Transboundary spread of pig diseases: the role of international trade and travel.

As globalization increases the interconnectedness between nations, economies, and industries, the introduction of diseases will continue to remain a prominent threat to the livestock sector and the trade of animals and animal products, as well as the livelihoods of farmers, food security and public health. The global pig sector, with its size and dichotomy between production type and biosecurity level, is particularly vulnerable to the transmission of transboundary animal diseases such as African and classical swine fever, foot and mouth disease, or porcine reproductive and respiratory syndrome. All of the above pose a constant threat to swine health, mainly as a result of both formal and informal international trade.Inspired in the risk assessment methodology, this paper classifies and provides an overview of the different pig disease introduction and exposure pathways, illustrated with abundant examples. Introduction pathways are classified as formal international trade (by product), informal international trade (by product), and spread through fomites. Formal trade of pigs and pork products is regulated by legislation and measures protecting animal populations from exotic diseases. Much more difficult to control is the transboundary swine disease transmission originating through informal trade, which entails illegal smuggling, but also the informal cross-border transfer of animals and products for personal use or within informal market chains. Meat products are most commonly mentioned, although fomites have also played a role in some cases, with live pigs, being more difficult to smuggle playing a role less frequently. The main exposure pathways are also described with the oral route playing a prominent role.Risk assessments can aid in the identification of pathways of pathogen introduction and exposure. However, quantitative information on informal disease introduction pathways remains very scarce and often incomplete, making it difficult to estimate the actual magnitudes of risks. Nevertheless, this knowledge is deemed essential to set up risk based awareness, prevention and surveillance programs that correspond to reality.

Cattle transhumance and agropastoral nomadic herding practices in Central Cameroon.

BACKGROUND: In sub-Saharan Africa, livestock transhumance represents a key adaptation strategy to environmental variability. In this context, seasonal livestock transhumance also plays an important role in driving the dynamics of multiple livestock infectious diseases. In Cameroon, cattle transhumance is a common practice during the dry season across all the main livestock production zones. Currently, the little recorded information of the migratory routes, grazing locations and nomadic herding practices adopted by pastoralists, limits our understanding of pastoral cattle movements in the country. GPS-tracking technology in combination with a questionnaire based-survey were used to study a limited pool of 10 cattle herds from the Adamawa Region of Cameroon during their seasonal migration, between October 2014 and May 2015. The data were used to analyse the trajectories and movement patterns, and to characterize the key animal health aspects related to this seasonal migration in Cameroon. RESULTS: Several administrative Regions of the country were visited by the transhumant herds over more than 6 months. Herds travelled between 53 and 170 km to their transhumance grazing areas adopting different strategies, some travelling directly to their destination areas while others having multiple resting periods and grazing areas. Despite their limitations, these are among the first detailed data available on transhumance in Cameroon. These reports highlight key livestock health issues and the potential for multiple types of interactions between transhumant herds and other domestic and wild animals, as well as with the formal livestock trading system. CONCLUSION: Overall, these findings provide useful insights into transhumance patterns and into the related animal health implications recorded in Cameroon. This knowledge could better inform evidence-based approaches for designing infectious diseases surveillance and control measures and help driving further studies to improve the understanding of risks associated with livestock movements in the region.

Corrigendum: Examination of critical factors influencing ruminant disease dynamics in the Black Sea Basin.

[This corrects the article DOI: 10.3389/fvets.2023.1174560.].

Comparison of vaccination schedules for foot-and-mouth disease among cattle and sheep in Mongolia.

Vaccines are a critical tool for the control strategy for foot-and-mouth disease (FMD) in Mongolia where sporadic outbreaks regularly occur. A two-dose primary vaccination course is recommended for most commercial vaccines though this can be logistically challenging to deliver among nomadic pastoralist systems which predominate in the country. Although there is evidence that very high potency vaccines can provide prolonged duration of immunity, this has not been demonstrated under field conditions using commercially available vaccines. This study compared neutralizing titres to a O/ME-SA/Panasia strain over a 6-month period following either a two-dose primary course or a single double-dose vaccination among Mongolian sheep and cattle using a 6.0 PD50 vaccine. Titers were not significantly different between groups except in sheep at six-months post vaccination when the single double-dose group had significantly lower titers. These results indicate the single double-dose regimen may be a cost-effective approach for vaccination campaigns supporting FMD control in Mongolia.

Examination of critical factors influencing ruminant disease dynamics in the Black Sea Basin.

Introduction: Ruminant production in the Black Sea basin (BSB) is critical for national economies and the subsistence of rural populations. Yet, zoonoses and transboundary animal diseases (TADs) are limiting and threatening the sector. To gain a more comprehensive understanding, this study characterizes key aspects of the ruminant sector in nine countries of the BSB, including Armenia, Azerbaijan, Belarus, Bulgaria, Georgia, Moldova, Romania, Türkiye, and Ukraine. Methods: We selected six priority ruminant diseases (anthrax, brucellosis, Crimean Congo haemorrhagic fever (CCHF), foot-and-mouth disease (FMD), lumpy skin disease (LSD), and peste des petits ruminants (PPR)) that are present or threaten to emerge in the region. Standardized questionnaires were completed by a network of focal points and supplemented with external sources. We examined country and ruminant-specific data such as demographics, economic importance, and value chains in each country. For disease-specific data, we analysed the sanitary status, management strategies, and temporal trends of the selected diseases. Results and discussion: The shift from a centrally planned to a market economy, following the collapse of the Soviet Union, restructured the ruminant sector. This sector played a critical role in rural livelihoods within the BSB. Yet, it faced significant challenges such as the low sustainability of pastoralism, technological limitations, and unregistered farms. Additionally, ruminant health was hindered by informal animal trade as a result of economic factors, insufficient support for the development of formal trade, and socio-cultural drivers. In the Caucasus and Türkiye, where diseases were present, improvements to ruminant health were driven by access to trading opportunities. Conversely, European countries, mostly disease-free, prioritized preventing disease incursion to avoid a high economic burden. While international initiatives for disease management are underway in the BSB, there is still a need for more effective local resource allocation and international partnerships to strengthen veterinary health capacity, protect animal health and improve ruminant production.

Peste des Petits Ruminants in Central and Eastern Asia/West Eurasia: Epidemiological Situation and Status of Control and Eradication Activities after the First Phase of the PPR Global Eradication Programme (2017-2021).

Peste des petits ruminants (PPR) is a highly contagious infectious disease of small ruminants caused by peste des petits ruminants virus (PPRV). PPR poses a significant threat to sheep and goat systems in over 65 endemic countries across Africa, the Middle East and Asia. It is also responsible for devastating outbreaks in susceptible wildlife, threatening biodiversity. For these reasons, PPR is the target of the Global Eradication Programme (PPR GEP), launched in 2016, which is aimed at eradicating the disease by 2030. The end of the first five-year phase of the PPR GEP (2017-2021) provides an ideal opportunity to assess the status of the stepwise control and eradication process. This review analyses 13 countries belonging to Eastern Europe, Transcaucasia, and Central and East Asia. Substantial heterogeneity is apparent in terms of PPR presence and control strategies implemented by different countries. Within this region, one country is officially recognised as PPR-free, seven countries have never reported PPR, and two have had no outbreaks in the last five years. Therefore, there is real potential for countries in this region to move forward in a coordinated manner to secure official PPR freedom status and thus reap the trade and socioeconomic benefits of PPR eradication.

Immunogenicity of Mycobacterium avium subsp. paratuberculosis proteins in Crohn's disease patients.

UNLABELLED: The association between Mycobacterium avium subsp. paratuberculosis (MAP) and Crohn's disease (CD) is supported by several studies reporting the detection or isolation of MAP from human tissues, but a direct association is still debatable. OBJECTIVE: To evaluate the survival of MAP in human intestinal cells and to measure the presence of antibodies against two mycobacterial proteins necessary for the survival of the bacterium in the sera of CD patients. MATERIAL AND METHODS: Human-derived intestinal cells were infected with three isolates of MAP and the survival of the microorganism was determined. The presence of antibodies against protein tyrosine phosphatase A (PtpA) and protein kinase G (two proteins secreted within the host in the early stages of the invasion) in the sera of CD patients was evaluated. Sera of 20 CD patients and 20 controls were collected and the presence of the antibodies was assayed using enzyme-linked immunosorbent assay (ELISA). Secretion of the PtpA in vivo was visualized by immunostaining. RESULTS: MAP survived in intestinal cells, and immunostaining of PtpA showed that the protein was secreted within these cells. Wilcoxon rank sum test revealed that CD patient sera had significantly higher titer of antibodies specific for both of these antigens compared to controls. ELISA results for either protein were not statistically different between men and women. CONCLUSIONS: The presence of specific antibodies against mycobacterial proteins essential for establishing an infection in the host suggests that MAP can potentially be active in CD patients, and a serological test can be developed for early detection of MAP in CD patients.

Seroprevalence of infectious bovine rhinotracheitis and bovine viral diarrhea virus type 1 and type 2 in non-vaccinated cattle herds in the Pacific Region of Central Costa Rica.

The objectives of this cross-sectional study were to estimate the seroprevalence of infectious bovine rhinotracheitis (IBR, BHV-1) and bovine viral diarrhea virus (BVDV) in a population of non-vaccinated, double purpose, dairy and beef herds in the Pacific Region of Central Costa Rica. Blood samples were collected from a total of 496 animals from 35 herds. Sera were tested for antibodies against BHV-1(IBR) and BVDV types 1 and 2 using serum neutralization test. The average number of animals tested in each herd for each of the viruses was 14. Overall individual seroprevalence was 48%, 27%, and 19% for IBR, BVDV type 1, and BVDV type 2, respectively. Median within-herd seroprevalence for IBR, BVDV type 1 and type 2 were 43%, 27%, and 24%, respectively.

African Swine Fever in Mongolia: Course of the Epidemic and Applied Control Measures.

African swine fever (ASF) is spreading rapidly in Asia and was confirmed in Mongolia on 10 January 2019. Following the outbreak confirmation, a state emergency committee was established with representation from municipal authorities and other relevant authorities including the General Authority for Veterinary Services, National Emergency Management Agency, General Agency for Specialized Inspection, and the Ministry of Environment and Tourism. The committee provided recommendations and coordinated closely with the State Central Veterinary Laboratory to ensure quick outbreak investigation and response. In addition to outbreak investigations, sampling took place at farms and food premises and suggests a link between the outbreaks and swill feeding practices among backyard pig farmers. Upon government request, the Food and Agriculture Organization of the United Nations (FAO) deployed an expert team to assist in identifying risk factors for the disease spread and provide recommendations as how to improve disease prevention and response. Following the control measures from the involved agencies, the epidemic was successfully controlled and declared over on 11 April 2019. In total, the epidemic affected 83 pig farming households and led to a total of 2862 dead or culled pigs in eleven districts of seven provinces in Mongolia.

Serologic survey for selected infectious diseases in raccoons (Procyon lotor) in Indiana, USA.

The objective of this study was to characterize the antibody prevalence of important livestock and domestic animal pathogens in raccoons (Procyon lotor) trapped and sampled in 39 forest patches in north-central Indiana, USA, between 2004 and 2005. A total of 459 serum samples were tested for antibodies to Leptospira serovars, 512 for Canine distemper virus (CDV) antibodies, and 340 for antibodies to Porcine reproductive and respiratory syndrome virus (PRRSV). In total, 47, 16, and 0% of the samples were seropositive for at least one Leptospira serovar, CDV, and PRRSV, respectively. Most of the positive Leptospira results were to serovars grippotyphosa (36%), autumnalis (22%), and hardjo (22%). No statistically significant correlation was found between antibody prevalence estimates for different Leptospira serovars. A significant association was found between body weight and antibodies for Leptospira serovars and CDV. In addition, age (adult vs. juvenile) was significantly associated with the presence of CDV antibody, with adults exhibiting a higher prevalence than juveniles. This study confirmed that raccoons in Indiana, USA, are exposed to different Leptospira interrogans serovars and CDV and that age and weight are associated with the presence of antibodies for both pathogens.

Prevalence and characterization of bovine viral diarrhea virus in the white-tailed deer population in Indiana.

Bovine viral diarrhea (BVD) is one of the economically important diseases of cattle. For many years, different types of vaccines have been commercially available, yet this disease is hard to control in high-density population areas. Detection and isolation of bovine viral diarrhea virus (BVDV) from any potential reservoir is vital, especially when considering virus eradication from a herd or locale. One potential source is wild ruminants. Ear notches and lymph nodes were collected from the wild population of white-tailed deer (Odocoileus virginianus) during deer hunting season in Indiana and tested for BVDV with a commercial BVD antigen capture enzyme-linked immunosorbent assay. Two samples out of 745 collected samples were positive, and subsequently cp and ncp BVDV was isolated from 1 ear notch and 1 lymph node. These isolates were genotyped as type 1a and 1b based on sequence analysis of the 5' untranslated region (UTR). The results of the present study indicate that the prevalence of BVDV in the white-tailed deer population of Indiana is about 0.3%. Wild ruminants infected with BVDV should be taken into consideration during an eradication program of BVDV from the livestock population.

Simulation model for evaluation of testing strategies for detection of paratuberculosis in midwestern US dairy herds.

We developed a stochastic simulation model to compare the herd sensitivity (HSe) of five testing strategies for detection of Mycobacterium avium subsp. paratuberculosis (Map) in Midwestern US dairies. Testing strategies were ELISA serologic testing by two commercial assays (EA and EB), ELISA testing with follow-up of positive samples with individual fecal culture (EAIFC and EBIFC), individual fecal culture (IFC), pooled fecal culture (PFC), and culture of fecal slurry samples from the environment (ENV). We assumed that these dairies had no prior paratuberculosis-related testing and culling. We used cost-effectiveness (CE) analysis to compare the cost to HSe of testing strategies for different within-herd prevalences. HSe was strongly associated with within-herd prevalence, number of Map organisms shed in feces by infected cows, and number of samples tested. Among evaluated testing methods with 100% herd specificity (HSp), ENV was the most cost-effective method for herds with a low (5%), moderate (16%) or high (35%) Map prevalence. The PFC, IFC, EAIFC and EBIFC were increasingly more costly detection methods. Culture of six environmental samples per herd yielded >or=99% HSe in herds with >or=16% within-herd prevalence, but was not sufficient to achieve 95% HSe in low-prevalence herds (5%). Testing all cows using EAIFC or EBIFC, as is commonly done in paratuberculosis-screening programs, was less likely to achieve a HSe of 95% in low than in high prevalence herds. ELISA alone was a sensitive and low-cost testing method; however, without confirmatory fecal culture, testing 30 cows in non-infected herds yielded HSp of 21% and 91% for EA and EB, respectively.

Immunogenicity of PtpA secreted during Mycobacterium avium ssp. paratuberculosis infection in cattle.

AIMS: Mycobacterium avium subsp. paratuberculosis (MAP) is the etiological agent of Johne's disease. To survive within host macrophages, the pathogen secretes a battery of proteins to interfere with the immunological response of the host. One of these proteins is tyrosine phosphate A (PtpA), which has been identified as a secreted protein critical for survival of its close relative M. tuberculosis within infected macrophages. METHODS AND RESULTS: In this study, the immune response to recombinant PtpA used as an antigen was investigated in a cohort of ∼1000 cows infected with MAP compared to negative control animals using ELISA. The sera from MAP-infected cows had significantly higher levels of antibodies against PtpA when compared to uninfected cows. CONCLUSIONS: The data presented here indicate that the antibodies produced against PtpA are sensitive enough to detect infected animals before the appearance of the disease symptoms. SIGNIFICANCE AND IMPACT OF STUDY: The use of PtpA as an antigen can be developed as an early diagnostic test. Moreover, PtpA is a candidate antigen for detection of humoral immune responses in cows infected with MAP.

The association between Mycobacterium avium subsp. paratuberculosis fecal shedding or clinical Johne's disease and lactation performance on two Minnesota, USA dairy farms.

Lactation performance of cows infected with Mycobacterium avium subsp. paratuberculosis (Map) was previously studied using only serum ELISA as a diagnostic method. This study evaluated on two dairy farms in Minnesota, USA the lactation performance (measures of health, production, reproduction, and survival) of cows shedding Map in feces before calving and of cows culled with clinical signs consistent with Johne's disease (JD) during the subsequent lactation. Fecal samples were collected from 1052 cows within 21 day before calving and tested for Map with bacterial culture. Producers' observed signs of clinical disease (milk fever, retained placenta, metritis, ketosis, displaced abomasum, lameness, mastitis, pneumonia, and JD) and production and reproduction data were recorded for each cow. The association between fecal shedding or clinical JD and lactation performance was evaluated. Logistic regression was used to evaluate the association with any clinical and subclinical diseases as the outcome. General linear model was used to evaluate the association with milk production, and survival analysis techniques were used to evaluate the association with days in the study before culling and days from calving to conception. In 84 cows (8% of 1052 cows) fecal samples were positive for Map (46% light, 26% moderate, and 28% heavy shedders). In multivariable analysis, light, moderate, and heavy fecal shedding cows produced on average 537, 1403, and 1534 kg, respectively, less milk per lactation and 1.4, 5.2, and 7.5 kg, respectively, less milk per day than fecal negative cows. Fecal culture positive cows were less likely to be bred and conceive. In the multivariable analysis the 56 cows culled with presumed JD produced approximately 1500 kg/lactation or 5 kg/day less than all other cows. The negative economic impact implied by decreased lactation performance in cows shedding Map or with clinical JD may motivate producers to implement programs to control Map infection and subsequent JD.

The associations between culling due to clinical Johne's disease or the detection of Mycobacterium avium subsp. paratuberculosis fecal shedding and the diagnosis of clinical or subclinical diseases in two dairy herds in Minnesota, USA.

Our objectives were to identify associations between clinical or subclinical diseases and subsequent culling because of clinical Johne's disease (JD) or the detection of fecal shedding of Mycobacterium paratuberculosis (Map) after 305 days in milk (DIM). A total of 1297 cows from two Minnesota dairies were enrolled in the study. From study cows, fecal samples were obtained prior to calving (close-up period) and after at least 305 DIM or at the time of leaving the herd (sold/dead). Between 3 and 21 DIM, blood samples were obtained for serum betahydroxybutyrate (BHB) and serum total-protein testing. Body-condition score (BCS) was evaluated during the close-up period, between 3 and 21 DIM, and at the end of lactation. The diagnosis time (DIM) of clinical disease events (culling because of JD clinical signs, ketosis, lameness, mastitis, displacement abomasum, injury, metritis, milk fever, pneumonia, and retained placenta) was recorded. Sixty-six cows were culled because of JD clinical signs (CCDJ) with average DIM of 209. CCDJ was associated with event of pneumonia (n=131) (OR=3.0, 95% CI=1.0-6.0) and level of fecal shedding (light: OR=13.0, 95% CI=5.3-30.0; moderate: OR=34.0, 95% CI=13.0-89.0; heavy: OR=66.0, 95% CI=26.0-171.0). Detection of fecal shedding at the end of the lactation (n=79) was associated only with event of pneumonia (OR=2.0, 95% CI=1.0-4.0).

Characterization of Minnesota dairy herds participating in a Johne's disease control program and evaluation of the program risk assessment tool.

The objectives of this study were to: (1) characterize Minnesota dairy herds participating in a Johne's disease control program (JDCP) based on herd size, milk production, and clinical Johne's disease (JD) history, (2) evaluate if change in farm management practices, expressed in risk assessment (RA) total score, is associated with the change between the first and most recent ELISA test herd seroprevalence or change in clinical JD culling rate, and (3) identify farm factors associated with ELISA seroprevalence. A total of 1234 RA, performed between January 2000 and February 2004, were available for analysis from 714 dairy herds. ELISA test results from herd sampling between 2000 and 2004 were obtained from the Minnesota board of animal health (MBAH) database, and were available for 474 herds. Both the first and the most recent ELISA test results for herds with more than one RA were available for 262 herds. Mean herd size and mean annual milk production per cow was higher in JDCP dairy herds (161 milking cows) than either all Minnesota dairy herds or Minnesota dairy herd improvement association (DHIA) herds. For herds with more than one RA available, the most recent RA total score was significantly lower (mean 11% less) than the first. The change in RA total score (and any RA subtotal scores) between the first and most recent RA was not associated with the change between the first and the most recent ELISA within-herd seroprevalence or the change in JD culling rate between the first and most recent RA. The most recent ELISA test results were positively associated with postweaned heifer score and JD culling rate. The RA score was not found to be an effective tool for the prediction of ELISA seroprevalence.

Mycobacterium avium subsp. paratuberculosis from free-ranging deer and rabbits surrounding Minnesota dairy herds.

The objectives of this study were to estimate the prevalence of Mycobacterium avium subsp. paratuberculosis (MAP) among deer and rabbits surrounding infected and noninfected Minnesota dairy farms using fecal culture, and to describe the frequency that farm management practices were used that could potentially lead to transmission of infection between these species. Fecal samples from cows and the cow environment were collected from 108 Minnesota dairy herds, and fecal pellets from free-ranging white-tailed deer and eastern cottontail rabbits were collected from locations surrounding 114 farms; all samples were tested using bacterial culture. In addition, a questionnaire was administered to 114 herd owners. Sixty-two percent of the dairy herds had at least 1 positive fecal pool or environmental sample. A total of 218 rabbit samples were collected from 90% of the herds, and 309 deer samples were collected from 47% of the herds. On 2 (4%) of the farms sampled, 1 deer fecal sample was MAP positive. Both farms had samples from the cow fecal pool and cow environment that were positive by culture. On 2 (2%) other farms, 1 rabbit fecal sample was positive by culture to MAP, with one of these farms having positive cow fecal pools and cow environmental samples. Pasture was used on 79% of the study farms as a grazing area for cattle, mainly for dry cows (75%) and bred or prebred heifers (87%). Of the 114 farms, 88 (77%) provided access to drylot for their cattle, mainly for milking cows (77/88; 88%) and bred heifers (87%). Of all study farms, 90 (79%) used some solid manure broadcasting on their crop fields. Of all 114 farms, the estimated probability of daily physical contact between cattle manure and deer or rabbits was 20% and 25%, respectively. Possible contact between cattle manure and deer or rabbits was estimated to occur primarily from March through December. The frequency of pasture or drylot use and manure spreading on crop fields may be important risk factors for transmission of MAP among dairy cattle, deer, and rabbits. Although the MAP prevalence among rabbits and deer is low, their role as MAP reservoirs should be considered.

Evaluation of horizontal transmission of bovine viral diarrhea virus type 1a from experimentally infected white-tailed deer fawns (Odocoileus virginianus) to colostrum-deprived calves.

OBJECTIVE: To assess the transmission of bovine viral diarrhea virus (BVDV) from experimentally infected white-tailed deer fawns to colostrum-deprived calves by use of a BVDV strain isolated from hunter-harvested white-tailed deer. ANIMALS: 5 white-tailed deer (Odocoileus virginianus) fawns and 6 colostrum-deprived calves. PROCEDURES: Fawns were inoculated intranasally with a noncytopathic BVDV-1a isolate (2 mL containing 10(6.7) TCID(50)/mL), and 2 days after inoculation, animals were commingled until the end of the study. Blood and serum samples were obtained on days -6, 0, 7, 14, and 21 after inoculation for reverse transcriptase PCR assay, virus neutralization, and BVDV-specific antibody ELISA. Nasal, oral, and rectal swab specimens were collected on days 0, 3, 7, 14, 17, and 21 for reverse transcriptase PCR testing. By 21 days after inoculation, all animals were euthanized and necropsied and tissues were collected for histologic evaluation, immunohistochemical analysis, and virus isolation. RESULTS: All fawns became infected and shed the virus for up to 18 days as determined on the basis of reverse transcriptase PCR testing and virus isolation results. Evidence of BVDV infection as a result of cohabitation with acutely infected fawns was detected in 4 of the 6 calves by means of reverse transcriptase PCR testing and virus isolation. CONCLUSIONS AND CLINICAL RELEVANCE: On the basis of these findings, BVDV transmission from acutely infected fawns to colostrum-deprived calves appeared possible.

Long-Term Survival of Mycobacterium avium subsp. paratuberculosis in Fecal Samples Obtained from Naturally Infected Cows and Stored at -18°C and -70°C.

The objective was to evaluate the survival of Mycobacterium paratuberculosis (Map) in naturally infected dairy cows feces under long periods of freezing at -18°C and -70°C. Samples were collected from cows previously tested positive with serum ELISA or fecal culture, or with clinical signs of Johne's disease. Samples were stored at -18°C and/or -70°C and recultured in Herrold's egg yolk media every 3-6 months. A proportional odds mixed model was used for data analysis. Sixty nine fecal samples were stored for different periods between September 2002 and January 2005. Of these, 45 (65%) were stored at -18°C and 24 (35%) at -70°C. Average number of days between repeated culture dates was 98 and 84 for -18°C and -70°C, respectively. Median number of repeated cultures was 6 and 4 for samples stored at -18°C and -70°C, respectively. After adjusting for initial sample bacterial load, the effects of temperature or number of thawing and refreezing cycles on Map viability were not significant. The probability that a sample decreases from high to moderate-low bacterial load and from moderate-low to negative bacterial load was 13.5% per month. Although this study found gradual reduction of Map concentration in stored fecal samples through time, overall survival in -18°C can ease fecal samples management in laboratories with low-processing capacity or lack of -70°C freezer.

Estimated within-herd prevalence (WHP) of Mycobacterium avium subsp. paratuberculosis in a sample of Minnesota dairy herds using bacterial culture of pooled fecal samples.

The objective of this study was to describe the estimated within-herd prevalence (WHP) of Mycobacterium avium subsp. paratuberculosis (Map) in a sample of infected dairy herds in Minnesota (N = 66) using test results from bacterial culture of pooled fecal samples. Fecal samples were collected from up to 100 cows in each herd and were tested using bacterial culture in pools of 5 cows based on age order. The mean herd size was 222 (44 to 1500) milking cows; the cows were predominantly Holstein. Using a frequentist approach, the within-herd mean individual fecal prevalence was 10% [95% confidence interval (CI) = 4% to 16%] assuming 70% test sensitivity and 99.5% test specificity. Using Bayesian methods, the estimated true within-herd individual cow prevalence was 14% (95% CI = 7% to 27%). Within-herd prevalence was higher in larger dairy herds than in herds with fewer cows. As Map is the causative agent of Johne's disease (JD), the results of this study could contribute to the success of a nationwide control program for this disease.