The effect of the tuberculin test and the consequences of a delay in blood culture on the sensitivity of a gamma-interferon assay for the detection of Mycobacterium bovis infection in cattle

The strategic use of the gamma-interferon (IFN-gamma) assay (Bovigam) can provide a means for the early identification of Mycobacterium bovis infected cattle, thus ensuring their removal from an infected herd. It has been reported that performance of the test can be influenced by various factors including a recent tuberculin skin test and the length of delay between collection and processing of blood samples. In this study, single intradermal comparative tuberculin test (SICTT) reactor and non-reactor cattle were recruited from herds infected with M. bovis and grouped according to their SICTT responses. Group 1 comprised reactor cattle selected on the basis of their SICTT response to PPD-bovine (purified protein derivative of tuberculin) exceeding that of PPD-avian by at least 12mm. Group 2 animals were selected from herds undergoing routine surveillance for bovine tuberculosis and contained standard SICTT reactor cattle (PPD-bovine exceeding that of PPD-avian by at least 4mm) and non-reactors. We investigated the effects of the SICTT on the assay results by measuring the in vitro IFN-gamma responses of Group 1 reactor cattle at time intervals pre- and post-skin test. No significant differences were measured in the IFN-gamma responses of the reactor animals to PPD-bovine and PPD-avian for up to 65 days. To investigate if a delay in processing of blood affected the performance of the assay, we compared results using duplicate blood samples from Group 1 and Group 2 cattle stimulated with PPD antigen at 8h and at 24h after collection. In both groups of animals the mean optical density (OD) values of the assay at 24h post-collection were significantly lower than those at 8h. Our results demonstrated that a delay in processing of the blood samples from cattle subjected to routine surveillance could significantly impact on the outcome of the IFN-gamma assay resulting in a change of the IFN-gamma status of the animals.     

Female caudal reproductive tract abnormalities

The report by McCarthy and coworkers (McCarthy et al. 2021) in this issue describes a case of vaginal septum with pyometra in a 10-week-old Westphalian filly foal. This is an interesting case report which evokes thought about the developmental abnormalities of the caudal female reproductive and urogenital tracts and their pathophysiology. Such anomalies are rare, reflected by the limited number of cases reported in the literature involving mares/fillies with uterine, cervical, vaginal and hymenal abnormalities.     

Immunological responses following experimental endobronchial infection of badgers (Meles meles) with different doses of Mycobacterium bovis

The Eurasian badger (Meles meles) is a wildlife reservoir for Mycobacterium bovis infection in Ireland and Great Britain and has been implicated in the transmission of tuberculosis to cattle. Vaccination of badgers is an option that could be used as part of a strategy to control the disease. In this study we used an endobronchial infection procedure to inoculate groups of badgers with three different doses (3x10(3), 2x10(2) and <10 Colony Forming Units (CFUs)) of M. bovis. After 17 weeks the disease status of each animal was determined by post-mortem pathology and culture for M. bovis. Each of the inoculum doses resulted in establishment of infection in the badgers. The cell-mediated immune (CMI) responses were measured by lymphocyte transformation assay (LTA) of peripheral blood mononuclear cells (PBMCs) cultured with bovine tuberculin (PPD-B). In each infected group the CMI responses increased with a kinetic profile corresponding to the delivered dose and the post-mortem pathology. The serological responses were measured by ELISA and a multi-antigen print immunoassay (MAPIA) in order to investigate any changes in the antigenic repertoire associated with different infective doses. In contrast to the CMI responses, the ELISA and MAPIA showed that the recognition of antigens by the badgers was intermittent and not strongly influenced by the dose of M. bovis.     

Bronchoalveolar lavage cytology from captive badgers

Background: Bronchoalveolar lavage (BAL) fluid is evaluated for the diagnosis and study of lung disease and airway inflammation. Cytologic profiles for BAL fluid have not been reported for badgers and may be useful in understanding the pathogenesis of pulmonary diseases such as Mycobacterium bovis. Objective: The aim of this study was to evaluate cytologic and microbial findings in BAL fluid from captive European badgers (Meles meles) and identify correlates with the results of concurrently collected blood and fecal samples. Methods: BAL fluid (by a nonbronchoscopic method) and jugular venous blood samples (for routine CBC) were obtained from 23 captive tuberculosis‐free anesthetized badgers on 2 occasions 4 weeks apart. Fecal samples were collected for routine parasitology. Morphologic evaluation and 100‐cell differentials were done on cytocentrifuged BAL specimens. Pellets from centrifuged BAL were aerobically cultured for bacteria. Results: With the 2 BAL samples from each of the 23 badgers combined, the median (range) cell percentages were 73.0% (5–95%) neutrophils, 7.5% (2–16%) macrophages, 8.0% (0–27%) lymphocytes, and 9.5% (0–92%) eosinophils. Macrophages frequently contained silica‐like crystals. Other findings included ciliated epithelial cells, goblet cells, mucus, and Aelurostrongylus sp. larvae. A light growth of Streptococcus, Pasteurella, or Escherichia coli was cultured in 6 badgers. Trypanosoma pestanai were identified in blood from 10 badgers and fecal parasites (mainly coccidia) were found in 20 badgers. No correlation was found between BAL and CBC results and the presence of parasites. Conclusions: The predominance of neutrophils in BAL fluid from badgers differs from the predominance of macrophages found in BAL from other species. This difference may reflect the burrowing lifestyle or the unique immune response of badgers.     

Tuberculosis in cattle herds are sentinels for Mycobacterium bovis infection in European badgers (Meles meles): the Irish Greenfield Study

In Ireland badgers are removed in response to tuberculosis (TB) breakdowns in cattle herds (focal culling). Prevalence studies, conducted using a detailed post mortem and bacteriological examination, showed that 36-50% of badgers were infected with Mycobacterium bovis. Focal culling forms part of the medium term national strategy for the control of bovine TB in cattle and is based on the premise that badgers in areas with herd breakdowns have a higher prevalence of infection than the badger population at large. However, the hypothesis that cattle can be used as sentinels for infection in the badger population has never been formally tested. In this study we tested the hypothesis by determining the infection prevalence in badgers in areas where there had been historically, a consistently low prevalence of infection in cattle. Low cattle TB prevalence areas were defined as those herds with ≤ 2 standard reactors in the annual round of skin testing over the preceding 5 years (Greenfield sites). Using GIS, and adjusting for variation in land use, previous culling and cattle density, 198 Greenfield sites were identified and surveyed, and 138 areas with badger setts or signs of badger activity were identified. A single badger was removed from 87 sites and all were examined using detailed post mortem and bacteriological procedures. A prevalence of M. bovis infection of 14.9% was found in the Greenfield site badgers. This prevalence was significantly lower (P<0.001) than in badgers removed during focal culling (36.6%). The results validate the use of cattle as sentinels for TB in badgers and support the medium term national strategy for the control of bovine TB. The geographic variation in M. bovis infection prevalence in the Irish badger populations will be used when devising strategies for the incorporation of badger vaccination into the long term bovine TB control programme.     

Trial design to estimate the effect of vaccination on tuberculosis incidence in badgers

The principal wildlife reservoir of Mycobacterium bovis in Ireland is the European badger. Studies in the Republic of Ireland (RoI) have shown that badgers culled in association with cattle herd tuberculosis breakdowns (focal culling) have a higher prevalence of infection than the badger population at large. This observation is one rationale for the medium term national strategy of focal badger culling. A vaccination strategy for the control of bovine tuberculosis (bTB) in badgers is a preferred long-term option. The Bacillus Calmette-Guérin (BCG) vaccine has been shown to decrease disease severity in captive badgers under controlled conditions. As the vaccine has been tested in a controlled environment with precise information on infection pressure, it cannot be assumed a priori that the effects of vaccination are similar in the wild, where other environmental and/or ecological factors prevail. For this reason we have designed a vaccine field trial to assess the impact of vaccination on the incidence of TB infection in a wild badger population. The selected study area for the vaccine trial (approximately 755 square kilometers) is divided into three zones each of which has similar characteristics in terms of size, number of main badger setts, cattle herds, cattle and land classification type. Three vaccination levels (100%, 50% and 0%) will be allocated to the three zones in a way that a gradient of vaccination coverage North to South is achieved. The middle zone (zone B) will be vaccinated at a 50% coverage but zone A and C will be randomly allocated with 100% or 0% vaccination coverage. Vaccination within zone B will be done randomly at individual badger level. The objective of this paper is to describe the design of a field tuberculosis vaccination trial for badgers, the epidemiological methods that were used to design the trial and the subsequent data analysis. The analysis will enable us to quantify the magnitude of the observed vaccination effect on M. bovis transmission in badgers under field conditions and to improve our knowledge of the biological effects of vaccination on susceptibility and infectiousness.     

Modelling daily catches of silver-phase European eel (Anguilla anguilla) in two hydropower-regulated rivers

Estimation of silver eel production, Anguilla anguilla L., is fundamental for the management of eel stocks. In the hydropower-regulated rivers Shannon and Erne, Ireland, production is calculated using catch data from a conservation trap and transport programme. However, in both rivers gaps in silver eel catch datasets tend to occur, which can lead to biases in production estimates. Generalised additive models (GAMs) were used to model daily catch in these rivers based on a variety of environmental variables. Final models for the Shannon and Erne explained 83.7% and 78.8% of deviance in daily catch, respectively. A second model on the Erne included catch from a nearby fishing site in an attempt to increase explanatory power and explained 91.7% of deviance. Although model accuracy was increased, reliance on catch from another site may limit the applicability of the model. Model predictions were combined with estimates of fishing efficiency to predict production for the Shannon (36,210 kg; 0.85 kg/ha) and Erne (66,899–67,047 kg; 2.55–2.56 kg/ha). These values represented a 9.3% and 2.8%–3.0% increase on production estimated from incomplete catch records alone for the Shannon and Erne, respectively.