牛副结核病防控

牛副结核病也称牛副结核性肠炎,由副结核分枝杆菌引起的以持续顽固性腹泻、渐进性消瘦为特征的慢性人兽共患传染病。该病呈全球性分布,在世界各国均有流行,牛场一旦发病很难根除,给畜牧业造成了巨大危害。对该病进行深入了解并进行有效防控,对我国畜牧业持续健康发展和公共卫生安全都有非常重要的意义。本文从该病的病原学、流行病学、危害、诊断、防控等方面进行了综述,基于此提出在未来的防控实践中,牛场要切实做好相关防控工作,政府要加强防控规划,从上至下形成科学有效的综合性防控体系,以减少该病的发生并严格控制该病的传播,从而将该病对畜牧业及公共卫生的危害降至最低。     

牛副结核病的诊断与防控

副结核病是由禽分枝杆菌副结核亚种引起的慢性疾病,会感染多种反刍动物.副结核病的流行病学十分复杂,该病在牛中的临床表现和经济影响可能因畜群管理、年龄、感染菌量和疾病流行情况等因素而异.此外,在控制牛副结核病方面还面临着相当大的挑战,如缺乏准确可靠的诊断检测方法.尽管如此,全球仍致力于控制牛副结核病,因为该病对养牛羊可造成...     

牛副结核病及其综合防控

牛副结核病是由副结核分枝杆菌引起的以牛产奶量下降、渐进性消瘦、慢性卡他性肠炎、持续性腹泻为特征的人兽共患传染病,给畜牧养殖业造成了巨大的经济损失,并且对公共卫生安全也造成了潜在的威胁,目前还没有特效的治疗药物。为提高养牛行业的经济效益,更好地满足人们对肉类、奶类食品的需求,促进牛养殖业的全面发展,现就该病的诊断及其防控措施作一阐述。     

肉牛副结核病的诊断及防控方法

肉牛副结核病由副结核分枝杆菌引起的牛的一种慢性消化道传染病,具有潜伏期长、发病初期不明显、治愈可能性低、传染性强等特点。在肉牛养殖过程中,养殖人员要明确该病的流行病学特点、临床症状,以及实验室诊断方法等,以便有效应对该病的发生。本文对该病进行了比较详细的分析,给出了多种实验室诊断方法和防控该病的具体措施,供参考。     

牛结核病及其防控措施

牛结核病是常见的慢性疾病,是由牛型结核分枝杆菌引起的人畜共患病,可导致病牛的组织器官出现结核结节性肉芽肿和干酪样、钙化的坏死病灶,严重制约我国养牛业的发展.本文主要就牛结核病的危害、流行特点、诊断方法和防控措施进行简述,为防控该病提供一定的参考,降低发病率.     

牛副结核病的科学防治措施

牛副结核病由副结核分枝杆菌感染所引起,主要经消化道传播,感染后潜伏期和病程均较长,病牛以慢性顽固性腹泻为特征,同时表现机体衰弱、脱水、生产性能下降;应激对本病的发生有促进作用,病原对环境不良因素抵抗力较强,发病牛场很难彻底净化;本病应以预防为主,加强牛场的生物安全建设,做好养殖环境的全方位消毒;无饲养价值的病牛不建议治疗,可直接淘汰销毁,饲养价值较高的牛可采用化药抗菌配合中药调理的方法尝试治疗。     

牛副结核病的诊断与防控

该文简要介绍牛副结核病的病原、流行病学特点、临床表现和剖检病变,重点分析了该病的鉴别诊断、实验室诊断方法与防控措施,以供参考。     

羊副结核病的致病特点及综合防控措施

羊副结核病是由副结核分枝杆菌引起的一种以顽固性腹泻、肠粘膜显著增厚为主要临床特征的慢性传染病,多呈现隐形感染,该病潜伏期长,传播迅速,呈全球性分布,对畜牧业的危害巨大。本文立足羊副结核病研究进展的实际情况,梳理总结了目前学术界对羊副结核病病原学、流行病学、致病机制、临床诊断和免疫综合防控等方面的研究讨论,为羊副结核病的诊断与防控提供了理论基础。在此基础上提出防控羊副结核病政府需制定有效政策,养殖场需加强饲养管理,形成科学有效的防控系统,为我国畜牧业的可持续健康发展提供了理论支撑。     

牛副结核病及其预防

副结核病是一种以潜延性腹泻为特征的慢性传染病,主要发生于牛,尤其是乳牛。本病分布广泛,一般养牛区都可能存在。1发病特点早期为间断性腹泻,时轻时重,排泄物稀薄、恶臭,并带有气泡和粘液块,以后渐渐变为顽固性拉稀,病牛逐渐消瘦、衰弱、脱水、眼窝下陷、精神萎靡、喜卧;下颌及垂皮水肿,皮肤粗糙,被毛粗乱。但体温一般无变化。一些常用药物如链霉素、青霉素以及对氨水杨酸、异烟肼等对这种病均无疗效。而止泻药只能短期起作用,停药时即刻复发。病牛最后因腹泻不止经3～4个月衰竭而死,染疫牛群的死亡     

浅析副结核病对奶牛场的影响及防控措施

牛副结核病是一种人畜共患慢性感染性疾病,世界动物卫生组织（OIE）将该病列为必须通报的动物疫病,我国农业部将其列为二类动物疫病。牛副结核病同样严重威胁着我国奶牛养殖业的健康发展,并且对公共卫生安全具有潜在的威胁。近年来我国逐渐意识到牛副结核病防控工作的重要性,相关动物疫控部门也对该病的流行特点、防控措施进行了多方调查研究。     

Evaluation of serum and milk ELISAs for paratuberculosis in Danish dairy cattle

A milk and a serum ELISA for detection of antibodies against Mycobacterium avium ssp. paratuberculosis (MAP) were evaluated against the complement-fixation test (CFT) and culture of faecal samples from 580 cows collected between August 1996 and December 1996. Milk and serum were obtained concurrently from six dairy herds infected with MAP and from two dairy herds without history of infection with MAP.

A cut-off value of 7 OD% was used in the ELISAs. At this cut-off value, all six culture-positive herds were positive in the serum ELISA but one was negative in the milk ELISA. All six culture-positive herds were positive in the CFT. In the two culture-negative herds, the serum and the milk ELISA deemed all serum samples negative at this cut-off value, whereas four serum samples from one of these herds were positive in the CFT. The highest cut-off value enabling the milk ELISA to record all six culture-positive herds as positive was 4 OD%. The highest cut-off value enabling the serum ELISA to record all six culture-positive herds as positive was 17 OD%. Individual-sample relative sensitivities of the ELISAs ranged from 49 to 64% and relative specificities were 80–96% at the cut-off values of 4, 7 and 17 OD%.     

副结核分枝杆菌MAP0862、MAP1345基因的串联表达及其在间接ELISA中的初步应用

为探索副结核特异性蛋白MAP0862与MAP1345在牛副结核病血清学诊断中的作用,将通过串联表达获得的融合蛋白MAP0862-1345纯化定量后包被酶标板,经过对反应条件的优化,初步建立了基于融合蛋白MAP0862-1345的间接ELISA诊断方法。使用建立的ELISA方法对牛副结核病阳性血清、牛结核病阳性血清、牛布病阳性血清、卡介苗免疫牛血清、健康牛血清检测的结果表明其具有较好的特异性;使用该方法与IDEXX副结核病抗体检测试剂盒共同对300份临床血清样本检测,总符合率为92.7%。     

Distribution of IS900 restriction fragment length polymorphism types among animal Mycobacterium avium subsp. paratuberculosis isolates from Argentina and Europe

Sixty-one Mycobacterium avium subsp. paratuberculosis isolates from cattle and deer from the Buenos Aires province, an important livestock region in Argentina, were typed by restriction fragment length polymorphisms (RFLP) analysis based on IS900. Four different RFLP patterns (designated ‘A’, ‘B’, ‘C’ and ‘E’) were identified in BstEII digests of genomic DNA. The most frequently observed type, pattern ‘A’, was found in 46 isolates (75%). The second, pattern ‘E’, included 8 isolates (13%), while the third, pattern ‘B’, included 6 isolates (10%). Pattern ‘C’ was found for only one isolate. All of the deer isolates were classified as pattern ‘A’, while cattle isolates represented all four RFLP patterns. Twenty-one isolates representing the four different BstEII-RFLP patterns were digested with PstI. Twenty isolates showed identical PstI-RFLP pattern. BstEII-RFLP patterns from Argentine cattle and deer were compared with patterns found in cattle, goat, deer, rabbit, and human isolates from Europe. The most common pattern in Argentina, pattern ‘A’, was identical to a less frequently occurring pattern R9 (C17) from Europe. The other Argentine patterns ‘B’, ‘C’ and ‘E’, were not found in the Europe. These results indicate that the distribution of M. avium subsp. paratuberculosis genotypes in the Buenos Aires province of Argentina is different from that found in Europe.     

Sensitivity and specificity of two serological tests for the detection of ovine paratuberculosis

OBJECTIVE: To determine the sensitivity and specificity of an absorbed ELISA and an AGID test for the detection of clinical and subclinical paratuberculosis in sheep. DESIGN: By testing a panel of sera from 1257 Australian Merino and crossbred sheep greater than 1 year of age, of which 1137 sheep were not infected with Mycobacterium avium subsp paratuberculosis and 120 sheep had paratuberculosis. PROCEDURE: Sera were collected from 457 sheep in Victoria and 800 sheep in Western Australia. Presence of M a paratuberculosis infection in Victorian sheep was determined by histological examination of intestinal tissues, whereas sheep from Western Australia were presumed to be free of Johne's disease. The ability of an absorbed ELISA to discriminate between infected and uninfected sheep was described by test sensitivity and specificity, the distribution of ELISA OD, and the area under a receiver operating characteristic curve. RESULTS: The absorbed ELISA had a specificity of 98.2 to 99.5% (CI) and a sensitivity of 35 to 54% (CI). In sheep from infected flocks in Victoria, the AGID test had a specificity of 99 to 100% (CI) and a sensitivity of 38 to 56% (CI). The sensitivity of serological tests was higher in sheep with a body condition representative of the lower quintile of their flock of origin. CONCLUSION: The AGID test and absorbed ELISA are useful tests for the detection of ovine paratuberculosis. Although the tests had a similar accuracy, they detected different subpopulations of infected sheep with only moderate overlap. The AGID test had a higher specificity than the absorbed ELISA.     

Evaluation of a high-throughput nucleic acid extraction method for the detection of Mycobacterium avium subsp. paratuberculosis in bovine fecal samples by PCR

Johne’s disease (paratuberculosis) is an economically important disease of cattle worldwide. The disease is caused by Mycobacterium avium subsp. paratuberculosis (MAP), a fastidious gram-positive bacterium. PCR is increasingly used in diagnostic laboratories for the detection of MAP in fecal samples given the rapid test turnaround time and sensitivity and specificity comparable to fecal culture. However, efficient extraction of DNA for sensitive detection of MAP by PCR is affected by the complex lipid-rich cell wall of MAP and the presence of PCR inhibitors in feces. We evaluated a high-throughput nucleic acid extraction method (MagMAX core nucleic acid purification kit with mechanical lysis module) in conjunction with an hspX gene PCR for the detection of MAP from bovine fecal samples, which resulted in correct identification of all negative (13 of 13) and positive (35 of 35) proficiency test samples obtained from the National Veterinary Services Laboratories. In addition, all 6 negative and 50 of 51 positive diagnostic specimens tested were categorized correctly.     

Subclinical paratuberculosis in goats following experimental infection: An immunological and microbiological study

An experimental oral infection of goats with a caprine isolate of Mycobacterium a. subsp. paratuberculosis was used to investigate immunological and bacteriological events during the subclinical phase of infection. Seven goats at 5–8 weeks of age were given a bacterial suspension in milk-replacement three times weekly for 9 weeks. Six animals were kept as controls.

Cellular recall responses against M. a. paratuberculosis were analysed by means of a lymphocyte proliferation test, an IFN-γ assay and an IL-2 receptor assay. All inoculated animals had detectable CMI responses from 9 weeks post-inoculation and through the 2 years of study, although the responses were highest during the first year. Antibodies against M. a. paratuberculosis could be detected from weeks 15–20 in four of the seven animals, and one additional animal became antibody positive at week 35, while two inoculated animals did not produce significant antibody titres during the experiment. At about 1-year post-inoculation, two animals became faecal shedders, while two others started to excrete bacteria into faeces about 2 years post-inoculation. The appearance of M. a. paratuberculosis in faeces was not associated with a decline in cellular responses as far as could be assessed using the current methods for measuring CMI.

Pathological lesions due to M. a. paratuberculosis infection and presence of bacteria were recorded in the intestine and/or mesenteric lymph nodes of five animals while lymph node changes suggestive of paratuberculosis were observed in one animal. Only the two animals with no signs of an active infection at necropsy showed a considerable decline in the cellular parameters during the last year of the study, particularly in the IFN-γ assay.

The two animals with the highest levels of M. a. paratuberculosis responsive CD8+ lymphocytes in the circulation about 1-year post-inoculation had no detectable lesions in the distal ileum and colon at necropsy, while high numbers of γδ T-cells responsive to M. a. paratuberculosis in the circulation were associated with disseminated lesions in the distal ileum and colon.     

DNA fingerprinting of Australian isolates of Mycobacterium avium subsp paratuberculosis using IS900 RFLP

OBJECTIVES: To evaluate additional restriction enzymes for IS900 RFLP of Mycobacterium avium subsp paratuberculosis and examine the genetic diversity among Australian isolates for epidemiological studies of Johne's disease. DESIGN AND PROCEDURE: Seventy-one isolates of M paratuberculosis from cattle, sheep, goat, alpaca and rhinoceros in six Australian States and the Northern Territory, reference strains and reference DNA from previously characterised strains were tested for genetic variation. Bst EII, Pvu II and Pst I restriction enzymes were used, and four others (Bam HI, Alu I, Xho I and Dra I) were assessed for their ability to detect polymorphisms. Multiple isolates from some animals were tested. RESULTS: Bam HI, was the most effective enzyme for identifying polymorphisms (12 types), followed by Bst EII (11 types). Both Pvu II and Pst I were relatively ineffectual. Fifteen different types were identified, 12 in clinical isolates. Most isolates were cattle (C) strains and fell into the C1 (n = 28) and C3 (n = 32) groupings. All isolates from alpaca were type C1, and bovine isolates were commonly C1 (n = 15) or C3 (n = 28). All of the sheep were infected with sheep (S) strains; no S strains were identified in cattle. Two of six isolates from one animal had single band differences. CONCLUSION: The epidemiological features of M paratuberculosis in Australia are similar to those reported in New Zealand, where cattle and sheep are commonly infected with different strains. However, because of the lack of polymorphism identified within the major groups, it is unlikely that DNA fingerprinting will have a significant role in epidemiological studies of Johne's disease, unless an unusual strain in being studied.     

Efficacy of spheroplastic and cell-wall competent vaccines for Mycobacterium avium subsp. paratuberculosis in experimentally-challenged baby goats

A Mycobacterium avium subspecies paratuberculosis (MAP) vaccine that reduced the incidence of clinical disease or reduced fecal shedding of MAP would aid control of Johne's disease (JD). The objective of the present study was to evaluate the efficacy of four MAP vaccine combinations, including cell-wall competent (CWC) alum adjuvant, CWC-QS21 adjuvant, cell-wall deficient (CWD) alum adjuvant and CWD-QS21 adjuvant vaccines. Eighty baby goats were vaccinated at 1 and 4 weeks of age with one of these vaccines or a sham control vaccine consisting of alum adjuvant. Kids were challenged orally with approximately 6.0 × 10⁹ organisms in four divided doses of 1.5 × 10⁹ organisms using a goat isolate of MAP. Vaccinated challenged and challenged control groups had 10 and 6 kids per group, respectively. Half of the kids within each group were necropsied at either 6 or 9 months post-challenge. Gross and microscopic lesions and relative number of acid-fast bacilli were evaluated and scored at necropsy. Results indicated all challenged kids had some lesions compatible with JD suggesting none of the vaccines prevented infection. Three vaccines (CWC-alum, CWC-QS21 and CWD-QS21) reduced lesion scores by 46–51% at 9 months. CWD-alum vaccine resulted in a more severe (+33.5%) lesion score than sham-vaccinated challenged control. Lesion scores were greater at 9 months than at 6 months post-challenge in the sham-vaccinated challenged group and CWD-alum vaccinated group, while lesion scores were generally stable with remaining vaccines. Mean fecal CFU/g were significantly different across time from challenge to 9-month necropsy (p = 0.043) and the CWC-QS21 vaccine group had a marked reduction in fecal CFU/g at all time points post-challenge. A reduction in MAP CFU/g was also detected in necropsy tissues from kids given the CWC-alum, CWC-QS21 and CWD-QS21 vaccines, and increased CFU/g were detected in tissues from kids given the CWD-alum vaccine. Immunological tests evaluated included, humoral response evaluation by AGID, ELISA and Western blot, and cell mediated response by comparative PPD skin testing (M. avium, Old Johnin, M. bovis and Lot 2 Johnin PPD's), and production of MAP induced γ-interferon. Vaccination also resulted in false-positive PPD skin test reactions for M. avium PPD, Old Johnin PPD and γ-interferon tests. When a 2-mm cutoff above normal skin thickness was used to define positive skin test reactions, false-positive reactions for M. bovis were detected in only 2 of 32 kids given a vaccine with QS21 adjuvant.     

Trends and advances in the diagnosis and control of paratuberculosis in domestic livestock

Paratuberculosis (pTB) is a chronic granulomatous enteritis caused by Mycobacterium avium subsp. paratuberculosis (MAP) in a wide variety of domestic and wild animals. Control of pTB is difficult due to the lack of sensitive, efficacious and cost-effective diagnostics and marker vaccines. Microscopy, culture, and PCR have been used for the screening of MAP infection in animals for quite a long time. Besides, giving variable sensitivity and specificity, these tests have not been considered ideal for large-scale screening of domestic livestock. Serological tests like ELISA easily detects anti-MAP antibodies. However, it cannot differentiate between the vaccinated and infected animals. Nanotechnology-based diagnostic tests are underway to improve the sensitivity and specificity. Newer generation diagnostic tests based on recombinant MAP secretory proteins would open new paradigm for the differentiation between infected and vaccinated animals and for early detection of the infection. Due to higher seroreactivity of secretory proteins vis-à-vis cellular proteins, the secretory proteins may be used as marker vaccine, which may aid in the control of pTB infection in animals. Secretory proteins can be potentially used to develop future diagnostics, surveillance and monitoring of the disease progression in animals and the marker vaccine for the control and eradication of pTB.     

Temporal patterns and quantification of excretion of Mycobacterium avium subsp paratuberculosis in sheep with Johne's disease

OBJECTIVES: To determine the frequency of excretion of Mycobacterium avium subsp paratuberculosis in Merino sheep with Johne's disease and to quantify excretion in a group of Merino sheep. DESIGN: A pen and laboratory experiment. PROCEDURE: Seven sheep selected from an affected flock on the basis of acid-fast bacilli in the sheep's faeces were housed and total daily faecal output was collected, weighed and subjected to culture for M avium subsp paratuberculosis. An end-point titration method was used to enumerate viable M avium subsp paratuberculosis in a 15 day pooled sample from five sheep that had acid-fast bacilli in their faeces while housed. RESULTS: Four sheep with subclinical multibacillary Johne's disease excreted M avium subsp paratuberculosis each day for 11 days of cultural observation. A further three sheep were intermittent excreters but lacked other evidence of infection with M avium subsp paratuberculosis. The average number of viable bacteria excreted was 1.09 x 10(8) per gram of faeces while total daily excretion was 8.36 x 10(10) viable M avium subsp paratuberculosis per sheep. Examination of faecal smears stained with Ziehl Neelsen was an unreliable means of assessing daily excretion in individual animals except in those with severe lesions. CONCLUSION: Excretion of M avium subsp paratuberculosis in Merino sheep with multibacillary Johne's disease occurred daily, proving that environmental contamination can be continuous on farms with endemic ovine Johne's disease. Faecal culture is a useful method for detecting infection as it does not appear to be affected by the timing of collection of a sample from sheep with multibacillary disease however, to maximise the sensitivity of disease surveillance using faecal culture, sampling rates should be adjusted to take account of the proportions of multibacillary and paucibacillary cases.