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A case-control study was conducted among commercial table-egg layer and pullet operations in Iowa and Nebraska, United States, to investigate potential risk factors for infection with highly pathogenic avian influenza (HPAI) H5N2. A questionnaire was developed and administered to 28 case farms and 31 control farms. Data were collected at the farm and barn levels, enabling two separate analyses to be performed-the first a farm-level comparison of case farms vs. control farms, and the second a barn-level comparison between case barns on case farms and control barns on control farms. Multivariable logistic regression models were fit using a forward-selection procedure. Key risk factors identified were farm location in an existing control zone, rendering and garbage trucks coming near barns, dead-bird disposal located near barns, and visits by a company service person. Variables associated with a decreased risk of infection included visitors changing clothing, cleaning and disinfecting a hard-surface barn entryway, and ceiling/eaves ventilation in barns.

Highlights • We conducted retrospective testing of clinical samples submitted to veterinary diagnostic laboratories for porcine deltacoronavirus. • Over 2000 samples were tested over three separate years with 3 positive samples detected prior to the PDCoV outbreak detection in February 2014. • We surveyed 42 early infected swine farms to determine various farm level characteristics, descriptive estimates of biosecurity practices and disease status over time of each operation. • Clinical signs of PDCoV were reported to be similar to those of PEDv. The average number of animals on each operation exhibiting clinical signs (morbidity) and the average number of case fatalities was greatest for suckling and weaned pigs. • The survey included questions regarding biosecurity practices for visitors and operation employees; trucks, equipment and drivers; and feed sources. These questions attempted to identify a likely pathway of introduction of PDCoV onto the operations surveyed., Porcine deltacoronavirus (PDCoV) was first reported in the United States (US) in February 2014. This was the second novel swine enteric coronavirus detected in the US since May 2013. In this study, we conducted retrospective testing of samples submitted to three veterinary diagnostic laboratories where qualifying biological samples were derived from previously submitted diagnostic case submissions from US commercial swine farms with a clinical history of enteric disease or from cases that had been previously tested for transmissible gastroenteritis virus, PEDV, or rotavirus. Overall, 2286 banked samples were tested from 27 States. Samples were collected in 3 separate years and in 17 different months. Test results revealed 4 positive samples, 3 collected in August 2013 and 1 collected in October 2013. In addition, a case series including 42 operations in 10 States was conducted through administration of a survey. Survey data collected included information on characteristics of swine operations that had experienced PDCoV clinical signs. Special emphasis was placed on obtaining descriptive estimates of biosecurity practices and disease status over time of each operation. Clinical signs of PDCoV were reported to be similar to those of PEDV. The average number of animals on each operation exhibiting clinical signs (morbidity) and the average number of case fatalities was greatest for suckling and weaned pigs. Average operation-level weaned pig morbidity was greatest in the first week of the outbreak while average operation-level suckling pig case fatality was greatest in the second week of the outbreak. The survey included questions regarding biosecurity practices for visitors and operation employees; trucks, equipment and drivers; and feed sources. These questions attempted to identify a likely pathway of introduction of PDCoV onto the operations surveyed.

This short communication summarizes the strengths and weaknesses of current organisational structures in government veterinary services, as well as future technical, financial and societal challenges and related necessary competencies for government veterinary services of the future as discussed by an expert panel at ISVEE2015. First, participating representatives of veterinary services of diverse geographical backgrounds and statements from the audience confirmed that non-technical skills such as ability to work in teams, adaptability to new environments and situations, social and communication skills are increasingly seen as important. The second challenge faced by veterinary services is related to capacity issues, i.e. to have enough manpower to plan and deliver according to legislation in a period of shrinking budgets. New and emerging diseases can result in sudden, massive increases in the workload of veterinary services. Technical complexity has also increased for some hazards. Staff skills in veterinary services therefore need to be continuously updated, and it is essential to establish cross technical collaboration with other sectors including food safety, public health and environment. ISVEE conferences were seen as an opportunity to provide a global platform to develop skills needed by veterinary services, now and in the future.

Vesicular stomatitis (VS) is caused by a contagious rhabdovirus that affects horses, cattle, and swine. Clinical signs of vesicular stomatitis virus (VSV) infection in pigs and cattle are indistinguishable from foot-and-mouth disease (FMD), a foreign animal disease and reportable disease in the United States (Rodriguez et al., 2000). A VS epidemic occurred in the Rocky Mountain region in 2014-15. A study was conducted in Colorado to evaluate horse- and management-level factors associated with VS. For a horse to be considered a clinical VS horse, there were two requirements. First, clinical VS horses had to have clinical signs consistent with VS, including one or more of the following: vesicles, ulcers, erosions or crusting on the muzzle, nares, lips, oral or nasal mucosa, ears, ventrum, udder or penile sheath, or coronary band lesions. Second, clinical VS horses had to have laboratory confirmation of VSV exposure via virus isolation from lesions or a positive complement fixation test performed on sera. All non-clinical horses residing on VSV-affected premises enrolled in the study were evaluated for exposure (i.e., seroconversion) to VSV. Overall, management and housing data were collected from 334 horses on 48 premises in Colorado. Approximately one-third (31.4%) of enrolled horses were clinical cases and two-thirds (68.6%) were controls. Three premises-matched logistic regression models were constructed in SAS using backward elimination (P-value < 0.05) after univariate screening of a priori-selected variables (P-value < 0.20). Model outcomes included differences in characteristics and management of 1) clinical and nonclinical horses, 2) exposed and unexposed horses, and 3) exposed nonclinical and unexposed nonclinical horses. Overall, factors most strongly associated with risk of being a VS clinical horse were access to pasture (P-value = 0.002), and pregnancy status (P-value = 0.001). Factors most strongly associated with VSV exposure among horses were access to pasture (P-value = 0.003) and lack of any insect control (P-value = 0.001). The only factor associated with VSV-exposed nonclinical horses compared with unexposed VSV horses was contact with clinical horses (P-value = 0.013). There were no associations identified regarding clinical horses compared with exposed nonclinical horses. With regard to severity of lesions (severe vs. moderate or mild), no variables met the criteria for inclusion in the multivariable model. Results of this study provide evidence that pasture access and fly control are important factors associated with VSV exposure.

SUMMARY We investigated the plausibility of aerosol transmission of H5N2 highly pathogenic avian influenza (HPAI) virus during the 2015 spring outbreaks that occurred in the U.S. midwest. Air samples were collected inside and outside of infected turkey and layer facilities. Samples were tested to assess HPAI virus concentration (RNA copies/m3 of air), virus viability, and virus distribution by particle size. HPAI virus RNA was detected inside and up to 1000 m from infected facilities. HPAI virus was isolated from air samples collected inside, immediately outside, up to 70 m from infected facilities, and in aerosol particles larger than 2.1 μm. Direct exposure to exhausted aerosols proved to be a significant source of environmental contamination. These findings demonstrate HPAI virus aerosolization from infected flocks, and that both the transport of infectious aerosolized particles and the deposition of particles on surfaces around infected premises represent a potential risk for the spread of HPAI.

Between December 2014 and June 2015, an outbreak of H5N2 HPAI caused the largest and most expensive agriculture emergency in U.S. Department of Agriculture-Animal and Plant Health Inspection Service history. The outbreak affected 21 states; 232 poultry farms (211 commercial and 21 backyard) were affected, and approximately 49.6 million birds were depopulated on poultry farms. The majority of affected farms were commercial turkey operations (n = 160). This report is a case series describing 104 H5N2 HPAI-affected turkey farms in Iowa, Minnesota, Missouri, North Dakota, South Dakota, and Wisconsin that had H5N2 HPAI virus detected between March 5 and June 1, 2015. The farm manager or farm personnel voluntarily completed an epidemiologic questionnaire administered by state and federal animal health officials. Equipment and vehicle sharing with other farms was common, particularly for feed trucks (77% of farms shared feed trucks with other farms), live haul loaders (90.4%), poult trailers (72.0%), and preloaders (80.7%). Many farms had water bodies in proximity to the farm, such as a pond (42.6%) or stream (21.8%). About one-third of farms (33.7%) reported seeing wild birds inside the turkey barns. Only 44.2% of farms reported that third-party biosecurity audits or assessments had been conducted. Because the newly introduced Asian H5N8 HPAI and two new HPAI viruses, H5N2 and H5N1, are now circulating in U.S. wild birds, primarily migratory waterfowl, a greater potential for reoccurrence exists with the spring and fall migratory seasons, representing higher risk periods for outbreaks of HPAI in commercial poultry farms in the future. Eliminating exposure to wild birds, especially waterfowl or environments contaminated by wild waterfowl, will reduce risk of reintroduction of H5N2 HPAI virus, and ensuring good on-farm biosecurity will help the poultry industry avoid introduction of influenza and lateral spread between farms.

The reporting and monitoring of swine enteric coronavirus diseases (SECD), including porcine epidemic diarrhea virus and porcine delta coronavirus, in the United States have been challenging because of the initial absence of a regulatory framework and the emerging nature of these diseases. The National Animal Health Laboratory Network, the Emergency Management and Response System, and the Swine Health Monitoring Project were used to monitor the disease situation between May 2013 and March 2015. Important differences existed between and among them in terms of nature and extent of reporting. Here, we assess the implementation of these systems from different perspectives, including a description and comparison of collected data, disease metrics, usefulness, simplicity, flexibility, acceptability, representativeness, timeliness, and stability. This assessment demonstrates the limitations that the absence of premises identification imposes on certain animal health surveillance and response databases, and the importance of federally regulated frameworks in collecting accurate information in a timely manner. This study also demonstrates the value that the voluntary and producer-organized systems may have in monitoring emerging diseases. The results from all three data sources help to establish the baseline information on SECD epidemiological dynamics after almost 3 years of disease occurrence in the country.

Five laboratories participated in a study to evaluate sources of variation in results from an enzyme-linked immunosorbent assay (ELISA) for antibodies against Mycobacterium avium subsp. paratuberculosis. Each laboratory repeatedly tested duplicates of a negative, positive (P), and high-positive (HP) serum sample, which were supplied by the United States Department of Agriculture: Animal and Plant Health Inspection Service: Veterinary Services, National Veterinary Services Laboratories, Ames, IA, on all 96-well microtiter plates when routinely testing other samples for M. avium subsp. paratuberculosis antibodies. These 3 sera were aliquoted and sent to the 5 participating laboratories. This study focused on variation in test results because of assay reagents and laboratory techniques and did not account for biologic variability associated with the time course of infection in cattle. Overall, results from 868 microtiter plates were used in the study. For each sample a sample-to-positive (S/P) ratio was calculated according to the manufacturer's directions. The S/P ratio for the P sample ranged from 0.06 to 1.039 (mean = 0.466 and 0.484 for wells 1 and 2, respectively) and those for the HP sample ranged from 2.446 to 8.727 (mean = 4.027 and 3.980 for wells 1 and 2, respectively). The majority of the variation in S/P ratio for the P sample was attributed to kit lot (37.5%), followed by random (unexplained) error (27.0%), laboratory (18.3%), and kit lot by laboratory (11.9%). By eliminating plates in which the separation between negative and positive control ODs was less than 0.4, the proportion of variation attributed to laboratory was reduced markedly. These results confirm that there is variability in M. avium subsp. paratuberculosis ELISA results and that several sources contribute to the observed variability. The study gives a relative estimate of the contribution of various sources to the overall variability observed in the M. avium subsp. paratuberculosis ELISA results with kit lot being a primary contributor. Similar data for other ELISA tests for antibodies to M. avium subsp. paratuberculosis or other antigens also should be developed.

A competitive enzyme-linked immunosorbent assay (C-ELISA) for the detection of antibodies against vesicular stomatitis virus New Jersey (VSV-NJ) and vesicular stomatitis virus Indiana (VSV-IN) was compared with the serum neutralization test (SNT) using 1,106 serum samples obtained from dairy cattle on sentinel study farms in the Poa´s region of Costa Rica. Kappa coefficients between the C-ELISA and the SNT were 0.8871 (95% confidence interval [CI]: 0.8587–0.9155) and 0.6912 (95% CI: 0.6246–0.7577) for the VSV-NJ and VSV-IN tests, respectively. These results indicate good to excellent agreement between the 2 tests under these conditions. Ensayo inmunoabsorbente ligado a enzimas competitivo (C-ELISA) para la detección de anticuerpos contra El virus de la estomatitis vesicular de Nueva Jersey (VSV-NJ) y el virus de la estomatitis vesicular de Indiana (VSV-IN) se compararon con la prueba de seroneutralización (SNT) utilizando 1106 muestras de suero obtenidas de ganado lechero en granjas de estudio centinela en la región del Poa de Costa Rica. Los coeficientes Kappa entre el C-ELISA y el SNT fueron 0,8871 (95% intervalo de confianza [IC]: 0,8587–0,9155) y 0,6912 (IC 95 %: 0,6246–0,7577) para las pruebas VSV-NJ y VSV-IN, respectivamente. Estos resultados indican una concordancia de buena a excelente entre las 2 pruebas en estas condiciones. Universidad Nacional, Costa Rica Escuela de Medicina Veterinaria

Milk and serum samples from 35 dairy herds in 17 states were evaluated for cow- and herd-level Mycobacterium avium subspecies paratuberculosis (MAP) antibody test agreement. Evaluation of 6,349 samples suggested moderate agreement between milk and serum enzyme-linked immunosorbent assay (ELISA) results, with a kappa value of 0.50. Cow-level sensitivity (Se) for 18 dairy operations with 1,921 animals was evaluated relative to fecal culture results. At the cow level, the milk ELISA relative Se was not significantly different from that of the serum ELISA (21.2 and 23.5%, respectively). Logistic regression models revealed a positive association between lactation number and milk ELISA status. Non-Holstein cows were more likely to test milk ELISA positive than Holstein cows. Cows in the first 2 weeks of lactation and after week 45 of lactation were more likely to test milk ELISA positive than cows between 3 and 12 weeks of lactation. Milk production > 80% of herd average was negatively associated with testing milk ELISA positive. Animals in the West and Midwest regions were less likely than animals in the Southeast region to test ELISA positive by either test. Estimates for herd-level sensitivity for the milk and serum ELISA, relative to fecal culture results, ranged from 56 to 83%. At the cow and herd levels, milk ELISA performed equivalent to serum ELISA using fecal culture as a reference for MAP infection and has the advantage of decreased labor costs on farms that use Dairy Herd Improvement Association testing.

The use of sentinel chickens in establishing the negative status of commercial poultry flocks depopulated due to exotic Newcastle disease (END) is considered to be an economically beneficial process. However, the costs and benefits of using sentinel chickens in noncommercial operations are in question. The objective of this study was to use sentinel chickens to evaluate whether adequate cleaning and disinfection coupled with an appropriate time period without susceptible poultry species on the premises would eliminate END virus from a noncommercial poultry operation and preclude the need for placement of sentinels in previously infected operations before declaring them free of virus. Noncommercial poultry operations were selected from the 2002 to 2003 END outbreak database. Operations included in the study had one or more isolations of END virus (ENDV) from cloacal or oropharyngeal swabs of birds on the premises. A total of 546 birds were placed on 53 premises. All sentinel birds sampled after placements were negative by virus detection methods and serologic tests. Results of this study indicate that time and the application of appropriate cleaning and disinfection procedures will adequately mitigate the risk of viable virus persisting in noncommercial poultry operations. In the future, this information may eliminate the need for sentinel bird placement to ensure virus free status of premises before repopulation, thereby decreasing the costs of END eradication.

A multiplex single-tube reverse transcription—polymerase chain reaction (RT-PCR) has been developed for the detection and differentiation of vesicular stomatitis viruses (VSV), Indiana 1 and New Jersey, from insect samples. Using this assay, detection of either or both viruses in as little as 20 fg of total RNA from tissue culture was achieved, along with detection of vesicular stomatitis (VS) RNA from macerates containing 2 infected mosquitoes in pools of 10—30 noninfected mosquitoes. Vesicular stomatitis virus was detected by RT-PCR in all culture-positive samples, and detection as low as 4 plaque forming units per milliliter was achieved. Comparison between RT-PCR and tissue culture revealed that RT-PCR was able to detect VSV in a volume of insect macerate averaging almost 100 times less than that required for detection by tissue culture. The reported RT-PCR is a potential valuable tool for rapid and sensitive detection and differentiation of VS in insects because intense work associated with viral isolation, the cytotoxicity of insect extracts, and separate virus identification steps can be avoided. Potential application to detection and differentiation of VSV serotypes from vertebrate hosts is addressed.

Ground beef has been implicated in most outbreaks of Shiga-toxin-producing Escherichia coli O157 infections in humans.1–3 No outbreaks of infection with this organism have been linked to lamb; however, ruminants such as sheep may contribute to transmission of this organism to humans. Shiga-toxin-producing E. coli (STEC) have been reported to be more prevalent in sheep and goats than in cattle and have been isolated from several retail meats, including lamb.4,5 Sheep can be colonized by E. coli O157 experimentally and through natural exposure, and this serotype has been isolated from retail lamb in the United States.6–8 Here, we describe a pilot study conducted to determine the prevalence of E. coli O157 and other STEC serotypes in market lambs just prior to slaughter and to evaluate specific factors that may be associated with a higher or lower prevalence of E. coli O157 in lambs at slaughter. Lambs were sampled at a slaughter facility that processed lambs only located in the midwestern United States. All samples were collected over 3 days in October 1995. Sixty lambs were sampled from lots that exceeded 100 lambs; otherwise, 30 lambs were sampled per lot. Each lot was categorized by source (single farm flock vs. commercial feedlot), distance shipped, and elapsed time from farm to slaughter. Fecal samples were collected by rectal extraction of fecal pellets or, if feces were not in pellet form, by rectal swab with a cotton-tipped swab. For detection of E. coli O157, a single fecal pellet or fecal swab was placed in a culture tube containing 3 ml of tryptic soy brotha to which had been added 40 mg/ml vancomycinb and 50 ng/ml cefiximec (TSBcv). From every 10 lamb in order of sampling, a larger volume of feces was obtained with a gloved hand for culture of 10 g of feces (in addition to the pellet or swab cultures on the same lambs). The larger samples were placed into a polypropylene tube containing 20 ml of TSBcv to which was also added 2.5 mg/ml potassium tellurited (TSBcvt). From each of the same systematically selected lambs, a single fecal pellet was placed into a culture tube containing 10 ml of EC broth containing 20 mg/ml of novobiocind (ECBn). Samples in ECBn were used to determine the presence of STEC