A simulation model of brucellosis spread in British cattle under several testing regimes

Brucellosis is a widespread, economically devastating and highly infectious zoonosis. In cattle, infection predominantly is caused by Brucella abortus, and is usually detected in pregnant females through abortions. Great Britain (GB) has been declared free from brucellosis (officially brucellosis free (OBF)) since 1993 and as such is required by European Union (EU) regulations to test > or =20% of both beef and dairy cattle >24 months old routinely. Currently, however, GB serologically tests more cattle than required and the issue of reducing the level of testing has come under consideration. We developed a simulation model to determine the rate of spread of brucellosis under a variety of testing regimes. For dairy herds, we found that reducing the level of testing would have a major effect on the rate of spread of infection, should it be imported. For beef herds, reducing the level of testing would have much less effect. We also found that abortion notification is a very-important additional means of surveillance. As a result of our predictions, policy-makers decided not to reduce the level of testing and actively to promote abortion notification.     

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

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

An all-island approach to mapping bovine tuberculosis in Ireland

This study used techniques in Geographical Information Systems (GIS) to explore the spatial patterns of bovine tuberculosis (TB) in the whole island of Ireland over an 11-year period. This is the first time that data pertaining to TB from the Republic of Ireland and Northern Ireland have been collated and examined in an all-Ireland context. The analyses were based on 198, 156 point locations representing active farms with cattle in Northern Ireland and the Republic of Ireland between the years 1996 and 2006. The results consist of a series of maps giving a visual representation of cattle populations and associated detected bTB levels on the island of Ireland over this time interval.     

Relative effectiveness of irish factories in the surveillance of slaughtered cattle for visible lesions of tuberculosis, 2005-2007

Background

In Ireland, every animal is examined at slaughter for its fitness for human consumption. The aim of this study was to determine the relative effectiveness of factories in submitting and subsequently in having suspect lesions confirmed as bovine tuberculosis (TB) lesions during the years 2005-2007. This work provides an update from previously published data for years 2003-2004. During 2005-2007 data were available on 4,401,813 cattle from attested herds (i.e. herds classified free of bovine TB), from which data for potential confounding factors were available for 3,344,057 slaughtered animals at one of the 37 export-licensed factories.

Findings

From these animals, 8,178 suspect lesions were submitted for laboratory confirmation. Lesions from 5,456 (66.7%) animals tested as positive, and 269 (3.2%) were inconclusive for bovine TB. Logistic regression was used to determine adjusted submission and confirmation risks for each factory while controlling for confounding factors. Factory rankings based on adjusted and crude risks were similar. The average crude submission risk for all the factories was 25 lesions per 10,000 animals slaughtered, ranging from 0 to 52. The crude confirmation risk varied between 30.3% and 91.3%.

Conclusions

Substantial variation in the effectiveness of lesion submission and subsequent confirmation as bovine TB was found among the 37 factories. Compared to previous years (2003-2004), there was an increased bovine TB lesion submission and confirmation risk. Continued monitoring of the effectiveness of slaughter surveillance in Ireland is recommended; emphasis should be placed on efforts to improve bovine TB surveillance in factories with lower rankings.     

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.     

Cattle transfers between herds under paratuberculosis surveillance in The Netherlands are not random

The rate and structure of cattle transfers between 206 Dutch cattle herds with a ‘Mycobacterium avium subsp. paratuberculosis (Map)-free’ status by November 2002, were analyzed over a 3-year period (November 1999–November 2002). Of the 206 ‘Map-free’ herds, 184 were closed herds during the period studied. In total, 280 cattle had been introduced into 22 herds at an average rate of 0.33 animals per year per 100 cattle present in the 206 herds. Assuming a random herd-contact structure, the observed rate of cattle transfers between certified ‘Map-free’ herds was sufficiently low to relax the surveillance scheme to biennial herd examinations by pooled fecal culture of all cattle ≥2 years of age.

The cattle transfers were not randomly distributed over the herds. Forty-four of the 280 cattle originated from 12 other ‘Map-free’ herds. The other 236 cattle did not originate from a ‘Map-free’ herd and were introduced into a herd before it obtained the ‘Map-free’ status. No cattle were introduced into any of the ‘Map-free’ herds from which cattle were transferred to other ‘Map-free’ herds. Thus, continued propagation of the infection by cattle transfers was impossible in the group of herds studied during the study period. Therefore the surveillance scheme may be further relaxed, and may be differentiated regarding the risk herds pose to other herds.     

Seroprevalence of chlamydial infection in cattle in Ireland

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

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.     

Cattle brucellosis in traditional livestock husbandry practice in Southern and Eastern Ethiopia,and its zoonotic implication

Background

Cattle brucellosis has significant economic and zoonotic implication for the rural communities in Ethiopia in consequence of their traditional life styles, feeding habits and disease patterns. Hence, knowledge of brucellosis occurrence in traditional livestock husbandry practice has considerable importance in reducing the economic and public health impacts of the disease.

Methods

A total of 1623 cattle sera were serially tested using the rose Bengal test as screening and complement fixation test as confirmatory tests. The Stata survey command was used to establish prevalences for the overall and individual variables, while potential risk factors for seropositivity were analyzed using a multivariable logistic regression analysis.

Results

The results showed that 3.5% (95% CI = 2.4, 4.5%) of the animals and 26.1% (95% CI = 18.6, 33.7) of the herds tested had antibodies against Brucella species. Village level seroprevalence ranged from 0% to 100%. A higher seroprevalence was observed in pastoral system than mixed farming although this variable was not significant in the final model. The final logistic regression model identified herd size; with large (odd ratio (OR) = 8.0, 95% CI = 1.9, 33.6) and medium herds (OR = 8.1, 95% CI = 1.9, 34.2) showing higher risk of Brucella infection when compared to small herds. Similarly, the odds of Brucella infection was higher in cattle aged above 4 years when compared to age groups of 1-2 (OR = 5.4, 2.1, 12.9) and 3-4 years (OR = 3.1, 95% CI = 1.0, 9.6). Herd level analysis of the risk factors revealed that large and medium herds as well as herds kept with multiple livestock species were at higher risk of acquiring Brucella infection. Brucellosis in traditional livestock husbandry practices certainly poses a zoonotic risk to the public, in consequence of raw milk consumption, close contact with animals and provision of assistance during parturition. Due to lack of diagnostic facilities and information on its occurrence, human brucellosis is most likely misdiagnosed for other febrile diseases prevailing in the areas and treated empirically.

Conclusions

The results of this study demonstrated that bovine brucellosis is widely prevalent in the study areas particularly in pastoral production system. Hence, the study suggests the need for implementing control measures and raising public awareness on prevention methods of brucellosis.     

The Animal and Plant Health Inspection Service (APHIS) brucellosis eradication program in the United States

Efforts to eradicate brucellosis caused by Brucella abortus in the United States began in 1934 as part of an economic recovery program to reduce the cattle population because of the Great Depression and concurrent severe drought conditions. A number of states saw this as an opportunity to reduce the level of brucellosis, which was the most significant livestock disease problem in the US at the time. In 1934 and 1935, the reactor rate in adult cattle tested was 11.5%. In 1954, the magnitude of the brucellosis problem in the United States in terms of economics to the cattle industry and human health prompted Congress to appropriate funds for a comprehensive national effort to eradicate brucellosis. The brucellosis eradication program was designed as a cooperative effort between the federal government, the states, and livestock producers. As the science and technology of brucellosis has developed over the years through research and experience, the eradication program has been modified many times.

As of 31 December 2000, there were no affected cattle herds in the United States. This was the first time in the history of the brucellosis program that the United States had no known brucellosis affected herds. However, brucellosis has a variable, sometimes quite lengthy incubation period, so it is expected that additional affected herds will be disclosed. It is likely that additional affected herds will be disclosed before brucellosis is finally eradicated from cattle. Animal health officials remain prepared to aggressively pursue any newly disclosed affected herds to eliminate the disease as quickly as possible.

The State-Federal Brucellosis Eradication Program has made tremendous progress since its inception. In an eradication program, it is critically important to recognize that, despite all the tools that are available to eliminate the disease, an effective surveillance system is the critical first step that must be in place in order to be successful. It is imperative, not only to be able to find the disease and eliminate it, but to find it before it spreads to susceptible herds. When brucellosis can be identified, contained, and eliminated before spread occurs, eradication can be achieved.     

An evaluation of Irish cattle herds with inconclusive serological evidence of bovine brucellosis

Since 1998, there has been a steady decline in herd restrictions and de-populations in Ireland due to bovine brucellosis. There is concern that the interpretation of laboratory results may become increasingly problematic, as brucellosis prevalence falls in Ireland. Therefore, the purpose of the current study was to evaluate the infection status of Irish herds and animals with inconclusive serological evidence of bovine brucellosis. During 12 months from September 1, 2004, laboratory and observational epidemiological data were collected from all Irish herds where animal testing identified at least one animal with a complement fixation test (CFT) reading greater than zero and/or a positive result to the indirect enzyme-linked immunosorbent assay (iELISA). Due to the observational nature of the study, we have robust estimates of the relative, but not the absolute, performance of the CFT, iELISA and brucellin skin test (BST). Herds were divided into three categories (Group A, B or C) on the basis of test results at initial assessment. A total of 639 herds were enrolled into the study, and observed for at least two years following enrolment. A rising CFT titre, with a CFT reading of 111 International CFT Units (IU) or greater at the subsequent blood test, was generally associated with herds where other evidence of infection was also available. Knowledge of the CFT reading at the initial and a subsequent blood test proved useful in distinguishing false-positive and true-positive brucellosis results. There was poor correlation between the CFT and iELISA results, and between the CFT and BST results. As a result of this study, national policy has been modified to include re-sampling of all animals with CFT readings of 20 IU or greater. This project has also led to a reduction in the number of herds restricted, as well as restriction duration. It has also contributed to a reduction in the number of herds listed for contiguous tests, and therefore the potential for contiguity testing of false positive results.     

Potential infection-control benefit for Ireland from pre-movement testing of cattle for tuberculosis

Pre-movement testing for bovine tuberculosis (BTB) was compulsory in Ireland until 1996. We determined the proportion of herd restrictions (losing BTB-free status) attributable to the recent introduction of an infected bovid; described events between restoration of BTB-free status (de-restriction) and the next herd-level test for BTB; estimated the proportion of undetected infected cattle present at de-restriction; identified high-risk movements between herds (movements most likely to involve infected cattle); and determined the potential yield of infected cattle discovered (or herds that would not lose their BTB-free status) by pre-movement testing, relative to the numbers of cattle and herds tested. We used national data for all 6252 herds with a new BTB restriction in the 12 months from 1 April 2003 and 3947 herds declared BTB-free in the 12 months from 1 October 2001. We identified higher-risk animals from our logistic generalized estimating-equation models. We attributed 6-7% of current herd restrictions to the recent introduction of an infected animal. There were considerable changes to herd structure between de-restriction and the next full-herd test, and infection was detected in 10% of herds at the first assessment (full-herd test or abattoir surveillance) following de-restriction. Following movement from a de-restricted herd, the odds of an animal being positive at the next test increased with increasing time in the source herd prior to movement, increasing time between de-restriction and the next full-herd test and increasing severity of the source herd restriction. The odds decreased with increasing size of the source herd. We estimated that 15.9 destination-herd restrictions per year could be prevented for every 10,000 cattle tested pre-movement and that 3.3 destination-herd restrictions per year could be prevented for every 100 source herds tested pre-movement. The yield per pre-movement test can be increased by focusing on high-risk movements; however, this would result in a substantial decrease in the total number of potential restrictions identified.     

Detection of brucellosis in diary herds after an outbreak of the disease using a delayed-type hypersensitivity test

A study of 14 herds was conducted to assess the skin delayed-type hypersensitivity (SDTH) test used in combination with the serum agglutination test and complement fixation test to curtail the spread of brucellosis. The tests were used to screen dairy herds suspected of brucellosis, and herds located near infected herds. The herds were tested every 5–6 weeks for 6 months. Cattle with a palpable skin swelling at the injection site of the allergen were considered as brucellosis suspects and were slaughtered. Supramammary lymph nodes of these cattle were collected and examined bacteriologically to determine their infection status.

Brucellosis was detected and confirmed bacteriologically in three seronegative herds in which cattle tested positive with the SDTH test. Brucellosis was also confirmed bacteriologically in five seropositive herds in which cattle tested positive with the SDTH test. Brucellosis was not confirmed bacteriologically in one seronegative and four seropositive herds in which cattle reacted positively with the SDTH test. One herd reacted negatively in all tests. Because the SDTH test is able to detect latent carriers of Brucella organisms in seronegative herds, we recommend that the SDTH test be used in combination with serologic tests to curtail the spread of brucellosis.     

The effects of warble fly dressing using an organophosphorus compound on bovine pediculosis

The added benefit of the national campaign in the Republic of Ireland to eradicate bovine warble flies using pour-on organophosphorus compounds annually, has been a significant reduction in the degree of pediculosis there compared to that in Northern Ireland. In Northern Ireland, only warbled cattle are dressed and the level of pediculosis was 72.8% compared to 56.2% in the Republic, 30.4% of 250 cattle examined in the former country had moderate to heavy infestation compared to 8.6% of 185 cattle examined in the latter. Four species of cattle lice were found and reported for the first time in both countries; these were Haematopinus eurysternus, Solenoptes capillatus, Linognathus vituli, and Bovicola bovis. The first named was far more common in Northern Ireland. Following the withdrawal of the national eradication scheme in the Republic in 1975, there are indications that pediculosis has increased in cattle. Of 100 adult cattle examined in the Republic in February, 1976, 19 had heavy infestations, whilst 78 were infested.     

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.     

Risk of tuberculosis cattle herd breakdowns in Ireland: effects of badger culling effort,density and historic large-scale interventions

Bovine tuberculosis (bTB) continues to be a problem in cattle herds in Ireland and Britain. It has been suggested that failure to eradicate this disease is related to the presence of a wildlife reservoir (the badger). A large-scale project was undertaken in the Republic of Ireland during 1997–2002 to assess whether badger removal could contribute to reducing risk of cattle herd breakdowns in four areas. During the period of that “four area” study, there was a significant decrease in risk in intensively culled (removal) areas relative to reference areas. In the present study, we revisit these areas to assess if there were any residual area effects of this former intervention a decade on (2007–2012). Over the study period there was an overall declining trend in bTB breakdown risk to cattle herds. Cattle herds within former removal areas experienced significantly reduced risk of breakdown relative to herds within former reference areas or herds within non-treatment areas (OR: 0.53; P < 0.001). Increased herd breakdown risk was associated with increasing herd size (OR: 1.92-2.03; P < 0.001) and herd bTB history (OR: 2.25-2.40; P < 0.001). There was increased risk of herd breakdowns in areas with higher badger densities, but this association was only significant early in the study (PD*YEAR interaction; P < 0.001). Badgers were culled in areas with higher cattle bTB risk (targeted culling). Risk tended to decline with cumulative culling effort only in three counties, but increased in the fourth (Donegal). Culling badgers is not seen as a viable long-term strategy. However, mixed policy options with biosecurity and badger vaccination, may help in managing cattle breakdown risk.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0109-4) contains supplementary material, which is available to authorized users.     

Quantification of the relative efficiency of factory surveillance in the disclosure of tuberculosis lesions in attested Irish cattle

In Ireland, factory surveillance of cattle for gross lesions is an important supplementary method for detecting herds infected with bovine tuberculosis (tb), and in recent years between 27 and 46 per cent of all new herd breakdowns in any year have been detected by this method. The aim of this study was to determine the relative efficiency of factories in detecting lesions among attested cattle slaughtered during 2003 and 2004. National databases were available on animal slaughter, programmes of tuberculin testing for bovine tb and laboratory confirmation of suspected lesions. Factories were ranked according to their submission risk (number of animals submitted with lesions/number of attested animals killed) and confirmation risk (number of animals with laboratory-confirmed lesions/number of animals submitted with lesions), adjusting for the risk profile of the animals slaughtered, including potential confounding factors such as their age and sex, whether they were purchased or homebred, the test history of their herd, the prevalence of bovine tb in the area and the season of slaughter. Approximately 3.7 million cattle were slaughtered in 42 Irish export-licensed factories during the two years. Complete data were available for 2,374,987 animals from 84,510 attested herds in 2845 District Electoral Divisions. Samples from 7398 animals with suspected tb lesions were submitted for laboratory examination; 4767 (64.4 per cent) were positive, 2011 were negative and 620 were inconclusive. The average unadjusted submission risk for all the factories was 22 per 10,000, ranging from 0 to 58 per 10,000. The unadjusted factory confirmation risk (excluding factories that had sent in fewer than 10 lesions) varied between 34.3 per cent and 86.3 per cent. The unadjusted and adjusted submission and confirmation risks were highly correlated, and animal-related factors (including their characteristics and origin) therefore did not contribute to the variations in factory-level submission and confirmation risks.     

Prevalence of and factors associated with Brucella sero-positivity in cattle in urban and peri-urban Gulu and Soroti towns of Uganda

Brucellosis is a key zoonosis of major public health, animal welfare and economic significance, and is endemic in livestock in Uganda. A cross-sectional epidemiological study was carried out to estimate the sero-prevalence of brucellosis and identify factors associated with sero-positivity in cattle in urban and peri-urban Gulu and Soroti towns of Northern and Eastern Uganda, respectively. A total of 1007 sera and data on biologically plausible risk factors from 166 herds and their spatial locations, were collected from cattle reared in urban and peri-urban Gulu and Soroti towns of Uganda. The sera were analyzed using indirect ELISA and sero-positive reactors confirmed by competitive ELISA. Multivariable models were used to investigate for risk factors. The overall animal-level and herd-level sero-prevalence was 7.5% (76/1007, 95% Confidence Interval (CI): 6.15–9.4%) and 27.1% (45/166, 95% CI: 20.9–34.3%), respectively. Herd-level sero-prevalence was significantly (P<0.001) higher in Soroti than Gulu. In Gulu town, sero-positivity increased with an increase in herd size (P=0.03) and age (P=0.002), and was higher in cattle brought in from western Uganda (P<0.0001). In Soroti town, introduction of new cattle into a herd was significantly (P=0.027) associated with herd sero-positivity. There was a geographically differential risk (clustering) of Brucella sero- positivity in herds in Soroti, while sero-positivity was homogeneously distributed in Gulu. The data highlight brucellosis occurrence and major risk factors for its transmission in cattle in urban and peri-urban areas.     

Brucellosis and associated risk factors in dairy cattle of eastern Ethiopia

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