Is it safe to use ovarian follicular fluid from cows seropositive for brucellosis in the production of in vitro embryos?

Animal reproduction biotechniques are important tools for the technological advancement of livestock, as they allow the selection of the reproductive potential of superior quality females and males; however, infectious diseases that have a predilection for the reproductive system can be a hindrance for the use of these technologies. Therefore, the present study aimed to detect Brucella spp. in the ovarian follicular fluid of brucellosis-positive bovine cows. A total of 47 bovine ovarian follicular fluid aspirates from cows, positive in tests for brucellosis and from Brucella-positive herd, were submitted to PCR. The primers used in the PCR were specific to the genus Brucella (bcsp31 gene). All 47 bovine aspirates were negative for Brucella spp. 0.00% (95% CI: 0.00-4.00%). Our results demonstrated that Brucella spp. was absent in the ovarian follicular fluid from seropositive cows, which indicates that Brucella spp.-infected cows could be used for reproductive biotechnologies carried out with follicular aspirates. Future studies are needed to more precisely evaluate the feasibility and safety of using these oocytes from brucellosis-seropositive cows to transfer embryos to heifers/cows not infected by Brucella, aiming to produce calves free of the infection.

Distribution, seroprevalence and risk factors for bovine brucellosis in Brazil: Official data, systematic review and meta-analysis.

Bovine brucellosis is an endemic disease in Brazil, and evidence-based assessments of the available literature on its seroprevalence and risk factors are limited. The aim of this study was to systematically review and summarize studies related to seroprevalence and risk factors of bovine brucellosis in the entire Brazil, in addition to comparing published data with the most recent official reports. Articles available in scientific databases and published between October 2006 and October 2021 were evaluated. Forty-five publications were included in the meta-analysis on the seroprevalence of brucellosis and 29 publications in the review on risk factors. The largest number of publications was found for the State of Mato Grosso do Sul (n=4), and the highest and lowest seroprevalences were observed in Acre (11%; 95% CI: 8.0-14.0%) and in the Federal District (0.4%; 95% CI: 0.2-0.7%). The main risk factors were the purchase of animals for breeding, vaccination, the number of heifers (female ≥2 years), the presence of calving paddocks and the occurrence of abortions. The need for new official studies has been suggested to determine the true prevalence of bovine brucellosis in Brazil, supported by the National Program for the Control and Eradication of Animal Brucellosis and Tuberculosis.

A spatiotemporal analysis of bovine brucellosis cases in Minas Gerais state, Brazil, from 2011 to 2018.

Our study explored the patterns of bovine brucellosis dissemination in Minas Gerais state, Brazil, by examining data on passive surveillance of bovine brucellosis cases from the Instituto Mineiro de Agropecuaria (IMA) (Animal Health Authority), as well as cattle population and bovine brucellosis testing, from 2011 to 2018 by means of a spatiotemporal analysis. We plotted cases, populations and testing distributions and performed spatial autocorrelation (Moran's I test) and local indicators of spatial autocorrelation (LISA) analyses. Moreover, we assessed the correlation of the spatial distribution and the compiled data (brucellosis cases, cattle populations, and brucellosis testing) by Lee's test. Our results showed that bovine brucellosis cases occurred mainly in the Triângulo Mineiro, Alto Paranaíba and Northwest regions, which reported cases in all analyzed years (2011 to 2018). The cattle population of Minas Gerais was concentrated in the same regions as bovine brucellosis cases, and the performed tests through the analyzed years (2011 to 2018). Moran's I test results of the case data showed significant spatial autocorrelation in 2011, 2015 and 2018 (p value < 0.05), and from 2011 to 2018, the population and testing data were also significant in Moran's I test (p value < 0.01). The results of cluster analysis (LISA) of cases showed clusters mainly in the Triângulo Mineiro, Alto Paranaíba, Northwest and South regions in 2011, 2015 and 2018. The local clusters for cattle populations and brucellosis testing were also observed in the same regions as bovine brucellosis cases in all years (2011 to 2018). The correlation results between clusters (Lee's test) were 0.22 (p value < 0.01) in 2011, 0.15 (p value < 0.01) in 2015 and 0.43 (p value <0.01) in 2018 between cases and populations, and 0.25 (p value <0.01) in 2011, 0.14 (p value <0.01) in 2015 and 0.38 (p value < 0.01) in 2018 for testing and cases. Therefore, our results showed that brucellosis cases were distributed together with cattle populations and brucellosis testing data, indicating that brucellosis in cattle in Minas Gerais state is being identified where there are more animals and where more tests are performed.

Accuracy of serological tests for bovine brucellosis: A systematic review and meta-analysis.

The direct methods for diagnosis of bovine brucellosis have several limitations, therefore serological tests are the basis for the diagnosis of the disease. However, a meta-analysis estimating the diagnostic sensitivity (DSe) and diagnostic specificity (DSp) on the main tests used in bovine brucellosis control programs worldwide has not been performed. This systematic review and meta-analysis aimed to estimate the DSe, DSp and thereby accuracy of serological tests individually used in the diagnosis of bovine brucellosis. The databases CABI, Cochrane Library, PubMed/MEDLINE, SciELO, Scopus and Web of Science were used to select articles. The search resulted in 5308 studies, of which 71 were selected for systematic review using quality assessment tools and 65 studies were included in the meta-analysis. For the meta-analysis, 178 assays and 11 different serological tests were considered. To estimate DSe and DSp of the tests, studies were divided according to animal selection for the studies: (1) studies that carried out a random or consecutive selection of participants (noncasecontrol studies) and (2) all studies, including casecontrol studies. Considering only the non-case-control studies to estimate the DSe, the tests that exhibited the best and worst performance were the iELISA test (indirect enzyme immunoassay - bacterial suspension as antigen - BS) (96.5%, 95% CI: 94.1-97.9%) and 2ME (2- mercaptoethanol test) (85.0%, 95% CI: 79.6-89.1%), respectively; while for DSp, the FPA (fluorescence polarization assay) (99, 7%, 95% CI: 99.5-99.8%) and PCFIA tests (protein concentration fluorescence immunoassay) (78.5%, 95% CI: 70.0-85.1%) showed better and worse performance, respectively. Overall, our results showed an overestimation in the DSe and DSp of the eleven serological tests assessed when casecontrol studies were included in the meta-analysis, which is a concern considering its impacts on the time and costs associated with populational diagnosis of the diseases, since several of these tests are routinely used in the control and eradication programs of bovine brucellosis worldwide. Furthermore, the tests that exhibited the best DSe and DSp, iELISA (BS) and FPA, respectively, are relatively easy to perform and interpret and the test which showed the best overall accuracy was FPA.

Use of recombinant malate dehydrogenase (MDH) and superoxide dismutase (SOD) [CuZn] as antigens in indirect ELISA for diagnosis of bovine brucellosis.

The objective of this study was to validate an indirect enzyme-linked immunoassay (iELISA) using the recombinant proteins, malate dehydrogenase (MDH) and superoxide dismutase (SOD) [CuZn], as antigens and to evaluate its ability to discriminate antibodies produced by vaccination from those induced by infection in the diagnosis of bovine brucellosis. Sera from six groups were evaluated: G1 - culture-positive animals (52 serum samples) (naturally infected); G2 - non-vaccinated animals (28 serum samples) positive in RBT (Rose Bengal test) and 2ME (2-mercaptoethanol test) selected from brucellosis-positive herds; G3 - animals from a brucellosis-free area (32 serum samples); G4 - S19 vaccinated heifers (114 serum samples); G5 - RB51 vaccinated heifers (60 serum samples); G6 - animals inoculated with inactivated Yersinia enterocolitica O:9 (42 serum samples). Diagnostic sensitivity (DSe) and diagnostic specificity (DSp) were estimated using the frequentist approach and the confidence interval (CI) (95%) calculated by the Clopper-Pearson (exact) method. The DSe for iELISA_MDH in the G1 group was 71.7% (CI 95%: 57.6-83.2%) and for the G2 100.0% (CI 95%: 87.7-100.0%), whereas the DSp was 84.4% in the G3 (CI 95%: 67.2-94.7%). For the iELISA_SOD the DSe was estimated 67.3% for the G1 (CI 95%: 52.9-79.7%) and 71.4% for G2 (CI 95%: 51.3-86.8%), while the DSp for G3 was 87.5% (CI 95%: 71.0-96.5%). iELISA_MDH and iELISA_SOD showed potential to be used in the diagnosis of infected animals, increasing the range of serological tests available for the diagnosis of bovine brucellosis, with the advantage of being S-LPS-free. However, none of the tests could differentiate between infection and vaccination.

Turning chaotic sample group clusterization into organized ones by feature selection: Application on photodiagnosis of Brucella abortus serological test.

Bovine brucellosis diagnosis is a major problem to be solved; the disease has a tremendous economic impact with significant losses in meat and dairy products, besides the fact that it can be transmitted to humans. The sanitary measures instituted in Brazil are based on disease control through diagnosis, animal sacrifice, and vaccination. Although the currently available diagnostic tests show suitable quality parameters, they are time-consuming, and the incidence of false-positive and/or false-negative results is still observed, hindering effective disease control. The development of a low-cost, fast, and accurate brucellosis diagnosis test remains a need for proper sanitary measures at a large-scale analysis. In this context, spectroscopy techniques associated with machine learning tools have shown great potential for use in diagnostic tests. In this study, bovine blood serum was investigated by UV-vis spectroscopy and machine learning algorithms to build a prediction model for Brucella abortus diagnosis. Here we first pre-treated the UV raw data by using Standard Normal Deviate method to remove baseline deviation, then apply principal component analysis - a clustering method - to observe the group formation tendency; the first results showed no clustering tendency with a messy sample score distribution, then we properly select the main principal components to improve clusterization. Finally, by using machine learning algorithms (SVM and KNN), the predicting models achieved a 92.5% overall accuracy. The present methodology provides a test result in an average time of 5 min, while the standard diagnosis, with the screening and confirmatory tests, can take up to 48 h. The present result demonstrates the method's viability for diagnosing bovine brucellosis, which can significantly contribute to disease control programs in Brazil and other countries.

Retrospective benefit-cost analysis of bovine brucellosis control in the state of Mato Grosso, Brazil.

From 2001, due to the launching of the National Program for the Control and Eradication of Brucellosis and Tuberculosis, Brazil began a series of studies aimed at the epidemiological characterization of bovine brucellosis in its federative units. In the state of Mato Grosso, studies conducted in 2003 and 2014 showed a reduction in the prevalence of infected herds from 41.2 % to 24 % and in the prevalence of seropositive animals from 10.2 % to 5.1 %. These results stemmed from the efficient heifer vaccination program with S19 implemented by the state. From this scenario, the present study quantified and compared the benefits and costs of bovine brucellosis control in the state through a financial analysis. When considering private and public costs, the Net Present Value (NPV) was estimated at US$ 37.5 million, the Internal Rate of Return (IRR) was calculated at 31 %, and the Benefit-Cost Ratio (BCR) was 2.3. When considering only the private costs, the NPV was US$ 51.3 million, the IRR was 46 % and the BCR was 3.43, indicating that the cattle producer in Mato Grosso had a return of US$ 3.43 for each US$ 1.00 invested. Therefore, the bovine brucellosis control actions carried out in the state of Mato Grosso between 2003 and 2014 were highly advantageous from the economic point of view. The most rational way to continue decreasing bovine brucellosis prevalences at low cost in the state is to maintain the heifer vaccination program, using the RB51 vaccine in addition to S19.

Role of the cGAS/STING pathway in the control of Brucella abortus infection acquired through the respiratory route.

Despite the importance of the respiratory route for Brucella transmission, the lung immune response to this pathogen is scarcely characterized. We investigated the role of the cGAS/STING pathway of microbial DNA recognition in the control of respiratory Brucella infection. After in vitro B. abortus infection, CFU numbers were significantly higher in alveolar macrophages (AM) and lung explants from STING KO mice than in samples from wild type (WT) mice, but no difference was observed for cGAS KO samples. CFU were also increased in WT AM and lung epithelial cells preincubated with the STING inhibitor H151. Several proinflammatory cytokines (TNF-α, IL-1ß, IL-6, IP-10/CXCL10) were diminished in Brucella-infected lung explants and/or AM from STING KO mice and cGAS KO mice. These cytokines were also reduced in infected AM and lung epithelial cells pretreated with H151. After intratracheal infection with B. abortus, STING KO mice exhibited increased CFU in lungs, spleen and liver, a reduced expression of IFN-ß mRNA in lungs and spleen, and reduced levels of proinflammatory cytokines and chemokines in bronchoalveolar lavage fluid (BALF) and lung homogenates. Increased lung CFU and reduced BALF cytokines were also observed in cGAS KO mice. In summary, the cGAS/STING pathway induces the production of proinflammatory cytokines after respiratory Brucella infection, which may contribute to the STING-dependent control of airborne brucellosis.

Economic analysis of bovine brucellosis control in the Rondônia state, Brazil.

Bovine brucellosis, mainly caused by Brucella abortus, is a worldwide distribution anthropozoonosis that causes great economic losses. In 2001, Brazil launched the National Program for the Control and Eradication of Brucellosis and Tuberculosis (PNCEBT). Contemporaneously, a great effort to characterize the epidemiology of the disease in Brazilian states was started. In the state of Rondônia, a first epidemiological study was carried out in 2004, revealing a prevalence of 35.2% of infected herds and 6.22% of seropositive females. In 2014, after a successful heifer vaccination program with strain 19 (S19), a second study detected a reduction in the prevalence of infected herds to 12.3% and of seropositive females to 1.9%. The present study aimed to quantify and compare the costs and benefits related to the control of bovine brucellosis in the state through an accounting analysis. Vaccinating heifers and performing serological tests to move animals were computed as private costs. The expenditures of the state official veterinary service for brucellosis control were considered public cost. The considered benefits of lowering prevalence were decreased cow replacement, decreased abortions, decreased perinatal and cow mortality, and increased milk production. Considering private and public costs, the net present value (NPV) was estimated at US$ 18.3 million, the internal rate of return (IRR) was calculated at 23%, and the benefit-cost ratio (BCR) was 1.7. When considering only the private costs, the NPV was US$34.9 million, the IRR was 49%, and the BCR was 3.0, meaning that the bovine producer had a return of 3 for each unit of currency invested. The results showed that the bovine brucellosis control measures implemented in the state of Rondônia, which had as its main strategy the vaccination of heifers with S19, produced highly advantageous economic results. The state should continue with its vaccination program, stimulating the use of the RB51 vaccine in addition to S19, to achieve further reductions in prevalence at low cost.

Deletion in KARLN intron 5 and predictive relationship with bovine tuberculosis and brucellosis infection phenotype.

Kalirin (gene: KALRN) is a Rho-GEF kinase linked to neurodegenerative diseases in humans. Unexpectedly, various polymorphisms in KALRN gene were previously associated with resistance to bacterial infections in ruminants. In this study, we evaluated the effect of the rs384223075 (RS-075) deletion in KALRN intron 5 on the occurrence of Mycobacterium bovis and Brucella abortus infections in cattle. We performed two separate case-control association analyses: one for bovine tuberculosis (bTB) using 308 Holstein and Jersey cows from three herds with prevalence between 5 and 15% for this infection; and another for brucellosis using 140 Holstein and beef crossbred cows from two herds with high prevalence for brucellosis (> 30%). In the bTB analysis, the RS-075 deletion frequency was higher among cases than controls (p = 0.0001), and the absence of the RS-075 deletion allele was associated with negative PPD-skin test results (p = 0.0009) at genotype level. On the contrary, RS-075 was not associated with Brucella spp. serological status (p = 0.72) but, unexpectedly, the deletion allele was more frequent among controls than cases in the beef crossbred herd (0.31 vs. 0.14, p = 0.02). In concordance with this observation, in vitro assays showed that the RS-075 deletion could be linked to an enhanced cellular response to bacterial antigens and unspecific stimulation in mononuclear cells derived from beef crossbred cows, specifically the reactive nitrogen species production (p = 0.008) and proliferation capacity (p = 0.018). This study is consistent with other reports that support an important role of the KALRN gene and its polymorphisms in the host response to intracellular pathogens.

Descriptive cross-sectional study on major bovine diseases and associated risk factors in north-eastern Ecuadorian Amazon / Estudo transversal descritivo sobre as principais doenças bovinas e fatores de risco associados no nordeste da Amazônia equatoriana

Cattle raising is a crucial element of production systems in the tropics and subtropics. However, in recent years, global public health security has been threatened by disease emergence. In Orellana Province, livestock is the most important activity to generate economic income. Nevertheless, there is no available data about Animal Health status. With this objective, a study was performed to describe the major Bovine diseases recorded between 2011 to 2019, and the main Risk factors associated. Data on main Bovine diseases were retrieved from the World Animal Health Information System database. Whereas Bovine population data used to calculate the prevalence rates and confidence intervals were obtained from Ecuador's Ministry of Agriculture. By contrast, the Risk factors identified with an epidemiological questionnaire were applied to 300 livestock farmers. As a result, from 2011 to 2019 in Orellana has been confirmed: 90 cases of Infectious Bovine Rhinotracheitis (31.58%), Bovine Rabies by hematophagous bats (Desmodus rotundus), 83 cases (29.12%), Bovine viral diarrhea with 43 cases (15.10%), Brucellosis by Brucella abortus 35 cases, which was (12.28%), and 34 cases related to Enzootic bovine leukosis (11.92%). Overall, the prevalence rates ranged from (0.24 to 15.37%). In addition, farm size, presence of forest, herd, and paddock sizes, cutting frequency of forages, and other animal species were involved as Risk factors (OR = 3.15 to 11.75; 95% CI, 0.01 to 0.69). In conclusion, there are animal diseases with reproductive and neurologic symptomology and high-Risk factors implicated in the transmission. Consequently, space-temporal and seroprevalence epidemiological studies should be performed in Orellana.

A criação de gado é um elemento crucial dos sistemas de produção nos trópicos e subtrópicos. No entanto, nos últimos anos, a segurança da saúde pública global tem sido ameaçada pelo surgimento de doenças. Na província de Orellana, a pecuária é a atividade mais importante para gerar renda econômica. Contudo, não há dados disponíveis sobre o estado de saúde animal. Com este objetivo, foi realizado um estudo para descrever as principais doenças dos bovinos registradas entre 2011 e 2019, além dos principais fatores de risco associados. Os dados sobre as principais doenças bovinas foram recuperados do banco de dados do World Animal Health Information System. Os dados da população bovina usados para calcular as taxas de prevalência e os intervalos de confiança foram obtidos do Ministério da Agricultura do Equador. Por outro lado, os fatores de risco identificados com um questionário epidemiológico foram aplicados a 300 criadores de gado. Como resultado, de 2011 a 2019 em Orellana foram confirmados: 90 casos (31,58%) de rinotraqueíte infecciosa bovina, 83 casos (29,12%) de raiva bovina por morcegos hematófagos (Desmodus rotundus), 43 casos (15,10%) de diarreia viral bovina, 35 casos (12,28%) de brucelose por Brucella abortus e 34 casos relacionados à leucose enzoótica bovina (11,92%). No geral, as taxas de prevalência variaram de 0,24 a 15,37%. Além disso, tamanho da fazenda, presença de floresta, tamanho do rebanho e dos piquetes, frequência de corte de forragens e outras espécies animais estiveram envolvidos como fatores de risco (OR = 3,15-11,75; IC 95% 0,01-0,69). Em conclusão, existem doenças animais com sintomatologia reprodutiva e neurológica e fatores de alto risco implicados na transmissão. Portanto, estudos epidemiológicos espaço-temporais e de soroprevalência devem ser realizados em Orellana.

Analysis of cattle movement networks in Paraguay: Implications for the spread and control of infectious diseases.

Beef exports represent a substantial part of Paraguay's agricultural sector. Cattle movements involve a high risk due to the possible spread of bovine diseases that can have a significant impact on the country's economy. We analyzed cattle movements from 2014 to 2018 using the networks analysis methodology at the holding and district levels at different temporal scales. We built two types of networks to identify network characteristics that may contribute to the spread of two diseases with different epidemiological characteristics: i) a network including all cattle movements to consider the transmission of a disease of rapid spread like foot and mouth disease, and ii) a network including only cow movements to account for bovine brucellosis, a disease of slow spread that occurs mainly in adult females. Network indicators did not vary substantially among the cattle and cow only networks. The holdings/districts included in the largest strongly connected components were distributed throughout the country. Percolation analysis performed at the holding level showed that a large number of holdings should be removed to make the largest strongly connected component disappear. Higher values of the centrality indicators were found for markets than for farms, indicating that they may play an important role in the spread of an infectious disease. At the holding level (but not at the district level), the networks exhibited characteristics of small-world networks. This property may facilitate the spread of foot and mouth disease in case of re-emergence, or of bovine brucellosis in the country through cattle movements. They should be taken into account when implementing surveillance or control measures for these diseases.

Levantamento sorológico de brucelose em bovinos monitorados no estado da Bahia / Serological survey of brucellosis in cattle monitored in the estate of Bahia

A brucelose é uma doença bacteriana de grande importância para a economia pecuária e para a saúde pública por se tratar de uma zoonose. É uma doença infecto-contagiosa que tem com agente etiológico bactérias do gênero Brucella. Em bovinos, as espécies do gênero é a Brucella abortus, que são cocobacilos gram negativo, intracelulares facultativos, imóveis e não esporulado. A infecção apresenta evolução crônica e acomete animais de todas as idades, sendo mais frequente em indivíduos sexualmente maduros. O objetivo desse trabalho é investigar, por meio da sorologia para brucelose bovina, utilizando a técnica do ELISA indireto, amostras de animais reagentes abatidos em frigoríficos inspecionados no estado da Bahia. Foram utilizados 666 animais, selecionados aleatoriamente no momento do abate. O sangue foi coletado com finalidade de obtenção de soro, todas as amostras foram submetidas à prova de triagem do Antígeno Acidificado Tamponado (AAT), prova do 2mercaptoetanol (2-ME) e ELISA Indireto. Das amostras reagentes no teste do AAT, obteve-se uma prevalência estimada em 1,2%. A prevalência no teste do ELISA foi de 13,21% (n=86). Esse resultado sugere a ocorrência de falsos negativos quando se utiliza a prova do antígeno acidificado tamponado.

Brucellosis is a bacterial disease of great importance to the livestock economy and to public health because it is a zoonosis. It is an infectious disease that has etiologic agent with bacteria of the genus Brucella. In cattle, the species of the genus Brucella is Brucella abortus that are gram negative, facultative intracellular, real estate and not sporulated. The infection presents chronic and affects animals of all ages, being more frequent in sexually mature individuals. This study aimed to investigate through serology for brucellosis, using the technique of indirect ELISA, samples from positive animals slaughtered in slaughterhouses inspected in the state of Bahia. A total of 666 animals were used, randomly selected at the time of slaughter. Blood was collected in order to obtain serum, all samples were subjected to a screening test Antigen Buffered Acidified (AAT), proof of 2-mercaptoethanol (2-ME) and Indirect ELISA. Of reagents in the test samples of AAT obtained an estimated prevalence of 1.2%. The prevalence in the ELISA test was 13.21% (n = 86). This result suggests the occurrence of false negatives when using the buffered acidified antigen test.

Análise da correlação dos índices de estrutura e recursos humanos do serviço veterinário oficial em 2019 e as incidências acumuladas (2017 ­ 2019) de doenças de notificação obrigatória em bovinos e equinos no Brasil / Analysis of the correlation between the structure and human resources indexes of the official veterinary service in 2019 and the accumulated incidences (2017 ­ 2019) of notifiable diseases in cattle and horses in Brazil

As doenças de notificação obrigatória em bovinos podem gerar impactos sociais e econômicos significativos na cadeia pecuária brasileira, além de consequências negativas no mercado internacional devido a embargos sanitários. Para auxiliar no entendimento de como um sistema de vigilância epidemiológica com mais recursos pode gerar mais credibilidade para o país, foram realizadas análises de correlação entre a notificação de doenças e a estrutura veterinária disponível nos Órgãos Executores de Sanidade Agropecuária (OESAs), a partir dos dados contidos no Sistema Nacional de Informação Zoossanitária (SIZ), entre os anos de 2017 e 2019. Com base nos dados do serviço veterinário, foram produzidos o Índice de Estrutura Física Oficial (IEFO) e o Índice de Recursos Humanos do Serviço Oficial (IRHSO). Foi realizada análise de correlação entre a notificação de doenças de bovinos com a capacidade de estrutura física e recursos humanos de vigilância epidemiológica disponíveis no Serviço Veterinário brasileiro. Os estados AP, RR e SC foram os que mais notificaram brucelose e tuberculose no período e estão entre os melhores índices de estrutura e recursos humanos do país. A análise dos índices mostrou que a raiva não possui correlação significativa com estrutura e recursos humanos do serviço, entretanto, brucelose e tuberculose possuem correlação positiva com estrutura veterinária oficial disponível para a vigilância em bovinos. Portanto, melhorias na estrutura podem refletir no incremento dos índices de notificação das doenças de bovinos, assim como na qualidade de suas informações.

Notifiable diseases in cattle can generate significant social and economic impacts on the Brazilian livestock chain, in addition to impacts on the international market due to sanitary embargoes. To help understand how an epidemiological surveillance system with more resources can generate more credibility for the country, correlation analyzes were carried out between the notification of diseases and the veterinary structure available in the Executing Bodies of Agricultural Health (OESAs), based on the data contained in the National System of Zoosanitary Information (SIZ), between the years 2017 to 2019. Based on public data from the veterinary service, the Official Physical Structure Index (IEFO) and the Official Service Human Resources Index (IRHSO) were produced. Correlation analysis was performed between the notification of bovine diseases with the capacity of physical structure and human resources for epidemiological surveillance available in the Brazilian Veterinary Service. AP, RR and SC were the states that most notified brucellosis and tuberculosis in the period and are among the best indices of structure and human resources in the country. The analysis of correlation indices showed that the rabies disease does not have a significant correlation with the structure and the human resources of the service, however, brucellosis and tuberculosis does have a positive correlation with the official veterinary structure available for surveillance in cattle. Therefore, Improvements in the structure can reflect in the increase of the notification rates, as well as in the quality of its information.

Development of a novel glycoprotein-based immunochromatographic test for the rapid serodiagnosis of bovine brucellosis.

AIMS: Bovine brucellosis is a worldwide zoonotic disease that causes important economic losses and public health concerns. Because control of the disease depends on vaccination, serodiagnosis and isolation of the infected animals, affordable, rapid and accurate point of care (POC) tests are needed. METHODS AND RESULTS: We developed and evaluated a novel glycoprotein-based immunochromatographic test for the detection of IgG antibodies against the O-polysaccharide of Brucella in bovine serum samples. Brucella GlycoStrip combines the power of immunochromatographic and bacterial glycoengineering technologies for the diagnosis of bovine brucellosis. The analysis of positive and negative reference samples indicated that the test has a diagnostic sensitivity and specificity of 96.9% (95% CI: 92.7%-100.0%) and 100%, respectively. CONCLUSIONS: Due to the recombinant glycoprotein-based antigen OAg-AcrA, which consists of the O-side chain of Brucella smooth lipopolysaccharide (sLPS) covalently linked to the carrier protein AcrA, the test is highly accurate, allows the differentiation of infected animals from those vaccinated with a rough strain or with a single dose of a smooth strain and fulfil the minimum diagnostic requirements established by the national and international regulations. SIGNIFICANCE AND IMPACT OF STUDY: This strip test could provide a rapid (10 min) and accurate diagnosis of bovine brucellosis in the field contributing to the control of the disease.

Determination of streptomycin and doxycycline using LC/MS towards an effective treatment against an experimental Brucella abortus infection in mice.

Herein we described a versatile liquid chromatographic method for detection and quantification of the total levels of two antimicrobials [i.e., streptomycin (STM) and doxycycline (DOX)], in mice plasma and selected tissues, with the aid of a single quadrupole as a detection method. The method included a few sample preparation steps, including freeze-drying and in situ triphasic solvent-assisted defatting, precipitation, and extraction, allowing easy and fast tissue sample processing and avoiding analyte loss. Using a murine model, we demonstrated that mass spectrometry detects simultaneously and with high specificity two of the most widespread antimicrobials used against Brucellosis. An accurate [recoveries varied from 75.23 (bone marrow) to 101.33% (liver)] and sensitive (LoD in the ng g-1 range) method to assess STM and DOX in murine tissue, including subtherapeutic and therapeutic doses of the antimicrobials, was achieved. This validated method can be successfully used to monitor the depletion of STM and DOX in several mice tissues and plasma during metabolism after administration.

Survey of bovine brucellosis on the island of Fernando de Noronha, Pernambuco, Brazil / Investigação sobre brucelose bovina na Ilha de Fernando de Noronha, Pernambuco, Brasil

Considering the lack of information about livestock diseases on Brazilian oceanic islands, the occurrence of bovine brucellosis was investigated on the island of Fernando de Noronha, state of Pernambuco, Brazil. Serum samples were collected in October 2009, from all the 105 cows raised on the island at that time. These were examined concurrently using the Rose Bengal test and the Complement Fixation Test. All the samples were negative in both tests, indicating that the cows on the island were likely free from infection by smooth forms of Brucella. These results can partly be explained by the prohibition of introduction and importation of both small and large-sized animals that had been implemented through District Decree 19 of February 28, 2004.(AU)

Tendo em vista a inexistência de informações sobre a ocorrência da brucelose bovina em ilhas oceânicas brasileiras, investigou-se a presença da infecção na ilha de Fernando de Noronha, Estado de Pernambuco, Brasil. Soros de todas as 105 fêmeas bovinas existentes, colhidos em outubro de 2009, foram examinados concomitantemente pelo teste do Antígeno Acidificado Tamponado e pela Reação de Fixação de Complemento. Todas as amostras foram negativas em ambos os testes, indicando que provavelmente os animais presentes na ilha encontravam-se livres da infecção por Brucella. Estes resultados podem ser explicados, em partes, pela proibição da introdução e importação de grandes e pequenos animais, implementada pelo Decreto Distrital 19, de 28 de fevereiro de 2004.(AU)

Evaluation of diagnostic tests' sensitivity, specificity and predictive values in bovine carcasses showing brucellosis suggestive lesions, condemned by Brazilian Federal Meat Inspection Service in the Amazon Region of Brazil.

Cervical bursitis is a suggestive lesion of bovine brucellosis. Diagnostic sensitivity and specificity of two brucellosis serological tests, Rose Bengal (RB) and serum agglutination test with 2-mercaptoethanol (SAT/2-ME), and of isolation and identification (bacteriology) were evaluated through Bayesian latent class analysis (BLCA). A total of 165 paired serum and cervical bursitis samples detected at inspection by Brazilian federal meat inspection services were analyzed. The best model fit to the data occurred when accounting for the conditional dependence between serological tests. According to this model, RB and SAT/2-ME had almost the same sensitivity, 0.960 [0.903 - 0.992] and 0.963 [0.906 - 0.994] with 95 % Credible Interval (95 %CrI), respectively. Specificities were 0.9068 [0.562 - 0.997] and 0.875 [0.546 - 0.990] for RB and SAT/2-ME, respectively, also with 95 %CrI. Bacteriology had lower sensitivity than serological tests, 0.594 (95 %CrI: [0.525 - 0.794]) and the highest specificity of all evaluated tests, 0.992 (95 %CrI: [0.961-1.00]). Prevalence of infected animals was 0.829 (95 %CrI: [0.700-0.900]). BLCA showed that both RB and SAT/2-ME fitted to the purpose of initial screening the brucellosis suspect in carcasses with cervical bursitis in a reliable way. The results of RB or SAT/2-ME can guide the sanitary actions for brucellosis control and help the implementation of a risk-based surveillance system in the meat production chain. This strategy is especially true in remote areas with large beef cattle herds, raised extensively, where in vivo tests are rarely performed due to logistic and management constraints, as in the Northern region of Brazil.

The endoplasmic reticulum stress sensor IRE1α modulates macrophage metabolic function during Brucella abortus infection.

Endoplasmic reticulum (ER) stress plays a major role in several inflammatory disorders. ER stress induces the unfolded protein response (UPR), a conserved response broadly associated with innate immunity and cell metabolic function in various scenarios. Brucella abortus, an intracellular pathogen, triggers the UPR via Stimulator of interferon genes (STING), an important regulator of macrophage metabolism during B. abortus infection. However, whether ER stress pathways underlie macrophage metabolic function during B. abortus infection remains to be elucidated. Here, we showed that the UPR sensor inositol-requiring enzyme 1α (IRE1α) is as an important component regulating macrophage immunometabolic function. In B. abortus infection, IRE1α supports the macrophage inflammatory profile, favoring M1-like macrophages. IRE1α drives the macrophage metabolic reprogramming in infected macrophages, contributing to the reduced oxidative phosphorylation and increased glycolysis. This metabolic reprogramming is probably associated with the IRE1α-dependent expression and stabilization of hypoxia-inducible factor-1 alpha (HIF-1α), an important molecule involved in cell metabolism that sustains the inflammatory profile in B. abortus-infected macrophages. Accordingly, we demonstrated that IRE1α favors the generation of mitochondrial reactive oxygen species (mROS) which has been described as an HIF-1α stabilizing factor. Furthermore, in infected macrophages, IRE1α drives the production of nitric oxide and the release of IL-1ß. Collectively, these data unravel a key mechanism linking the UPR and the immunometabolic regulation of macrophages in Brucella infection and highlight IRE1α as a central pathway regulating macrophage metabolic function during infectious diseases.