Seroprevalence of Mycobacterium bovis infection in warthogs (Phacochoerus africanus) in bovine tuberculosis-endemic regions of South Africa.

Bovine tuberculosis (bTB), caused by Mycobacterium bovis (M. bovis), has been reported in many species including suids. Wild boar are important maintenance hosts of the infection with other suids, that is domestic and feral pigs, being important spillover hosts in the Eurasian ecosystem and in South Africa, warthogs (Phacochoerus africanus) may play a similar role in M. bovis-endemic areas. However, novel diagnostic tests for warthogs are required to investigate the epidemiology of bTB in this species. Recent studies have demonstrated that serological assays are capable of discriminating between M. bovis-infected and uninfected warthogs (Roos et al., ). In this study, an indirect ELISA utilizing M. bovis purified protein derivative (PPD) as a test antigen was used to measure the prevalence and investigate risk factors associated with infection in warthogs from uMhkuze Nature Reserve and the southern region of the Greater Kruger National Park (GKNP). There was a high overall seroprevalence of 38%, with adult warthogs having a higher risk of infection (46%). Seroprevalence also varied by geographic location with warthogs from Marloth Park in the GKNP having the greatest percentage of positive animals (63%). This study indicates that warthogs in M. bovis-endemic areas are at high risk of becoming infected with mycobacteria. Warthogs might present an under-recognized disease threat in multi-species systems. They might also serve as convenient sentinels for M. bovis in endemic areas. These findings highlight the importance of epidemiological studies in wildlife to understand the role each species plays in disease ecology.

Effect of a Pheromone on Stress-Associated Reactivation of Feline Herpesvirus-1 in Experimentally Inoculated Kittens.

BACKGROUND: Stress contributes to reactivation of feline herpesvirus-1 (FHV-1). The usage of pheromones to decrease stress in FHV-1 experimentally inoculated kittens has not previously been investigated. HYPOTHESIS/OBJECTIVES: To determine whether a feline pheromone would lessen stress, resulting in decreased recurrence of FHV-1-associated illness in kittens. ANIMALS: Twelve 5-month-old, purpose-bred kittens. METHODS: Randomized, double-blind, placebo-controlled clinical trial. Kittens previously infected with the same dose of FHV-1 were randomized into 2 separate but identical group rooms. After a 2-week equilibration period, a diffuser containing either the pheromone or placebo was placed in each of the rooms, and the kittens acclimated for an additional 2 weeks. Every 2 weeks thereafter, for the 8-week study period, housing was alternated between kennel- and group housing. Blinded observers applied a standardized clinical and behavioral scoring rubric daily. After each 2-week period, serum cortisol concentrations and quantitative PCR for FHV-1 and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) ratios were evaluated. Clinical, behavioral, and laboratory test results were compared between groups within individual and combined study periods. RESULTS: Sneezing occurred more frequently in the placebo group during individual (P = 0.006) and combined study periods (P = 0.001). Sleep at the end of observation periods occurred more frequently in the pheromone group during individual (P = 0.006) and combined study periods (P < 0.001). CONCLUSIONS AND CLINICAL IMPORTANCE: The findings suggest that the pheromone decreased stress, and the decrease in stress response may have resulted in decreased sneezing associated with FHV-1.

Tuberculosis serosurveillance and management practices of captive African elephants (Loxodonta africana) in the Kavango-Zambezi Transfrontier Conservation Area.

Transfrontier conservation areas represent an international effort to encourage conservation and sustainable development. Their success faces a number of challenges, including disease management in wildlife, livestock and humans. Tuberculosis (TB) affects humans and a multitude of non-human animal species and is of particular concern in sub-Saharan Africa. The Kavango-Zambezi Transfrontier Conservation Area encompasses five countries, including Zimbabwe, and is home to the largest contiguous population of free-ranging elephants in Africa. Elephants are known to be susceptible to TB; thus, understanding TB status, exposure and transmission risks to and from elephants in this area is of interest for both conservation and human health. To assess risk factors for TB seroprevalence, a questionnaire was used to collect data regarding elephant management at four ecotourism facilities offering elephant-back tourist rides in the Victoria Falls area of Zimbabwe. Thirty-five working African elephants were screened for Mycobacterium tuberculosis complex antibodies using the ElephantTB Stat-Pak and the DPP VetTB Assay for elephants. Six of 35 elephants (17.1%) were seropositive. The risk factor most important for seropositive status was time in captivity. This is the first study to assess TB seroprevalence and risk factors in working African elephants in their home range. Our findings will provide a foundation to develop guidelines to protect the health of captive and free-ranging elephants in the southern African context, as well as elephant handlers through simple interventions. Minimizing exposure through shared feed with other wildlife, routine TB testing of elephant handlers and regular serological screening of elephants are recommended as preventive measures.

Anti-inflammatory drugs decrease infection of brain endothelial cells with EHV-1 in vitro.

BACKGROUND: Equine herpesvirus-associated myeloencephalopathy is the result of endothelial cell infection of the spinal cord vasculature with equine herpesvirus-1 (EHV-1) during cell-associated viraemia. Endothelial cell infection requires contact between infected peripheral blood mononuclear and endothelial cells. Inflammation generated during viraemia likely upregulates adhesion molecule expression on both cell types increasing contact and facilitating endothelial cell infection. OBJECTIVES: Evaluating the role of anti-inflammatory drugs in decreasing endothelial cell infection with EHV-1. STUDY DESIGN: In vitro assay, crossover design, multiple drug testing. METHODS: In vitro modified infectious centre assay using immortalised carotid artery endothelial cells or primary brain endothelial cells with plaque counts per well as outcome. Cells were either anti-inflammatory drug treated or left untreated. RESULTS: Significant reduction of plaque count when cells were treated compared with untreated cells. No dose-dependent effect when drug concentrations were increased to 10× dose. Treatment of both peripheral blood mononuclear cells (PBMC) and endothelial cells (EC) is required for significant plaque count reduction. MAIN LIMITATIONS: In vitro study. CONCLUSIONS: Anti-inflammatory drugs decrease infection of endothelial cells likely by reducing contact between EHV-1 infected PBMC and endothelial cells in vitro. The role of adhesion molecules in this process needs further investigation. In vitro results suggest anti-inflammatory drug therapy during EHV-1 infection and viraemia in horses could be clinically relevant.

Rhodococcus equi pneumonia in foals: an assessment of the early diagnostic value of serum amyloid A and plasma fibrinogen concentrations in equine clinical practice.

Early diagnosis and prevention of Rhodococcus equi pneumonia in foals represent important goals for equine clinicians. Recent protocols for diagnosis and treatment of Rhodococcosis in foals typically rely on a multimodal approach based on sonographic evidence suggestive of pyogranulomas, sonographic abscess scores and laboratory findings including plasma fibrinogen concentrations, blood biochemistry testing and platelet and leukocyte counts. The aim of this study was to assess the utility of weekly testing of serum amyloid A (SAA) and plasma fibrinogen concentrations in foals to achieve early diagnosis of R. equi pneumonia prior to the onset of clinical signs. This testing was used to simulate a clinically practical screening procedure and compared with thoracic ultrasonography performed in parallel. The present study suggests that SAA does not represent a reliable early marker of Rhodococcosis when plasma concentrations are tested weekly. However, when clinical signs of R. equi pneumonia are present, SAA concentrations may allow clinicians to obtain 'real-time' indications concerning both the progress of infection and the effectiveness of therapy. This study raises the possibility that plasma fibrinogen monitoring starting at 1 week of age and repeated on a weekly basis, could serve as a screening test allowing clinicians to identify foals as suspected of R. equi infection. Future investigations regarding both physiological plasma fibrinogen concentrations in foals as well as fibrinogen kinetics in foals affected with R. equi pneumonia, including the establishment of appropriate reference intervals for the test method employed in this study, will be necessary in order to clarify this possibility.

Dairy cow handling facilities and the perception of Beef Quality Assurance on Colorado dairies.

A survey was conducted on Colorado dairies to assess attitudes and practices regarding Dairy Beef Quality Assurance (DBQA). The objectives were to (1) assess the need for a new handling facility that would allow all injections to be administered via DBQA standards; (2) establish if Colorado dairy producers are concerned with DBQA; and (3) assess differences in responses between dairy owners and herdsmen. Of the 95 dairies contacted, 20 (21%) agreed to participate, with a median herd size of 1,178. When asked to rank the following 7 traits--efficiency, animal safety, human safety, ease of animal handling, ease of operation, inject per Beef Quality Assurance (BQA) procedures, and cost--in order of priority when designing a new handling facility, human and animal safety were ranked highest in priority (first or second) by the majority of participants, with ease of animal handling and efficiency ranked next. Interestingly, the administration of injections per BQA standards was ranked sixth or seventh by most participants. Respondents estimated the average annual income from the sale of cull cows to be 4.6% of all dairy income, with 50% receiving at least one carcass discount or condemnation in the past 12 mo. Although almost all of the participating dairy farmers stated that the preferred injection site for medications was the neck region, a significant number admitted to using alternate injection sites. In contrast, no difference was found between responses regarding the preferred and actual location for intravenous injections. Although most participating producers are aware of BQA injection guidelines, they perceive efficiency as more important, which could result in injections being administered in locations not promoted by BQA. Dairy owners and herdsmen disagreed in whether or not workers had been injured in the animal handling area in the last 12 mo. Handling facilities that allow for an efficient and safe way to administer drugs according to BQA guidelines and educational opportunities that highlight the effect of improved DBQA on profitability could prove useful. Dairy producers play a key role in ensuring that dairy beef is safe and high quality, and just as they are committed to producing safe and nutritious milk for their customers, they should be committed to producing the best quality beef.

Evaluation of strategies for the eradication of Pseudorabies virus (Aujeszky's disease) in commercial swine farms in Chiang-Mai and Lampoon Provinces, Thailand, using a simulation disease spread model.

Several strategies for eradicating Pseudorabies virus (Aujeszky's disease) in Chiang-Mai and Lampoon Provinces, Thailand, were compared using a computer simulation model, the North American Animal Disease Spread Model (NAADSM). The duration of the outbreak, the number of affected herds and the number of destroyed herds were compared during these simulated outbreaks. Depopulation, zoning for restricted movement and improved detection and vaccination strategies were assessed. The most effective strategies to eradicate Pseudorabies as per the findings from this study are applying depopulation strategies with MOVEMENT RESTRICTIONS in 3-, 8- and 16-km ZONES surrounding infected herds and enhancing the eradication with vaccination campaign on 16-km radius surrounding infected herds.

Using temperature-sensing reticular boluses to aid in the detection of production diseases in dairy cows.

The objective of this study was to investigate associations between increases in reticular temperature (RT) in dairy cows and the diagnosis of metritis, mastitis, lameness, and pneumonia by dairy personnel. A prospective case-control study was conducted on a 2,175-cow dairy operation in Colorado from May 2010 to April 2011. Each cow received an orally administered temperature sensing reticular bolus after parturition and RT measurements were recorded 3 times per day as lactating cows exited the milking parlor. A cow was identified as having an increased RT when a deviation of 0.8°C above baseline (average of readings of previous 10d) was recorded by the TempTrack software (DVM Systems, LLC, Greeley, CO). During the same study period, dairy personnel without access to RT data recorded health events and classified them according to clinical signs observed. A total of 201 health events (cases) were included in the data analysis. Cows with clinical mastitis and pneumonia had significantly higher odds (6.7 and 7.5 times higher, respectively) of having an increased RT of 0.8°C above their baseline within 4d preceding diagnosis when compared with control cows. Specificity and sensitivity for an increase of 0.8°C above baseline RT within 4d of disease diagnosis was 76.85 and 66.97% for mastitis, and 69.23 and 76.92% for pneumonia, respectively. No significant difference in RT was found for cows diagnosed with lameness or metritis. Results of this study suggest that RT monitoring can be a useful tool in the early detection of mastitis and pneumonia in dairy cows.

Equine bone marrow-derived mesenchymal stromal cells (BMDMSCs) from the ilium and sternum: are there differences?

REASONS FOR PERFORMING STUDY: The 2 sites of bone marrow harvest for isolation of mesenchymal stromal cells (MSC) in the horse are the sternum and ilium. The technical procedure is based on practitioner preference, but no studies have compared MSC concentrations and growth rates between the sites in horses aged 2-5 years. OBJECTIVES: The objective of this study was to compare nucleated cell counts and growth rates between the sternum and ilium and between consecutive 5 ml bone marrow aspirates. We hypothesised that there would be a higher concentration of MSCs in the sternum than the ilium, and that the first sequential aspirate from either site would yield the greatest concentration of MSCs. We hypothesised that growth rates of cells from each site would not differ. METHODS: Seven horses, aged 2 to 5 years, had 2 sequential 5 ml marrow aspirates taken from the sternum and ilium. Nucleated cell counts (NCCs) were obtained before and after marrow processing. Cells were expanded in culture for 3 passages and growth rate characteristics compared for all aspirates. RESULTS: The NCCs of the first 5 ml aspirate were higher than those of the second 5 ml aspirate for both sites (P<0.05). There was no difference between growth rates for any of the groups (P>0.05). CONCLUSIONS: The NCCs and growth rates of progenitor cells in the ilium and sternum are similar for horses in the 2-5 year age category. The first 5 ml bone marrow aspirate has a higher concentration of NCCs and resulting bone marrow-derived MSC population than subsequent aspirates. POTENTIAL RELEVANCE: The first 5 ml aspirates from the sternum and ilium offer a rich supply of bone marrow-derived MSCs with similar growth rate characteristics. The harvesting procedure of only a 5 ml draw from either the sternum or ilium should result in adequate numbers of MSCs.

Risk of bovine tuberculosis for cattle sold out from herds during 2005 in Ireland.

A retrospective cohort study was conducted to determine the risk of bovine tuberculosis (TB) among animals sold out from herds that were free to trade animals during the year 2005 according to their bovine TB testing history during the year 2005. The present study sample comprised of 338,960 animals, of which 124,360 animals were sold out from herds that were restricted from trading at some stage during 2005 (bovine TB 'exposed') and 214,600 animals that were sold from herds which did not have their trading status withdrawn in 2005 (bovine TB 'non-exposed'). The overall risk of a diagnosis of bovine TB during the two-year period after the animals were sold out was 0.69 per cent. The odds of bovine TB were 1.91 higher for animals sold out from bovine TB 'exposed' herds compared with animals sold out from bovine TB 'non-exposed' herds (OR 95 per cent CI: 1.76 to 2.07, P<0.0001). Ten per cent of animals identified during field surveillance with bovine TB did so less than two months after being sold out in 2005, and similarly, 10 per cent of the animals classified as bovine TB positive by finding a bovine TB lesion at slaughter did so within 25 days (or less) of being sold out in 2005.

Intestinal parasites of dogs on the Galapagos Islands.

Dogs on the Galapagos Islands are a unique population created by isolation from the mainland and regulations prohibiting further importation. The effect of infectious agents of these domestic dogs on the indigenous fauna is largely unknown. The purpose of this study was to determine the prevalence of intestinal parasites in dogs on the Galapagos Islands. Fecal samples were collected from 97 dogs presented during neutering campaigns on Santa Cruz (n=51), San Cristobal (n=17), and Isabela (n=29) islands. Feces were evaluated for parasites by microscopic examination after zinc sulfate centrifugation flotation as well as by a commercially available IFA for Cryptosporidium spp. and Giardia spp. Polymerase chain reaction for Cryptosporidium spp. DNA and Giardia spp. DNA was performed on all positive samples to provide the infecting genotypes. Ancylostoma caninum (57.7%) and Toxocara canis (16.5%) were most commonly detected, followed by Giardia spp. (5.2%), Isospora canis (4.1%), Sarcocystis canis (3.1%), and Cryptosporidium spp. (1%). Adequate DNA for sequencing was available for one Giardia spp. which was shown to be assemblage D. Despite being isolated, the dogs on the Galapagos have many of the same enteric parasites detected on the mainland of South America. These dogs are not routinely administered anthelmintics or other drugs, but are often allowed to roam the streets and live in close proximity to humans. Parasite prophylaxis is necessary to decrease the parasite burden within the population and to lessen the risk of spread to humans or other animals also inhabiting the islands.

Targeted badger removal and the subsequent risk of bovine tuberculosis in cattle herds in county Laois, Ireland.

We investigated the impact of targeted removal of badgers on the subsequent bovine tuberculosis (BTB) risk in cattle herds in county Laois, Ireland. The study period was 1989-2005. For each of 122 targeted badger-removal licenses (permit to remove badgers in the proximity of cattle herds undergoing a serious BTB episode), the herd number (index herd) for which the license was given was obtained. The herds in the proximity of the index herd were identified from another database. The main "exposure" in our study was the geographical location of herds relative to the area in which targeted badger removal was conducted. We categorized herds into five different exposure groups: herds were classified as non-exposed and denoted as group 0 (reference group) if they were located 500 m or more from the edge of any parcel of land of the index herd; group 1, was the index herds, group 2 the immediate (contiguous) neighbors of the index herd, group 3 herds were not immediate neighbors but within 150 m and group 4 herds were between 150 m and 500 m distance from the edge of any parcel of land of the index herd, respectively. We conducted a survival analysis (allowing multiple failures per herd) to compare the hazard of having a BTB episode in any of the four groups of exposed herds vs. the hazard in herds in the reference group. We controlled for other known risk factors as well taking into account a temporal component. Our analysis showed that the hazard ratio for the index herds (group 1) were non-significantly increased, indicating that there was no difference in the hazard of failing a BTB test (after the targeted badger removal was conducted) between index herds and reference herds. For the rest of the herds farther away from badger removal activities the hazards were lower than herds in areas not under badger removal. The hazard in the reference group decreased over the study period.

Risk factors for disclosure of additional tuberculous cattle in attested-clear herds that had one animal with a confirmed lesion of tuberculosis at slaughter during 2003 in Ireland.

All the Irish cattle herds considered "clear" of bovine tuberculosis (BTB) having a single animal with a tuberculous lesion at slaughter during 2003 were identified. We performed a descriptive and logistic regression analysis to investigate whether selected risk factors had an association with the result of the herd test immediately after the tuberculous lesion was found ("Factory Lesion Test", FLT). At the FLT, only 19.7% (n=338) of these 1713 herds had 1 or more standard reactors. The lesioned animal was home-bred in 46% of the "source" herds; these herds had an increased risk (23.4%) of having at least 1 standard reactor animal relative to herds with a purchased-lesioned animal (16.6%) (RR=1.41). Our logistic models identified a number of important risk factors; two that appeared most important in predicting the FLT outcome were the time spent (residency) by the lesioned animal in the "source" herd, and the presence, or not, of the lesioned animal in a previous BTB episode in either the "source" herd, or the seller's herd in the case the lesioned animal was purchased. Our models fit the data well based on the Hosmer-Lemeshow test, however their sensitivity and specificity were very low (57% and 61% respectively). Surveillance of the cattle population for BTB using lesions found at slaughter is an essential component of an overall control program. Nonetheless, due to the poor predictability of the variables we measured, complete herd investigations are needed to help explain the FLT outcome of a herd.

Quantifying badger exposure and the risk of bovine tuberculosis for cattle herds in county Kilkenny, Ireland.

The objectives of the study were to quantify the levels of badger exposure for cattle and to test the hypothesis that increased badger exposure does not increase the risk of bovine tuberculosis (BTB) in a herd. Information that became available from the targeted removal of badgers over the study period, and from a badger-removal project in county Kilkenny, during 1996-1999 was used. The specific location of cattle within each farm, and the length of time that cattle spent in each farm field during the grazing season, and in the barnyard during winter, was used to build an exposure coefficient to quantify the amount of badger exposure that cattle encountered either on pasture or in the barn. The study design was a matched case-control study in which the control herds were selected using incidence density sampling. During the 4-year study period, 543 badgers were removed and of these 96 badgers were classified as tuberculosis positive; 96 BTB herd breakdowns occurred. There was a significant association between case herds and having a higher badger sett exposure coefficient during 1996-1998. No significant association between case herds and having a higher exposure coefficient based on the number of badgers, or the number of tuberculous badgers, during September 1997-December 1999 was found.

Spatial relationship between Mycobacterium bovis strains in cattle and badgers in four areas in Ireland.

We investigated whether strains (restriction fragment length polymorphism, RFLP-types) of Mycobacterium bovis isolated from badgers and from cattle clustered among and within four areas in Ireland. The spatial scan test and nearest-neighbor analysis were used as the spatial cluster-detection techniques. In addition, for each of the major strains, associations between the distance to badger setts and the "centroid" of the cattle farm were assessed in a logistic model. Overall, between September 1997 and May 2000, 316 and 287 M. bovis samples, from badgers and cattle, respectively, were strain-typed. The distribution of strains in badgers, and separately in cattle, differed among areas. Within each of the four large areas, badgers and cattle tended to have similar strains; this is consistent with the sharing of M. bovis strains within an area. In more detailed within-area analyses, some spatial clusters of M. bovis strains were detected, separately, in both cattle and badgers. Almost half of the infected badger setts with a specific strain were located outside of the "detected" clusters. There was no association between the number of infected badgers with a specific M. bovis strain within 2 or 5 km distances to cattle herds, and the risk of the same strain in cattle. We speculate about the dynamic nature of badger movements, as an explanation for the absence of more clusters of most of the strains of M. bovis isolated from badgers, and its impact on trying to study transmission of M. bovis between cattle and badger.

Breakdown severity during a bovine tuberculosis episode as a predictor of future herd breakdowns in Ireland.

A retrospective cohort study of Irish cattle herds investigated whether the severity of a herd's bovine tuberculosis (BTB) breakdown was a predictor of the hazard of a future BTB breakdown in that herd. Data on 10,926 herds not having had BTB in 1995 (the "non-exposed" group) were obtained using a 10% random sample from all herds without BTB in 1995. Data on 6757 herds that had a new BTB breakdown in 1995 (the "exposed" group) were obtained and categorized into five increasing exposure-severity classes based on the total number of standard reactors (to the single intra-dermal comparative cervical tuberculin test) detected during the breakdown. Exposed herds were deemed to be free of BTB after they passed a 6-month check test; non-exposed herds were deemed free as of the date of the first negative herd-test in 1995. In the 5 years after 1995, 18% of the non-exposed herds had a BTB breakdown, whereas 31% of the exposed herds had a subsequent breakdown. Relative to the hazard for non-exposed herds, the hazard for the first future singleton standard reactor breakdown, was 1.6-times higher for exposed herds with only 1 standard reactor in 1995, and 1.8-times higher for those exposed herds with 4-8 standard reactors during the 1995 episode. When the outcome for future breakdowns was 2 or more standard reactors, the hazard ratios ranged from 1.6 for exposed herds with only 1 standard reactor in 1995 up to 2.9 in exposed herds with 8 or more standard reactors during the 1995 episode. The latter hazard ratio varied over time, decreasing to 1.7 after 3 years of risk. The hazard of a future BTB breakdown increased directly with number of cattle in the herd, a positive history of previous BTB in the herd, and the local herd prevalence of BTB. The presence of confirmed BTB lesions in reactor cattle was not predictive of the future breakdown hazard when the effects of other factors were controlled.

Bovine tuberculosis in badgers in four areas in Ireland: does tuberculosis cluster?

We described the distribution of badger populations in four different areas in the Republic of Ireland. The data came from periodic targeted badger-removal and subsequent post-mortem examinations conducted between 1989 and September 1997, and from a formal badger-removal project in the same areas from 1997 through 1999. Records were complete for 2292 badgers regarding the date of capture, tuberculosis status, geographical area and specific sett from where the badgers were snared. Of 3187 setts, 2290 had no badgers recorded against them (i.e. were inactive). The badger-level prevalence of tuberculosis differed among areas (range 13-29%). Badger populations were highly clustered by sett, and this result was similar over the four study areas. The median number of badgers per active sett was 2. Tuberculous badgers also clustered within a sett. The third quartile of tuberculous badgers was 1 per active sett. The prevalence of tuberculous badgers within a sett was not related to the total number of badgers. There was little evidence of spatial clustering with only one local cluster of tuberculous setts in each of three areas, and none in the fourth area. After adjusting for the number of badgers per sett, only one area had spatial clusters identified.