A mixed methods study of ruminant brucellosis in central-eastern Tunisia

In this study, we conducted an investigation to determine the true prevalence of bovine and ovine brucellosis in central-eastern Tunisia. A total of 1134 veterinary samples taken from 130 ruminant herds were screened for brucellosis using IS711-based real-time PCR assay. Sera collected from the ruminants were tested using the Rose Bengal test and indirect enzyme-linked immunosorbent assay. Based on serological and molecular results, the true adjusted animal population level prevalence was 23.5 % in cattle, against 13.5 % in sheep. In addition, the true adjusted herd level prevalence of brucellosis was 55.6 % in cattle and 21.8 % in sheep. A statistically significant association was found between vaginal and milk shedding for ruminants. In addition, our results showed that Brucella abortus could be responsible for bovine and ovine brucellosis. Multivariable logistic regression analysis at the animal population level indicated that age and origin variables were important risk factors for cattle. However, age and abortion variables were found to be associated with ovine brucellosis. At the herd level, risk factors for Brucella positivity were as follows: abortion and herd composition for cattle against herd composition, mortality rates, and hygiene for sheep. Animal hygiene, food quality, and sanitary practices on the farm should be applied as strategies to control brucellosis in herds.     

布鲁氏菌病的致病机理、流行特征及其防控和防治

布鲁氏杆菌病是由布鲁氏菌属细菌侵入机体引起的牛羊易感的一种人畜共患传染病。布鲁氏杆菌病的发生不仅降低了养殖户的经济效益,还对人的生命健康产生极大威胁。观察最近15年的发病趋势,发现布氏杆菌病疫情在世界范围内呈反弹趋势,我国尤甚。应在之前的防控、防治的基础上,进一步研究布氏杆菌病的发病机理,寻求更加有效的措施降低发病率,并探寻能够降低人感染率的有效方法。本文概述了牛羊布氏杆菌病的发病机理和临床特征,并结合近几年布氏杆菌病的流行情况和国内外已有的防治措施,探寻新背景下能够有效降低感染率的方法。     

The use of skin delayed‐type hypersensitivity as an adjunct test to diagnose brucellosis in cattle: A review

Summary

Brucellosis, caused by bacteria of the genus Brucella, is a contagious disease that causes economic loss to owners of domestic animals due to loss of progeny and milk yield. Because cattle, sheep, goats, and to a lesser extent pigs are considered to be the source of human brucellosis, serological tests have been used to screen domestic animals for antibodies against Brucella. Although the serological tests helped to eradicate brucellosis in many countries, serological tests are not always adequate to detect latent carriers of Brucella. Therefore, the use of the skin delayed‐type hypersensitivity (SDTH) test, which is independent of circulating antibodies, might improve the diagnosis of brucellosis. In the literature, however, there are conflicting reports as to the value of the SDTH test for the diagnosis of brucellosis. Some studies consider the test unreliable, whereas others advocate its use because it detects brucellosis earlier than serological tests. The objectives of this study were therefore to assess the characteristics of the SDTH test, to select a Brucella strain that will yield a suitable brucellin for use in the field, and to determine whether the use of serological tests in combination with the SDTH test improves the detection of brucellosis. The results of this study clearly show that the SDTH test detects latent carriers of Brucella and confirms brucellosis in cattle with ambiguous serological test results. Brucellins prepared from smooth or mucoid strains of Brucella are better suited for use in the field than brucellins prepared from rough strains because they detect brucellosis in cattle with acute as well as chronic infection. The SDTH test is highly specific (99.3% specificity), and repeated testing of naive cattle or cattle infected with microorganisms that serologically cross‐react with Brucella does not sensitize cattle to subsequent SDTH tests. However, it is possible that some naive cattle may serologically react to the injection of brucellin. The effect of these serological reactions on the sero‐diagnosis of brucellosis is limited, because cattle may only now and then react serologically either with the serum agglutination test (SAT) or the complement fixation test (CFT). Nevertheless, cattle infected with microorganisms that serologically cross‐react with Brucella may test seropositive for brucellosis 4 to 7 weeks after injection of brucellin, depending on the cross‐reacting microorganism. The value of the SDTH test for the diagnosis of brucellosis was demonstrated after an outbreak of brucellosis. When the SDTH test was used in combination with SAT and CFT at diagnostic threshold ≥2 mm or ≥1 mm (increase in skinfold thickness), respectively, 39/44 (88%) or 42/44 (95%) of the infected cattle were detected compared with only 27/44 (61%) when SAT and CFT were used. When cattle in areas of low prevalence or in areas free from brucellosis are tested with the SDTH test an increase ≥2 mm in skinfold thickness should be considered indicative of infection. When the control and eradication of brucellosis is based on test‐and‐slaughter, an increase of ≥1 mm in skinfold thickness should be considered indicative of infection. Repeated serological testing complemented with the SDTH test in this programme will shorten the quarantine (movement control) period of a suspect herd, limiting the financial loss incurred during outbreaks of the disease. Consequently, since the SDTH test usually does not interfere with the serological diagnosis and can safely be used to establish the infection status of cattle in a suspect herd, it is opportune to consider adding the SDTH test to the procedure currently used to diagnose brucellosis in individual animals.     

Risk associated with animals moved from herds infected with brucellosis in Northern Ireland

The movement of cattle from herds infected with Brucella abortus was investigated in order to assess the control measures for eradication of brucellosis from the cattle population of Northern Ireland. Using recorded cattle movement data, a historical cohort study was designed and carried out to quantify the risk of seropositivity in bovine animals moved from herds infected with brucellosis. The study found that 3.1% of animals, moved in the 6-month period prior to disclosure of infection in the source herd and subsequently tested, were interpreted as seropositive in their destination herds. The odds of seropositivity were approximately 19 (95% confidence interval: 7.8-46.4) times higher in this cohort compared with animals from herds with no history of infection. A multivariate logistic regression model was constructed to examine factors influencing the risk of seropositivity in the exposed cohort of animals, identifying maternal status (whether the dam had been a brucellosis reactor) and age at leaving the infected herd as the main risk factors.     

The use of skin delayed-type hypersensitivity as an adjunct test to diagnose brucellosis in cattle: a review

Brucellosis, caused by bacteria of the genus Brucella, is a contagious disease that causes economic loss to owners of domestic animals due to loss of progeny and milk yield. Because cattle, sheep, goats, and to a lesser extent pigs are considered to be the source of human brucellosis, serological tests have been used to screen domestic animals for antibodies against Brucella. Although the serological tests helped to eradicate brucellosis in many countries, serological tests are not always adequate to detect latent carriers of Brucella. Therefore, the use of the skin delayed-type hypersensitivity (SDTH) test, which is independent of circulating antibodies, might improve the diagnosis of brucellosis. In the literature, however, there are conflicting reports as to the value of the SDTH test for the diagnosis of brucellosis. Some studies consider the test unreliable, whereas others advocate its use because it detects brucellosis earlier than serological tests. The objectives of this study were therefore to assess the characteristics of the SDTH test, to select a Brucella strain that will yield a suitable brucellin for use in the field, and to determine whether the use of serological tests in combination with the SDTH test improves the detection of brucellosis. The results of this study clearly show that the SDTH test detects latent carriers of Brucella and confirms brucellosis in cattle with ambiguous serological test results. Brucellins prepared from smooth or mucoid strains of Brucella are better suited for use in the field than brucellins prepared from rough strains because they detect brucellosis in cattle with acute as well as chronic infection. The SDTH test is highly specific (99.3% specificity), and repeated testing of naive cattle or cattle infected with microorganisms that serologically cross-react with Brucella does not sensitize cattle to subsequent SDTH tests. However, it is possible that some naive cattle may serologically react to the injection of brucellin. The effect of these serological reactions on the sero-diagnosis of brucellosis is limited, because cattle may only now and then react serologically either with the serum agglutination test (SAT) or the complement fixation test (CFT). Nevertheless, cattle infected with microorganisms that serologically cross-react with Brucella may test seropositive for brucellosis 4 to 7 weeks after injection of brucellin, depending on the cross-reacting microorganism. The value of the SDTH test for the diagnosis of brucellosis was demonstrated after an outbreak of brucellosis. When the SDTH test was used in combination with SAT and CFT at diagnostic threshold > or =2 mm or > or =1 mm (increase in skinfold thickness), respectively, 39/44 (88%) or 42/44 (95%) of the infected cattle were detected compared with only 27/44 (61%) when SAT and CFT were used. When cattle in areas of low prevalence or in areas free from brucellosis are tested with the SDTH test an increase > or =2 mm in skinfold thickness should be considered indicative of infection. When the control and eradication of brucellosis is based on test-and-slaughter, an increase of > or =1 mm in skinfold thickness should be considered indicative of infection. Repeated serological testing complemented with the SDTH test in this programme will shorten the quarantine (movement control) period of a suspect herd, limiting the financial loss incurred during outbreaks of the disease. Consequently, since the SDTH test usually does not interfere with the serological diagnosis and can safely be used to establish the infection status of cattle in a suspect herd, it is opportune to consider adding the SDTH test to the procedure currently used to diagnose brucellosis in individual animals.     

Active surveillance for scrapie in the Netherlands: effect of a breeding programme on the prevalence of scrapie in sheep (2002-2010)?

The susceptibility of sheep to scrapie is modulated by the prion protein (PrP) genotype of the animal. An ambitious voluntary scrapie control programme was started in the Netherlands in 1998, based on selection of rams with theARR/ARR genotype for breeding. This programme was followed by an obligatory programme in 2004; the programme has been voluntary since 2007. We monitored the prevalence of PrP genotype frequencies and the prevalence of scrapie in the Dutch sheep population between 2002 and June 2010. Results showed that selection for scrapie-resistant sheep resulted in an increase in the ARR allele frequency in the Dutch national flock from 37.5% in 2005 to 61.4% in 2009. Moreover, surveillance data showed that there was a significant decrease in the prevalence of scrapie a few years after the start of the obligatory breeding programme, from more than 0.2% in 2004 to 0.015% in 2009. This decrease is a consequence of the increased number of scrapie-resistant sheep in the Dutch sheep population. To date, the results and the models based on the data show that the selective breeding programme should be continued for several years in order to successfully eradicate scrapie. It will be important to monitor the PrP frequency and scrapie prevalence in the Dutch sheep population in the coming years.     

肉牛布鲁氏杆菌病防控现状及措施

布鲁氏菌病是由布鲁氏菌引起的一种急性或慢人畜共患传染病,该病除了导致养牛企业直接经济损失外,还可能发生严重食品安全问题和因人被感染导致的严重公共卫生安全问题。多年来,各级动物防疫部门主要关注奶牛的布病防控,对肉牛的布病防控介入很少,造成肉牛布病感染情况不明、底子不清,防控力度不够。随着规模化肉牛养殖业的不断兴起,肉牛布鲁氏菌病防控可不容缓。因此,必须采取综合措施有效防控肉牛布鲁氏菌病。     

Epidemiology,zoonotic aspects,vaccination and control/eradication of brucellosis in India

In India, brucellosis was first recognised in 1942 and is now endemic throughout the country. The disease is reported in cattle, buffalo, sheep, goats, pigs, dogs and humans. B. abortus biotype-1 in cattle and buffaloes and B. melitensis biotype-1 in sheep, goats and man are the predominant infective biotypes. The long-term serological studies have indicated that 5% of cattle and 3% of buffaloes are infected with brucellosis. Economic losses due to brucellosis in livestock are considerable in an agrarian country like India. There is no organised and effective brucellosis control programme in the country. With the indigenous development of serum and milk based ELISA kits, the population survey of the disease has been undertaken on a large scale in several states and plans for the control of the disease through calf-hood vaccination are being worked out. An innovative approach—Bovine Brucellosis Progressive Control Programme (BBPCP) is targeted to overcome the basic problems of ban on cow slaughter, distress sale of animals following the positive serological diagnosis of brucellosis and absence of a disease control strategy. The work plan for the implementation of BBPCP is presented.     

Systematic review of the prevalence of paratuberculosis in cattle,sheep, and goats in Latin America and the Caribbean

Mycobacterium avium subsp. paratuberculosis causes paratuberculosis or Johne’s disease (JD) in domestic ruminants and wild species. The aim of the present study was to systematically review the prevalence of paratuberculosis among farmed animals (cattle, sheep, and goats) in Latin America and the Caribbean. The initial search for existing publications reporting systematic reviews and primary studies was carried out by searching the available databases. For the final selection of studies, an initial screen for basic eligibility and a detailed appraisal of quality were performed. After study selection, the relevant data were extracted. The detailed appraisal generated 24 publications that reported 52 studies, of which 73.1, 11.5, and 15.4 % were from cattle, sheep, and goats, respectively. Thirty-three (63.5 %) of the studies were animal level studies, while 19 (36.5 %) were herd-/flock-level studies. No flock-level studies on prevalence in sheep were found. Studies in Latin American and Caribbean countries revealed an overall prevalence of 16.9 (95 % CI (confidence interval) 13.2–20.5) and 75.8 % (95 % CI 50.1–101.5) in cattle at the animal and herd levels, respectively; the prevalence was 16 % (95 % CI 7.9–24.1) in sheep at the animal level and 4.3 % (95 % CI 1.9–6.8) and 3.7 % (95 % CI 0.1–7.4) in goats at the animal and flock levels, respectively. In general, prevalence results reported by the studies were insufficient to accurately determine the prevalence of paratuberculosis in farmed animals in Latin America and the Caribbean. Several flaws in the design of studies limit the quality of evidence regarding the prevalence of paratuberculosis in the region.     

Simulation analysis of the effect of herd immunity and age structure on infection of a cattle herd with bluetongue viruses in Queensland, Australia

A state-transition model based on Leslie matrix formulation was used to investigate the effects of herd immunity and age structure on the infection of a simulated cattle herd with bluetongue viruses under Australian climatic conditions. Increasing duration of immunity decreased the prevalence of infection. A duration of immunity of 33 months was consistent with prevalence estimates made from previous serological studies of bluetongue virus. Herd prevalence displayed slowly dampening cyclical variation over time (most pronounced when a short duration of immunity was simulated). Increasing calving and mortality risk rates in the simulated herd increased prevalence, whereas increasing age at first calving decreased prevalence. Manipulation of calving rates had the greatest effect on the predicted prevalence of infection in the herd. Simulation of a number of herd-management scenarios suggested that management systems in which cattle are bred early and where high calving rates are achieved are likely to contribute to high levels of infection with bluetongue viruses. Results confirm the importance of management factors in influencing the prevalence of infectious diseases in animal populations.     

Breeding programmes for TSE resistance in British sheep. II. Assessing the impact on the prevalence and incidence of scrapie

By establishing a breeding programme for transmissible spongiform encephalopathie (TSE) resistance, there are plans to eradicate sheep TSEs from member states of the European Union (EU). In this paper, we used a simple age- and genotype-structured model to assess the impact of four breeding strategies on the prevalence and incidence of scrapie in the British sheep flock. The strategies ranged from the minimum EU requirements to compulsory implementation of the current National Scrapie Plan for Great Britain (NSP). All four strategies were predicted to reduce the prevalence and incidence of disease, though there was likely to be a delay of several years between the implementation of a breeding programme and the reduction in incidence. There were differences in the efficacy of the strategies, with the most stringent resulting in the greatest reduction in prevalence and incidence. However, the magnitude of the differences was not great, largely because all four strategies eliminated the VRQ allele, which is associated with a markedly higher risk of disease than any of the other alleles. Sensitivity analyses indicated that the model results were robust to selection bias when estimating the risk of infection; and that the efficacy of a breeding programme was unlikely to be compromised, unless the risk of infection is substantially underestimated by data on clinical disease.     

山西省大同市灵丘县养殖户布鲁菌病知识宣传干预调查

[目的]了解山西省大同市灵丘县养殖户对布鲁菌病相关知识知晓情况、可能的感染危险行为,为针对性开展布鲁菌病健康宣教提供参考。[方法]采用简单随机抽样方法,在灵丘县选取70户牛养殖户,通过设计的调查问卷对养殖户的布病防控知识和高危行为情况开展现状调查。[结果]在70名被调查对象中,有77.14%的养殖户没有对新购入牛进行单独饲喂的防疫隔离意识。有45.71%的养殖户对牛群未免疫布病疫苗。虽然灵丘县近年来多次开展过布鲁菌病的宣传干预,但该次调查结果显示,宣传干预仍有较大的改善空间。[结论]灵丘县养殖户对布鲁菌病的知识知晓率相对较高,但仍距离规划目标还存在较大差距,仍存在不小的感染布鲁菌病的风险。因此,在接下来的工作中应加强针对养殖户进行布鲁菌病宣传干预,提高该部分人群对布鲁菌病的防护意识。     

Seroprevalence of chlamydial infection in cattle in Ireland

Although few studies have investigated the prevalence of chlamydial infections in cattle, reported prevalence rates vary hugely. In order to assess the prevalence of this infection in cattle in Ireland, serum samples (100 herds, 20 samples/herd) collected for statutory screening for brucellosis were examined by soluble chlamydial antigen indirect ELISA. The assay detects antibodies to the two most common Chlamydiaceae spp. affecting cattle, namely Chlamydia abortus and Chlamydia pecorum. A total of 95 samples from 57 herds were seropositive, representing an observed prevalence rate of 4.75%. The parametric bootstrap estimate of the mean disease prevalence in the population was 6.04% (95%, CI 4.70-7.50). The results suggest the prevalence of chlamydial infection is low in cattle in Ireland.     

Brucellosis in the European Union and Norway at the turn of the twenty-first century

Control and eradication programs of brucellosis in cattle, sheep, goats and pigs have been more or less successfully implemented within the Member States (MS) of the European Union (EU) and Norway after Word War II. As a result, the epidemiological situation of animal brucellosis is extremely diverse among different MS or regions within a MS and among the different animal species. Some MS, mainly North European countries, and Norway are declared “officially bovine brucellosis free” and/or “officially ovine and caprine (Brucella melitensis) free”. The situation is less favorable in Southern European countries, particularly as far as sheep and goat brucellosis are concerned. This situation has important zoonotic consequences as reflected in the number of human brucellosis cases due to B. melitensis that are still encountered in those countries. Brucellosis in swine has re-emerged as a result of spillover from the wild boar brucellosis (Brucella suis biovar 2) reservoir, particularly in outdoor reared pigs. Besides the actual challenge to eradicate brucellosis, further issues have to be addressed: (1) the management of false positive serological results that occur in the course of brucellosis testing, particularly in cattle; (2) the impact of wildlife brucellosis, particularly wild boar brucellosis in domestic animals; and (3) the importance of B. melitensis infection in cattle that are in contact with infected sheep.     

云南建水地区2017-2019年牛羊布鲁氏菌病的流行病学调查分析

为了解建水地区近几年牛羊布鲁氏菌病的流行情况,采用虎红平板及试管凝集试验对2017-2019年期间采集的150份牛血清、450份羊血清进行检测,并对数据进行统计分析。结果显示,2017-2019年牛布鲁氏菌病的阳性率为0.67%,羊布鲁氏菌病的阳性率为0.22%。其中2017年牛羊布鲁氏菌病的阳性率均为0%;2018年牛羊布鲁氏菌病的阳性率均为0%;2019年牛羊布鲁氏菌病的阳性率分别为2%、0.67%。本研究为促进本地区畜牧业健康稳定发展、养殖户效益增收以及保障畜产品质量安全提供了参考依据。     

Brucellosis and associated risk factors in dairy cattle of eastern Ethiopia

Knowing the status of Bovine brucellosis and associated risk factors is a crucial step in formulating evidence based control scheme. In this study, a total of 967 dairy cows from 307 dairy farms in eastern Ethiopia were serologically tested for Brucella antibodies. The screening was done first using RBPT and positive samples were subsequently subjected to CFT for confirmation. A pre-tested structured questionnaire was used to collect relevant data from 307 dairy cattle owners or attendants to assess their awareness and routine practice. The data were run using univariable logistic regression analysis using STATA version 11.0 for Windows. Accordingly, herd and individual animal seroprevalence were found to be 6.8% (95% CI?=?4.28–10.28) and 1.3% (95% CI?=?0.72–2.29), respectively. The prevalence of sero-reactors among local breeds was observed to be higher compared to cross breed (p?<?0.05). Herd level analysis of the risk factors indicated that in farms with large herd size (>20 animals), the odds ratio (OR)?=?9.13, p?=?0.00, CI?=?3.01–27.69 of having brucellosis was 9.13 times higher than smaller size herds (<20 animals). Intensively managed herds had shown the highest seroprevalence (20.8%) than extensive (6.7%) and semi-intensive (4.2%). Experience of dairy handlers about the disease that cause abortion in late pregnancy was significantly associated (p?<?0.001) with the occurrence of brucellosis in the herds. However, about 91% of the dairy cattle owners/attendants lack awareness about disease(s) that causes abortion in late pregnancy. Similarly risk of having brucellosis in those herds experiencing abortion was 6.3 times higher (OR?=?6.3, p?<?0.001, CI?=?2.50–15.92). This study identified some of the handling practices for aborted and retained fetal materials to be risky. Therefore, the study highlights the need of comprehensive brucellosis surveillance in animal and human and institutions of public education and on farm biosafety measures in shaping proper disease control scheme.

     

Seroprevalence and risk factors for leptospirosis in cattle,sheep, and goats at consorted rearing from the State of Piauí, northeastern Brazil

Leptospirosis is an endemic disease in Latin America, caused by pathogenic bacteria of the genus Leptospira. It is considered one of the main causes responsible for the negative economic impact on global livestock by causing reproductive problems. The research aimed to determine the prevalence of leptospirosis in cattle, sheep, and goats at consorted rearing in the micro-region of Teresina, Piauí state, northeastern Brazil, as well as to identify prevalent serovars and risk factors associated with seroprevalence. Serum samples were analyzed in 336 sheep, 292 goats, and 253 cattle using microscopic agglutination test (MAT). Overall, 378 samples were positive to MAT, with seroprevalence of 42.9%. The prevalences in cattle, sheep, and goats were 50.5, 40.5, and 34.6%, respectively. All herds presented at least one seropositive animal; the Hardjo/Wolffi serovar association was the most common in cattle and Icterohaemorrhagiae in goats and sheep. Beef production (OR?=?4.9), cattle herd over 35 animals (OR?=?4.0), feeding on pasture (OR?=?6.4), weir and/or stream as water source (OR?=?2.1), and no veterinary services (OR?=?2.9) were risk factors for cattle infection. For sheep, intensive management system (OR?=?5.3), suspended slatted facilities (OR?=?2.2), more than 20 sheep in reproductive age (OR?=?1.9), and absence of deworming (OR?=?3.5) were the risk factors, while for goats, the identified risk factors were sheep herd over 52 animals (OR?=?1.9) and no veterinary services (OR?=?1.8). We conclude that the infection was spreading in consorted herds in this region. Thus, it would be interesting and important to conduct educative activities to farmers on the economic impacts of this disease and the need of preventive and control strategies mainly focused on sanitary measures and animal handling.     

Modelling the transmission dynamics of Echinococcus granulosus in sheep and cattle in Kazakhstan

Cystic echinococcosis, caused by Echinococcus granulosus, is an emerging disease in many parts of the world and, in particular, in eastern Europe and the former Soviet Union. This paper examines the abundance and prevalence of infection of E. granulosus in cattle and sheep in Kazakhstan. Observed data are fitted to a mathematical model in order to determine if the parasite population is partly regulated by intermediate host immunity and to define parameters in the model. Such data would be useful to develop simulation models for the control of this disease. Maximum likelihood techniques were used to define the parameters and their confidence limits in the model and the negative binomial distribution was used to define the error variance in the observed data. The results indicated that there are significant variations in the infection pressure to sheep depending on their location. In particular sheep from Almaty Oblast and from central and northern Kazakhstan appeared to have a greater exposure than sheep from Jambyl or South Kazakhstan Oblasts. The infection pressure to cattle was somewhat lower in comparison. In common with other similar studies, there was no evidence of parasite-induced immunity in sheep or cattle in Kazakhstan due to natural infection. The highest abundance and prevalence were seen in the oldest age classes of animals.     

Recovery of dual serotypes of bluetongue virus from infected sheep and cattle

Dual serotypes of bluetongue virus (BTV) were recovered from field-collected samples of sheep and cattle blood. Two sheep, each infected with both BTV serotypes 10 and 17, were found in a flock with bluetongue disease associated with these two serotypes. One sheep infected with BTV serotypes 11 and 17 was found in a second flock; it was the only viremic sheep detected and was clinically ill. Dual serotype infections of one beef and two dairy cattle were found in three geographically separate herds; mixtures recovered were of BTV serotypes 10 and 17 and serotypes 11 and 17. Clinical signs of illness were absent in the cattle in two herds, but severe conjuctivitis was seen in several cows in a third herd, including the cow with a dual serotype infection (BTV 11 and 17). Two of the cattle with dual infections had no serological evidence of BTV as determined by the agar gel precipitin test; serum was not available from the other cow with a dual serotype infection. The significance of dual infections and immune tolerance are discussed.