Optical mapping compendium of structural variants across global cattle breeds.

Structural variants (SV) have been linked to important bovine disease phenotypes, but due to the difficulty of their accurate detection with standard sequencing approaches, their role in shaping important traits across cattle breeds is largely unexplored. Optical mapping is an alternative approach for mapping SVs that has been shown to have higher sensitivity than DNA sequencing approaches. The aim of this project was to use optical mapping to develop a high-quality database of structural variation across cattle breeds from different geographical regions, to enable further study of SVs in cattle. To do this we generated 100X Bionano optical mapping data for 18 cattle of nine different ancestries, three continents and both cattle sub-species. In total we identified 13,457 SVs, of which 1,200 putatively overlap coding regions. This resource provides a high-quality set of optical mapping-based SV calls that can be used across studies, from validating DNA sequencing-based SV calls to prioritising candidate functional variants in genetic association studies and expanding our understanding of the role of SVs in cattle evolution.

Sero-prevalence of Bovine Brucellosis and associated risk factors in mbeya region, Southern highlands of Tanzania.

A cross-sectional study was conducted to establish the seroprevalence of brucellosis and associated risk factors in indigenous and exotic breeds of cattle from 178 farms in Mbeya region. A total of 1211 cattle (929exotic cattle from 108 commercial farms and 282 indigenous cattle from 70 traditional farms) were tested for Brucella antibodies using the Rose Bengal Plate Test (RBPT) and competitive Enzyme Linked Immunosorbent Assay (c-ELISA) as screening and confirmatory tests, respectively. The overall animal-level seroprevalence was 9.3%; 11.3% (95% CI: 9.4-13.5) in indigenous cattle and 2.8% (95% CI:1.4-5.6) in exotic cattle. Further, the overall herd level seroprevalence was 32.0%; 50.5% (95% CI: 40.9-59.9) in indigenous cattle and 4.2% (95% CI: 1.3-12.4) in exotic cattle. Infections were higher in cattle aged 6-10 years old, (39.8%; 95% CI: 31.2-49.1) followed by those aged 1-5 years (5.8%; 95% CI: 4.8-6.6) and 11-15years old (2.7%; 95% CI: 0.8-8). When compared to cattle sampled from herds size of 1-50, those sampled from the herd sizes of 51-100 and 101-150 had higher odds of brucellosis seropositivity [(OR=3.6, CI: 1.76-7.16, p<0.001) and (OR=3.0, CI: 1.09-8.04, p=0.033). The odds of seropositivity in animals which calved on pasture was 3.0 (CI: 1.1-7.8, p=0.028) compared to those that calved at home. Brucella seroprevalence was also observed to vary according to districts, with Mbarari district recording the highest (45.4%). It is evident from the study that Brucellosis is present in Mbarari, Mbeya and Momba districts of Mbeya Region. The findings of this study provide some baseline data that could contribute to the design and implementation of brucellosis control measures in the study areas.

Challenges and economic implications in the control of foot and mouth disease in sub-saharan Africa: lessons from the zambian experience.

Foot and mouth disease is one of the world's most important livestock diseases for trade. FMD infections are complex in nature and there are many epidemiological factors needing clarification. Key questions relate to the control challenges and economic impact of the disease for resource-poor FMD endemic countries like Zambia. A review of the control challenges and economic impact of FMD outbreaks in Zambia was made. Information was collected from peer-reviewed journals articles, conference proceedings, unpublished scientific reports, and personal communication with scientists and personal field experiences. The challenges of controlling FMD using mainly vaccination and movement control are discussed. Impacts include losses in income of over US$ 1.6 billion from exports of beef and sable antelopes and an annual cost of over US$ 2.7 million on preventive measures. Further impacts included unquantified losses in production and low investment in agriculture resulting in slow economic growth. FMD persistence may be a result of inadequate epidemiological understanding of the disease and ineffectiveness of the control measures that are being applied. The identified gaps may be considered in the annual appraisal of the FMD national control strategy in order to advance on the progressive control pathway.

Taenia spp. infections in wildlife in the Bangweulu and Kafue flood plains ecosystems of Zambia.

Taenia spp. have an indirect life cycle, cycling between a definitive and an intermediate host with zoonotic species causing public health problems in many developing countries. During the course of 2 separate surveys in Zambia (2004 and 2009), the presence of Taenia larval stages (cysticerci) was examined in Kafue lechwe (Kobus leche kafuensis), Black lechwe (Kobus leche smithermani) and other wildlife species from the Kafue and Bangweulu flood plains. Examinations involved post-mortem inspection and serum specific antigen detection. The recovered cysts from seven carcasses were characterised using PCR and DNA sequence analysis. The overall proportion of infection in wildlife on post-mortem examination was 19.0% (95% CI: 9.1-29.0%). The proportion of infected wildlife based on post-mortem examinations in the Kafue flood plains was estimated at 28.6% (95% CI: 13.3-43.9%), while the seroprevalence was estimated at 25.0% (95% CI: 2.9-47.1%). The seroprevalence for cattle in the Kafue flood plains was estimated at 61.5% (95% CI: 42.0-81.0%) while that of Kafue lechwe in the same ecosystem was estimated at 66.6% (95% CI: 45.6-85.7%). Infection rates were higher in Kafue lechwe than in Black lechwe suggesting differences in the exposure patterns. The sequencing results indicated that none of the recovered cysts were either Taenia solium or Taenia saginata. We therefore conclude they most likely belong to a less studied (wildlife) Taenia species that requires further characterisation.

A qualitative assessment of the risk of introducing peste des petits ruminants into northern zambia from Tanzania.

A qualitative risk assessment was performed to evaluate the risk of introducing Peste des petits ruminants virus into northern Zambia from Tanzania via live goat trade. Data was collected during a mission to Tanzania and northern Zambia and also from literature and interviews with experts. The risk of PPRV introduction was evaluated as a function of the probability of hazard (PPRV) release, exposure of susceptible hosts, and the consequences of spread using the following parameters: prevalence of infection, volume of trade, C-ELISA and quarantine screening missing an infected animal, PPRV viability (remaining infective) in transit, and the virus potential for infection. The magnitude of the consequences was derived from the probability of transmission and spread and the impact of PPRV introduction and establishment. Accordingly, the probability of occurrence of PPRV in northern Zambia from Tanzania was rated as "high" and the economic consequences were also rated as "high." Finally, the overall risk of introducing PPRV into northern Zambia from Tanzania at the time of the assessment was rated "high." It was concluded that import of goats and sheep be prohibited until efficient and adequate measures to reduce the risk have been put in place.

Bovine tuberculosis and brucellosis in traditionally managed livestock in selected districts of southern province of zambia.

A study was performed in 2008 to estimate the prevalence of tuberculosis and brucellosis in traditionally reared cattle of Southern Province in Zambia in four districts. The single comparative intradermal tuberculin test (SCITT) was used to identify TB reactors, and the Rose Bengal test (RBT), followed by confirmation with competitive enzyme-linked immunosorbent assay (c-ELISA), was used to test for brucellosis. A total of 459 animals were tested for tuberculosis and 395 for brucellosis. The overall prevalence of BTB based on the 4 mm and 3 mm cutoff criteria was 4.8% (95% CI: 2.6-7.0%) and 6.3% (95% CI: 3.8-8.8%), respectively. Change in skin thickness on SCITT was influenced by initial skin-fold thickness at the inoculation site, where animals with thinner skin had a tendency to give a larger tuberculin response. Brucellosis seroprevalence was estimated at 20.7% (95% CI: 17.0-24.4%). Comparison between results from RBT and c-ELISA showed good agreement (84.1%) and revealed subjectivity in RBT test results. Differences in brucellosis and tuberculosis prevalence across districts were attributed to type of husbandry practices and ecological factors. High prevalence of tuberculosis and brucellosis suggests that control programmes are necessary for improved cattle productivity and reduced public health risk.

Field evaluation of urine antigen detection for diagnosis of Taenia solium cysticercosis.

(Neuro)cysticercosis is an important zoonotic disease caused by infection with Taenia solium metacestode larvae. Existing immunodiagnostic techniques detect antibodies and circulating antigens (Ag) in serum and cerebrospinal fluid (CSF). Blood/CSF collection is an invasive procedure associated with blood-borne infections and is often not well accepted by communities. Detection of circulating Ag in urine has been suggested as an alternative, however this has been evaluated in clinical settings only. The aim of the present study was to evaluate the performance of a urine Ag-ELISA under field conditions. Paired serum and urine samples were obtained from participants in endemic areas of Ecuador (n=748) and Zambia (n=690) and were subjected to a monoclonal antibody-based Ag-ELISA. Calculation of positive and negative agreement indices (AI) showed better agreement in the negative direction both for Ecuadorian and Zambian samples (AI of 93.1 and 86.8, respectively). Using a Bayesian approach to determine the test characteristics, similar sensitivities were obtained for serum and urine Ag detection, whereas a decreased specificity was determined for the urine Ag-ELISA with a lower specificity (78.6%) for Zambian samples than for Ecuadorian samples (88.4%). This study indicates a higher specificity for the serum test under field conditions and promotes further research to improve the urine test.

Brucella seroprevalence of the Kafue lechwe (Kobus leche kafuensis) and Black lechwe (Kobus leche smithemani): exposure associated to contact with cattle.

We investigated Brucella seroprevalence in Kafue (Kobus leche kafuensis) and Black (Kobus leche smithemani) lechwe antelopes to assess Brucella infections in relation to presence/absence of cattle interaction on the wetlands. Accordingly, two study populations based on cattle interaction were assesed: Kafue lechwe from Kafue flats which interact with cattle; and the Black lechwe with no known interaction with cattle from the Bangweulu swamps. Fourteen Kafue lechwe and thirty Black lechwe were slaughtered between October and December 2009 using special research licenses obtained from the Zambia wildlife authority to investigate diseases in lechwe antelope. For the purpose of this study, blood was collected and sera separated for Rose Bengal and indirect ELISA tests. Seroprevalence of Brucella in the Kafue lechwe was estimated at 42.9% [95% CI: 15.2-70.5] while that in Black lechwe was 0% [95% CI:0.0-11.6]. On the Kafue flats, cattle were spotted grazing in the same areas as lechwe while there was no evidence of cattle presence on the Bangweulu swamps. These differences in seroprevalence between Kafue lechwe and Black lechwe were assumed to be associated with interaction between Kafue lechwe and Brucella infected cattle, and no such contact existed between cattle and the Black lechwe. Our study suggests that brucellosis in the Kafue lechwe may have originated from cattle but has now established a reservoir in wild animals. It is also important to keep in mind that the Black lechwe can easily become infected with Brucella spp. once cattle are introduced in the surrounding areas.

Brucellosis at the animal/ecosystem/human interface at the beginning of the 21st century.

Following the recent discovery of new Brucella strains from different animal species and from the environment, ten Brucella species are nowadays included in the genus Brucella. Although the intracellular trafficking of Brucella is well described, the strategies developed by Brucella to survive and multiply in phagocytic and non-phagocytic cells, particularly to access nutriments during its intracellular journey, are still largely unknown. Metabolism and virulence of Brucella are now considered to be two sides of the same coin. Mechanisms presiding to the colonization of the pregnant uterus in different animal species are not known. Vaccination is the cornerstone of control programs in livestock and although the S19, RB51 (both in cattle) and Rev 1 (in sheep and goats) vaccines have been successfully used worldwide, they have drawbacks and thus the ideal brucellosis vaccine is still very much awaited. There is no vaccine available for pigs and wildlife. Animal brucellosis control strategies differ in the developed and the developing world. Most emphasis is put on eradication and on risk analysis to avoid the re-introduction of Brucella in the developed world. Information related to the prevalence of brucellosis is still scarce in the developing world and control programs are rarely implemented. Since there is no vaccine available for humans, prevention of human brucellosis relies on its control in the animal reservoir. Brucella is also considered to be an agent to be used in bio- and agroterrorism attacks. At the animal/ecosystem/human interface it is critical to reduce opportunities for Brucella to jump host species as already seen in livestock, wildlife and humans. This task is a challenge for the future in terms of veterinary public health, as for wildlife and ecosystem managers and will need a "One Health" approach to be successful.

Evaluation of sensitivity and specificity of RBT, c-ELISA and fluorescence polarisation assay for diagnosis of brucellosis in cattle using latent class analysis.

The sensitivity (Se) and specificity (Sp) of the Rose Bengal test (RBT), competitive ELISA (c-ELISA), serum (sFPA) and blood (bFPA) fluorescence polarisation assay for brucellosis were evaluated using latent class analysis using sera and whole blood collected from infected cattle reared in smallholder dairy farms of Zimbabwe. The latent class model allowed estimation of Se and Sp in the absence of a gold standard test. The c-ELISA had the highest Se (99.0%; 95% credible posterior interval (CPI): 94.8; 100%), while the RBT and sFPA had the highest Sp (99.0%; 95% CPI: 98.0; 99.6%). The bFPA had the lowest Se (71.3%; 95% CPI: 56.2, 83.5%), while its Sp (96.3%; CPI: 93.9; 98.0%) was marginally higher than that of the c-ELISA (95.4% CPI: 93.7; 96.8%). Therefore based on these data, test regimen using the RBT and c-ELISA could be suitable for diagnosis of brucellosis in smallholder dairies in Zimbabwe. Based on cost and ease of performance, the sFPA may be adopted as a confirmatory test, but its performance may be optimised by altering cut-off points to suit the Zimbabwean conditions. Thus, latent class models provide an alternative method for evaluating Se and Sp of diagnostic tests, which could be used to optimise test performance in different cattle populations.

Rabies status in Zambia for the period 1985-2004.

Rabies has been present in Zambia since the early years of the 20th century. It is a significant public health problem in Zambia. Domestic dogs accounted for 69.7% (1348/1935) of the samples received for rabies diagnosis for the period 1985-2004. Of the 1069 positive cases confirmed by the fluorescent antibody test, 747 (69.9%) were from domestic dogs, 139 (13.0%) from cattle and 98 (9.2%) from humans. Wildlife samples accounted for 4.5% (87/1935) of the samples tested with the jackal (Canis adustus) being the predominant species. Cases of rabies were highest in Lusaka Province followed by the Copperbelt, Southern and Central Provinces. The monthly distribution of canine rabies showed an average of 2.93 (95% CI 2.59-3.29) dog positive cases per month. The study confirms that rabies is endemic in Zambia and that the domestic dog is the principal maintenance host. The epidemiology and control measures currently used in Zambia are herein discussed highlighting their limitations and successes. Based on the findings obtained from this study we advocate for strengthening the delivery of public health services and that steps must taken to reduce the incidence of rabies in Zambia.

Risk factors for Brucella spp. infection in smallholder household herds.

Risk factors for Brucella infection, the association and impact of Brucella seropositivity on abortions were investigated in cattle (n=1291) reared in smallholder household herds (n=203) from six geographical areas of Zimbabwe between September 2004 and 2005. Data on management, abortion and herd structure were collected. Sera were tested for Brucella antibodies using the Rose Bengal test and a competitive enzyme-linked immunosorbent assay. Data were analysed by generalized estimating equation and logistic regression models. Brucella antibodies were estimated at 5·5% and 22·9% for individual cattle and herds, respectively. Abortions were reported in 3·2% of cows and 22·0% herds. The age of cows and Brucella seropositivity predicted abortion. For herds, Brucella seropositivity, geographical area, purchase of cattle and large herd size were independently associated with increased odds of abortion. Exposure to Brucella had a significant impact on abortion. These results highlight the important risk factors for Brucella spp. infection in smallholder herds. Thus, brucellosis control programmes which take these factors into consideration will be beneficial.

Helminth parasites of the Kafue lechwe antelope ( Kobus leche kafuensis): a potential source of infection to domestic animals in the Kafue wetlands of Zambia.

The Kafue lechwe antelope (Kobus leche kafuensis), a medium-sized, semi-aquatic antelope, grazes extensively on pastures accessed by livestock in and around Lochinvar and Blue Lagoon national parks in the Kafue wetlands of Zambia. This interaction has a potential for bi-modal transmission of a wide range of parasitic helminths between lechwe and domestic ruminants. A survey was conducted to investigate the status of helminths in the Kafue lechwe during the 2008 (July-December) hunting season, involving 65 animals hunted under special research licences. Worm identification was based on morphological features using standard identification keys. Eleven different types of helminths were identified in the animals studied; namely, Oesophagostomum, Bunostomum, Cooperia, Dictyocaulus, Marshallagia, Stilesia, Setaria, Trichuris, Fasciola, amphistomes and Schistosoma. Amphistomes (100%) and Oesophagostomum (60.9%) were the most common while Fasciola (7.8%) and Stilesia (1.6%) were the least of the identified helminths. There was no evidence that helminths, at intensities observed, adversely affected the health of the lechwe. The degree of worm infection was observed to vary between the two study areas, with Blue Lagoon recording higher infection levels compared to Lochinvar. The host range of many of the helminths found in the Kafue lechwe is broad and could serve as a potentially stable source of infection to domestic animals such as goats and cattle. Therefore, issues concerning livestock management and conservation may arise.

Tuberculosis in Kafue lechwe antelopes (Kobus leche Kafuensis) of the Kafue Basin in Zambia.

Tuberculosis (TB) has been reported in the Kafue lechwe antelopes (Kobus leche Kafuensis) of Zambia. However, previous reports are restricted to the southern parts in Lochinvar, where only old male animals were investigated. This study was conducted to gather epidemiological information on TB in Lechwe antelopes across sexes and age groups in relation to other explanatory variables of disease occurrence in the Kafue Basin. Animals were hunted under a special licence to investigate diseases in the Kafue Basin during the 2004, 2005 and 2008 hunting seasons. Histopathology, acid-fast staining and mycobacterial culturing from tissue samples were conducted. A total of 119 animals were slaughtered with an estimated age range of 2.5-20 years. Of these, 29 (24.3% [95% CI: 16.5, 32.3%]) had necropsy lesions suggestive of tuberculosis, of which 21 (17.6% [95% CI: 10.7, 24.6%]) tested positive on acid-fast staining while 33 (27.7% [95% CI: 19.6, 35.9%]) showed culture and colony morphological characteristics suggestive of Mycobacterium species. On univariate analysis, animals with poor body condition were twice as likely to have tuberculosis associated lesions as those having good body conditions (OR=2.3, 95% CI: 0.6, 9.3%). Based on lesion distribution, a respiratory route of mycobacterial infection is intimated.

Herd-level factors for Brucella seropositivity in cattle reared in smallholder dairy farms of Zimbabwe.

A cross-sectional study was conducted to investigate factors for Brucella seropositivity in smallholder dairy cattle herds from Gokwe, Marirangwe, Mushagashe, Nharira, Rusitu and Wedza areas located in different agro-ecological regions of Zimbabwe between September 2004 and November 2005. Sera were collected from cattle aged > or = 2 years from 203 herds. Data on herd-level and management variables were collected using a structured questionnaire. Sera were screened for anti-Brucella antibodies using the Rose Bengal test (RBT) and confirmed by competitive ELISA (c-ELISA). A herd was classified as Brucella seropositive if at least one animal tested seropositive on both tests. The herd-level factors for Brucella seropositivity were tested using multivariable logistic model with herd infection status as dependent variable while the levels of exposure in individual animals within-herds were analysed by negative binomial regression using the number of positive animals as the outcome. Of the 203 herds tested, 52 (25; 95% CI: 18.1, 31.9%) tested seropositive for brucellosis. Using the logistic regression model we identified area, with both Rusitu (OR=0.26; 95% CI: 0.07, 1.03) and Wedza (OR=0.07; 95% CI: 0.01, 0.49) having lower Brucella seropositivity compared to Gokwe. Keeping mixed cattle breeds (OR=8.33; 95% CI: 2.70, 25.72) compared to single breed herds, was associated with increased herd seropositivity. The farmer's knowledge of brucellosis was associated with lower odds (OR=0.17; 95% CI: 0.05, 0.55) of farms testing seropositive. The odds of Brucella seropositivity were progressively higher with increasing stocking density and herd size. Using the negative binomial regression model we identified area, keeping mixed breed herds, stocking density and herd size as independently associated with increased counts of seropositive cattle in a herd.

Prevalence of bovine tuberculosis and animal level risk factors for indigenous cattle under different grazing strategies in the livestock/wildlife interface areas of Zambia.

A cross-sectional study was conducted to investigate the prevalence and animal level risk factors for bovine tuberculosis (BTB) in indigenous cattle of the livestock/wildlife interface areas in Zambia. A total of 944 cattle from 111 herds were investigated. The comparative intradermal tuberculin test (CIDT) was used to identify reactor animals for BTB. Animal level data on sex, age, parity and body condition score were registered. The overall animal prevalence of BTB as determined by the CIDT was 6.8% (95% CI: 4.2, 9.5%). In Lochinvar and Blue Lagoon areas, animal level prevalence were observed at 5.2% (95% CI: 2.2, 8.2%) and 9.6% (95% CI: 6.1, 13.2%), respectively. Kazungula, an area outside the livestock/wildlife interface, had a prevalence of only 0.8% (95% CI: 0.0, 2.3%). The age of the animal, its body condition score and the type of management system, were predictive of its BTB status. The study revealed that BTB was relatively high in the livestock/wildlife interface areas of Lochinvar and Blue Lagoon compared to Kazungula. These findings should raise a serious public health concern considering the extent to which the communities of the study areas are in contact with their animals and the levels at which they use untreated milk.

Effectiveness of Rose Bengal test and fluorescence polarization assay in the diagnosis of Brucella spp. infections in free range cattle reared in endemic areas in Zambia.

The effectiveness of Rose Bengal test (RBT) and fluorescence polarization assay (FPA) in diagnosing cattle brucellosis in endemic areas was assessed and RBT and FPA test agreement was compared (n = 319). The sensitivity of RBT and FPA in detecting low Brucella titres were evaluated in paired sera (n = 34). A logistic regression model was constructed to predict cattle test result in FPA using RBT as the main predictor and incorporating bio-data and animal history. There was 79.3% agreement between the RBT and FPA (Kappa = 0.59; Std error = 0.05; p = 0.000) and a high correspondence between high RBT scores and positive FPA results suggesting that sera with high RBT score may not require confirmation with tests such as competitive-ELISA or CFT. High FPA cut-off points were more likely to miss animals with low antibody titres. The RBT had a reduced ability in detecting low antibody titres compared to the FPA. FPA test interpretation was improved if a priori information, such as sex and age was used. Under the challenging disease surveillance conditions prevailing in rural Africa, field-testing methods that are sensitive and specific; allow single animal contact, low technical skills in data interpretation are suitable.

Mortality and commercial off-take rates in adult traditional cattle of Zambia.

A cohort study was conducted in the livestock-wildlife interface areas of Zambia to determine cattle mortality and commercial off-take rates among adult cattle as well as factors influencing them. A total of 416 animals from 43 herds were followed up for one year and animals were individually identified and their fate was indicated as sold, slaughtered, dead or present as appropriate. The overall mortality incidence risk was estimated at 7.5%. Cattle in Kazungula were at a greater risk of dying compared to those in Blue Lagoon and Lochnivar. Annual off-take was estimated at 13.7% (8.1-19.3%), unadjusted values, and 16.4% (8.1-24.5%) after adjusting for sampling fraction in primary sampling units (herds) and area stratification. Area variations were observed with Kazungula recording the highest in both instances, which was attributed to a contagious bovine pleural pneumonia (CBPP) outbreak. Herd size and gender were observed to influence cattle mortality rates. Cattle in the middle-sized herds (50-150 cattle) recorded high mortality rates (OR = 3.91) compared to smaller herds (10-50) and so were females compared to males (OR = 4.16). The logistic regression model showed that cattle death was influenced by managerial factors and that off-take rates tend to increase in the face of disease outbreaks.

Risk factors associated with bovine tuberculosis in traditional cattle of the livestock/wildlife interface areas in the Kafue basin of Zambia.

We conducted a cross-sectional study from August 2003 to February 2004 to identify risk factors for bovine tuberculosis (BTB) in the Kafue basin of Zambia. We investigated a total of 106 herds of cattle for presence of BTB using the comparative intradermal tuberculin test (CITT) while an interviewer-administered questionnaire was used to gather epidemiological data on herd structure, management and grazing strategies. BTB prevalence at herd level was estimated and possible risk factors were investigated using the multiple logistic regression model. The true herd level prevalence of BTB was estimated at 49.8% (95% CI: 37.9, 61.7%). The logistic regression model showed that cattle herd BTB status was highly associated with area and husbandry practices. When compared to Kazungula, cattle herds in Blue Lagoon were more likely to test positive for BTB when other factors such as management practices were controlled (OR=10.5). In terms of grazing strategies, transhumant herds (TH) had higher odds (OR=3.0) of being positive compared to sedentary herds (OR=1.0). The results in this study provide preliminary information about potential risk factors that were found to be associated with BTB status in cattle.

Evaluation of three serological tests for brucellosis in naturally infected cattle using latent class analysis.

Serological methods are traditionally used in diagnosis of brucellosis. However, the comparative performance of these tests and their accuracy under the local environment in Zambia has not been assessed. Thus, the objective of our study was to evaluate the diagnostic performance of three serological tests for brucellosis; Rose Bengal Test (RBT), competitive ELISA (c-ELISA) and Fluorescence Polarisation Assay (FPA) in naturally infected cattle in Zambia without an appropriate reference test to classify animals into truly infected and non-infected. Serological test results from a study to determine sero-prevalence were used to compare the performance of RBT, c-ELISA and FPA in diagnosing brucellosis in traditional cattle. Since none of the tests can be seen as a perfect reference test or gold standard, their performance in a population of naturally infected cattle was evaluated using latent class analysis which allows the sensitivity (Se) and specificity (Sp) to be estimated in the absence of a gold standard. The highest Se was achieved by the c-ELISA (97%; Credible Posterior Interval (CPI)=93-100%) and the highest Sp by the FPA (93%; CPI=85-99%), conversely these tests also had the lowest Sp and Se, respectively, with the RBT performing well in both the Se (93%; CPI=84-98%) and Sp (81%; CPI=61-97).