Survey of the seroprevalence of brucellosis in ruminants in Kosovo

A cross-sectional survey of the seroprevalence of brucellosis in sheep, goats and cattle in Kosovo was made in January 2001. A total of 12,000 serum samples, from 7941 cattle, 3548 sheep and 511 goats, were screened using the Rose Bengal test. Doubtful and positive results were further tested with competitive and indirect ELISAS. The overall serological prevalences derived from the samples positive to all three tests, were 6.26 per cent (95 per cent confidence intervals [CI] 5.5 to 7.1 per cent) for sheep, 7.24 per cent (5.3 to 9.8 per cent) for goats and 0.58 per cent (0.43 to 0.77 per cent) for cattle. The survey covered 26 of the 29 municipalities and showed that brucellosis was widely but unevenly distributed throughout the province. Seropositive animals were found in 25 per cent (19 to 32 per cent) of 162 villages surveyed. The risk of cattle being infected on holdings where both cattle and sheep were kept was greater, with a risk ratio of 4.6 (2.2 to 9.6), than on holdings where only cattle were kept. Brucella melitensis probably predominates as the cause of brucellosis in ruminants in the province of Kosovo.     

Diagnosis of bovine brucellosis by skin test: conditions for the test and evaluation of its performance.

Brucellergene OCB (Rh?ne-Mérieux) was used as an allergen to define the intrinsic parameters of a skin test and to compare its properties with serology for the diagnosis of bovine brucellosis. The skin test was also evaluated for its capacity to solve problems associated with false positive reactions in serological tests. The optimal reading delay for the skin test was 72 hours. The brucellosis allergic reaction was two to three times less intense than the tuberculosis allergic reaction. An increase of 1.1 mm or more in the skin thickness was therefore considered to be an adequate cut-off. The specificity calculated for 1192 brucellosis-free animals (including animals from brucellosis-free herds in which false positive serological reactions had been reported) was 99-83 per cent (95 per cent confidence interval [CI] 99-40 to 99-98 per cent). The sensitivity determined from 27 experimentally infected heifers ranged from 93 per cent (95 per cent CI 76 to 100 per cent) to 78 per cent (95 per cent CI 58 to 91 per cent) when measured respectively one and six months after the infection. Allergic reactions could be detected in vaccinated animals up to four-and-a-half years after the vaccination. On the other hand, no sensitisation was recorded in na?ve animals after up to eight monthly injections of the allergen. The skin test gave valuable information, in combination with the serological tests, in both acute and chronic brucellosis. The skin test discriminated brucellosis clearly from false positive serological reactions due to infections with Yersinia enterocolitica O9.     

Brucellosis control in Tajikistan using Rev 1 vaccine: change in seroprevalence in small ruminants from 2004 to 2009

Cross-sectional serological surveys of 13,006 small ruminants were conducted in 2003 to establish baseline levels of seropositivity to brucellosis and of 12,102 again in 2009 to evaluate the efficacy of controls based on biannual conjunctival vaccination with Rev 1. Seroprevalence dropped by 80 per cent in five years in eight pilot programme districts where vaccination was generally well implemented, and the prevalence of households with evidence of infection in their animals dropped from 25.1 to 7.5 per cent. Seroprevalence was reduced by 40 per cent in 10 districts where vaccination was intermittent with low coverage during some seasons. There were no changes in 19 districts where no vaccinations were carried out. Sheep vaccinated one or more times were found to be 2.5 times more likely to be serotest positive than were non-vaccinated sheep, whereas vaccinated goats were 6.4 times more likely to be serotest positive than non-vaccinated goats.     

Factors associated with the wastage and achievements in competition of event horses registered in the United Kingdom

The aims of this study were to estimate the wastage of horses registered for eventing in Britain, to investigate the reasons for this wastage and to evaluate factors affecting the horses' achievement of grade I status (at least 61 points) while registered. An analysis of the database of the British Eventing register found that 33.7 per cent of horses registered for the first time in 1999 were not re-registered for eventing in subsequent years. By using multivariable logistic regression analysis, it was shown that horses that were kept at an event yard were more likely to be re-registered than those kept on other premises (odds ratio [or] 2.0, 95 per cent confidence interval [ci] 1.2 to 3.2), and those that took part in showjumping while registered were also more likely to be re-registered (or 1.5, 95 per cent ci 1.1 to 2.2). Horses that took part in unaffiliated eventing while registered were less likely to be re-registered the following year (or 0.7, 95 per cent ci 0.5 to 0.9), as were those that were not insured (or 0.7, 95 per cent ci 0.5 to 1.0) and those from outside the British Isles (or 0.6, 95 per cent ci 0.3 to 1.0). Veterinary problems were the most commonly cited explanation (35.1 per cent) why horses that remained in their original ownership were not re-registered with British Eventing the following year. Horses from Australia were more likely to achieve grade I status than horses from the British Isles (or 9.7, 95 per cent ci 7.1 to 13.2), as were horses from New Zealand (or 6.4, 95 per cent ci 5.0 to 8.2), the usa (or 5.2, 95 per cent ci 3.8 to 7.2) and France (or 2.8, 95 per cent ci 2.1 to 3.7), but horses from the Netherlands (or 0.5, 95 per cent ci 0.3 to 0.9) and Belgium (or 0.3, 95 per cent ci 0.1 to 0.9) were less likely to achieve grade I status. Mares were less likely to achieve grade I status than geldings (or 0.4, 95 per cent ci 0.4 to 0.5).     

Prevalence of mixed trematode infections in an abattoir receiving cattle from northern Portugal and north-west Spain

Between February 2007 and January 2009, a total of 776 slaughtered animals were examined for the presence of Trematoda in the liver, gall bladder, oesophagus and stomach (rumen and reticulum). Data collected were analysed by the age and breed of the animals. The percentage of cattle from which Trematoda were found was 38 per cent (95 per cent confidence interval [CI] 35 to 41 per cent); 28 per cent (95 per cent CI 25 to 31 per cent) had Fasciola hepatica, 12 per cent (95 per cent CI 10 to 14 per cent) Calicophoron daubneyi and 6 per cent (95 per cent CI 4 to 8 per cent) Dicrocoelium species. A significantly high prevalence of fasciolosis and dicrocoeliosis was observed in cattle over 10 years of age. Autochthonous Rubia Gallega cattle had the highest prevalence of fasciolosis and crossbred cattle had the highest prevalence of dicrocoeliosis (P<0.05). Twenty per cent (95 per cent CI 15 to 25 per cent) of the cattle positive for Fasciola also had Calicophoron species; 10 per cent (95 per cent CI 6 to 14 per cent) also had small liver flukes (Dicrocoelium species).     

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.     

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.     

Prevalence of classical swine fever, Aujeszky's disease and brucellosis in a population of wild boar in Switzerland

During two survey rounds of a national surveillance system for infectious diseases in wild boar in Switzerland, each lasting four months from November to February, between 2001 and 2003, 1949 blood samples and 62 tissue samples from the spleen and 50 from the reproductive organs were collected from hunted wild boar. The survey was designed so that freedom from infection could be detected with a probability of 95 per cent at a threshold prevalence of less than 1 per cent for classical swine fever and Aujeszky's disease and less than 1.5 per cent for brucellosis. There was no serological evidence of classical swine fever or Aujeszky's disease, but brucellosis due to Brucella suis biovar 2 was confirmed serologically and by bacterial isolation.     

Brucellosis in Ontario: a case control study.

Data from cattle herds infected with brucellosis and from control (noninfected) herds were collected and analyzed using case control techniques. It appeared that herds located close to other infected herds and those herds whose owners made frequent purchases of cattle had an increased risk of acquiring brucellosis, particularly those who made purchases from other herds or from cattle dealers. Infected herds had a lower level of vaccination than noninfected herds. However, the percentage vaccinated was highly variable in each group. Vaccination per se did not appear to adversely influence the interpretation of serological test results nor did it appear to protect the individual animal. Once infected, the time required to become free of brucellosis was increased by large herd size and/or loose housing. Closed herds also took longer to become brucellosis free than more open herds. The percentage of animals removed from the herd was increased by active abortion. Those herds with multiple serological reactors (positives and questionables) at the first herd test after the imposition of quarantine had the highest percentage of cattle removed.     

Prevalence of faecal excretion of verocytotoxigenic Escherichia coli O157 in cattle in England and Wales

During the decade to 1999, the incidence of human infections with the zoonotic pathogen verocytotoxin-producing Escherichia coli O157 (VTEC O157) increased in England and Wales. This paper describes the results of a survey of 75 farms to determine the prevalence of faecal excretion of VTEC O157 by cattle, its primary reservoir host, in England and Wales. Faecal samples were collected from 4663 cattle between June and December 1999. The prevalence of excretion by individual cattle was 4.2 per cent (95 per cent confidence interval [CI] 2.0 to 6.4) and 10.3 per cent (95 per cent CI 5.8 to 14.8) among animals in infected herds. The within-herd prevalence on positive farms ranged from 1.1 to 51.4 per cent. At least one positive animal was identified on 29 (38.7 per cent; 95 per cent CI 28.1 to 50.4) of the farms, including dairy, suckler and fattening herds. The prevalence of excretion was least in the calves under two months of age, peaked in the calves aged between two and six months and declined thereafter. The phage types identified most widely were 4, 34 and 2, which were each found on six of the 29 positive farms.     

Impact of geographical factors on the spread of animal brucellosis in the Republic of Kazakhstan

In Latin and Central America and in most Asian countries, brucellosis remains an insufficiently studied disease. This study aims to determine the national and regional incidence of brucellosis among cattle (cows) and small ruminants (sheep, goats) in the Republic of Kazakhstan, as well as to identify the effect of climatic and geographical factors on the incidence rates. Thematic maps were created in an open geographic information system QGIS version 2.8. in order to identify the natural and socio-economic factors that influence the spread of the disease overlay method was used. Local cluster analysis was used in order to identify additional causes of the disease. Findings show the following values of Pearson correlation between the overall population and the number of animals infected: 0.68 for cows, p ≤ 0.005, and 0.56 for sheep and goats, p ≤ 0.03. Thus, the larger the heard in a given area, the greater likelihood of having brucellosis. Data processing reveals that Kazakhstan has almost twice as many regions good for cattle breeding as regions that are good for the small ruminants farming. The correlation variables for cattle and small ruminants are approximately the same. On the basis of the performed research the author proposes to amend the accepted methodology of epidemiology surveillance by the methods based on spatial (geographical) analysis. It is also proposed to adjust the process of breeding cattle and small ruminants considering the additional health recommendations that take into account the geographical aspects of the spread of the disease.     

Experience with simple ELISA test systems for Brucella serology in cattle, sheep and goats

The objective of this work was to use the ELISA technique for the serological surveillance for freedom of brucellosis of cattle, sheep and goats. By comparing 28 cattle sera taken after a brucellosis outbreak, 15 bovine sera supplied by the Federal Institute for Health Protection of Consumers and Veterinary Medicine (BgVV) and 497 serum slow agglutination test (SSAT) and complement fixation test (CFT) negative bovine sera from herds officially declared free of brucellosis, the ELISA technique not only shows higher sensitivity as compared to SSAT and CFT but also distinguishes clearly between positive and negative reactions. The serological comparison by SSAT, CFT and ELISA of 615 cattle, 624 sheep and 630 goat sera from herds acknowledged as brucellosis free showed equivalent specificities for both CFT and ELISA. The specificity of the SSAT was much lower, 81.1% in cattle and 96.2% in goat sera. The examination of 5796 cattle, 1337 calf, 5031 sheep and 1796 goat sera demonstrates the advantage of the ELISA technique as routine method. The possible application of the ELISA technique as a screening method for serological brucellosis tests in sheep, goats and possibly also in pigs is discussed.     

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.     

The vaginal mucus agglutination test in the diagnosis of bovine brucellosis

Of 1140 vaginal mucus agglutination tests (VMAT) on specimens obtained in 1971-72 from 663 dairy cows in seven herds infected with brucellosis, 97 were positive. When the VMAT was positive one or more serological tests were also positive. Of the 97 corresponding serum agglutination tests 80 sera had titres of more than 533 international units. Only 69.8 per cent of VMAT from serologically positive cows were positive. No evidence was found of non-specific agglutinins in vaginal mucus and positive VMAT reactions appeared to be specific for field infection. Three cows showed evidence of local agglutinins in the vagina. Hence herd testing by VMAT has no advantage over tests of blood serum but the test could be an aid in establishing whether individual cattle are infected.     

Serological, bacteriological and molecularbiological survey of paratuberculosis (Johne's disease) in Austrian cattle

Paratuberculosis (Johne's disease) is a chronic infectious disease of ruminants, caused by Mycobacterium avium subspecies paratuberculosis (MAP). Because of its long incubation period, high economic losses, difficulties in diagnosis and possible links to Morbus Crohn in humans, paratuberculosis is one of the most important diseases of ruminants today. An abattoir-based nationwide survey on the occurrence of MAP in the Austrian cattle population was performed using serology (SVANOVIR-ELISA) as well as culture, ZN-stain and IS900-PCR on faeces and lymph node samples. A total of 756 Austrian slaughter cattle were serologically, bacteriologically and molecularbiologically tested for the occurrence of MAP and specific antibodies respectively. Samples were collected following a statistical plan to obtain balanced specimens from the whole country. Nineteen per cent of the animals tested were serological positive, 10.1% gave an inconclusive result and 70.9% showed no specific antibodies against MAP. Only in four individuals MAP could be detected by stain, bacteriology or Polymerase Chain Reaction. The calculated prevalence of 19.0% positive cattle, each representing one farm, showing specific antibodies against MAP is rather high in terms of animal-level but low in herd level prevalence compared with other countries. When this study is compared with a similar study performed in Austria 1999, a significant increase of positive cattle and farms could be seen in Austria.     

Evaluation of three LPS-specific monoclonal antibodies for the immunodiagnosis of bovine brucellosis

Bovine sera collected during the Australian brucellosis eradication campaign were used to assess the value of three monoclonal antibodies (MAb Bruce 1, 4 and 7) for the immunodiagnosis of bovine brucellosis in a competitive enzyme immunoassay (CEIA). Each MAb reacted to a different epitope of lipopolysaccharide molecules on the cell surface of Brucella abortus. When the sensitivity of the CEIA was set at 100 per cent so that all infected animals were identified, the specificity of the test using MAb Bruce 1 and Bruce 7 was 69 per cent and 52 per cent, respectively. On the other hand, a quarter of the sera from infected cattle did not inhibit the binding of MAb Bruce 4 to the antigen. With a maximum sensitivity of 75 per cent, the specificity of the CEIA using MAb Bruce 4 was 94 per cent. However, all three MAb cross reacted with sera from sheep infected with Bovis, Histophilus ovis and Actinobacillus seminis.     

Serological status of Canadian cattle for brucellosis,anaplasmosis, and bluetongue in 2007–2008

A national bovine serological survey was conducted to confirm that the prevalence of brucellosis, bluetongue, and anaplasmosis does not exceed 0.02% (95% confidence) in live cattle in Canada. Sampling consisted of a systematic random sample of 15 482 adult cattle slaughtered in federally inspected abattoirs, stratified by province. Samples were tested to detect antibodies for brucellosis, bluetongue, and anaplasmosis. All samples were negative for brucellosis. Three samples were seroreactors to bluetongue, 2 of which originated from the Okanagan Valley in British Columbia and 1 from Ontario, which after follow-up, was considered an atypical result. A total of 244 samples were seroreactors to Anaplasma and follow-up identified infection in Saskatchewan, Manitoba, and Quebec. In conclusion, the Canadian cattle population remains free of brucellosis and free of bluetongue outside the Okanagan Valley. Canada is no longer free of anaplasmosis and will be unable to claim freedom until eradication measures are completed.     

Bluetongue serotype 8 vaccine coverage in northern and south-eastern England in 2008

A postal survey of all registered cattle and sheep farmers in East Anglia was carried out from July 2008 to determine bluetongue virus serotype 8 (BTV-8) vaccine uptake in the region. The vaccine was available to farmers in this region from May 2008. The survey was repeated in Cumbria and Northumberland at the beginning of 2009. In these regions, the vaccine was not available until September 1, 2008. Holding-level vaccine uptake was estimated to be 85 per cent (95 per cent confidence interval [CI] 83 to 87 per cent, n=1623) in East Anglia and 36 per cent (95 per cent CI 32 to 40 per cent, n=633) in northern England. A telephone follow-up of non-responders reduced these estimates to 79 and 29 per cent in East Anglia and northern England, respectively. In both regions, vaccine coverage was higher in sheep than in cattle, with 92 per cent of sheep in East Anglia having been vaccinated. The proportion of holdings that had applied the vaccine or were intending to apply the vaccine in 2009 in the northern region was 51 per cent (95 per cent CI 47 to 54 per cent, n=664), with a further 37 per cent undecided at the time of response.     

A comparison of three serological tests in the diagnosis of caprine brucellosis.

The serum agglutination test (SAT), the Rose Bengal plate test (RBPT) and the milk ring test (MRT) were used in the diagnosis of caprine brucellosis. There was a close correlation between the SAT and RBPT when both tests were negative but the RBPT failed to detect 79.82 per cent of sera in excess of 50 iu. Also, owing to the relatively poor milking potential of the Nigerian goat and the false positive results with the MRT, it is concluded that the SAT offers a better serological diagnostic tool for caprine brucellosis in this locality.