Porcine Brucellosis Research Articles

Estimation of sensitivity and specificity of five serological tests for the diagnosis of porcine brucellosis

While serological tests are essential in surveillance and control programs of animal diseases, to date none of the common serological tests approved in the EU (complement fixation test or Rose-Bengal test) has been shown to be reliable in routine individual diagnosis of porcine brucellosis, and some more recent tests like ELISA have not been fully evaluated yet. In the absence of a gold standard, this study allowed the estimation of sensitivities and specificities of these tests with a Bayesian approach using Markov Chain Monte Carlo algorithms.The pig population that was tested included 6422 animals from Metropolitan France. Serum samples were collected from a large population of pigs, representative of European swine population and tested with five brucellosis serological tests: Rose-Bengal test (RBT), fluorescence polarization assay (FPA), indirect ELISA (I-ELISA) and two competitive ELISAs (C-ELISA). The sensitivity and the specificity of each test were estimated. When doubtful results were excluded, the most sensitive and specific test was C-ELISA2 (Se C-ELISA2=0.964, [0.907; 0.994], 95% credibility interval (CrI); Sp C-ELISA2=0.996, [0.982; 1.0], 95% CrI).When doubtful results were considered as negative, C-ELISA2 was still the most sensitive and specific test (Se C-ELISA2=0.960, [0.896; 0.994], 95% CrI and Sp C-ELISA2=0.994, [0.977; 0.999], 95% CrI). The same conclusions were reached when doubtful results were considered as positive (Se C-ELISA2=0.963, [0.904, 0.994], 95% CrI and Sp C-ELISA2=0.996, [0.986; 1.0], 95% CrI).

Revised Genome Sequence of Brucella suis 1330

Brucella suis is a causative agent of porcine brucellosis. We report the resequencing of the original sample upon which the published sequence of Brucella suis 1330 is based and describe the differences between the published assembly and our assembly at 12 loci.

Serological Survey of Porcine <i>Brucella</i> Infection in SouthEast, Nigeria

Porcine brucellosis, also called contagious abortion of swine is an infectious and zoonotic disease of swine caused by bacteria of the genus Brucella (Young, 1995). Brucella suis is the species found primarily in pigs. It is a zoonotic infection of domesticated and wild animals which humans(especially occupationally exposed workers), acquire by ingestion of improperly cooked contaminated pork, direct contact with infected animals or inhalation of infected aerosols (Radostits, 1995). Brucella infection in pigs is of wide spread occurrence, but the prevalence is low in most countries with the exception of South America and Southeast-Asia where the prevalence is high (Starnes et al., 1999). Pig production is drasticallyaffected by Brucella infection through abortion, birth of weak/unthrifty piglets, infertility and orchitis in the boar and these constitute serious economic losses (Ogundipe et al., 2001). Productivity of pigs in most countries is generally low due to high piglet mortality, slow growth rate,lack of adequate feed stuffs, ectoparasitism, trypanosomosis, helminthosis, lack of government' incentives, poor housing, lack of veterinary services and poor feed conversion ratio (Rekwot, 2003).This study was undertaken to determine the prevalence of Brucella infection in swine in southeast Nigeria.

돼지 brucellosis에 대한 항체가 조사

In order to investigate serum antibodies for detection of brucellosis in pigs, a total of 1208 sera were tested by Rose Bengal test (RBT), the standard tube agglutination test (STAT) and competitive ELISA (cELISA). The sera were collected from pigs of Gyeonggi, Chungnam, Chungbuk, Jeonnam and Jeonbuk, provinces during the period 2002 to 2004. All the sera were screened by RBT, and were confirmed by STAT and cELISA. Among 1208 sera, 26 sera (2.2%) were positive in screening test. All the 26 positive sera were positive by STAT, while all the sera were negative by cELISA. On the basis of this study, farmed pigs may be exposed to Brucella species. Furthermore, these results suggest that establishment of diagnoses for detection of porcine brucellosis is necessary.

A novel isolation method of Brucella species and molecular tracking of Brucella suis biovar 2 in domestic and wild animals

Brucella suis biovar 2 is the most common aetiological agent of porcine brucellosis in Europe. B. suis biovar 2 is considered to have low zoonotic potential, but is a causative agent of reproductive losses in pigs, and it is thus economically important. The multilocus variable-number of tandem repeats genotyping analysis of 16 loci (MLVA-16) has proven to be highly discriminatory and is the most suitable assay for simultaneously identifying B. suis and tracking infections. The aim of this study was to investigate the relatedness between isolates of B. suis biovar 2 obtained during a brucellosis outbreak in domestic pigs and isolates from wild boars and hares collected from proximal or remote geographical areas by MLVA-16. A cluster analysis of the MLVA-16 data revealed that most of the isolates obtained from Switzerland clustered together, with the exception of one isolate. The outbreak isolates constituted a unique subcluster (with a genetic similarity >93.8%) distinct from that of the isolates obtained from wild animals, suggesting that direct transmission of the bacterium from wild boars to domestic pigs did not occur in this outbreak. To obtain a representative number of isolates for MLVA-16, alternative methods of Brucella spp. isolation from tissue samples were compared with conventional direct cultivation on a Brucella-selective agar. We observed an enhanced sensitivity when mechanical homogenisation was followed by host cell lysis prior to cultivation on the Brucella-selective agar. This work demonstrates that MLVA-16 is an excellent tool for both monitoring brucellosis and investigating outbreaks. Additionally, we present efficient alternatives for the isolation of Brucella spp.

돼지에서 브루셀라병 항체조사 및 Yersinia enterocolitica O:9의 분리

Ten farrow-finish farms participated in this seromonitoring that was conducted to investigate the porcine brucellosis situation in Korea. In total, eight (80.0%) of the 10 farms and 139 (24.0%) of 578 pigs tested showed a positive response in the Rose Bengal test (RBT). Seroprevalence levels were determined using RBT according to age; 35 (14.6%) of 239 piglets, 36 (31.3%) of 115 growing pigs, and 68 (30.4%) of 224 finishing pigs and sows were positive, respectively. All positive samples in RBT were tested with the tube agglutination test (TAT) and competitive ELISA (C-ELISA), simultaneously. Although 48 samples came up positive in the TAT, all samples tested with C-ELISA were negative. Among 26 rectal swab samples from the TAT positive-pigs, Yersinia enterocolitica O:9 was isolated from seven samples (26.9%). Therefore, we speculated that the positive reaction of RBT and TAT in this study might be induced by the serologically cross-reacting bacteria with Brucella abortus.

Porcine brucellosis (Brucella suis)

Porcine brucellosis (Brucella suis)

Low reproductive performance and high sow mortality in a pig breeding herd: a case study

Sow performance is a key component of the productivity of commercial pig farms. Reproductive failure in the sow is common in pig production. For every 100 sows served, 89 should farrow. In absence of specific diseases such as porcine parvovirus, pseudo-rabies, swine fever, leptospirosis and brucellosis, management failures are the most important causes of loss. A syndrome associated with reproductive inefficiency, and post-service vaginal discharge and high sow mortality in a commercial pig farm is described. Pregnancy failures exceeded 20% and sow mortality exceeded 12% for two consecutive years. The abnormal post-service vaginal discharge rate was 1.7% during the period of investigation.An investigation involving an analysis of farm records, a review of breeding management practices, clinical examinations, laboratory analysis and examination of urogenital organs was conducted.The main contributing factors found were a sub-optimal gilt breeding management, an inadequate culling policy in combination with a sub-optimal culling rate and the presence of cystitis in more than 1% of the urogenital organs examined. The high sow mortality rate was related to an aged breeding herd.A control programme was recommended based on management changes involving oestrus detection, movement of gilts post-service, hygiene in the service area, boar exposure post-service and urinary acidification. This programme failed to increase the farrowing rate due to incomplete implementation of the recommendations made. The farrowing rate increased to 86.5% subsequent to a farm manager change in January 2005, which resulted in complete implementation of the control programme.

Improving the specificity of immunodiagnosis for porcine brucellosis

This report describes the use of cell mediated immunity to improve specificity of current diagnosis for Brucella suis. Diagnosis is problematic due to cross reactions that lead to false positive serological reactions (FPSR) in the standard diagnostic tests. A common cause of this cross reactivity is infection with the organism Yersinia enterocolitica O:9. Gottingen mini-pigs were experimentally infected with B. suis biovar I field strain or Y. enterocolitica serotype O:9 biotype 3. Infection was followed for 70 days. During this time whole blood stimulation assays were set up using Brucella specific antigen. IFNgamma was measured in the supernatants (SN) from these assays by ELISA. Concurrent standard serological tests were carried out. The results indicate that the IFNgamma assay is specifically able to distinguish Y. enterocolitica O:9 infection from a B. suis infection in experimentally infected mini-pigs. These results represent an improvement in diagnostic specificity compared to currently used serological tests. Thus suggesting that in a surveillance setting this test could be applied as a confirmatory test in the face of FPSR.

Serosurveillance for selected infectious disease agents in wild boars (Sus scrofa) and outdoor pigs in Switzerland

The large abundance of free-ranging wild boars (Sus scrofa) and a trend towards animal friendly outdoor management of domestic pigs lead to an increasing probability of disease transmission between those animal populations. In 2001, an active monitoring was started for classical swine fever (CSF), Aujeszky’s disease (AD) and porcine brucellosis (PB) in wild boars in Switzerland. The objective of this programme was to document the serological status of wild boars regarding the selected pathogens. To continue this serosurveillance, 1,060 wild boar samples were collected during two regular hunting seasons in 2004–2005. Furthermore, in a pilot study, 61 outdoor pigs from 14 farms located in areas with high wild boar densities were sampled in 2004 and serologically tested for AD and PB. All wild boar samples were negative for CSF. Seroprevalence for AD was 2.83% (95% CI 1.91–4.02%). Seroprevalence for PB was 13.5% (95% CI 10.7–16.7%) for the Rose Bengal test and 11.05% (95% CI 8.82–13.61%) for the indirect ELISA. There was no serological evidence for AD in domestic pigs. All tested animals from 13 piggeries were seronegative for PB, but three pigs from the same farm showed doubtful results. Further investigations on the farm did not indicate the presence of PB in the herd. These findings urge the need for better diagnostic tools to obtain reliable results concerning PB prevalence. Since contact and following transmission of infectious agents between infected wild boars and outdoor pigs might occur in the future, it is advisable to include outdoor pigs in areas at risk in routine surveillance programmes.

Les brucelloses humaine et animale en France en l'an 2000. Situation épidémiologique — programmes de contrôle et d'éradication

The epidemiological situation of human and animal brucellosis in 2000, and the control and surveillance programmes in place for domestic ruminants for more than 30 years in France are presented. Nowadays, human brucellosis is rare and the consumption of foreign dairy products has become a more important source of contamination. In cattle, sheep and goat populations, the national control programmes appear successful and the situation is at present close to eradication in these species. However, a new form of brucellosis has emerged in pigs bred extensively in open-air farms. Pigs are infected by Brucella suis biovar 2 and it is assumed that wild boars and maybe hares could be the reservoir. The public health consequences of porcine brucellosis in France are still not known. In other domestic and wild animal species, brucellosis remains accidental.

Validation of the fluorescence polarization assay as a serological test for the presumptive diagnosis of porcine brucellosis

Sera from Canadian pigs (brucellosis free, n = 14 037) and sera from pigs infected with Brucella suis ( n = 401) were tested by the buffered antigen plate agglutination test, the complement fixation test, an indirect and a competitive enzyme immunoassay and a fluorescence polarization assay. The results were analysed and assay sensitivity and specificity estimates were calculated. The sensitivity and specificity of the tests were as follows: the buffered antigen plate agglutination test, 77.1 and 96.9%; the complement fixation test (considering anticomplementary sera as negative), 93.3 and 95.5%; the complement fixation test (considering anticomplementary sera as positive), 58.1 and 99.9%; the indirect enzyme immunoassay, 94.0 and 97.9%; the competitive enzyme immunoassay, 90.8 and 96.6%; and the fluorescence polarization assay, 93.5 and 97.2%; respectively. It was concluded that the fluorescence polarization assay was a valuable asset to the diagnosis of porcine brucellosis because of its accuracy, ease of performance and relative cost.

Immunoblot studies in the differential diagnosis of porcine brucellosis: an immunodominant 62 kDa protein is related to the mycobacterial 65 kDa heat shock protein (HSP-65)

Smooth Brucella spp. share certain lipopolysaccharide antigens with other bacteria, resulting in serological cross-reactions which can prevent the definitive diagnosis of brucellosis. To identify other antigens with serodiagnostic potential, immunoblot studies following sodium dodecyl sulphate - polyacrylamide gel electrophoresis were carried out. Sera from pigs experimentally infected with Brucella suis and naturally infected feral pigs, sera from pigs from a farm with a known history of Yersinia enterocolitica 0:9 infection, Brucella Complement Fixation Test (CFT) reactor pigs (aetiology unknown) and pigs from consistently Brucella CFT negative farms were examined. Although B. suis infected pigs recognized a total of nine B. melitensis antigens, individual pigs rarely recognized more than three antigens in the range. A 62 kDa antigen was recognized by the majority (73%) of the Brucella infected pigs, but only by 10 to 23% of pigs from the other groups. This antigen was shown to be the Brucella homologue of the ubiquitous 65 kDa heat shock protein (HSP-65) family by immunoblot studies with 14 monoclonal antibodies to the Mycobacterium leprae HSP-65. Only four of these monoclones (Y1.2, ML-30, D7C and IIIC8) identified the B. melitensis 62 kDa protein suggesting that unshared, potentially Brucella specific, regions exist. Sera from Y. enterocolitica 0:9 infected pigs, CFT reactor pigs (aetiology unknown), CFT negative pigs and hyperimmune pig serum raised to Y. enterocolitica 0:9 also recognized B. melitensis antigens, most notably a 17 kDa protein. This antigen appears to be a common cross-reactive protein.