrhoea Duration Sample type Preliminary Diagnosis/History (Treatments, severity of scour etc) Reports to be sent Vet Property Manager Other: Name Other: Name Signed ………………………………………………………… Fax No Fax No Date………………………… Treated? (Y/N)   NATIONAL INSTITUTE OF VETERINARY RESEARCH ENTERIC DISEASES DIAGNOSTIC REQUEST FORM ID No: Characteristics of faecal specimens: Sample No: pH: Colour: Date: Parasitology: Consistency (Scale Isospora suis (Faecal float): 1-4): Cryptosporidium (MZN): (1 = thick custard; = pure water) Bacteriology: Other: Virology (TGE) Virology (Rotavirus) DIPSTICK (Herd only): ELISA: ELISA: Immunoflourescence: Diagnosis Signed ………………………………………………………… Date…………………………   NATIONAL INSTITUTE OF VETERINARY RESEARCH ENTERIC DISEASES DIAGNOSTIC REQUEST FORM Signed ………………………………………………………… Date………………………… CAUSES OF PRE-WEANING DIARRHOEA IN COMMERCIAL AND VILLAGE PIGS IN VIETNAM Do NT1, Nguyen XH1, Au XT1, Cu HP 1, Fahy VA2 , Cargill C3, and Trott DJ4 1National Institute of Veterinary Research, Hanoi, Vietnam 2National E coli Reference Laboratory, Department of Natural Resources and Environment, Epsom, VIC 3554, Australia 3The South Australian Research and Development Institute (SARDI), Adelaide SA 5001, Australia 4School of Veterinary Science, The University of Queensland, Brisbane, QLD 4072 INTRODUCTION Table 1: The contribution of different infectious agents to PrWD in commercial and village pigs  Pre-weaning diarrhoea in pigs (PrWD) is a complex problem involving a large number of causative agents: transmissible gastroenteritis virus (TGEV), rotavirus (RV), enterotoxigenic E coli (ETEC), Clostridium perfringens (C per.), coccidiosis due to Isospora suis (Cocci), and Cryptosporidium (Crypto) [1] Agent(s) detected Commercial (n=117) Cocci (2.5) RV  A comprehensive survey of the major causative agents Total single infections RV Cocci ETEC 17 (14.4) RV C per TGEV Cocci RV Cocci Crypto collected from cases of PrWD fro 6/2005 to 3/2008 (1.7) (6.7) (3.4) (6.7) ETEC (1.7) (4.4) (0.8) (2.2) ETEC (4.2) (4.4) ETEC TGEV RV Crypto  117 (from CP) and 45 (from VP) faecal specimens were (5.9) (0.8) (6.7) (0.8) (2.2) RV Crypto TGEV RV C per TGEV RV TGEV  All samples were tested for the presence of: 1) Cocci; RV and 2) Crypto oocysts by standard faecal flotation and modified Ziehl-Neelsen staining of faecal smears, respectively; 3) RV; and 4) TGEV using an ELISA kit (Institut Pourquier, France); 5) ETEC by aerobic culture and PCR for enterotoxins and fimbriae; and 6) C perfringens by anaerobic culture Results were interpreted according to manufacturer’s recommendations (ELISA kit) or as described [3] RV (2.5) TGEV  Detection of multiple infectious agents from a single specimen was common, with TGEV and RV being endemic to all piggeries (Table 1)  In village pigs, agents were always found together with one or more additional agents, while 25 cases from CP were infected with only causative agent  ETEC occurred in older (>4 day-old) piglets; most probably due to effective vaccination programs  These results and observations from farm audits suggest that environmental conditions and husbandry practices may be predisposing piglets to intestinal infections (0.8) ETEC TGEV TGEV Crypto C per (2.2) (4.4) ETEC RV Cocci (2.2) (0.8) (5.1) TGEV TGEV (4.4) (2.5) ETEC (4.4) (0.8) Crypto Crypto (6.7) (0.8) Crypto MATERIAL AND METHODS (5.1) TGEV Crypto Cocci (1.7) 25 (21.2) RV OBJECTIVES RESULTS (3.4) C per Crypto pre-weaning diarrhoea in commercial (CP) and village pigs (VP) in Vietnam 11 (9.3) ETEC of PrWD has not been attempted previously in Vietnam  To determine the prevalence of the six major causes of (2.5) TGEV Cocci Village (n=45) (1.7) Crypto  The predominant causes of PrWD and their relative contribution to morbidity vary between countries, and have continuously changed over time, due to the adoption of new management practices and the emergence of new diseases In addition, some pathogens are restricted to certain geographical locations [2] # of positive specimens (%) ETEC 14 (11.9) (15.6) RV Crypto Crypto RV Cocci Crypto (2.2) ETEC C per (1.7) (6.7) ETEC C per (1.7) TGEV ETEC RV Cocci (0.8) TGEV Crypto C per Cocci (36) TGEV RV TGEV Crypto RV RV TGEV (97) (111) (2.2) (0.8) (4.4) (2.2) C per (0.8) (3.4) (2.2) C per TGEV (50) C per TGEV Crypto Cocci (1.7) (0.8) ETEC (0.8) (2.2) C per (0.8) ETEC ETEC (76) (23) Total multiple infections 92 (78.8) 45 (100.0) REFERENCES Straw, B E., et al Diseases of Swine 1999: 41-59 Tzipori, S British Veterinary Journal 1988; 144: 521-523 Diagnostic Manual of the Pig Health and Research Unit (Bendigo, Victoria, Australia) ACKNOWLEDGMENTS This work was supported by Ministry of Agriculture & Rural Development (Vietnam), National Institute of Veterinary Research (NIVR), Australian Government (AusAID) under Collaboration for Agriculture and Rural Development (CARD) Program 1 Virulence characterizations of Vietnamese strains of coli causing diarrhoea in pigs in Vietnam E Do NT1, Trott DJ2, Desautels C3 , Fairbrother JM3 1Department 2School 3The of Bacteriology, National Institute of Veterinary Research, Hanoi, Vietnam of Veterinary Science, The University of Queensland, Brisbane, QLD 4072, Australia Escherichia coli Laboratory, Faculte de Medicine Veterinaire, Universite de Montreal, 3200 Sicotte, Saint-Hyacinthe, QC, Canada J2S 2M2 INTRODUCTION Table 2: The prevalence of different pathotypes  Escherichia coli is one of the most important enteric pathogens causing diarrhoea in pigs [1]  Pathogenic E coli often colonize the small intestine by means of adhesion factors and produce one or several disease-causing toxins [1]  Detection of virulence factors by molecular techniques such as DNA hybridization and PCR has been shown to be the most effective tool to evaluate if an E coli isolated obtained from diseased pigs is pathogenic and to provide suitable measures of control and prevention [2] Pathotype Source of isolates PrWD (n=18) PWD (n=41) F4/STa/STb F4/ Paa/STa/STb/LT/EAST1 F4/Paa/STb/LT/EAST1 F4/STa/STb/EAST1 F4/STa/STb/Aero F5/Paa/STa OBJECTIVES F4/Paa/STb/LT/EAST1 MATERIAL AND METHODS  E coli strains (n=18 from pre-weaning and n=41 from post-weaning piglets with diarrhoea) from different provinces in Vietnam  ETEC or ETEC/VTEC were confirmed by primary multiplex PCR (F4, F5, F6, F41, F18, STa, STb, LT, VT2e)  DNA hybridization and PCR were further applied to detect for the presence of 19 virulence factors according to the protocol of The Escherichia coli Laboratory, Faculte de Medicine Veterinaire, Universite de Montreal F18/AIDA-I/STa/STb F18/Paa/AIDA-I/STa/Stx2 F18/LT/Stx2 (F4, F5, F6, F17, F18, F41, EAE, P factor, Paa, AFA, AIDA-I, STa, STb, LT EASTI, Stx1, Stx2, CNF, Aero) in ETEC and ETEC/VTEC strains obtained from pig with diarrhoea in Vietnam F18/STa/EAST1 F18/AIDA-I/STb/Stx2  To screen for the presence of 19 virulence factors F18/STa/STb F18/AIDA-I/STa/STb/Stx2 F18/Paa/AIDA-I/STa/STb/Stx2 F18/Paa/STa/LT/Stx2 13 Paa/STa/LT/Stx2 Paa/STa/STb/LT/EAST1 AIDA-I/STb/EAST1 AIDA-I/STb/LT/EAST1 STa/STb STb/EAST1 RESULTS Table 1: Prevalence of individual virulence gene Source of isolates (%) Virulence factor PrWD (n=18) PWD (n=41) F4 (50.0%) (14.6%) F5 (11.1%) LT/Stx2 REFERENCES Bertschinger HU, Fairbrother JM Escherichia coli infections In: Straw BE, D'Allaire S, Mengeling WL, Taylor DJ, editors Diseases of Swine Ames, Iowa: Iowa State University Press; 1999 p 431-468 13 (72.2%) 25 (61.0%) 2 Wray C, Woodward MJ Laboratory diagnosis of Escherichia coli infections In: Gyles CL, editor Escherichia coli in Domestic Animals and Humans Wallingford, England: CAB International; 1994 p 595-628 AIDA-I (11.1%) 13 (31.7%) ACKNOWLEDGMENTS STa 14 (77.8%) 31 (75.6%) STb 16 (88.9%) 20 (48.8%) LT 12 (66.7%) 23 (56.1%) 30 (73.2%) F18 Paa STx2 This work was supported by Ministry of Agriculture & Rural Development (Vietnam), National Institute of Veterinary Research (NIVR), Australian Government (AuAID) under Collaboration for Agriculture and Rural Development (CARD) Program 28 (68.3%) EAST1 14 (77.8%) Aero (5.6%) (17.1%) 2 DEVELOPMENT AND EFFICACY TESTING OF A VACCINE FOR THE CONTROL OF PRE-WEANING COLIBACILLOSIS IN VIETNAM Cu HP1, VA2 , Fahy Driesen SJ2, Moore K2, Vanderfeen A2, Do NT1, and Trott DJ3 1National Institute of Veterinary Research, Hanoi, Vietnam 2National E coli Reference Laboratory, Department of Natural Resources and Environment, Epsom, VIC 3554, Australia 3School of Veterinary Science, The University of Queensland, Brisbane, QLD 4072 INTRODUCTION  The cost of pork production in Vietnam could be significantly reduced by the widespread use of locally made efficacious vaccines to control endemic diseases such as neonatal colibacillosis VACCINE DEVELOPMENT Table 1: E coli strains used for the preparation of vaccine Designation of strains Virulence determinants CARD-VN1 O8:5F-/STa/STb/LT CARD-VN2 O149: K91:F4/STa/STb/LT CARD-VN3 O64: F5/STa/STb/LT ml of inactivated whole cell vaccine contains 7.5 x 109 bacteria SAFETY TESTING  Naïve EFFICACY TESTING sows each received ml of vaccine Table 2: Summary of ELISA results on pre- and post- (approximately 1.5 x 1010 bacteria) at and 12 weeks vaccination sera samples of gestation Treatment group Mean (OD value) Control 0.2260a Litterguard 0.8129b EcoVac 0.9406b NIVR 0.8695b Table 1: Recorded criteria on safety study Group (8 sows) (Litterguard) Group (8 sows) (NIVR) Piglets born alive 80 82 Stillbirths Mummies (or died before 2 Recorded criteria a, b: p0.1) compared to an unvaccinated control group (p