AHSI magazine - University of Guelph

Animal Health 

Laboratory 

Laboratory Services Division 

ntario Ministry o Aricltre an oo Ministry o ral Aairs 

University of Guelph 

Animal Health Strategic Investment 

MAGAZINE 

 

Pg 5 

New and improved test methods 

Pg 12 

Disease surveillance 

Other collaborators 

Emergency preparedness 

Animal health tests 

Pg 13 Pg 19 

Pg 20

Contents 

EMERGENCY 

PREPAREDNESS 

07 

18 

DISEASE SURVEILLANCE 

Ear p necrosis in swine. 

Surveillance for enteric parvoviruses in 

poultry. 

Maedi-visna virus in Ontario sheep ocks. 

04 

05 

07 

08 

09 

Impact of partnership projects. 

Achievements and results. 

Equipment purchased. 

14 

Disease surveillance in swine. 

Network based disease spread model. 

Cyscercus ovis infecon in sheep carcasses. 

Chlamydia-like organism in rainbow trout . 

Q fever in sheep, goats, and farm workers. 

Ontario farm-call surveillance program. 

0 

 

 

 

5 

Viral and bacterial pathogens in broiler chickens. 

Novel toxin of Clostridium perfringens in calves. 

Chlamydophila spp. and Coxiella burne in caprine 

and ovine aborons. 

Inventory data and GIS locaon data into the AHL 

surveillance capacity. 

Bacterial hazards in the upper respiratory tract of 

swine by T-RFLP. 

Summer student assistantships. 

Johne’s disease in small ruminant dairy industries. 

Salmonella Enteridis baseline study in broilers. 

Surveillance of anmicrobial-resistant enteric bacteria 

in Ontario broilers. 

Harvesng surveillance data from AHL. 

Genotyping and detecon of Leptospira spp. in 

wildlife reservoir. 

Expanded data transfer from AHL to CAHSN. 

Regional PRRS eliminaon trial. 

Risk-based surveillance of respiratory infecons in 

growing pigs. 

Surveillance of disease agents in wildlife living on 

farms.

7 

20 

2 

22 

2 

24 

28 

0 

 

Associaons between the incidence of Cryptosporidium 

parvum and selenium status in neonatal dairy calves in Ontario. 

Inclusion body hepas in broiler breeders. 

Surveillance system for anmicrobial resistance in food 

animal pathogens in Ontario. 

Is the treatment of exudave epidermis in pigs becoming a 

problem because of the emergence of anmicrobial resistance 

Ontario swine veterinarian-based surveillance pilot study. 

Epidemiological analysis of Mycoplasma bovis isolated from 

Canada in the past 0 years. 

Molecular typing of Coxiella burnei by MLVA. 

Mycoplasma speciaon by molecular biology assays. 

Equivalency of strong posive Johne’s disease serum ELISA and 

interpretaon of repeated tesng for Johne’s disease in Ontario. 

Leporid herpesvirus 4 in Ontario rabbits. 

Validaon of MALDI-TOF for bacterial speciaon. 

ELISA detecon of Clostridium perfringens Cpb2 toxins in 

swine and cale. 

Evaluaon of massively parallel sequencing for detecon and 

characterizaon of viruses. 

Haemophilus parasuis detecon and typing. 

Coccidial DNA bar-coding. 

Development of rapid idencaon of Mycoplasma. 

Avian Chlamydophila and Coxiella infecon in Ontario. 

New PCR and ELISA tests for mammalian viruses. 

Validaon of serum amyloid A in horses. 

Development of immunohistochemical tests for small ruminant 

lenviruses. 

Brachyspira in dirty egg syndrome. 

Genotyping of fowl adenoviruses. 

Comparing four dierent sample types and three dierent culture 

methods for Salmonella spp. detecon in poultry. 

Anthelminc resistance in ocks with indicators of 

gastrointesnal parasism. 

Molecular typing method for characterizaon of Giardia 

duodenalis. 

AHSI-funded projects 2008-20. 

Theses. 

Summer students. 

AHSI presentaons and posters. 

AHSI publicaons. 

Supported by the OMAF and MRA - UofG 

Agreement through the Animal Health 

Strategic Investment fund (AHSI) managed 

by the Animal Health Laboratory of the 

University of Guelph. 

ASSOCIATE VICE-PRESIDENT, RESEARCH 

ANIMAL HEALTH LABORATORY, DIRECTOR 

ANIMAL HEALTH AND WELFARE BRANCH, 

DIRECTOR - OMAF and MRA 

VETERINARY SCIENCE AND POLICY, 

MANAGER - OMAF and MRA 

LEAD VETERINARIAN - PREPAREDNESS & 

PLANNING - OMAF and MRA 

COORDINATOR, ANIMAL HEALTH LAB 

PROGRAM - OMAF and MRA 

PROJECT ADMINISTRATORS 

GRAPHIC DESIGN AND PHOTO COORDINATOR 

COPY EDITOR 

STAFF SUPPORT

AHL 

H E R E 

IMPACT OF PARTNERSHIP PROJECTS 

S Inn n Ana Ha 

hen we are dealing with Avian Inuenza, Salmonellosis, and 

other potenally serious animal diseases, the stakes are high. 

Protecng the health of Ontario’s animals and, by extension, 

people, is one of the most important jobs we do, and the 

University of Guelph’s Pathobiology – Animal Health Laboratory 

is one of the best places to do it. 

For the past ve years, the lab has used a 7.5 million investment 

from the Ontario Ministry of Agriculture and Food and Ministry of 

Rural Aairs named the Animal Health Strategic Investment – to 

develop new tests and improve current 

tests, monitor disease prevalence, and 

develop and update emergency plans 

and procedures. 

The results are impressive. Animal 

disease surveillance protocols have been 

enhanced. There is more and stronger 

data on which to base decision-making, 

policy and programs. And, because of 

ghtened protocols and simulaons, 

Ontario’s livestock industries are far 

beer prepared to handle a future outbreak of disease. 

Even more impressive are the strengthened es that have 

resulted from the collaboraon that took place over the course 

of the research projects – among producers, veterinarians, the 

laboratory, the university and the government. The relaonships 

that have been built over the past ve years will serve all of us, 

and the Ontario public, well into the future. 

Congratulaons to everyone involved in this program – you have 

much to celebrate. 

Deborah Stark 

Deputy Minister of Ministry of Agriculture and Food 

and Ministry of Rural Aairs 

Pn Ana and Han Ha 

- Tda and n F 

Keeping our food safe and ensuring that our animals are healthy is 

essenal to the connued success of Ontario’s agri-food industry. 

The University of Guelph’s partnership with the Ontario Ministry 

of Agriculture and Food and Ministry of Rural Aairs is key to this 

endeavour, especially through its support of the Animal Health 

Laboratory (AHL). The AHL houses a crical mass of experse that 

monitors animal health across the province. AHL’s accurate and 

rapid detecon and diagnosc facilies help protect the health 

of Ontarians while promong animal welfare and maintaining 

producon performance. 

The OMAF and MRA - U of G Partnership 

has helped make this possible. For 

example, in 2008 the partnership 

provided addional funding for 5 years 

through the Animal Health Strategic 

Investment program. This crical support 

resulted in important innovaons, 

including new tests for emerging 

diseases; established a baseline for 

the early detecon of new hazards; 

and developed and tested emergency 

conngency plans for managing disease outbreaks. 

Ontario’s capacity to ensure public and animal health and wellbeing 

has been greatly increased through the addion of such 

sophiscated tesng and high technology analycal equipment, 

along with the highly qualied personnel who perform this work. 

As well, the Animal Health Strategic Investment program has 

supported valuable research and development projects. 

The AHL, University of Guelph and OMAF and MRA partnership is 

having a posive impact on Ontario’s agri-food system. This unique 

collaboraon has enabled advancements in animal health tesng 

and surveillance that would not have been possible without a joint 

eort. The resulng research and innovaon is contribung to 

evidence-based policy in Ontario, which ulmately contributes to 

improved public health. 

I am pleased to share with you the AHSI Magazine, which 

highlights the important work of the Animal Health Strategic 

Investment Program. 

Richard Moccia 

Associate Vice-President Research, Strategic Partnerships 

University of Guelph

Achievements and results 

The OMAF and MRA - AHL Animal Health Strategic Investment has supported over 

50 projects since 2008 and has developed or improved numerous tests for emerging 

pathogens, enhanced health surveillance activities, and improved emergency 

preparedness. 

AHL 

H E R E 

ACTH (adrenocorcotropic hormone) - chemiluminescence 

Acnobacillus pleuropneumoniae - PCR 

AEV (Avian encephalomyelis virus) - ELISA 

Aleuan disease virus - PCR 

Anaplasmosis anbody - cELISA 

Avian adenovirus - PCR 

Avian bornavirus - RT-PCR 

Avian paramyxovirus (APMV-) - RT-PCR 

Avian paramyxovirus - ELISA 

Avian reovirus - PCR 

Bacterial ID, MALDI-TOF 

Bartonella spp - PCR 

Brachyspira hyodysenteriae - PCR 

Brachyspira hyodysenteriae - real-me PCR 

Brachyspira hyodysenteriae and pilosicoli - real-me PCR 

Brachyspira intermedia - PCR 

Brachyspira pilosicoli - real-me PCR 

Brachyspira spp - PCR 

Bovine respiratory syncyal virusinfecous bovine rhinotracheis 

virus Parainuenza virus mulplex - real-me PCR 

Bovine viral diarrhea virus (BVDV) - real-me RT-PCR 

BVDV - real-me RT-PCR typing 

Canine distemper virus - real-me RT-PCR 

Caninefelinemink parvovirus - real-me PCR 

Chicken anemia virus - ELISA 

Chicken anemia virus - PCR 

Chlamydophila (Chlamydia) spp - PCR 

Chlamydophila abortus - real-me PCR 

Chlamydophila psiaci - real-me PCR 

Classical swine fever virus - real-me RT-PCR 

Corsol, ACTH smulaon prole - chemiluminescence 

Corsol, dexamethasone suppression prole - chemiluminescence 

Corsol, dexamethasone suppression prole, equine - 

chemiluminescence 

Corsol, single sample - chemiluminescence 

Coxiella burnei - real-me PCR 

E. coli, VTEC (verotoxigenic) typing, O57O (r), companion 

other - PCR 

E. coli, VTEC (verotoxigenic) typing, O57O (r), food animal - 

PCR 

E. coli, VTEC, companionother - PCR 

E. coli, VTEC, food animal - PCR 

Elephant herpesvirus -7 - PCR 

Equid herpesvirus - real-me PCR 

Equine arteris virus - real-me RT-PCR 

Equine arteris virus - virus neutralizaon assay 

Equine encephalis virus (EEEVEEV) - RT-PCR 

Foot-and-mouth disease virus - real-me RT-PCR 

Fowl adenovirus - real-me PCR 

Giardia duodenalis - genotyping 

IBDV (Infecous bursal disease virus) - R ELISA 

IBV (Infecous bronchis virus) - ELISA 

IDE BVDV PI - angen ELISA 

Immunohistochemistry, CD 

Immunohistochemistry, CD 

Immunohistochemistry, Chlamydophila 

Immunohistochemistry, cell markers 

Immunohistochemistry, chromogranin 

Immunohistochemistry, Feline coronavirus 

Immunohistochemistry, melan A 

Immunohistochemistry, S00 

Immunohistochemistry, synaptophysin 

Immunohistochemistry, TGEV 

Immunohistochemistry, Toxoplasma, companionother 

Infecous bronchis virus (IBV) - RT-PCR 

Infecous bursal disease virus (IBDV) - real-me RT-PCR 

Infecous laryngotracheis virus (ILTV) - PCR 

Inuenza A virus, avian, H5, food animal - real-me RT-PCR 

Inuenza A virus, avian, H7, food animal - real-me RT-PCR 

Inuenza A virus, avian, matrix, real-me RT-PCR 

Inuenza A virus, canine - real-me RT-PCR 

Inuenza A virus, swine - PCR typing 

Inuenza A virus, swine - RT-PCR, gel 

Inuenza A virus, swine - RT-PCR, gel, virus typing 

Insulin - chemiluminescence 

Lawsonia intracellularis - PCR 

Mycobacterium paratuberculosis - real-me PCR 

Mycoplasma bovis - real-me PCR 

Mycoplasma bovis - AFLP typing 

Mycoplasma bovis - MLST typing 

Mycoplasma gallisepcum - PCR 

Porcine circovirus 2 (PCV-2) - PCR detecon 

PCV-2 - real-me PCR 

PCV-2 - real-me PCR typing 

PCV-2 - PCRRFLP, 902bp 

Porcine parvovirus - real-me PCR 

PAGE 

5

AHL 

H E R E 

Porcine reproducve and respiratory syndrome virus (PRRSV), 

09 bp - RT-PCRRFLP 

PRRSV, 7 bp - RT-PCRRFLP 

PRRSV, 9 bp - RT-PCRRFLP 

PRRSV, European strain - rt-RT-PCR (Tetracore) 

PRRSV, Next Generaon NorthEur, saliva - rt-RT-PCR 

PRRSV, NorthAm strain - rt-RT-PCR (Tetracore) 

PRRSV, NorthAmEur, same day tesng - RT-PCR 

Progesterone (P4) - chemiluminescence 

Protein electrophoresis - agarose gel 

Rotavirus group AC - real-me RT-PCR 

Rotavirus group B - RT-PCR 

Salmonella Enteridis - PCR 

Salmonella spp - PCR 

Salmonella Typhimurium DT04 - PCR 

Salmonella Typhimurium - PCR 

Scrapie resistance PrP genotyping, 

codons , 54, 7 - real-me PCR 

Streptococcus equi, companionother - SeM gene PCR 

Swine cytomegalovirus - real-me PCR 

Testosterone - chemiluminescence 

Tetracore EZ-PRRSV MP - mulplex real-me RT-PCR 

Thyroid, total T4 (TT4) - chemiluminescence 

Tritrichomonas foetus - PCR 

West Nile virus, real-me RT-PCR 

The AHSI fund has helped to expand the services at 

the Animal Health Lab by increasing diagnostic testing, 

validations, and interpretation of tests for the animal 

industries in Ontario. 

Present 

With support from the 

Animal Health Strategic 

Investment fund, the AHL has 

developed improved methods 

to detect, monitor, and recover 

from an animal health crisis 

in Ontario. Over the 5-year 

period, new test methods 

have been developed and 97 

methods have been improved 

to test for emerging disease 

hazards. The development 

of these tests has involved 

high-throughput equipment, 

such as, the LightCycler 480 

real-time PCR, GS Junior, 

Luminex and the ELISA robot, 

and Ventana autostainer, in 

order to expand services for 

PCR, ELISA , and IHC at the AHL. 

These enhanced test methods 

contribute to the global impact 

of Ontario agriculture by 

providing cost- effective and 

accurate methods to detect 

and control potential animal 

health events. 

Future 

AHSI has been a very useful 

program for developing 

new tests, increasing our 

surveillance base, and 

preparing for emergencies. 

Next steps include full 

implementation of all of the 

new and revised tests that have 

been developed, and bringing 

all new equipment on-line. The 

ELISA robot has proved to be a 

boon to our testing throughput, 

and is being upgraded to 

next generation software 

and reconfigured for greater 

efficiency. The new MALDI- 

TOF is at full capacity, and has 

improved turnaround times on 

bacterial speciation. We plan 

to further enhance integration 

of AHL lab surveillance results 

into the national picture, and 

would like to expand cluster 

analysis and other surveillance 

tools developed through 

AHSI. Annual emergency 

exercises will continue, as 

initiated by AHSI, with the aim 

of continuous improvement 

of this vital function, both 

with respect to postmortem 

room capabilities as well as 

laboratory surge capacity for 

recovery activities. 

PAGE 

6

AHL 

H E R E 

Equipment Purchased 

Lab Equipment 

Approximately . million has been 

invested into the AHL to improve efficiency 

and to achieve high-throughput of 

automated testing. 

Selected examples 

Roche LC480 thermocycler 

General Electric Nanovue spectrophotometer 

Ventana Benchmark T 

Bionumerics soware 

Luminex 200 

LifeSep 9F Cell separator 

MagMax Express 9 MME-9DW 

QIAcube extractor (2) 

Siemens Immulite 000 

Somagen Sebia coagulaon analyzer 

Hydrasys 2 and phoresis scanner E-400 4C 

Thermosher ELISA automaon system 

MALDI-TOF MS 

CBG Solvent recycler 

Eppendorf centrifuge 540 

ThermoScienc centrifuge CL-2 

Concept 400 anaerobic chamber 

PAGE 

7

AHL 

H E R E 

Disease Surveillance 

Surveillance studies and epidemiologic investigations have improved our understanding of 

animal health and diseases in Ontario. 

Disease surveillance in swine with minimal exposure to veterinary diagnostic services 

Tim Blackwell 

Disease impacts the Ontario swine industry by lowering financial 

returns, decreasing animal welfare and increasing antimicrobial 

use. This study demonstrated that an effective swine disease 

surveillance system could be established at the farm level 

utilizing a weekly mortality reporting format. This producerbased 

program could serve as a model for a province-wide 

on-farm disease surveillance network in the future. An effective 

on-farm surveillance system would benefit the Ontario swine 

industry by detecting and containing new disease outbreaks 

thereby improving economic returns for swine producers while 

establishing confidence in our trading partners regarding the 

purchase of pigs or pork originating from Ontario. 

Contact network based disease spread model in Ontario - Javier Sanchez 

The movement of animals and other related risk goods amongst farms is recognized as the main mechanism of spread of infectious 

diseases on farms. The main goal of this project was to carry out a pilot study to assess the implications of contact network structures 

and production systems on disease spread and on the effect of different control measures through simulation modelling. This study 

developed a more realistic disease simulation model that could be applied to other diseases, such as foot-and-mouth disease, PRRS, 

swine influenza, and will aid in developing sound disease contingency policies and plans for Ontario regulatory officials and swine industry 

practitioners. 

A survey of risk factors associated with condemnation of sheep carcasses due to 

Cysticercus ovis infection - Paula Menzies 

As Cysticercus ovis continues to be a significant cause of sheep carcass condemnation at slaughter, we 

set out to determine the prevalence and distribution of farms in Canada with carcass condemnations 

due to Cysticercus ovis infection and the risk factors associated with this parasitic disease in slaughter 

animals. Proper disposal of deadstock and prevention of scavenging of deadstock are important 

to reducing the risk of this infection. A C. ovis trace-back system was successfully created and the 

sheep industry is encouraged to continue using it in provincially inspected abattoirs across Ontario. 

Understanding the factors that might put a farm at higher risk for infection helps to develop an effective 

control and eradication program for Ontario sheep flocks. 

PAGE 

8

AHL 

H E R E 

The OMAF & MRA-AHL Animal Health Strategic Investment has supported 

31 surveillance projects and has invested $3.4 million to help provide 

innovative strategies to improve detection of animal diseases. 

Chlamydia-like organism in Ontario 

rainbow trout - John Lumsden 

In Ontario, a Chlamydia-like organism (CLO) has been 

identified as the cause of necrotizing gill disease that 

has affected arctic char. The objective of this project 

was to provide a tool to rapidly diagnose an emerging, 

production-limiting disease affecting rainbow trout 

aquaculture in Ontario. Contrary to published results and 

original hypothesis, Burkholderiales sp. was correlated 

with the disease rather than CLO. For these reasons, it is 

likely that the disease has been present in the province for 

quite some time. A molecular assay to detect both types 

of organism has been developed and has been applied to 

tissues from multiple fish species, and from both archived 

material and ongoing disease epizootics. The tools 

developed from this project provide answers to the impact 

of the disease, the range of species affected, and potential 

control measures in Ontario. 

Q fever (Coxiella burnetii) in Ontario sheep, goats 

and farm workers - Paula Menzies 

Coxiella burnetii has been known to be an important cause of abortion in 

small ruminants and contributes to human illness in Canada. This project 

aimed to determine the seroprevalence of Q fever (Coxiella burnetii) in 

sheep, goats and their farm workers in Ontario. Researchers provided 

additional examination of risk factors and protective measures that are 

associated with this disease. With 7.4 of humans and approximately 

0 of farms tested positive for this infection, it suggests that infection 

from C. burnetii is common in people who care for sheep and goats in 

Ontario. Factors associated with an increased risk of infection are being 

explored. The project benefits Ontario by developing methods to protect 

people from this serious pathogen. 

Ontario farm-call surveillance program - Kathy Zurbrigg 

The Ontario farm-call surveillance programs provided a unique way to determine data recording methods to ensure a sustainable 

livestock disease surveillance system among private veterinary practices. The quality of livestock laboratory data at the Animal Health 

Lab was enhanced and veterinarians were encouraged to conduct postmortems on moribunddead animals; surveillance on high-risk 

animals was monitored. This project provides Ontario with a model of a more extensive and rapid surveillance system to detect and 

analyze unusual trends in clinical syndromes among farmed animals, demonstrates excellent coverage of the province, and provides 

surveillance for zoonotic disease in animals. 

PAGE 

9

AHL 

H E R E 

Viral and bacterial pathogen surveillance in broiler chickens - Michele Guerin 

Lile is known about the prevalence of viral pathogens or associated risk factors in commercial broiler ocks in Canada. This project 

determined the baseline prevalence of 2 viral and bacterial pathogens of poultry health importance among 2 randomly selected 

commercial broiler chicken ocks in Ontario. Results indicate that Ontario broiler ocks were exposed to Avian adeno-associated viruses, 

Avian reovirus, Chicken anemia virus, Fowl adenovirus, Infecous bronchis virus, and Infecous bursal disease virus during the growing 

period, but not to Avian encephalomyelis virus or Newcastle disease virus. These potenally pathogenic genotypes of FAdV and IBDV can 

inform vaccine development and disease control eorts in Ontario. 

Identification of novel toxin gene(s) associated with type A Clostridium perfringensassociated 

hemorrhagic abomasitis of calves in Ontario - John Prescott 

In Canada, clostridial abomastitis has been reported as a sporadic disease that causes sudden death in calves. This project aimed to 

identify unique characteristics including novel toxin(s) of Clostridium perfringens associated with fatal bovine necrotizing abomasitis 

of calves, and to improve diagnosis and understanding of this infection. Neither genomic nor proteomic approaches identified novel 

toxin(s) in an isolate from this disease, but an alternate and unexpected picture of virulence emerged suggesting that anomalous 

virulence gene regulation might contribute to pathogenicity in this isolate. We looked for the novel genes identified in this genome 

sequence, as well as other existing known virulence genes in C. perfringens, in a collection of C. perfringens isolates from calves with 

necrotizing abomasitis, from adult cattle with jejunal hemorrhage syndrome, and from diarrheic calves with undifferentiated diarrhea. 

We did not find genes unique to the abomasitis isolates. The annotated genome of the Clostridium perfringens isolate sequenced from 

a case of fatal necrotizing abomasitis is now available in the international gene library, GenBank, for reference. This project benefits 

Ontario’s agriculture by providing in-depth genome analysis of an organism associated with a commonly fatal disease of young calves atfoot 

or on milk replacer. 

PAGE 

10

Investigation of the role 

of Chlamydophila spp. 

and Coxiella burnetii in 

caprine and ovine abortion 

Hugh Cai 

Abortion in sheep and goats has 

become a growing concern in Ontario 

and this project investigated the role 

of Chlamydophila abortus and Coxiella 

burnetii in caprine and ovine abortion in 

Ontario and determined if abortions are 

related to pathogen loads. This was done 

by testing Cp. abortus and C. burnetii in 

abortion and non-abortion samples using 

quantitative real-time PCR. Shedding 

routes of Cp. abortus and C. burnetii were 

determined by testing different tissue 

and fluid samples for the pathogens. 

Results show that the leading cause of 

ovine abortion was Toxoplasma gondii. 

C. burnetii was identified by rtPCR in 

of (9.0 ) sheep and 72 of 9 (75) 

of goat abortion submissions, but was 

considered to be significant in causing 

abortion in only 5 of () sheep 

and of 72 (5) goat submissions that 

tested positive. C. abortus was identified 

by rtPCR in 42 of 2 (2) sheep and 

54 of 92 (59) goat submissions, but was 

considered the cause of the abortion in 

of 42 (8) sheep and 4 of 54 () 

goat submissions which tested positive. 

Optimal sensitivity and specificity cutpoints 

for the rtPCR copy number for C. 

abortus and C. burnetii were determined 

using the final pathology diagnosis as the 

reference test. The information collected 

assists veterinarians and pathologists in 

selecting samples and interpreting test 

results in diagnosis of abortion caused 

by Chlamydophila abortus and Coxiella 

burnetii. In addition, the prevalence 

information is useful for prevention and 

treatment for these particular diseases. 

Inventory data and GIS 

location data into the 

Animal Health Laboratory 

surveillance capacity 

David Kelton 

Due to data limitaons that the AHL can 

provide regarding the numerator data 

in the form of diseased animal counts, 

this surveillance project evaluated the 

feasibility of combining animal health, 

animal inventory and animal locaon 

data with exisng AHL data to enhance 

the passive surveillance capacity of the 

AHL and the OAHSN, and; the ulity of 

bulk tank tests (PCR and ELISA) for the 

purposes of contribung to the passive 

and acve surveillance capacity of the AHL 

and the OAHSN. It also served as a pilot 

project to combine the AHL’s surveillance 

capacity to bring the cow milk test data 

and the dairy cale inventory data from 

CanWest DHI and the GIS locaon data 

from Dairy Farmers of Ontario into the 

surveillance system. From this study, 

there were major limitaons to moving 

this iniave forward because of the 

lack of a common herd idener among 

the three major sources of data used 

to complete this project. It is strongly 

recommended that the AHL work with 

veterinary praconers and animal 

industry organizaons to establish a 

common protocol for herd idencaon, 

such as Premises ID, to enhance the 

passive surveillance capacity in Ontario. 

At this point, the contribuon of bulk tank 

tests (PCR and ELISA) to the passive and 

acve surveillance capacity of the AHL 

and the OAHSN is poor, and alternaves 

need to be sought. This project benets 

the province by informing us of what 

needs to be done in order to enhance 

tesng capacity and services of the two 

laboratories and the surveillance coverage 

within Ontario. 

Evaluation of bacterial hazards in the upper respiratory tract of swine by terminal 

restriction fragment length polymorphism analysis - Janet MacInnes 

AHL 

H E R E 

There is growing recognition of the need to characterize the microbial communities of the upper respiratory tract of swine. Accordingly, 

this project sought to compare information obtained by routine bacterial culture with that obtained using terminal-restriction fragment 

length polymorphism (T-RFLP) analysis. The researchers also determined whether the presence or absence of a particular bacterial 

community was associated with a certain clinical condition. The results of this study indicated that T-RFLP analysis is more sensitive, but 

less specific, than classical culture and biochemical characterization. That said, known bacterial pathogens such as Actinobacillus spp., 

Campylobacter sp., Escherichia coli, Haemophilus parasuis, Lawsonia intracellularis, Mycoplasma spp., Salmonella spp., Staphylococcus 

spp., Streptococcus spp. as well as numerous non-pathogenic genera were presumptively identified by T-RFLP. Significant associations 

between T-RFLP clusters and the presence of anemia, abscesses, PRRS virus, and Mycoplasma spp. were also demonstrated. This 

project lays the foundation to determine whether T-RFLP, a relatively simple molecular method, can aid clinicians and producers in the 

identification of important bacterial pathogens and in the understanding of possible roles of commensal organisms. 

PAGE

AHL 

H E R E 

Disease Surveillance 

invesgaons detecon enhancements follow-up 

Summer student assistantships, AHL 

Mammalian Virology and Bacteriology 

Durda Slavic / Susy Carman 

Renewal of the veterinary diagnostic laboratory workforce 

continues to be a challenge, especially in the area of veterinary 

microbiology, and very few veterinarians enter this specialty 

because of lack of familiarity with this discipline. The project 

objective was to employ 2 second-year DVM undergraduate 

students for the summer of 200, and rotate them through 

the AHL mammalian virology and bacteriology laboratories. 

Each student was involved in day-to-day activities in both lab 

sections (tissue preparation, case setup, etc), and molecularbased 

testing, such as PCR. Both DVM undergraduate students 

were trained in the separate disciples of bacteriology and 

mammalian virology, and completed their case report and 

assessment of the program. The project’s intent was to 

provide these students with sufficient exposure and technical 

background experience to pursue graduate training in 

veterinary microbiology, and eventually become veterinary 

laboratorians in Ontario. 

Johne’s disease in Ontario’s 

small ruminant dairy 

industries - Andria Jones 

Johne’s disease, caused by Mycobacterium 

avium subspecies paratuberculosis (MAP) 

affects both domestic and wild ruminants. 

This study looked at determining the herd 

level prevalence, potential risk factors, level of agreement between 

various diagnostic methods, and validation against reference tests 

for Johne’s disease in Ontario dairy sheep flocks and dairy goat 

herds. Results indicate that Johne’s disease is common in goat 

herds and sheep flocks. A diagnostic test was performed using fecal 

cultures as a reference standard and it was the most sensitive test 

in goats and sheep; none of the other tests changed their ranking 

with respect to the calculations. The results from the validations 

help inform reliable diagnostic test recommendations for small 

ruminants and help to identify a faster, convenient and economical 

test for Ontario veterinarians and producers to use in surveillance 

of Johne’s disease on-farm. 

Salmonella Enteritidis baseline study in Ontario commercial broilers - Michele Guerin 

Salmonella enterica serovar Enteritidis (SE) has been a growing concern in Ontario and has become the top Salmonella serovar 

isolated from humans. The main objective of this project was to determine the current flock level prevalence of SE among a 

random selection of commercial broiler flocks in Ontario at the end of the growing period. In total, 0 samples were tested by an 

immunomagnetic separation method and 2,20 (4.9) were positive for Salmonella spp. of the positive isolates (0.5) were 

confirmed positive for SE using a polymerase chain reaction technique and all isolates were phage type 8. Results from this project 

have benefited Ontario agriculture by filling a gap in our current knowledge of the prevalence of SE among commercial broiler 

chicken flocks in Ontario. 

PAGE 

12 

Surveillance of antimicrobialresistant 

enteric bacteria in 

Ontario broilers - Michele 

Guerin 

Antimicrobial-free production is regarded 

as an important industry intervention 

to reduce antimicrobial residue and 

resistance (AMR). The purpose of this 

pilot study was to evaluate the utility 

of the sampling and data collection 

methodologies for the national farmlevel 

surveillance program in broilers to 

address knowledge gaps in understanding 

the epidemiology of AMR in broiler 

chickens. The study design allowed for 

a comparison of the microbial quality 

and AMR profiles in conventional (CON), 

antimicrobial-free (AMF), and organic 

(ORG) broiler production systems, as 

well as a determination of baseline AMR 

levels of these pathogens in domestic 

and U.S.-sourced day-old chicks as a 

surrogate for AMR at the breeder and 

hatchery levels. To date, Salmonella has 

been isolated from flocks raised under 

each production system type (CON, AMF, 

ORG), at both chick placement and preharvest, 

and that Salmonella Kentucky 

was the most frequently isolated serovar 

regardless of production system type. 

Escherichia coli was frequently isolated 

from all production system types at both 

sampling times, regardless of sample 

type. Campylobacter was only isolated 

from samples obtained at pre-harvest, 

and Campylobacter jejuni was the most 

commonly found species. This study 

helps to initiate an integrated on-farm 

assessment that can be used to further 

improve these operations in Ontario and 

Canada.

Outside contributors to AHSI 

University of Prince Edward Island, Fleming College, Canadian Food Inspecon Agency, and Ontario Pork Industry Council 

AHL 

H E R E 

Harvesting additional surveillance 

information from existing AHL 

diagnostic laboratory data 

Javier Sanchez / Crawford Revie 

Surveillance has been a growing concern for the 

laboratory and for the provincial government. This 

study aimed at developing and implementing advanced 

analytical techniques for OMAF and the AHL to harvest 

additional knowledge from passive animal health 

surveillance information on an ongoing basis. A 

syndromic surveillance system was created to generate 

an alarm concerning an elevated number of submissions 

for respiratory diseases – i.e., based on abnormal 

increases in the number of submissions for specific 

clinical syndromes. The tool developed by this project 

has been put into use, and benefits Ontario agriculture 

by improving surveillance analyses and information 

harvested from routinely collected AHL case submission 

data. 

Genotyping and detection of Leptospira spp. in 

wildlife reservoir hosts in Ontario - Karen Shearer 

Leptospira infection causes disease in humans and domestic livestock 

and this project sampled whole kidneys from potential wildlife reservoirs 

collected from hunters, trappers, fatalities from wildlife rehabilitation 

facilities, and motor vehicles across Ontario. In total, 40 wildlife samples 

were evaluated for the presence of Leptospira spp. bacteria through both 

immunohistochemistry and PCR and samples were plotted geographically 

using a global positioning system (GPS). Further analysis by genotyping 

was conducted on the Leptospira bacteria on PCR positive samples, in an 

attempt to determine if the type of leptospiral organisms found in the 

wildlife samples were consistent with those known to affect domestic 

animals and humans. This project benefits Ontario by acquiring current 

knowledge of the status of the distribution of Leptospira spp. in wildlife 

living in proximity to humans and livestock across Ontario. 

Expanded data transfer from 

AHL to CAHSN - Harold Kloeze 

Transferring Ontario diagnostic laboratory data to the Canadian Animal 

Health Surveillance Network (CAHSN) for surveillance is a concept that is 

fully supported by the AHL. This project identified information necessary 

to allow the data transfer to support notifiable disease reporting to 

the Canadian Food Inspection Agency, the World Organization for 

Animal Health (OIE), and OMAF. It also looked at initiating frequent 

routine transfer of selected information to CAHSN and ensured the 

capability to turn on and off such transfers if needed during an animal 

health emergency. This project benefits the Ontario agriculture sector 

by developing the process to automatically transfer numerator and 

denominator data of selected federally immediately notifiable diseases 

from the AHL data warehouse to the CAHSN database. 

Approved Ontario ARC&E projects. 

Regional PRRS elimination trial - Lori Moser 

To stay competitive with trading partners and to avoid any possible future 

disease outbreaks, this project sought to implement a voluntary regional porcine 

reproductive and respiratory syndrome (PRRS) ARC&E (area regional control and 

elimination) pilot project in Ontario. This project advanced the Ontario pork 

industry in disease surveillance, control and biosecurity. It also assessed the 

applicability of PRRS control and elimination approaches as executed in the U.S. 

to date, which added to the control and elimination strategies for Ontario. This 

project has benefited Ontario producers by informing them of their PRRS virus 

status, reducing disease prevalence and transmission risks in their areas, and 

increasing open communication and awareness among producers in the Niagara 

region. Producers are communicating with each other concerning change of site 

status and placement of positive pigs in the area. The success of this project has 

led to the initiation of other producer-driven projects in southern Ontario. 

PAGE

AHL 

H E R E 

- Robert Friendship 

Ear necrosis is a syndrome 

observed in pigs ranging in age 

from to weeks and has 

become an emerging problem 

that is difficult to control on 

affected farms. The overall 

goal of this project was to 

determine the cause of ear 

tip necrosis in swine and to 

investigate the associated risk 

factors. This work has helped 

to clarify that ear necrosis is 

widespread in Ontario and 

that it is possibly caused by 

certain toxin-producing strains 

of Staphylococcus aureus and 

or S. hyicus. In addition, there is 

an association between severe 

ear necrosis lesions and ear 

biting and, in these cases, the 

economic cost associated with 

the disease can be substantial. 

This project benefits Ontario 

by developing monitoring 

techniques to help stop the 

spread of this disease and to 

help reduce the impact of the 

disease on affected farms. 

PAGE 

14 

-Eva Nagy 

In Ontario, there is currently 

no surveillance data for 

the presence of the enteric 

parvovirus and this project 

sought to validate diagnostic 

tests for this virus, demonstrate 

the presence of parvovirus 

in Ontario poultry, in both 

broiler and layer flocks, and 

compare the phylogeny of the 

Canadian parvoviruses with 

the North American viruses. 

We completed the testing of 

2 samples using a NS gene 

PCR which had originated 

from 52 broiler and broiler 

breeder flocks and 2 layer 

flocks in Ontario. Out of the 

2 samples, 47 (29) tested 

positive for the presence of 

parvovirus DNA. The direct 

benefit to Ontario’s agriculture 

is that it provides a quick and 

reliable test for the detection of 

chicken parvovirus and disease 

control and protection across 

Ontario. 

- Paula Menzies 

Due to the Canadian Food Inspecon Agency (CFIA) ELISA being 

privazed to a provincial laboratory outside Ontario and the cost 

of an individual test for maedi-visna virus (MVV) having risen from 

2.50 to 7.50, this project compared test performance of the 

CFIA ELISA, VMRD ELISA, IDE ELISA and Pourquier ELISA, and PCR 

with respect to accuracy using sera and whole blood from naturally 

infected Ontario sheep ocks. In addion, this project also sought to 

isolate and sequence the MVV strains from naturally infected sheep 

ocks. Results indicated that the best comparave sensivity and 

excellent agreement (kappa) to the CFIA maedi-visna test was the 

Elitest (HYPHEN Biomed). Use of a PCR to detect virus found that 

85.8 of seroposive samples and .7 of seronegave samples 

were posive which meant that there was excellent specicity and 

the presence of a posive PCR in the face of a negave serological 

result, likely represenng infecon without seroconversion. 

Sequencing of the virus as idened in the PCR posive samples 

revealed a diverse phylogenec analysis within ocks and between 

ocks within Ontario, but dierent from strains of caprine arthrisencephalis 

virus found in goats. The results from this project 

helped the AHL to decide upon an accurate and aordable test that 

is how oered to Ontario veterinarians and sheep producers to 

detect and eradicate MVV.

AHL 

H E R E 

Zvonimir Poljak 

Porcine reproductive and respiratory syndrome (PRRS) has been considered 

primarily a production-limiting disease, and as such, it is of primary interest for 

individual producers and herd veterinarians. This project determined herd-level 

prevalence of infection and exposure to several respiratory pathogens in swine, 

evaluated sensitivity and specificity of tests applied in high-risk groups for PRRS 

virus and PCV2, compared classification of PRRS viruses on the basis of open 

reading frame (ORF)5 and ORF7 phylogenetic analysis of nucleotide sequences; 

established a baseline list of Porcine circovirus 2 sequences currently present in 

Ontario finisher pigs; determined which individual factors increase the likelihood 

of detection of pathogens included in the proposed study; and estimated whether 

groups of close-out pigs exist with respect to positivity to certain pathogens. 

Results indicated that in order to increase the sensitivity of detection of PRRS 

virus in a finisher barn, finisher pigs with dyspnea or rough hair coat, and without 

lameness should be selected. Further work on test accuracy should be done since 

assay accuracy could be different between specimens obtained for diagnostic and 

for monitoring purposes. This work provides current epidemiological information 

about prevalence of several important pathogens at the herd-level and their more 

precise genetic grouping. Also, it could be directly used to inform producers and 

herd veterinarians, and provides key pieces of methodological information about 

risk-based surveillance as the most efficacious method of herd-level surveillance. 

- Claire Jardine 

Wild mammals have been implicated as reservoirs 

for several important zoonotic disease agents 

associated with production animals. The purpose 

of this research was to determine if enteric 

pathogenic and antimicrobial-resistant bacteria 

isolated from farm wildlife and the environment 

are phenotypic and genotypic matches to those 

found in production animals on the same farm. 

It also looked at demonstrating the feasibility of 

routinely testing wildlife living on or near farms for 

pathogens of animal and public health significance. 

The samples that were positive for parvovirus came 

from 2 farms; which indicates that the parvovirus 

was present on 48 of the farms at the time of 

testing. Understanding the role of wildlife in the 

dynamics of these disease agents is essential for 

the development of effective surveillance and 

control measures to deal with these diseases agents 

now and in the future. 

PAGE 

5

AHL 

H E R E 

More results... 

Associations between the incidence of Cryptosporidium parvum 

and selenium status in neonatal dairy calves in Ontario - Ken Leslie 

Cryptosporidiosis is an ongoing calf health problem for the dairy industry. The costs associated with 

diarrhea due to Cryptosporidium parvum are manifested though the loss of calves, as well as the failure 

to thrive following infecon. The intent of this study was to establish a surveillance program to evaluate 

the prevalence of C. parvum in dairy calves in Ontario, and to study the associaon of this infecon with 

selenium status, as well as the potenal for selenium supplementaon on newborn calf selenium status 

and the prevalence of C. parvum. Results indicated that supplemenng calves with injectable selenium 

(Dystosel) does increase circulang selenium levels and has a protecve eect against rotavirus infecon. 

The results from this project established a baseline for the rates of the infecon and disease caused by C. parvum in calves on dairy 

farms in Ontario. In addion, supplementaon of calves with selenium at birth would reduce the risk of rotavirus infecon and improve 

general health, but not necessarily reduce the risk of C.parvum infecon. 

Surveillance for inclusion body hepatitis 

in broiler breeders - Davor Ojkic 

Fowl adenoviruses (FAdVs) are widespread in domestic fowl and have 

been known to be associated with inclusion body hepatitis (IBH). The 

objective of the project was to investigate the prevalence of FAdV 

infection in Ontario broiler breeder flocks by examining serological 

status related to FAdV serotypes known to circulate in Ontario, and 

shedding of FAdVs. A serotype-specific microneutralization assay 

was developed to examine the level of flock exposure to FAdV08 and 

FAdV in order to determine the need for vaccination. Flocks that 

carried a sufficient level of antibodies need not be vaccinated, which 

decreases the cost associated with vaccination of Ontario flocks. This 

project benefited Ontario by allowing better assessment of the role of 

horizontal and vertical FAdV transmission. More detailed knowledge 

of virus distribution in IBH and non-IBH flocks allowed development 

of a vaccination strategy to prevent the occurrence of the disease and 

associated losses. 

Surveillance system for antimicrobial 

resistance in food animal 

pathogens in Ontario - David Pearl 

Antimicrobial resistance (AMR) has emerged as a major 

concern for animal and human health worldwide. Selection 

for resistance has been linked to therapeutic antimicrobial 

use in veterinary medicine, as well as prophylactic and 

growth promotion in agricultural production. This project 

focused on developing a surveillance program for AMR 

in animal pathogens of livestock in Ontario based on 

laboratory submissions to the AHL. The AMR surveillance 

system has been designed and has the ability to provide 

various stakeholders in animal health with regularly 

updated information concerning temporal changes in 

AMR patterns in various bacterial pathogens affecting 

Ontario livestock. Currently, the system has been used 

to retrospectively examine temporal trends in AMR in 

three swine pathogens, but the system can be expanded 

to include additional pathogens and commodity groups 

depending on data availability and quantity. 

PAGE 

16

AHL 

H E R E 

Ontario swine veterinarian 

based surveillance pilot study 

Robert Friendship 

Is the treatment of exudative epidermitis 

in pigs becoming a problem because of 

emergence of antimicrobial resistance? 


Exudave epidermis (EE), or greasy pig disease, is caused 

by Staphylococcus hyicus. The goal of this project was to 

invesgate whether treatment failure in cases of greasy pig 

disease was the result of emerging anmicrobial resistance. 

Anmicrobial resistance tesng revealed that almost all skin 

isolates of S. hyicus and S. aureus were resistant to penicillin 

and ampicillin, and the majority of isolates were also resistant 

to ceiofur. The results from this research can inform how 

Ontario pig farmers and veterinarians treat greasy pig disease, 

secondary infecons, and increase animal welfare by not using 

ineecve anbiocs. 

Given Ontario’s concern for recognizing and 

responding to emerging infections from domestic 

livestock of significance to food safety and to 

animal and public health, this project investigated 

the feasibility of conducting veterinary practicebased 

syndromic 

surveillance using 

the swine industry 

as a model. 

Participation and 

cooperation from 

swine veterinarians 

helped provide 

a large amount 

of surveillance 

data that often involved activities that went 

beyond their regular clinical duties. This 

project provided active surveillance within the 

Ontario swine industry based on this industry’s 

interest in enhanced surveillance following its 

experience dealing with the health and economic 

consequences from the introduction of novel 

strains of swine influenza virus, PRRS virus, and 

Porcine circovirus (PCV) 2b in recent years. 

Hugh Cai 

Mycoplasma bovis is the most important bovine mycoplasmal pathogen in North America, causing mass, pneumonia, arthris, decubital 

abscesses, os media, and other diseases in both dairy and beef cale. The overall objecve of this project was to perform molecular 

characterizaon, anmicrobial resistance proling, and epidemiological analysis on Mycoplasma bovis isolated from veterinary clinical samples. 

This study developed an M. bovis mul-locus sequence (MLS) typing scheme using the dnaN, hsp70, metS genes. This scheme produced 2 MLS 

types among the 2 M. bovis isolates collected in Ontario from 978-200. The MLS types revealed temporal distribuon of the isolates; over 

the course of three decades, some MLS types disappeared while new types appeared. Recent isolates had a greater variety of types, which may 

indicate that new strains are emerging more rapidly now than in the past. There was no obvious clustering by geographic locaon or breed or by 

ssue type. All M. bovis isolated in Ontario from the last three decades demonstrated suscepbility to enrooxacin, specnomycin and amulin, 

and over 7 of isolates were suscepble to danooxacin and gentamicin. Almost all isolates (99) were resistant to neomycin, and only 2 

isolates were suscepble to orfenicol. The suscepbility to chlortetracycline, oxytetracycline, lmicosin and tylosin tartrate decreased over 

me; more than of the isolates in 980s were suscepble to these anbiocs; the number of suscepble isolates dropped to less than 4 

and 7 in the 990s and 2000s, respecvely. Over 92 of the isolates from the 980s were suscepble to clindamycin and tulathromycin. The 

number decreased to less than 7 in the 990s, then increased back to over 8 in the 2000s. The isolates were clustered into two groups by 

MIC prole analysis, one consists mostly of isolates from the 990s (n58; 990s45, 2000s0, and 980s), and the other consists mainly of 

980s and 2000s (n55). Within the 980s-2000’s cluster, the isolates were further separated by years. It appears that anmicrobial resistance 

was not related to geographic locaon, animal breed (dairy or beef) or ssue sample type. This informaon provides Ontario with data that 

helps with the presentaon, treatment and management of bovine M. bovis infecon.

AHL 

H E R E

AHL 

H E R E 

Emergency 

Preparedness 

AHL necropsy laboratories emergency 

preparedness exercises and incident 

command system training - Maria Spinato 

Emergency planning has become an important operational goal for the AHL because of the 

risk of a foreign animal disease outbreak. The two primary objectives of this project were 

to train AHL supervisory staff in the principles of the Incident Command System (ICS), and 

to conduct emergency preparedness exercises in the postmortem laboratory sections. Seven AHL 

veterinarians and two OVC Department of Pathobiology faculty members achieved ICS certification 

and familiarized themselves with procedures used to interact with animal health partners at the 

front line of an emergency. The two foreign animal disease (FAD) simulation exercises held in 20 

and 202 were deemed successful at both Guelph and Kemptville postmortem laboratories. These 

full-scale operational simulations and ICS training have supported AHL’s emergency preparedness 

strategy, and offered opportunities for interactions with public stakeholders and animal health 

partners, thus facilitating coordination during an FAD outbreak. 

PAGE 

9

AHL 

H E R E 

Animal health tests 

Molecular typing of Coxiella burnetii 

 

variable number of tandrem repeat analysis 

(MLVA) - Shu Chen 

Q fever continues to be an on-going issue in Ontario,causes 

abortion and stillbirth in animals, and poses risks to human 

health. This Coxiella burnetii project established a rapid 

diagnostic method for genotyping of C. burnetii and to obtain 

surveillance information on Ontario’s historical samples to inform 

emergency preparedness. A multiplexed VNTR-PCR approach was 

developed and is able to directly type C. burnetii in complex field 

samples with rapid, reproducible, portable and cost-effective 

results. The genotype information will allow animal health 

and food safety professionals to know the risks associated with 

Ontario strains, facilitate more accurate identifications and early 

detection of an outbreak, and track sources of infection. 

Mycoplasma speciation by molecular biology 

assays - Hugh Cai 

Since Mycoplasma antibodies are not commercially available and 

are difficult to produce, alternative methods were needed to 

replace this traditional method that was produced in the early 

990s. This project was aimed at developing a cost-effective and 

rapid assay to identify Mycoplasma species. A real-time PCR 

followed by a high resolution melting curve analysis (rt-PCR-HRM) 

was developed using one set of universal primers specific for the 

spacer region between the S rRNA and the 2S rRNA genes. The 

rt-PCR-HRM assay was evaluated by testing field isolates of M. 

bovis, M. arginini, M. bovirhinis, M. bovigenitalium, M. iowae and 

M. spumans with results consistent with those of the fluorescent 

antibody test. This project provides Ontario with an inexpensive 

and rapid assay for the differentiation of Mycoplasma species in 

pure cultures. 

Equivalency of strong positive Johne’s disease serum ELISA results and interpretation of 

repeated testing for Johne’s disease in Ontario herds - David Kelton 

In collaboration with CanWest Dairy Herd Improvement and Ontario dairy herds participating in the Ontario Johne’s Education and 

Management Assistance Program (OJEMAP), this project sought to establish an equivalent high test (HT) positive samplepositive 

(sp) cut-point for the IDE Johne’s ELISA for both serum and milk. HT-positive cows were originally defined as having an antibody 

test result of .0 or higher in milk using the Prionics Parachek Milk ELISA. These cows are of particular concern because they are 

reliably classified as infected and a very high proportion of these cows (over 90) are actively shedding Mycobacterium avium ssp. 

paratuberculosis (MAP), the organism causing Johne’s disease, in their manure. HT positive cows must be culled from the herd as part 

of OJEMAP compliance. In order to be able to use the IDE test for program testing, an equivalent HT positive cut-point was needed. 

Based on results of this project, the HT cut-points for the milk and serum for the IDE test should be 2.0 and 2.50, respectively. These 

equivalent cut-points are important for the Ontario dairy industry in order to continue to remove cows at high risk of fecal shedding of 

MAP into the environment from the provincial dairy herd. 

Leporid herpesvirus in Ontario rabbits - Patricia Turner 

Herpesviruses are important agents of disease in humans and animals and can have a significant adverse economic impact on 

production herds, as well as adverse effects on animal welfare. This project investigated the prevalence of a novel, highly pathogenic 

herpesvirus, Leporid herpesvirus 4 (LHV-4), in commercial rabbitries in Ontario. The objectives of this research project were five-fold 

) to develop and validate a standard microtiter virus neutralization assay that can be used for screening diagnostic sera for antibody 

to LHV-4, 2) to develop and validate a specific LHV-4 ELISA that can be used for rapid and high-throughput screening of rabbit serum 

for antibody to LHV-4, ) to upgrade and validate a confirmatory real-time PCR assay for LHV-4, 4) to characterize the nature and time 

course of LHV-4 infection in rabbits following experimental challenge, and 5) to survey Ontario commercial rabbitries to determine 

seroprevalence of this virus. As a result, a VN assay, a qPCR assay, and a subunit ELISA have been developed to enhance the diagnostic 

capabilities for detecting this virus. These novel diagnostic tests for LHV-4 can be used to identify LHV4 in various rabbit populations, 

while protecting the safety of lab workers who must conduct the testing. Time course assays have demonstrated that virus is only shed 

within the first week of infection, although other diagnostic features, including the presence of neutralizing antibodies, can be used to 

diagnose more chronic cases or animals that have been exposed. The prevalence studies have led to the conclusion that this virus is not 

enzootic within the Ontario commercial rabbit population, as no known positive animals were detected. The project provides Ontario 

agriculture with new information about this virus, including clinical course of disease, viral shedding patterns, and development of 

protective antibodies. In addition we have refined or developed and validated new sensitive assays for detecting the virus, which can be 

used either in-house or commercially across the province and across Canada.

AHL 

H E R E 

Evaluation of massively parallel sequencing 

for detection and characterization of viruses 

Davor Ojkic 

Validation of MALDI-TOF for bacterial 

speciation - Durda Slavic 

Validation of the MALDI-TOF MS for bacterial identification 

as a new tool in routine clinical microbiology was 

supported through the hiring of a part-time technician. 

Validation of the instrument for identification of 

Enterobactericeae and major mastitis pathogens was 

completed, and the MALDI-TOF is in daily use for bacterial 

identification for these two groups. With this system in 

production, bacterial identification can be provided in less 

than 2 minutes per isolate, which provides clients with a 

one day shorter turnaround time, particularly for timesensitive 

cases. 

ELISA detection of Clostridium 

perfringens Cpb2 toxins in swine and 

cattle - John Prescott 

Clostridium perfringens is commonly known to cause 

diarrhea in neonatal swine, and this project sought to 

develop a diagnostic enzyme-linked immunosorbent 

assay (ELISA) for C. perfringens Cpb2 toxin based on 

antigen captured in intestinal content. It also provided an 

assessment of the sensitivity of the ELISA in comparison to 

Cpb2 positive C. perfringens bacterial counts in diarrheic 

and healthy swine and calves. A sensitive capture ELISA 

was developed for detection and quantification of C. 

perfringens Cpb2 in the neonatal piglet intestine. The 

results from this research provides a diagnostic ELISA that 

could be used commercially in Ontario. 

Due to limitations in detecting viruses in patients with clinical signs of 

lower respiratory tract infections, an etiological agent cannot be identied 

in 0 to 45 of cases by laboratory diagnosis because methods routinely 

used for detection of viruses are limited in various aspects. This project 

evaluated massively parallel sequencing for detection and characterization 

of viruses in tissues, swabs in virus transport medium (VTM), inactivated 

samples (Tripure), formalin-fixed paraffin-embedded samples, and Flinders 

Technology Associates (FTA) cards. Untreated nucleic acid extractions from 

cultures provided sufficient product to enable sequencing of the entire 

FAdV genome using the Roche Rapid Library Procedure and the GS Junior 

sequencing system. Utilizing this technology benefits Ontario by reducing 

the turnaround time for sequencing data for identifying and characterizing 

viruses. 

Haemophilus parasuis detection and typing 

Hugh Cai 

Haemophilus parasuis has traditionally been considered as a sporadic 

cause of stress-associated disease of swine. The intent of the project was 

to provide Ontario producers with a PCR-based rapid detection method 

for identification and molecular typing of H. parasuis, which will improve 

the diagnosis of Glasser’s disease. The project evaluated three published 

PCR assays and developed an OMP P2 PCR assay for the diagnosis of H. 

parasuis infection. Different molecular typing assays were also evaluated. 

The assays developed and evaluated would aid in controlling this disease 

in Ontario. 

Coccidial DNA bar-coding - John Barta 

Coccidian parasites infect chickens and other fast-growing meat production 

animals such as meat turkeys, feeder calves, goats and lambs. The research 

project objective was to create and evaluate a quantitative PCR method 

for the identification and enumeration of coccidian parasites (Eimeria 

species) that infect domestic poultry, based on sequences found in the 

mitochondrial cytochrome c oxidase subunit I (COI) gene. A real-time 

PCR primer pair assay was developed, qPCR-400F and qPCR-05R that 

quantify single or mixed parasite samples and amplifies COI sequences 

from 7 poultry Eimeria species. Species identification was determined 

by conducting melting point analysis of the products generated during 

the qPCR reaction. This project provides in-depth information on how to 

implement control measures that would prevent outbreaks of coccidiosis 

and also help in ameliorating gastroenteritis as a disease syndrome in 

Ontario poultry. In addition, this test could serve as both a live vaccine 

production tool as well as a method of ensuring that production flocks 

have been vaccinated successfully.

AHL 

H E R E 

 

Mycoplasma - Hugh Cai 

Since Mycoplasma spp. are difficult to identify and mixed 

infections are very common, we developed a rapid and low 

cost test to identify Mycoplasma spp. using MALDI-TOF MS. As 

a result, a database of 4 veterinary Mycoplasma species has 

been established. A large number of veterinary field isolates 

were tested to evaluate the assay. The success of this project 

has provided the AHL with an accurate, rapid and low-cost 

identification of Mycoplasma spp. 

New PCR and ELISA tests for 

mammalian viruses - Susy Carman 

The Virology laboratory at the AHL provides a number of real-me 

PCR assays and anbody ELISAs and this project was undertaken to 

expand services. To date, we have validated an addional 20 real- 

me PCR tests (7 swine; 9 cale; 4 equine), 8 anbody ELISAs ( 

swine, cale; 5 sheepgoats), and virus neutralizaon assay for 

horses. The standard operang procedures and validaon reports 

were wrien and tests implemented. Roune tesng is ongoing. 

The Tetracore PRRSV real-me RT-PCR, the Inuenza A virus – swine 

matrix real-me RT-PCR, and the IDE PRRSV HerdChek PRRSV 

anbody ELISA were ISO-7025 accredited by the Standards Council 

of Canada, and are currently in use for tesng swine for export. 

Avian Chlamydophila and Coxiella infection 

in Ontario - Marina Brash / Hugh Cai 

In Canada, Chlamydophila psittaci is a well-known zoonotic 

pathogen which affects humans, small ruminants, 

poultry and wild birds. Recently, a new avian Coxiella-like 

bacterium has been identified in psittacine birds and the 

gross and histologic lesions closely resemble those of avian 

Chlamydophila infections. This project determined the 

prevalence of Chlamydophila psittaci, C. burnetii and the 

avian Coxiella-like bacterium in psittacine birds submitted for 

routine postmortem to the AHL. Different diagnostic tools 

and different sample types including fecal samples were 

evaluated. Either liver or liver and spleen from three separate 

submissions of psittacine birds tested positive for the avian 

Coxiella-like organism using a S rRNA gene sequencing 

method. However, all of the birds tested negative for 

Chlamydophila psittaci by PCR and for organisms suspicious for 

Chlamydophila organisms using modified acid-fast impression 

smears of liver or liverspleen. As a result of this project, 

a commercial test for the diagnosis of avian Coxiella-like 

infection has been developed. This test is a valuable addition 

to the AHL testing capabilities because of the newly emerging 

disease closely resembling avian chlamydiosis and its potential 

of being zoonotic and immediately notifiable across Ontario 

and Canada. 

Validation of serum amyloid A in horses 

Brent Hoff 

Serum amyloid A (SAA) has been used as a diagnosc and prognosc 

marker for diseases. This project validated an automated SAA for the 

use of a diagnosc tool for sick horses. It also looked at examining ways 

to provide an ecient and cost eecve diagnosc tool to provide 

prognosis and monitoring of unhealthy horses. Results indicate that 

clinically well horses had a very low SAA value whereas aected animals 

exhibited dramac, acute increases, which mirrored the magnitude of 

disease or ssue trauma present. This study demonstrated that the 

automated Eiken SAA TIA measures equine SAA in a reliable manner. The 

assay is automated and commercially available for roune use on equine 

diagnosc proles in diagnosc laboratories and has been added to AHL 

equine biochemistry proles. 

Development of immunohistochemical 

tests for small ruminant lentiviruses 

Josepha DeLay 

There were no tests available for immunohistochemical (IHC) 

staining for small ruminant lentiviruses (SRLV) in Canada; SRLV 

include Caprine arthritis-encephalitis virus (CAEV) and Maedivisna 

virus (MVV). This project sought to develop and validate 

an IHC test for SRLV. A successful IHC test was developed using 

a commercial anti-CAEV MVV monoclonal antibody that can be 

used to confirm infection in postmortem samples from goats and 

sheep. Using histologic lesions of SRLV infection in lung as a gold 

standard, sensitivity of the IHC test was 70 for CAEV and 80 of 

MVV, with 00 specificity for both viruses. This test fills a void 

among test methods, and provides a new, previously unavailable 

in Canada method for postmortem confirmation of CAEV and MVV 

infections. 

PAGE 

22

Brachyspira in dirty 

egg syndrome 

Durda Slavic / Michele Guerin 

For more than 2 decades, diarrhea 

resulting in fecal staining of eggshells 

(dirty egg syndrome) has been well 

known for causing reduction in egg 

production. This project aimed at 

developing a diagnostic test for detection and speciation of 

Brachyspira spp. in chicken samples, and determining both the 

overall prevalence of Brachyspira spp. in Ontario layer flocks, 

and the prevalence of pathogenic Brachyspira spp. Results from 

this project have helped Ontario to gain a better understanding 

of whether or not pathogenic Brachyspira spp. are present in 

both flocks with and without dirty eggs, and identifies farm 

management practices that contribute to the clinical onset of 

avian intestinal spirochetosis. 

Genotyping of fowl adenoviruses - Davor Ojkic 

P 

oultry producers in Ontario and Canada-wide have 

struggled with non-pathogenic and pathogenic strains of 

Fowl adenovirus (FAdV) and Infectious bursal disease virus 

(IBDV) for quite some time. This project determined genotypes of 

FAdVs and IBDVs from randomly selected broiler flocks in Ontario. 

Genotyping of FAdVs and IBDVs from randomly selected samples 

removed the collection bias experienced in previous studies. This 

project benefited Ontario by allowing appropriate selection of field 

viruses for further investigations related to antigenic properties, 

pathogenicity and vaccine development. 

Comparing four different sample types and 

three different culture methods for Salmonella 

spp. detection in poultry - Durda Slavic 

Salmonella spp. are zoonotic pathogens that can cause diseases 

in humans and animals and reduction of Salmonella presence in 

primary food production sites. This project sought to compare 

4 different methods for isolation of Salmonella spp. using 

4 different sample types in poultry environments. Common 

trends of higher recovery of Salmonella spp. from boot covers 

and fluff samples were observed. Based on statistical analyses 

of results, if boot covers are used for sampling they will be 

5.98 times more likely to result in Salmonella isolation than 

swabs. Similarly, fluff samples from hatcheries are .7 times 

more likely to result in Salmonella isolation than egg shells. 

These sample types and their results will need to be taken into 

consideration when updating the Ontario Hatchery and Supply 

Flock Policy to establish standard samples type for Salmonella 

testing. 

 

indicators of gastrointestinal parasitism 

Andrew Peregrine 

AHL 

H E R E 

The anthelmintic resistance in Ontario sheep project had 4 

objectives ) to establish and validate a larval development 

test for anthelmintic susceptibility in Ontario, 2) to determine 

the prevalence of ivermectin drench failure in Ontario flocks, 

) to determine the prevalence of anthelmintic resistance in 

Ontario flocks using a fecal egg count reduction test (FECRT) 

and the larval development test, and 4) to identify risk factors 

associated with drench failure and anthelmintic resistance. 

Ivermectin drench failure was demonstrated on 540 (87) 

farms. On the basis of the FECRT, ivermectin resistance was 

confirmed on 2729 (9) farms; fenbendazole resistance 

was confirmed on 2020 (00) farms; and levamisole 

resistance was confirmed on 7 () farms. Culture of 

fecal samples indicated that most resistance was associated 

with Haemonchus. The in vitro larval development test was 

established at the Ontario Veterinary College and performed on 

20 farms that reached the threshold mean fecal egg count of 

200 eggs per gram of feces. Teladorsagia sp., Haemonchus sp. 

and Trichostrongylus spp. were recovered from all treatment 

groups; however, Haemonchus sp. was the most commonly 

isolated parasite on the majority of the farms. In summary, 

anthelmintic resistance appears to be a common problem on 

Ontario sheep farms and appears to be primarily associated 

with Haemonchus. By validating the performance of in vivo 

and in vitro tests for quantifying anthelmintic susceptibility 

in gastrointestinal parasites, this project has developed a 

tool that enables Ontario producers to regularly monitor the 

anthelmintic susceptibility of parasites on their farms. 

Molecular typing method for characterization 

of Giardia duodenalis - Hugh Cai 

Giardia duodenalis, which is known to infect humans and 

other mammalian hosts, has been a great concern in Ontario. 

The purpose of this study was to establish and evaluate 

procedures for G. duodenalis detection and molecular typing. 

An ssu-rRNA PCR sequencing method, in conjunction with 

the bg PCR sequencing method, typed the greatest number 

of samples and identified all mixed infections, whereas the 

gdh and tpi PCR sequencing methods had little value. The 

established assays were employed to identify the prevalence 

of G. duodenalis assemblage types among canine and feline 

fecal samples in Ontario, and therefore determine the zoonotic 

potential of these infections. This study benefits Ontario by 

detecting mixed G. duodenalis assemblage types, including the 

potentially zoonotic assemblages. 

PAGE 

2

AHL 

H E R E 

F aa 

AHSI - funded projects 2008-2013 

P ad P a P 

Surveillance 

Tim Blackwell Franklin Kains, Kathy Zurbrigg Tesng the eecveness of producer-based mortality 

reporng system in commercial swine herds in Ontario 

Animal 

health test 

Andrew Peregrine 

Jocelyn Jansen, Andria Jones, Jane 

Learmount, Ralph Marn, Paula 

Menzies, Mike Taylor, John Van 

Leeuwen 

Determinaon of prevalence of anthelminc 

resistance in Ontario sheep ocks with indicators of 

gastrointesnal parasism 

Surveillance 

Michele Guerin Davor Ojkic, Durda Slavic Enhanced surveillance for viral and bacterial pathogens 

in commercial broiler chicken ocks in Ontario 

Surveillance 

Claire Jardine 

Ian Barker, Patrick Boerlin, Bob 

Friendship, Sco McEwen, Davor 

Ojkic, Jane Parmley, Richard Reid- 

Smith, Jan Sargeant 

Surveillance of disease agents of public health and 

agricultural signicance in wildlife living on farms in 

Ontario 

Animal 

health test 

John Presco 

Valeria Parreira, Durda Slavic, Glenn 

Songer 

Development and assessment of a diagnosc ELISA 

for Clostridium perfringens Cpb2 toxin, with special 

reference to swine and cale 

Surveillance 

Paula Menzies 

Jocelyn Jansen, Andria Jones, Andrew 

Peregrine 

A survey of risk factors associated with condemnaon 

of sheep carcasses due to Cyscercus ovis infecon 

Surveillance 

David Kelton 

Karen Hand, Davor Ojkic, George 

MacNaughton, Beverly McEwen, Dave 

McKeen, Neil Petreny, Ian Rumbles, 

Durda Slavic, Deb van de Water 

Integrang milk based cow and bulk tank test data, 

dairy cale inventory data and GIS locaon data into 

the Animal Health Laboratory surveillance capacity 

Animal 

health test 

Hugh Cai 

Mycoplasma speciaon by molecular biology assays 

Surveillance 

Surveillance 

Animal 

health test 

Surveillance 

Surveillance 

Animal 

health test 

Hugh Cai 

Susy Carman, Josepha DeLay, Murray 

Hazle, Beverly McEwen, Durda Slavic 

Invesgaon of infecous eology of small ruminant 

aboron in Ontario with emphasis on Chlamydophila 

spp. and Coxiella burnei 

Hugh Cai Jinzhong Fu Epidemiological analysis of Mycoplasma bovis 

isolated from Canada in the past 0 years 

Hugh Cai Andrew Peregrine Development of molecular typing method for the 

characterizaon of Giardia duodenalis 

Kathy Zurbrigg Beverly McEwen, Bruce McNab Ontario farm-call syndromic surveillance 

Robert Friendship Rocio Amezcua, David L. Pearl Rening the Ontario swine veterinarian-based 

surveillance (OSVS) system for real me surveillance 

and integraon with laboratory-based surveillance 

John Barta 

Hugh Cai, Julie Cobean, Robert 

Hanner, Joseph Ogedengbe, Mosun 

Ogedengbe 

DNA bar-coding of coccidial parasites of producon 

animals development and eld tesng of a 

LightCycler-based quantave test


AHL 

H E R E 

F aa 

P ad P a P 

Surveillance 

Animal 

health test 

Surveillance 

Surveillance 

Surveillance 

Surveillance 

Animal 

health test 

Surveillance 

Robert Friendship Jeonghwa Park Is the treatment of exudave epidermis in pigs 

becoming problemac because of the emergence of 

anmicrobial resistance 

Murray Hazle 

Ken Leslie 

Davor Ojkic 

David Pearl 

Javier Sanchez 

Crawford Revie 

Durda Slavic 

Michele Guerin 


Hugh Cai, Josepha DeLay, Beverly 

McEwen 

Todd Dueld, Brent Ho, Brian 

McBride, Brian Nelson, Andrew 

Peregrine 

Cheryl Firby, Sco Gillingham, Doug 

McGhee, Rachel Ouckama, Cynthia 

Philippe, Babak Sanei 

Agnes Agunos, Jim Fairles, Shiona 

Glass, David Léger, Beverly McEwen, 

Sco McEwen, Jane Parmley, Richard 

Reid-Smith, Durda Slavic 

Fernanda Dórea, David Kelton, Bruce 

McNab 

Marina Brash, Hugh Cai, Leanne 

Cooley, Sco Houghton, Babak Sanei, 

Lloyd Weber 

George Charbonneau, Janet 

MacInnes, Jeonghwa Park, Zvonimir 

Poljak, Durda Slavic 

Development of in-situ hybridizaon (ISH) for 

diagnosis of Coxiella burnei and Toxoplasma gondii 

in small ruminants 

Associaons between the incidence of 

Cryptosporidium parvum and selenium status in 

neonatal dairy calves in Ontario 

Surveillance for inclusion body hepas in broiler 

breeders 

The development and design of a surveillance system 

for anmicrobial resistance in agriculture animal 

pathogens in Ontario 

Developing and implemenng techniques to harvest 

addional surveillance informaon from exisng AHL 

diagnosc laboratory data 

Opmizaon and validaon of real-me PCR to 

determine the prevalence of pathogenic and nonpathogenic 

Brachyspira spp. in Ontario layer ocks 

and to idenfy risk factors associated with dirty egg 

syndrome 

Ear p necrosis in swine 

Surveillance 


Agnes Agunos, David Léger, Sco 

McEwen, Rachel Ouckama, Cynthia 

Philippe, Richard Reid-Smith, Babak 

Sanei, Durda Slavic 

Surveillance of anmicrobial-resistant enteric 

bacteria in anmicrobial-free and convenonal 

broiler in Ontario 

Animal 

health test 

Brent Ho 

Luis Arroyo, Dan Kenney, Helen 

Kocmarek, Beverly McEwen, Krisina 

Ruotsalo 

Validaon of serum amyloid A (SAA) an acute-phase 

protein as marker of inammaon in horses 

Surveillance 

Paula Menzies 

Jocelyn Jansen, Andria Jones 

Prevalence of Q fever (Coxiella burnei) in Ontario 

sheep ocks, goat herds and their farm workers 

Surveillance 

Eva Nagy 

Marina Brash, Laszlo Zsák 

Surveillance for enteric parvoviruses in poultry 

Surveillance 


Susy Carman, Enrique Castro, George 

Charbonneau, Robert Friendship, 

Durda Slavic 

Risk-based surveillance of respiratory infecons in 

growing pigs

AHL 

H E R E 

Focus area 


Project lead Project team members Project tle 

Surveillance 

John Presco 

Vicki Nowell, Valeria Parreira, Durda 

Slavic 

Idencaon of novel toxin gene(s) associated 

with type A Clostridium perfringens-associated 

hemorrhagic abomasis of calves in Ontario 

Surveillance 

Karen Shearer 

Doug Campbell, Josepha DeLay, 

Davor Ojkic 

Genotyping and detecon of Leptospira spp. 

in wildlife reservoir hosts in Ontario through 

comparison of immunohistochemical and 

polymerase chain reacon genotyping methods 

Surveillance 

Michele Guerin/ 


Rob Deardon, Davor Ojkic 

Case series of pandemic HN in Ontario turkey 

breeder ocks 

Surveillance 

Janet Alsop 

Suzanne Burlatschenko, Susy 

Carman, Jim Fairles, Hank Harris, 


Using glycantyping to predict outcome in Ontario 

sow herds experiencing porcine reproducve and 

respiratory syndrome virus (PRRSV) outbreaks 

Animal 

health test 

Susy Carman 

Validang and implemenng new real-me PCR tests 

and ELISAs for mammalian viruses 

Animal 

health test 

Shu Chen 

Hugh Cai, Murray Hazle, Bruce 

Keown 

Molecular typing of Coxiella burne idened in 

Ontario by mulple-locus variable number of tandem 

repeat analysis (MLVA) 

Surveillance 

Andria Jones 

Jocelyn Jansen, David Kelton, 

Paula Menzies, Davor Ojkic, 

Durda Slavic 

Johne’s disease in Ontario’s small ruminant dairy 

industries Prevalence, potenal risk factors, and 

performance comparison of serum, milk and fecal 

diagnosc methods 

Surveillance 

Janet MacInnes 

Shu Chen, Stefan C. Kremer, Zvonimir 

Poljak, Durda Slavic 

Comprehensive evaluaon of bacterial hazards in 

the upper respiratory tract of swine by terminal 

restricon fragment length polymorphism (T-RFLP) 

analysis 

Surveillance Lori Moser Doug MacDougald, Marn Misener, 


Regional PRRS eliminaon trial 

Animal 

health test 

Animal 

health test 

Hugh Cai 

Durda Slavic 

Janet MacInnes, Durda Slavic, Patricia 

Bell-Rogers, Rita Rebelo, Rebeccah 

Travis 

Teresa Cereno, Keith Harron, Edward 

Malek, Beverly McEwen, Davor Ojkic, 

Babak Sanei 

Rapid detecon and typing of Haemophilus parasuis 

Comparison of four dierent sample types and 

three dierent culture methods for Salmonella spp. 

detecon in poultry environmental samples 

Animal 

health test 

Davor Ojkic 

Elizabeth Hillyer, Sarah Hoyland 

Evaluaon of massively parallel sequencing for 

detecon and characterizaon of viruses 

Surveillance 

Crawford Revie/ 

Javier Sanchez 

Caroline Dube, Bruce McNab, 

Zvonimir Poljak, Crawford Revie 

Contact network based disease spread model in 

Ontario


AHL 

H E R E 

Focus area 

Focus area 

Project lead Project team members Project tle 

Surveillance 

Durda Slavic/ Susy 

Carman 

Summer student assistantships-AHL 

Emergency 

Preparedness 

Maria Spinato Jan Shapiro AHL necropsy emergency preparedness and ICS 

training 

Animal 

health test 

Patricia Turner 

Marina Brash, Susy Carman, Éva 

Nagy, Brian Tapsco 

Prevalence and characterizaon of Leporid 

herpesvirus-4 (LHV-4) in Ontario rabbits, an 

emerging rabbit pathogen 

Surveillance 


Agnes Agunos, Dean Middleton, 

Babak Sanei, Csaba Varga, Gwen 

Zellen 

Salmonella Enteridis baseline study in Ontario 

commercial broiler layer ocks and to idenfy risk 

factors associated with dirty egg syndrome 

Animal 

health test 

Hugh Cai 

Marina Brash, Emily Marn, 

Margaret Stalker 

Avian Chlamydophila and Coxiella infecon in 

Ontario molecular diagnosis method development 

Surveillance 

Paula Menzies 

Susy Carman, Jocelyn Jansen, 

Beverly McEwen, Sarah Wooon 

Detecon of maedi-visna virus infecon in Ontario 

sheep ock 

Animal 

health test 

Josepha DeLay Murray Hazle, Susan Lapos Development of immunohistochemical tests for 

small ruminant lenviruses (caprine arthrisencephalis 

virus, maedi-visna virus) 

Animal 

health test 

Durda Slavic 

Validaon of MALDI-TOF for bacterial speciaon 

Animal 

health test 

Animal 

health test 

Davor Ojkic 

Hugh Cai 

Michael Eregae, Michele Guerin, 

Thelma Marnez 

Genotyping of fowl adenoviruses and infecous 

bursal disease viruses from Ontario broiler breeder 

ocks 

Development of rapid idencaon assays for 

veterinary mycoplasma using matrix-assisted 

laser desorpon ionizaon-me of ight mass 

spectrometry and polymerase chain reacon 

genotyping methods 

Surveillance 

John Lumsden 

Spencer Russell, Lincoln Tubbs 

Diagnosis and surveillance of Chlamydia-like 

organisms in Ontario rainbow trout 

Animal 

health test 

David Kelton 

Richard Cann, Jim Fairles, Ann 

Godkin, Davor Ojkic, Durda 

Slavic, Deb van de Water 

Equivalency for strong posive serum ELISA Johne’s 

disease test results and interpretaon of repeated 

tesng for Johne’s disease in Ontario dairy herds 

Surveillance 

Harold Kloeze 

Gaston Annamunthodo, Beverly 

McEwen, Shamir Nizar Mukhi, 

Bruce McNab, Andre Vallieres 

Expanded data transfer from AHL to CAHSN In 

support of improved noable disease reporng 

to CFIA, emergency preparedness, mely disease 

detecon and data analysis

AHL 

H E R E 

Theses 

Bauman C. Johne’s disease in the Ontario dairy sheep and dairy 

goat industries prevalence, test comparison, and evaluaon 

of risk factors. University of Guelph, Guelph, ON. PhD thesis. 

July 20. 

Bondo K. Prevalence of zoonoc enteric pathogens in wildlife, 

livestock, and soil on beef, dairy, and swine farms in Ontario. 

University of Guelph, Guelph, ON. PhD thesis. (In progress) 

Dodds H. Anmicrobial resistance in Escherichia col isolated 

from wild small mammals and soil in southern Ontario. 

University of Guelph, Guelph, ON. MSc thesis. 20. 

Dórea F. Syndromic surveillance based on laboratory test 

requests to the AHL. University of Prince Edward Island, 

Charloetown, PEI. PhD thesis. March 20. 

Eregae M. Epidemiology of fowl adenovirus and associated 

viruses among commercial broiler chicken ocks in Ontario. 

University of Guelph, Guelph, ON. PhD thesis. August 20. 

Falzon L. Gastro-intesnal nematodes in Ontario sheep ocks An 

epidemiological study of over-wintering and 

anthelminc resistance. University of Guelph, Guelph, ON. 

PhD thesis. December 202. 

Glass- Kaastra S. Surveillance system for anmicrobial resistance 

in Ontario swine. University of Guelph, Guelph, ON. PhD 

thesis. (In progress) 

Kircanski J. Development of an angen-capture ELISA for 

Clostridium perfringens Cpb2 (beta2) toxin in the neonatal 

piglet intesne. University of Guelph, Guelph, ON. MSc 

thesis. 20. 

Le H. Epidemiological aspects of transmission and control of 

porcine reproducve and respiratory syndrome virus 

infecon and associated diseases. University of Guelph, 

Guelph, ON. December 20. 

Meadows S. Determining the prevalence and factors associated 

with Coxiella burnei (Q fever) exposure in sheep, goats and 

their farm workers in Ontario. University of Guelph, Guelph, 

ON. MSc thesis. 20. 

Nowell V. Genomic and proteomic analysis of a bovine 

hemorrhagic isolate of type A Clostridium perfringens. 

University of Guelph, Guelph, ON, Canada. MSc thesis. 20. 

Summer students 

Allen S. Surveillance of disease agents of public health and agricultural signicance in wildlife living on farms in Ontario. 

Baker D. Determinaon of the prevalence of anthelminc resistance in Ontario sheep ocks with indicators of gastrointesnal 

parasism. 

Beck E. Surveillance of disease agents of public health and agricultural signicance in wildlife living on farms in Ontario. 

Davis B. Surveillance of disease agents of public health and agricultural signicance in wildlife living on farms in Ontario. 

Flockhart L. Surveillance of disease agents of public health and agricultural signicance in wildlife living on farms in Ontario. 

Fountain J. Surveillance for enteric parvoviruses in poultry – a pilot study. 

Gauthier N. Determinaon of the prevalence of anthelminc resistance in Ontario sheep ocks with indicators of gastrointesnal 

parasism. 

Harris C. Determinaon of the prevalence of anthelminc resistance in Ontario sheep ocks with indicators of gastrointesnal 

parasism. 

Hok M. Risk-based surveillance of respiratory infecons in growing pigs.

AHL 

H E R E 

Innes C. Integrang milk based cow and bulk tank test data, dairy cale inventory data and GIS locaon data into the Animal Health 

Laboratory surveillance capacity. 

Kozlowski K. Determining the prevalence of Q fever (Coxiella burnei ) in Ontario sheep ocks, goat herds and their farm workers. 

Myers E. Enhanced Surveillance for viral and bacterial pathogens in commercial broiler chicken ocks in Ontario. 

Painter K. Associaons between the incidence of Cryptosporidium parvum and selenium status in neonatal dairy calves in Ontario. 

Puskas K. Determinaon of the prevalence of anthelminc resistance in Ontario sheep ocks with indicators of gastrointesnal 

parasism. 

Rosborough A. Associaons between the incidence of Cryptosporidium parvum and selenium status in neonatal dairy calves in Ontario. 

Schlegel B. Determining the prevalence of Q fever (Coxiella burnei) in Ontario sheep ocks, goat herds and their farm workers. 

Siertsema L. Determinaon of the prevalence of anthelminc resistance in Ontario sheep ocks with indicators of gastrointesnal 

parasism. 

Sinclair J. Determinaon of the prevalence of anthelminc resistance in Ontario sheep ocks with indicators of gastrointesnal 

parasism. 

Sinclair C. Equivalency for strong posive serum ELISA Johne’s disease test results and interpretaon of repeated tesng for Johne’s 

disease in Ontario dairy herds. 

Sippel K. Determinaon of the prevalence of anthelminc resistance in Ontario sheep ocks with indicators of gastrointesnal 

parasism. 

Socha R. Determinaon of the prevalence of anthelminc resistance in Ontario sheep ocks with indicators of gastrointesnal 

parasism. 

Stewart H. Determinaon of the prevalence of anthelminc resistance in Ontario sheep ocks with indicators of gastrointesnal 

parasism. 

Tatone E. Associaons between the incidence of Cryptosporidium parvum and selenium status in neonatal dairy calves in Ontario. 

Tenbergen R. Determinaon of the prevalence of anthelminc resistance in Ontario sheep ocks with indicators of gastrointesnal 

parasism. 

Thompson A. Integrang milk based cow and bulk tank test data, dairy cale inventory data and GIS locaon data into the Animal Health 

Laboratory surveillance capacity. 

Thompson G. Risk-based surveillance of respiratory infecons in growing pigs. 

Toon S. Surveillance of disease agents of public health and agricultural signicance in wildlife living on farms in Ontario. 

Yates D. Determinaon of the prevalence of anthelminc resistance in Ontario sheep ocks with indicators of gastrointesnal parasism.

AHL 

H E R E 

AHSI presentations and posters 

AHSI rncal nesators ae arcated n oer oral resentaons and contrbuted to oer 

ublcaons trou Arl Selected eamles 

Amezcua R, Pearl DL, Friendship RM, Marnez A. A retrospecve analysis of the temporal paerns of condemnaon rates from 

2005 – 2007 with reference to a provincial wide outbreak of PCVAD. Conference of Research Workers in Animal Diseases. 

Chicago, IL. December -8, 2009. 

Barrere V, Falzon L, Shakya K, Peregrine A, Prichard RK. Detecon of benzimidazole resistance in the sheep nematode, Haemonchus 

contortus, using pyrosequencing on pools of eggs collected from sheep on farms in Ontario, Canada. Internaonal Conference, 

World Associaon for the Advancement of Veterinary Parasitology. Buenos Aires, Argenna. Aug 2-24, 20. 

Bell-Rogers P, Parker L, Rosenbusch RF, Cai H. Development of mul-locus sequence typing assay for Mycoplasma bovis molecular 

typing and preliminary analysis of M. bovis isolated from 980-200 in Ontario. Canadian Animal Health Laboratorians Network 

Annual Conference. Guelph, ON. June 8, 20. 

Chen S. A molecular subtyping approach for rapid diagnoscs and enhanced risk assessment, management and control. University of 

Guelph - Department of Pathobiology Seminar Series. Guelph, ON. October 20-2, 20. 

Contador E, LePage V, Blackburn K, Lumsden J. Epitheliocyss disease in rainbow trout (Onchorhynchus mykiss). OMAF Animal 

Research Expo. Guelph, ON. June 2, 202. 

De Wolf B. Ovine cyscercosis An emerging problem in Canada. Ontario Veterinary College Bovine Club. Guelph, ON. February 

200. 

De Wolf B. Emerging tapeworm issues in Canada. Ontario Veterinary College Pathobiology Seminar Series. Guelph, ON. May 20. 

Dodds H, Boerlin P, Janecko N, Reid-Smith R, Pearl D, Jardine C. Wildlife as indicators of anmicrobial resistance in the 

environment. Am Soc Microbiol, Conf Anmicrobial Resistance in Zoonoc Bacteria and Foodborne Pathogens in Animals, 

Humans and the Environment. Toronto, ON. 200. 

Falzon L, Jones-Bion A, Menzies P, Peregrine A, Vanleeuwen J, Jansen J, Shakya K, Avula J. Anthelminc resistance in Ontario sheep 

ocks. Canadian Associaon of Veterinary Epidemiology and Prevenve Medicine. St. Hyacinthe, Quebec. May 2-27, 20. 

Flockhart L, Janecko N, Reid-Smith R, Parmley J, Jardine C. Occurrence of Campylobacter in wildlife living on livestock farms in Ontario 

Canada. (Poster, st prize of 5). Graduate Student Research Symposium. Guelph, ON. 200. 

Friendship RM, Pearl DL, Amezcua R. Ontario swine veterinarian-based surveillance (OSVS) Project - Update. Centralia Swine Research 

Update. Kirkton, ON. Jan 28, 2009. 

Glass-Kaastra SK, Pearl D, McEwen SA, McEwen B, Reid-Smith R, Parmley J, Léger D, Slavic D, Agunos A, Fairles J. Mulclass resistance 

in Streptococcus suis, Escherichia coli, and Pasteurella multocida in Ontario swine, 998-200. Anmicrobial Stewardship in 

Canadian Agriculture and Vet Med Conf. Toronto, ON. Oct 0-Nov 2, 20. 

Hazle M, van Dreumel T, DeLay J, Stalker M, Spinato M, McEwen B, Binnington B, Fairles J, Cai H. The small ruminant aboron project 

A pathologist’s perspecve. Canadian Associaon of Veterinary Pathologists, Annual Meeng. Guelph, ON. June 20. 

Jardine C, Flockhart L, Janecko N, Parmley J, Cook A, Reid-Smith R. Prevalence of zoonoc enteric pathogens in wildlife, livestock, soil 

on beef, dairy, and swine farms in Ontario. Internaonal Wildlife Disease Associaon Meeng. Quebec City, QC. 20. 

Kelton D. Equivalency of strong-posive serum ELISA Johne’s disease test results and interpretaon of repeated tesng for Johne’s 

disease in Ontario dairy herds. OMAF-AHL Animal Health Strategic Investment wrap-up event. Guelph, ON. Nov 8, 202. 

Kircanski J, Hodgins D, Pei Y, Parreira VR, Presco JF. Development of a Cpb2 diagnosc ELISA for Clostridium perfringens Cpb2 

(beta2) toxin and preliminary evaluaon of the role of Cpb2 in enteric disease of neonatal piglets. OMAF Animal Health 

Forum. Guelph, ON. Nov 200. 

Lumsden JS. Chlamydia-like organism in Lake trout (Salvelinus namaycush). Wildlife Dis Assoc Conf. Quebec, QC. Aug 4-9, 202. 

Meadows S, Menzies P, Jones-Bion A, Jansen J, McEwen S. Determining the prevalence of Q fever (Coxiella burnei) in Ontario 

sheep ocks, goat herds and their farm workers. AHSI Wrap-up event. Guelph, ON. 202. 

Menzies P. Cyscercus ovis. The Sheep Veterinary Society. Jersey Island, United Kingdom. June, 200. 

Nelson B, Godden SM, McBride BW, Dueld TF, Leslie KE. The eect of oral supplementaon of selenium on passive transfer of 

immunoglobulins in dairy calves. ADSA-ASAS Joint Annual Meeng. Denver, CO. July 2, 200. 

Ogedengbe ME, Naaum A, Hanner R, Barta JR. Real-me qPCR for idencaon and enumeraon of Eimeria species of chickens 

based on the DNA barcoding target (mitochondrial cytochrome c oxidase subunit I sequences). Ontario Veterinary College, 

Graduate Research Symposium. Guelph, ON. Nov, 202. 

PAGE 

30

AHSI presentations and posters, continued 

Ojha S, Cai H, Hazle M, Chen S. Molecular typing of Coxiella burne idened in Ontario by mulple-locus variable number of 

tandem repeat analysis (MLVA). OMAF Food Safety Research Forum 20. Guelph, Ontario. May 5, 20. 

Ojkic D, Guerin M, Eregae M. Surveillance of Ontario broiler ocks for viral agents. Western College of Veterinary Medicine 

departmental seminar. Saskatoon, SK. October , 202. 

Park J, Weese S, Dewey C, Poljak Z, Friendship R. Exudave epidermis is dicult to treat because of anmicrobial resistance 

associated with Staphylococcus hyicus. Internaonal Pig Vet Society Congress. Vancouver, Brish Columbia. July 200. 

Peregrine A. Managing internal parasites in sheep – know the enemy Sustainable parasite management dealing with parasites – 

take control. Small Ruminant Veterinarians of Ontario. Guelph, ON. May , 2009. 

Puskas K, Shakya K, Falzon L, Peregrine A, Menzies P, Jansen J, Avula J, Stewart H, Siertsema L. Evaluaon of drug suscepbility of 

gastrointesnal nematodes on sheep farms using an in vitro larval development assay. OVC Summer Leadership and Research 

Program. Guelph, ON. Aug , 20. 

Russell S, Tubbs L, Huber P, LePage V, Al-Hussinee L, Reid A, Contador E, Allen S, Young K, Lumsden J. Diagnosc sh health; new 

agents, new challenges, new models. Can Comp Immunol Pathol Workshop. Waterloo, ON. May 2-28, 202. 

Sippel K, Falzon L, Shakya K, Avula J, Menzies P, Jansen J, Jones A, Peregrine A. The prevalence of anthelminc resistance in Ontario 

sheep ocks. OVC Summer Leadership and Research Program. Guelph, ON. Aug 0, 200. 

Spinato M, Shapiro J, Stalker MJ, Brooks A. Emergency preparedness in the necropsy laboratory Designing and conducng a fullscale 

operaonal exercise. Annual meeng American Associaon of Veterinary Laboratory Diagnoscians. Greensboro, NC. Oct 

8-24, 202. 

AHSI publications 

AHL 

H E R E 

Cai H, Hazle M. A prospecve study of sheep and goat aboron using real-me PCR and cut point esmaon shows Coxiella 

burnei and Chlamydophila abortus infecon concurrently with other major pathogens. J Vet Diagn Invest, 20; 25, . 

Contador E, Russell S, Young K, Di Natale A, Wright E, Lumsden J. Epitheliocyss in Lake trout (Salvelinus namaycush). 7th 

Eastern Fish Health Workshop, Lake Placid, NY. April 202. Abstract. 

Falzon L, Jones-Bion A, Menzies P, Peregrine A, Vanleeuwen J, Jansen J, Shakya K, Avula J. Anthelminc resistance in Ontario sheep 

ocks. Proc, Can Assoc Vet Epidemiol Prevenve Med, St. Hyacinthe, QC May 2-27, 20; p 48. 

Guerin M. Research summary of the baseline study of the prevalence of Salmonella Enteridis among broiler chicken ocks in Ontario. 

Annual Review for the Centre for Public Health and Zoonoses 20; p 9. 

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prospecve study of sheep and goat aboron using real-me polymerase chain reacon and cut point esmaon shows Coxiella 

burnei and Chlamydophila abortus infecon concurrently with other major pathogens. Published online 9 April 20 J Vet Diagn 

Invest -0. DOI 0.77/04087484729 . 

LePage V, Contador E, Frasca S, Lumsden J. Invesgaon of bacterial branchis and dermas in capve syngnathids. 7th 

Eastern Fish Health Workshop. Lake Placid, NY. April 202. Abstract. 

Meadows S, Menzies P, Jones-Bion A, Jansen J, McEwen S. Determining the prevalence of Q fever (Coxiella burnei) in Ontario 

sheep ocks, goat herds and their farm workers. Int Sheep Vet Congr, Rotorua, New Zealand. 20;8-22. 

Nagy E, Brash M, Zsak L. Surveillance of enteric paroviruses in poultry - a pilot study. 0 th CAHLN meeng. Guelph, ON. June 20. 

Abstract. 

Ogedengbe JD, Hanner RH, Barta J. DNA barcoding idenes Eimeria species and contributes to the phylogenecs of coccidian 

parasites (Eimeriorina, Apicomplexa, Alveolata). Int J Parasitol 20; 484-850. 

Park J, Friendship RM, Poljak Z, DeLay J, Slavic D, Dewey C. Invesgaon of ear necrosis syndrome in pigs. Centralia Swine Research 

Update, Kirkton, ON 20; pp 5-. hp//www.centraliaswineresearch.ca 

Peregrine A. Managing internal parasites in sheep – know the enemy Proceedings, Sustainable parasite management dealing with 

parasite – take control. Small Ruminant Vets of Ontario. Guelph, ON. May 2009; p.8. 

Sippel K, Falzon L, Shakya K, Avyla J, Menzies P, Jansen J, Jones A, Peregrine A. The prevalence of anthelminc-resistant gastrointesnal 

nematodes in Ontario sheep ocks. Studies by Undergraduate Researchers at Guelph (SURG). Sept 200. Abstract. 

Zurbrigg K. The use of diagnosc tesng as an incenve for veterinarian parcipaon in an animal health surveillance program and 

its eect on increasing laboratory submission rates. Ann Mtg Can Animal Health Laboratorians Network 20; p 2. 

PAGE

OMAF & MRA- AHL Animal Health Strategic Investment 

2008-2013 

SURVEILLANCEANIMAL HEALTH TESTSEMERGENCY PREPAREDNESS 

Director of the 

Animal Health 

Laboratory 

Animal Health Laboratory 

Laboratory Services Division 

49 Gordon Street, NW Corner 

Gordon / McGilvray 

Guelph, Ontario NG 2W 

59.824.420 x5450 

hp//www.ahl.uoguelph.ca 

Photos Ontario Farm Animal Council Agriculture Photo Library 

Photography Mark York and AHSI project leads

AHSI magazine - University of Guelph

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