Proteomics-based screening and antibiotic resistance assessment of clinical and sub-clinical Brucella species: An evolution of brucellosis infection control.

Brucellae are intracellular sneaky bacteria and they can elude the host's defensive mechanisms, resulting in therapeutic failure. Therefore, the goal of this investigation was to rapid identification of Brucella species collected from animals and humans in Saudi Arabia, as well as to evaluate their resistance to antibiotics. On selective media, 364 animal samples as well as 70 human blood samples were cultured. Serological and biochemical approaches were initially used to identify a total of 25 probable cultured isolates. The proteomics of Brucella species were identified using the MALDI Biotyper (MBT) system, which was subsequently verified using real-time polymerase chain reaction (real-time PCR) and microfluidic electrophoresis assays. Both Brucella melitensis (B. melitensis) and Brucella abortus (B. abortus) were tested for antimicrobial susceptibility using Kirby Bauer method and the E-test. In total, 25 samples were positive for Brucella and included 11 B. melitensis and 14 B. abortus isolates. Twenty-two out of 25 (88%) and 24/25 (96%) of Brucella strains were recognized through the Vitek 2 Compact system. While MBT was magnificently identified 100% of the strains at the species level with a score value more than or equal to 2.00. Trimethoprim-sulfamethoxazole, rifampin, ampicillin-sulbactam, and ampicillin resistance in B. melitensis was 36.36%, 31.82%, 27.27%, and 22.70%, respectively. Rifampin, trimethoprim-sulfamethoxazole, ampicillin, and ampicillin-sulbactam resistance was found in 35.71%, 32.14%, 32.14%, and 28.57% of B. abortus isolates, correspondingly. MBT confirmed by microfluidic electrophoresis is a successful approach for identifying Brucella species at the species level. The resistance of B. melitensis and B. abortus to various antibiotics should be investigated in future studies.

Whole-genome sequencing of rough Brucella melitensis in China provides insights into its genetic features.

Brucella spp. can cause the zoonosis brucellosis, which affects public health and safety and even economic development. B. melitensis has a smooth phenotype, while 28 B. melitensis isolates had a rough phenotype in 2018. In this study, rough phenotype detection and whole genome sequencing methods were used to analyze the genetic features of rough B. melitensis. A drug susceptibility test was also performed. The results showed that the rough B. melitensis strains originated from strains isolated in China rather than from foreign strains. Furthermore, an MS tree showed that 9 complexes to be epidemic in China. For the rough B. melitensis strains, expression of the metabolic function genes varied in the earlier stages of evolution compared to the cellular process and signalling function genes. Expression of some transcriptional regulatory factors also varied in the later stages of evolution, and compared to MFS transporter genes, ABC transporter genes varied in the earlier stages. Moreover, as there was no significant difference in rifampicin, doxycycline and streptomycin susceptibility between the smooth and rough B. melitensis strains, treatment of brucellosis was not affected by strain type. This study provided important information for understanding the genetics and evolution of rough B. melitensis in China.

The cycle of distrust in health policy and behavior: Lessons learned from the Negev Bedouin.

BACKGROUND: Over the last decades, health systems worldwide have faced a decline in public trust. For marginalized minority populations, who generally suffer from poverty and political exclusion, the roots of this trend go much deeper, establishing a state of bi-directional distrust between them and health institutions. Although studied to a lesser extent compared to trust, distrust does impede health initiatives, such as infectious diseases prevention programs, mostly of so-called Neglected Zoonotic Diseases (NZDs). Where distrust prevails, even trust building actions such as defining rights and obligations, prioritizing "the greater good" and increasing transparency, are prone to failure. In this study, we deepen the understanding of the concept of distrust through a unique case study of Brucellosis, a prevalent bacterial zoonotic disease endemic to disadvantaged Bedouin communities in southern Israel. METHODS: In the years 2015-2019, we qualitatively studied socio-political aspects in a governmental Brucellosis control campaign in southern Israel. We used in-depth interviews with 38 governmental and private health workers, agriculture and nature preservation workers, livestock owners and community leaders. Further, we conducted participant observation in 10 livestock pens and in policymaking meetings, and collected policy and media documents in order to triangulate the results. RESULTS: We conceptualize three different types of distrust between authorities and marginalized communities-"intention-based distrust", "values-based distrust" and "circular distrust"-to better explain how distrust originates and reinforces itself, reproducing the endemicity of NZDs. Based on that, we portray a practical framework to reduce distrust in health policies, by reframing local discourses, reshaping disease monitoring schemes from enforcement-based to participation-based, and promoting political inclusion of disadvantaged communities. CONCLUSIONS: The suggested analysis and framework redirect health policy objectives to not only acknowledge, contain and reduce the consequences of distrust, but also to strive for societal justice as a tool for health promotion.

Preliminary Comparative Assessment of Brucellergene Skin Test for Diagnosis of Brucellosis in Dromedary Camels (Camelus dromedarius).

This study was conducted to evaluate the use of Brucellergene skin test (BST) for the diagnosis of Brucellosis in camels (Camelus dromedarius) in comparison with Rose Bengal test (RBT) and competitive enzyme-linked immunosorbent assay (c-ELISA). A total of 68 apparently healthy adult dromedary camels of either gender from three different geographical locations of Abu Dhabi Emirate, United Arab Emirates (UAE), were included in the study. The skin test was applied on two shaved areas at the middle of the neck: one for the test and the other area was injected with normal saline as a control. Reading was done 72 h postinjection. Results were subjected to Bayesian analysis to assess the test performances in camels. The model estimated the following sensitivity and specificity median values: BST: Se = 70.72%, Sp = 98.82%; RBT: Se = 93.27%, Sp = 97.79%; and c-ELISA: Se = 94.78%, Sp = 98.48%. As the BST investigated in this study proved to be a highly specific test, we propose using it as a confirmatory test in camels particularly when the serological tests give doubtful results on individual animals.

Brucellosis: A rapid risk assessment by a regional outbreak team and its coordinated response with the Directorate-General for Food and Veterinary, North region of Portugal, 2019.

We report a Brucella outbreak with seven cases in the Northern Region of Portugal in 2018-2019, associated with the consumption of fresh cheese. This outbreak has implications for risk assessment in Portuguese migrants related to this area, and it is an example of cooperation between public institutions, in a One Health based approach.

Status and influencing factors of farmers' private investment in the prevention and control of sheep brucellosis in China: A cross-sectional study.

BACKGROUND: Brucellosis is one of the most common zoonoses worldwide, causing direct losses to the livestock industry and threatening human health. Little is known about the status and factors affecting farmers' private investment in the prevention and control of sheep brucellosis in China. METHODOLOGY/PRINCIPAL FINDINGS: From April to October 2017, a cross-sectional, house-based study was conducted in 7 Chinese provinces. A total of 1037 households included in the study were analyzed. The average amount of private investment in the prevention and control of brucellosis was $0.73±0.54 per sheep. Multivariable analysis showed that factors facilitating private investment included older age of householder (OR = 1.07, 95%CI: 1.03-1.11), herd size >100 (OR = 2.49, 95%CI: 1.38-4.51), a higher percentage of income from sheep farming comparing to the total household income (OR = 1.14, 95%CI: 1.11-1.16), higher score of brucellosis knowledge (OR = 3.85, 95%CI: 1.40-10.51), actively learning related knowledge (OR = 2.98, 95%CI: 1.55-5.74), actively participating in related training courses (OR = 3.07, 95%CI: 1.52-6.18), care about other people's attitudes (OR = 1.75, 95%CI: 1.35-2.28), concern about the health of neighbors' livestock (OR = 1.75, 95%CI: 1.23-2.51). The analysis found a discouraging factor for private investment, supporting culling policy (OR = 0.67, 95%CI: 0.49-0.91). CONCLUSIONS/SIGNIFICANCE: In addition to providing interventions related to farmers' knowledge, attitudes and practices, guidance must be offered to help farmers understanding the importance of private investment in the prevention and control of brucellosis.

Emergency response to occupational brucellosis in a pharmaceutical manufacturing enterprise.

OBJECTIVES: Human brucellosis, as a zoonosis, is a serious public health problem in many developing countries, including China. To date, there has been no case report on occupational brucellosis and the emergency response in the pharmaceutical manufacturing industry. The aim of this report is to describe an emergency brucellosis outbreak in this unusual setting and the associated treatment methods from an occupational health perspective. METHODS: The emergency response included a field epidemiological investigation, a hierarchy of control strategy, and validation of control measures. RESULTS: Seven workers in the defrosting, grinding, and hydrolysis departments suffered from human brucellosis during two periods. The main symptoms were whole-body joint pain and undulant fever for several weeks, and Brucella-specific serum antibody was found in the patients. A hierarchy of exposure control strategy was implemented, including elimination, engineering control, administrative control, and the use of personal protection equipment. Raw material and environmental monitoring, measurement of placental temperature, Escherichia coli inactivation tests, and health examinations demonstrated that these measures were effective. CONCLUSIONS: This report describes an emergent case and the response to two consecutive outbreaks of occupational brucellosis among workers in a pharmaceutical manufacturing enterprise. The hierarchical control strategy used in this case may prevent new outbreaks, and several lessons can be learned from this event.

Brucella Infection in Asian Sea Otters (Enhydra lutris lutris) on Bering Island, Russia.

Infection with Brucella spp., long known as a cause of abortion, infertility, and reproductive loss in domestic livestock, has increasingly been documented in marine mammals over the past two decades. We report molecular evidence of Brucella infection in Asian sea otters (Enhydra lutris lutris). Brucella DNA was detected in 3 of 78 (4%) rectal swab samples collected between 2004 and 2006 on Bering Island, Russia. These 78 animals had previously been documented to have a Brucella seroprevalence of 28%, markedly higher than the prevalence documented in sea otters (Enhydra lutris) in North America. All of the DNA sequences amplified were identical to one or more previously isolated Brucella spp. including strains from both terrestrial and marine hosts. Phylogenetic analysis of this sequence suggested that one animal was shedding Brucella spp. DNA with a sequence matching a Brucella abortus strain, whereas two animals yielded a sequence matching a group of strains including isolates classified as Brucella pinnipedialis and Brucella melitensis. Our results highlight the diversity of Brucella spp. within a single sea otter population.

Assessment of community awareness and risk perceptions of zoonotic causes of abortion in cattle at three selected livestock-wildlife interface areas of Zimbabwe.

A study was conducted to assess the awareness of cattle abortions due to brucellosis, Rift Valley fever (RVF) and leptospirosis, and to compare frequencies of reported abortions in communities living at the periphery of the Great Limpopo Transfrontier Conservation Area in southeastern Zimbabwe. Three study sites were selected based on the type of livestock-wildlife interface: porous livestock-wildlife interface (unrestricted); non-porous livestock-wildlife interface (restricted by fencing); and livestock-wildlife non-interface (totally absent or control). Respondents randomly selected from a list of potential cattle farmers (N = 379) distributed at porous (40·1%), non-interface (35·5%) and non-porous (26·4%), were interviewed using a combined close- and open-ended questionnaire. Focus group discussions were conducted with 10-12 members of each community. More abortions in the last 5 years were reported from the porous interface (52%) and a significantly higher per cent of respondents from the porous interface (P < 0·05) perceived wildlife as playing a role in livestock abortions compared with the other interface types. The odds of reporting abortions in cattle were higher in large herd sizes (odds ratio (OR) = 2·6; 95% confidence interval (CI) 1·5-4·3), porous (OR = 1·9; 95% CI 1·0-3·5) and non-porous interface (OR = 2·2; 95% CI 1·1-4·3) compared with livestock-wildlife non-interface areas. About 21·6% of the respondents knew brucellosis as a cause of abortion, compared with RVF (9·8%) and leptospirosis (3·7%). These results explain to some extent, the existence of human/wildlife conflict in the studied livestock-wildlife interface areas of Zimbabwe, which militates against biodiversity conservation efforts. The low awareness of zoonoses means the public is at risk of contracting some of these infections. Thus, further studies should focus on livestock-wildlife interface areas to assess if the increased rates of abortions reported in cattle may be due to exposure to wildlife or other factors. The government of Zimbabwe needs to launch educational programmes on public health awareness in these remote areas at the periphery of transfrontier conservation areas where livestock-wildlife interface exists to help mitigate the morbidity and mortality of people from some of the known zoonotic diseases.

Colony-level assessment of Brucella and Leptospira in the Guadalupe fur seal, Isla Guadalupe, Mexico.

The relatively small population size and restricted distribution of the Guadalupe fur seal Arctocephalus townsendi could make it highly vulnerable to infectious diseases. We performed a colony-level assessment in this species of the prevalence and presence of Brucella spp. and Leptospira spp., pathogenic bacteria that have been reported in several pinniped species worldwide. Forty-six serum samples were collected in 2014 from pups at Isla Guadalupe, the only place where the species effectively reproduces. Samples were tested for Brucella using 3 consecutive serological tests, and for Leptospira using the microscopic agglutination test. For each bacterium, a Bayesian approach was used to estimate prevalence to exposure, and an epidemiological model was used to test the null hypothesis that the bacterium was present in the colony. No serum sample tested positive for Brucella, and the statistical analyses concluded that the colony was bacterium-free with a 96.3% confidence level. However, a Brucella surveillance program would be highly recommendable. Twelve samples were positive (titers 1:50) to 1 or more serovars of Leptospira. The prevalence was calculated at 27.1% (95% credible interval: 15.6-40.3%), and the posterior analyses indicated that the colony was not Leptospira-free with a 100% confidence level. Serovars Icterohaemorrhagiae, Canicola, and Bratislava were detected, but only further research can unveil whether they affect the fur seal population.

First evaluation of an influenza viral vector based Brucella abortus vaccine in sheep and goats: Assessment of safety, immunogenicity and protective efficacy against Brucella melitensis infection.

Previously we developed and evaluated a candidate influenza viral vector based Brucella abortus vaccine (Flu-BA) administered with a potent adjuvant Montanide Gel01 in cattle, which was found safe and highly effective. This study was aimed to establish a proof-of-concept of the efficacy of Flu-BA vaccine formulation in sheep and goats. We vaccinated sheep and goats with Flu-BA vaccine and as a positive control vaccinated a group of animals with a commercial B. melitensis Rev.1 vaccine. Clinically, both Flu-BA and Rev.1 vaccines were found safe. Serological analysis showed the animals received Flu-BA vaccine did not induce antibody response against Brucella Omp16 and L7/L12 proteins during the period of our study (56days post-initial vaccination, PIV). But observed significant antigen-specific T cell response indicated by increased lymphocyte stimulation index and enhanced secretion of IFN-γ at day 56 PIV in Flu-BA group. The Flu-BA vaccinated animals completely protected 57.1% of sheep and 42.9% of goats against B. melitensis 16M challenge. The severity of brucellosis in terms of infection index and colonization of Brucella in tissues was significantly lower in the Flu-BA group compared to negative control animals group. Nevertheless, positive control commercial Rev.1 vaccine provided strong antigen-specific T cell immunity and protection against B. melitensis 16M infection. We conclude that the Flu-BA vaccine induces a significant antigen-specific T-cell response and provides complete protection in approximately 50% of sheep and goats against B. melitensis 16M infection. Further investigations are needed to improve the efficacy of Flu-BA and explore its practical application in small ruminants.

Molecular assessment of the transplacental transmission of Toxoplasma gondii, Neospora caninum, Brucella canis and Ehrlichia canis in dogs.

Given the fact that numerous microbial species can be detected in pregnant female dogs, the objective of this study was to assess the transplacental transmission of Brucella canis, Ehrlichia canis, Neospora caninum and Toxoplasma gondii in stillborn puppies. This study involved 41 stillborn puppies, 78.6% of which were positive for T. gondii, 52.4% for N. caninum and 59.5% for B. canis. E. canis was not detected in any of the analyzed puppies. Pregnancy is an important physiological condition for the transmission of infectious agents to puppies and transplacental transmission may be epidemiologically relevant in the spread of these opportunistic agents.

Economic losses occurring due to brucellosis in Indian livestock populations.

Brucellosis is a serious public health issue in India. Estimation of economic losses occurring due to brucellosis is required to help formulate prevention and control strategies, but has not been done in India. We estimated economic losses due to brucellosis by sourcing prevalence data from epidemiological surveys conducted in India. Data for livestock populations were obtained from official records. Probability distributions were used for many of the input parameters to account for uncertainty and variability. The analysis revealed that brucellosis in livestock is responsible for a median loss of US $ 3.4 billion (5th-95th percentile 2.8-4.2 billion). The disease in cattle and buffalo accounted for 95.6% of the total losses occurring due to brucellosis in livestock populations. The disease is responsible for a loss of US $ 6.8 per cattle, US$18.2 per buffalo, US $ 0.7 per sheep, US $ 0.5 per goat and US $ 0.6 per pig. These losses are additional to the economic and social consequences of the disease in humans. The results suggest that the disease causes significant economic losses in the country and should be controlled on a priority basis.

Assessment of genetic diversity of zoonotic Brucella spp. recovered from livestock in Egypt using multiple locus VNTR analysis.

Brucellosis is endemic in most parts of Egypt, where it is caused mainly by Brucella melitensis biovar 3, and affects cattle and small ruminants in spite of ongoing efforts devoted to its control. Knowledge of the predominant Brucella species/strains circulating in a region is a prerequisite of a brucellosis control strategy. For this reason a study aiming at the evaluation of the phenotypic and genetic heterogeneity of a panel of 17 Brucella spp. isolates recovered from domestic ruminants (cattle, buffalo, sheep, and goat) from four governorates during a period of five years (2002-2007) was carried out using microbiological tests and molecular biology techniques (PCR, MLVA-15, and sequencing). Thirteen strains were identified as B. melitensis biovar 3 while all phenotypic and genetic techniques classified the remaining isolates as B. abortus (n = 2) and B. suis biovar 1 (n = 2). MLVA-15 yielded a high discriminatory power (h = 0.801), indicating a high genetic diversity among the B. melitensis strains circulating among domestic ruminants in Egypt. This is the first report of the isolation of B. suis from cattle in Egypt which, coupled with the finding of B. abortus, suggests a potential role of livestock as reservoirs of several zoonotic Brucella species in the region.

A comparative assessment of culture and serology in the diagnosis of brucellosis in dairy cattle.

To investigate the usefulness of culture for the confirmation of brucellosis in cattle, a comparison of culture and serology was undertaken on 248 animals in four dairy herds where the disease was active. Paired supramammary (SM), retropharyngeal (RP), and internal iliac (IL) lymph nodes were cultured, and five serological tests were deployed: the microserum agglutination test (MSAT), complement fixation test (CFT), the indirect (iELISA) and competitive ELISA, and the fluorescence polarisation assay (FPA). Brucella abortus was isolated from 86.8% of animals on combined culture of all three lymph nodes. Individually, the highest isolation rate was from the RP (90.5% of culture positives). Of culture positive animals, 13.7% and 6.2% were positive from the RP and SM alone, respectively. Approximately half of the positive cultures yielded <10 colonies/culture plate. Although 80.9% of animals were positive in at least one serological test, only 45.2% were positive in all five. For culture-positive animals, the MSAT was the most sensitive test (71.8%). Of the culture-negative animals 67.7% were positive in at least one test, while 12.9% were positive in all five. Titres were higher in animals culture-positive from the SM, and there was a direct correlation between higher titres and higher colony counts in SM cultures. Only 8.9% of animals were both culture-negative and seropositive (in at least one test), while 16.5% were culture-positive and seronegative in all five tests. The results highlight and validate the sensitivity of bacteriological culture in confirming a diagnosis of bovine brucellosis. While the MSAT and FPA were the most sensitive serological tests, a significant percentage of infected animals were undetectable using these standard serological assays.

Exact alpha-error determination for two-stage sampling strategies to substantiate freedom from disease.

Sampling strategies to substantiate freedom from disease are important when it comes to the trade of animals and animal products. When considering imperfect tests and finite populations, sample size calculation can, however, be a challenging task. The generalized hypergeometric formula developed by Cameron and Baldock (1998a) offers a framework that can elegantly be extended to multi-stage sampling strategies, which are widely used to account for disease clustering at herd-level. The achieved alpha-error of such surveys, however, typically depends on the realization of the sample and can differ from the pre-calculated value. In this paper, we introduce a new formula to evaluate the exact alpha-error induced by a specific sample. We further give a numerically viable approximation formula and analyze its properties using a data example of Brucella melitensis in the Austrian sheep population.

Evaluation of five serological tests for the diagnosis of porcine brucellosis in French Polynesia.

Porcine brucellosis due to Brucella suis biovar 1 raises important issues for pig breeders in French Polynesia. In this region, the disease is enzootic, spreads silently and engenders economic losses in infected farms as well as sporadic human cases. While serological tests are essential in surveillance and control programmes of animal diseases, to date none of the available tests have been shown to be reliable enough to be used as a gold standard in routine individual diagnosis of porcine brucellosis. Few studies about the estimation of the sensitivity and the specificity of porcine brucellosis screening tests have been published, none of them dealing with French Polynesia. The studied population included 1,595 pigs from French Polynesia. Five tests were evaluated: Rose Bengal test, fluorescence polarisation assay, indirect ELISA, and two competitive ELISAs (C-ELISA). The sensitivity and the specificity of each test were estimated. C-ELISA2 was the most sensitive test (Se C-ELISA2=0.954 [0.889; 0.992] 95% credibility interval (CrI)) while both C-ELISA and Rose Bengal test (RBT) were the most specific ones (Sp C-ELISA1=0.856 [0.806; 0.915] 95% CrI; Sp C-ELISA2=0.849 [0.817; 0.879] 95% CrI; Sp RBT=0.853 [0.812; 0.898] 95% CrI).

Cattle producers' economic incentives for preventing bovine brucellosis under uncertainty.

Cattle in the Greater Yellowstone Ecosystem occasionally contract bovine brucellosis from free-ranging elk and bison. Cattle producers use a variety of brucellosis prevention activities to reduce their herds' risk of contracting brucellosis, such as: (1) having state agency personnel haze elk off private land, (2) fencing haystacks, (3) administering adult booster vaccination, (4) spaying heifers, (5) altering the winter-feeding schedule of cattle, (6) hiring riders to prevent cattle-elk commingling, and (7) delaying grazing on high-risk allotments. Their brucellosis prevention decisions are complicated, however, by several sources of uncertainty, including the following: a cattle herd's baseline risk of contracting brucellosis, the inherent randomness of brucellosis outbreaks, the cost of implementing prevention activities, and the activities' effectiveness. This study eliminates one source of uncertainty by estimating the cost of implementing brucellosis prevention activities on a representative cow/calf-long yearling operation in the southern GYE. It then reports the minimum level of effectiveness each prevention activity must achieve to justify investment by a risk-neutral producer. Individual producers face different levels of baseline risk, however, and the US government's brucellosis-response policy is constantly evolving. We therefore estimate breakeven levels of effectiveness for a range of baseline risks and government policies. Producers, animal health experts, and policymakers can use this study's results to determine which brucellosis prevention activities are unlikely to generate sufficient expected benefits to cover their cost of implementation. Results also demonstrate the influence of government policy on producers' incentives to prevent brucellosis. Policies that increase the magnitude of economic loss a producer incurs when their herd contracts brucellosis subsequently decrease prevention activities' breakeven levels of effectiveness, and increase producers' incentives to implement those activities. Producers' incentives to implement prevention activities also increase as activities' costs decrease. Policymakers can easily adapt the results of this analysis to help target cost-share agreements to producers and prevention activities most likely to generate positive expected net benefits. Epidemiologists can also use our results to help prioritize future research on the technical effectiveness of various brucellosis prevention activities.

Assessment of the diagnostic sensitivity and specificity of an indirect ELISA kit for the diagnosis of Brucella ovis infection in rams.

BACKGROUND: Brucella ovis causes an infectious disease responsible for infertility and subsequent economic losses in sheep production. The standard serological test to detect B. ovis infection in rams is the complement fixation test (CFT), which has imperfect sensitivity and specificity in addition to technical drawbacks. Other available tests include the indirect enzyme-linked immunosorbent assays (I-ELISA) but no I-ELISA kit has been fully evaluated.The study aimed to compare an I-ELISA kit and the standard CFT. Our study was carried out on serum samples from 4599 rams from the South of France where the disease is enzootic. A Bayesian approach was used to estimate tests characteristics (diagnostic sensitivity, Se and diagnostic specificity, Sp). The tests were then studied together in order to optimise testing strategies to detect B. ovis. RESULTS: After optimising the cut-off values in order to avoid doubtful results without deteriorating the concordance between the results of the two tests, the I-ELISA appeared to be slightly more sensitive than CFT (Se I-ELISA=0.917 [0.822; 0.992], 95% Credibility Interval (CrI) compared to Se CFT=0.860 [0.740; 0.967], 95% CrI). However, CFT was slightly more specific than I-ELISA (Sp CFT=0.988 [0.947; 1.0], 95% CrI) compared to Sp I-ELISA =0.952 [0.901; 1.0], 95% CrI).The tests were then associated with two different interpretation schemes. The series association increased the specificity of screening and could be used for pre-movement testing in rams from uninfected flocks. The parallel association increased sequence sensitivity, thus appearing more suitable for eradicating the disease in infected flocks. CONCLUSIONS: The high sensitivity and acceptable specificity of this I-ELISA kit support its potential interest to avoid the limitations of CFT. The two tests could also be used together or combined with other diagnostic methods such as semen culture to improve the testing strategy. The choice of test sequence and interpretation criteria depends on the epidemiological context, screening objectives and the financial and practical constraints.

Assessment of performance of selected serological tests for diagnosing brucellosis in pigs.

Swine brucellosis due to Brucella suis is considered an emerging zoonotic disease whose control is based on serological testing and the subsequent culling of seropositive animals or the full depopulation of affected flocks. Here we assessed the performance of several serological tests (Rose Bengal Test [RBT], indirect ELISA [i-ELISA], blocking ELISA [b-ELISA], and two competitive ELISAs [c-ELISA]) for diagnosing swine brucellosis caused by B. suis biovar 2. Both frequentistic and Bayesian statistical inference were used. A frequentistic analysis, using sera from known gold standard (GS) populations (i.e., from truly infected or brucellosis free animals), resulted in maximum (100%) diagnostic sensitivity (Se) and specificity (Sp) in the RBT, i-ELISA and b-ELISA tests. However, c-ELISAs resulted in lower diagnostic Se (ranging from 68.5% to 92.6%, according to the different cut-offs selected). A Bayesian analysis of tests yielding the best diagnostic performance with GS sera (RBT, i-ELISA and b-ELISA), but using a large collection of field sera, resulted in similar Se among tests but markedly lower (≈ 80%) than that resulting from the frequentistic analysis using the GS serum populations. By contrast, the estimated Sp in the Bayesian analysis was only slightly lower than 100%, thus similar to that obtained frequentistically. Our results show that adequate diagnostic tests for brucellosis in swine are available, but also emphasize the need for more extensive validation studies before applying these tests under field conditions.