Animal-side serologic assay for rapid detection of Mycobacterium bovis infection in multiple species of free-ranging wildlife

Numerous species of mammals are susceptible to Mycobacterium bovis, the causative agent of bovine tuberculosis (TB). Several wildlife hosts have emerged as reservoirs of M. bovis infection for domestic livestock in different countries. In the present study, blood samples were collected from Eurasian badgers (n=1532), white-tailed deer (n=463), brushtail possums (n=129), and wild boar (n=177) for evaluation of antibody responses to M. bovis infection by a lateral-flow rapid test (RT) and multiantigen print immunoassay (MAPIA). Magnitude of the antibody responses and antigen recognition patterns varied among the animals as determined by MAPIA; however, MPB83 was the most commonly recognized antigen for each host studied. Other seroreactive antigens included ESAT-6, CFP10, and MPB70. The agreement of the RT with culture results varied from 74% for possums to 81% for badgers to 90% for wild boar to 97% for white-tailed deer. Small numbers of wild boar and deer exposed to M. avium infection or paratuberculosis, respectively, did not cross-react in the RT, supporting the high specificity of the assay. In deer, whole blood samples reacted similarly to corresponding serum specimens (97% concordance), demonstrating the potential for field application. As previously demonstrated for badgers and deer, antibody responses to M. bovis infection in wild boar were positively associated with advanced disease. Together, these findings suggest that a rapid TB assay such as the RT may provide a useful screening tool for certain wildlife species that may be implicated in the maintenance and transmission of M. bovis infection to domestic livestock.The authors are grateful to Peter Andersen and Jim McNair for kindly providing certain antigens used in C this study. Badger samples were taken under projects funded by the Department for Environment, Food, and Rural Affairs (Defra), UK. The authors acknowledge the support of staff from CSL, VLA Starcross, Defra Wildlife Unit, and permission from the Independent Scientific Group for use of sera from the RBCT. Spanish wild boar samples were obtained with support from MEC Plan Nacional AGL2005-07401 and Santander - Fundacion M. Botin

Investigation on the role of red fox in tuberculosis maintenance community ¿ second opus: experimental infection with a virulent field Mycobacterium bovis strain

Trabajo presentado al: 69th Wildlife Disease Association and 14th European Wildlife Disease Association Conference. Cuenca, Spain. p. 135. 31 agosto-2 septiembre

АНАЛИЗИРУЮЩИЙ МАГНИТ ДЛЯ ТЕХНОЛОГИЧЕСКОГО УСКОРИТЕЛЯ ЭЛЕКТРОНОВ ЛУ-10М

Основной проблемой при использовании магнитов из редкоземельных материалов в ускорительных технологиях является изменение магнитных свойств материала под действием излучения. Особенно актуальна эта задача для мощных технологических ускорителей электронов с энергией до 10 МэВ. Для выбора наиболее стойкого материала были проведены исследования радиационной стойкости образцов магнитов из Sm2Co17 и Nd-Fe-B сплава. Постоянные магниты типа Sm2Co17 и Nd-Fe-B были изгтовлены методом порошковой технологи с использованием PLP процесса в производстве последних. Магнитные образцы подвергались прямому воздействию электронного пучка с энергией 10 МэВ и воздействию тормозного излучения этого пучка. Поглощённая доза от электронов для образцов составляла 16 Град (общий поток электронов, попавший на 1 см2 образца, был равен 1,4х1017) и 160 Град. Активность образцов после облучения изменилась в пределах допустимых норм. Этот факт существенно упрощает использование редкоземельных магнитных материалов в готовых изделиях ускорителей. В процессе облучения магниты охлаждались водой с температурой 38 °С для избежания перегрева. Для оценки изменения величины поля, создаваемого вокруг образца, использовался интеграл интерполированной нормальной к поверхности образца составляющей магнитного поля S по области интерполяции данных в относительных единицах. Для образцов из Nd-Fe-B сплава магнитный поток вокруг образца уменьшился и составил 0,92 и 0,717 от начального значения для указанных доз облучения. Магнитное поле вокруг образцов из Sm2Co17 сплава не изменилось в пределах точности измерений для тех же доз. На основе образцов из Sm2Co17 сплава размерами 30х24х12 мм было проведено моделирование и конструирование магнита для анализа пучка электронов технологического ускорителя на энергию до 10 МэВ. Наибольшее значение поля в медианной плоскости магнита равно 0,3110 Тл. Расстояние между полюсами магнита равно 25,25 мм. Эффективная длина магнита – 33,53 мм. Измеренные параметры поля магнита удовлетворяют заданным при разработке величинам. Магнит также может быть использован для настройки ускорителя в диапазоне энергий до 10 МэВ

ЭКСПЕРИМЕНТАЛЬНОЕ ОБОСНОВАНИЕ МИКРОХИРУРГИЧЕСКОГО ПИЩЕВОДНО-ТОНКОКИШЕЧНОГО АНАСТОМОЗА

In the experimental study on 6 mongrel dogs the comparative morphometry of layers of the wall of abdominal section of esophagus and small intestine was carried out. Tworowed microsurgical esophago-enteroanastomosis “end-to-end” was formed in 12 animals. From the surplus of the outer case of the esophagus a sphincter-like structure was created. The dynamics of anastomosis adhesion in the postoperative period was studied macroscopically and on longitudinal histotopograms. A good epithelization has been recorded by the 7th day as well as a formation of thin and firm cicatrix without a rough fibrillar tissue by the 21st day.В экспериментальном исследовании на 6 беспородных собаках выполнена морфометрия слоёв стенок абдоминального отдела пищевода и тонкой кишки. У 12 животных сформирован двухрядный микрохирургический эзофагоэнтероанастомоз конец в конец. Из избытка наружного футляра пищевода сформирована сфинктероподобная структура. Изучена динамика заживления анастомоза в послеоперационном периоде. Показана хорошая эпителизация к 7 суткам и формирование тонкого прочного рубца на 21 сутки после операции.

Bovine Tuberculosis in a Nebraska Herd of Farmed Elk and Fallow Deer: A Failure of the Tuberculin Skin Test and Opportunities for Serodiagnosis

In 2009, Mycobacterium bovis infection was detected in a herd of 60 elk (Cervus elaphus) and 50 fallow deer (Dama dama) in Nebraska, USA. Upon depopulation of the herd, the prevalence of bovine tuberculosis (TB) was estimated at ∼71–75%, based upon histopathology and culture results. Particularly with elk, gross lesions were often severe and extensive. One year ago, the majority of the elk had been tested for TB by single cervical test (SCT), and all were negative. After initial detection of a tuberculous elk in this herd, 42 of the 59 elk were tested by SCT. Of the 42 SCT-tested elk, 28 were TB-infected with only 3/28 reacting upon SCT. After SCT, serum samples were collected from the infected elk and fallow deer from this herd at necropsy and tested by three antibody detection methods including multiantigen print immunoassay, cervidTB STAT-PAK, and dual path platform VetTB (DPP). Serologic test sensitivity ranged from 79 to 97% depending on the test format and host species. Together, these findings demonstrate the opportunities for use of serodiagnosis in the rapid detection of TB in elk and fallow deer

Potential for rapid antibody detection to identify tuberculous cattle with non-reactive tuberculin skin test results

Abstract Background Bovine tuberculosis (TB) control programs generally rely on the tuberculin skin test (TST) for ante-mortem detection of Mycobacterium bovis-infected cattle. Results Present findings demonstrate that a rapid antibody test based on Dual-Path Platform (DPP®) technology, when applied 1-3 weeks after TST, detected 9 of 11 and 34 of 52 TST non-reactive yet M. bovis-infected cattle from the US and GB, respectively. The specificity of the assay ranged from 98.9% (n = 92, US) to 96.0% (n = 50, GB) with samples from TB-free herds. Multi-antigen print immunoassay (MAPIA) revealed the presence of antibodies to multiple antigens of M. bovis in sera from TST non-reactors diagnosed with TB. Conclusions Thus, use of serologic assays in series with TST can identify a significant number of TST non-reactive tuberculous cattle for more efficient removal from TB-affected herds

Bovine Tuberculosis at the Wildlife-Livestock-Human Interface in Hamer Woreda, South Omo, Southern Ethiopia

Bovine tuberculosis (BTB) is endemic in cattle in the Ethiopian Highlands but no studies have been done so far in pastoralists in South Omo. This study assessed the prevalence of bovine tuberculosis (BTB) at an intensive interface of livestock, wildlife and pastoralists in Hamer Woreda (South Omo), Ethiopia. A cross-sectional survey including a comparative intradermal skin testing (CIDT) was conducted in 499 zebu cattle and 186 goats in 12 settlements. Sputum samples from 26 symptomatic livestock owners were cultured for TB. Fifty-one wildlife samples from 13 different species were also collected in the same area and tested with serological (lateral flow assay) and bacteriological (culture of lymph nodes) techniques. Individual BTB prevalence in cattle was 0.8% (CI: 0.3%–2%) with the >4 mm cut-off and 3.4% (CI: 2.1%–5.4%) with the >2 mm cut-off. Herd prevalence was 33.3% and 83% when using the >4 and the >2 mm cut-off respectively. There was no correlation between age, sex, body condition and positive reactors upon univariate analysis. None of the goats were reactors for BTB. Acid fast bacilli (AFB) were detected in 50% of the wildlife cultures, 79.2% of which were identified as Mycobacterium terrae complex. No M. bovis was detected. Twenty-seven percent of tested wildlife were sero-positive. Four sputum cultures (15.4%) yielded AFB positive colonies among which one was M. tuberculosis and 3 non-tuberculous mycobacteria (NTM). The prevalence of M. avium-complex (MAC) was 4.2% in wildlife, 2.5% in cattle and 0.5% in goats. In conclusion, individual BTB prevalence was low, but herd prevalence high in cattle and BTB was not detected in goats, wildlife and humans despite an intensive contact interface. On the contrary, NTMs were highly prevalent and some Mycobacterium spp were more prevalent in specific species. The role of NTMs in livestock and co-infection with BTB need further research

Diagnosis of Tuberculosis in the Wild Boar (Sus scrofa): A Comparison of Methods Applicable to Hunter-Harvested Animals

To obtain robust epidemiological information regarding tuberculosis (TB) in wildlife species, appropriate diagnostic methods need to be used. Wild boar (Sus scrofa) recently emerged as a major maintenance host for TB in some European countries. Nevertheless, no data is available to evaluate TB post-mortem diagnostic methods in hunter-harvested wild boar. METHODOLOGY/PRINCIPAL FINDINGS: Six different diagnostic methods for TB were evaluated in parallel in 167 hunter-harvested wild boar. Compared to bacteriological culture, estimates of sensitivity of histopathology was 77.8%, gross pathology 72.2%, PCR for the MPB70 gene 66.7%, detection of acid-fast bacilli (AFB) in tissue contact smears 55.6% and in histopathology slides 16.7% (estimated specificity was 96.7%, 100%, 100%, 94.4% and 100%, respectively). Combining gross pathology with stained smears in parallel increased estimated sensitivity to 94.4% (94.4% specificity). Four probable bacteriological culture false-negative animals were identified by Discriminant Function Analysis. Recalculating the parameters considering these animals as infected generated estimated values for sensitivity of bacteriology and histopathology of 81.8%, gross pathology 72.7%, PCR for the MPB70 gene 63.6%, detection of AFB in tissue contact smears 54.5% and in histopathology slides 13.6% (estimated specificity was 100% for gross pathology, PCR, bacteriology and detection of AFB in histopathology slides, 96.7% for histopathology and 94.4% for stained smears). CONCLUSIONS/SIGNIFICANCE: These results show that surveys for TB in wild boar based exclusively on gross pathology considerably underestimate prevalence, while combination of tests in parallel much improves sensitivity and negative predictive values. This finding should thus be considered when planning future surveys and game meat inspection schemes. Although bacteriological culture is the reference test for TB diagnosis, it can generate false-negative results and this should be considered when interpreting data.This study was funded by laboratory funds from Life and Health Sciences Research Institute (ICVS), School of Health Sciences, University of Minho. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript