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Research concerning leptospirosis in donkeys and mules has been neglected around the world. Therefore, the aim of this study was to investigate the epidemiological situation of the prevalence of anti-Leptospira spp. antibodies in donkeys and mules from the state of Minas Gerais, Brazil. Blood serum samples were collected from 180 animals (109 donkeys and 71 mules) in two rural properties from the state of Minas Gerais, Brazil, and then submitted to a microscopic agglutination test (MAT). Urea and creatinine values were also quantified. Epidemiological variables such as age, breeding system, contact with other animal species, source of water and food, vaccination against leptospirosis, presence of reproductive alterations, and rodent control were also investigated. From 180 samples collected, 39 (21.67%) showed positive results in the MAT, at a dilution ≥ 1:100. Some animals were reactive for more than one serovar. The serovar Tarassovi was the most frequent (14.07%), followed by Hardjo (11.85%) and Wolffi (11.11%). There was a statistically significant difference between animals from 0 to 3 years of age reactive in the MAT in comparison to the other age groups. Most of the animals had urea and creatinine concentrations within the acceptable reference limit; however, there was a significant increase in creatinine levels in some of the test animals. The studied properties showed differences in some epidemiological aspects such as vaccination of the animals, presence of reproductive problems in the herd, and rodent control. Such aspects pointed as risk factors that may influence the frequency of positive serological results in property 1. The present study demonstrated that the prevalence of leptospirosis in donkeys and mules is high and several serovars are being maintained by these animals, representing a potential public health risk.

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Leptospirosis is a globally important zoonosis that causes death in humans and animals, especially unvaccinated dogs. Dogs are infected through contact with urine or water contaminated by pathogenic spirochaete bacteria of the genus Leptospira. This study aimed to assess the performance of two diagnostic tests in characterizing dogs with leptospirosis and associate it with the main clinical-pathological alterations in dogs with suspected infection. From September 2019 to September 2020, 24 dogs with suspected canine leptospirosis were treated at the Veterinary Hospital of the Federal University of Uberlândia (UFU). Complementary tests such as a complete blood count (CBC) and serum creatinine, ALT (alanine transaminase) and ALP (alkaline phosphatase) levels were prescribed. In order to confirm the leptospirosis diagnosis, serum and molecular tests were performed using the microscopic agglutination test (MAT) and real-time quantitative polymerase chain reaction (qPCR). Of the 24 suspected cases of canine leptospirosis, six (25%) were positive in the MAT and three (12.5%) in the qPCR, with one (4.17%) returning positive results in both diagnostic tests. Positive results were obtained for 8/24 (33.3%) for combined MAT + qPCR. The predominant reactive serogroups were Icterohaemorrhagiae (33.33%) and Djasiman (33.33%), followed by Ballum (16.60%). The sensitivity and specificity of the qPCR in relation to the gold standard test (MAT) were 16.7 and 88.9%, respectively. Changes were observed in serum levels of creatinine, urea and ALP in the group that tested positive (MAT/qPCR); however, only urea and ALP levels were high in the negative group. Comparison of biochemical parameters in the positive (MAT/qPCR) and negative groups showed no statistically significant difference between the elements assessed (p>0.05). It was concluded that in order to diagnose acute canine leptospirosis, veterinarians should combine the clinical-pathological findings with paired sample MAT and qPCR urine test results.(AU)

A leptospirose é uma zoonose de importância mundial que causa mortes tanto em humanos, como em animais, especialmente em cães não vacinados. A infecção em cães ocorre pelo contato com urina ou água contaminada pela bactéria espiroqueta patogênica do gênero Leptospira. Objetivou-se avaliar dois testes diagnósticos na caracterização de cães com leptospirose, e associar com as principais alterações clínicos-patológicas de cães suspeitos. Foram atendidos no Hospital Veterinário da Universidade Federal de Uberlândia no período de setembro de 2019 a setembro de 2020, 24 cães com suspeita clínica de leptospirose canina. Exames complementares como hemograma completo, dosagens séricas de creatinina, ureia, ALT (alanina aminotransferase), FA (fosfatase alcalina) foram solicitados. Para o diagnóstico confirmatório de leptospirose foi realizado teste sorológico pelo teste de aglutinação microscópica (MAT) e também teste molecular pela reação em cadeia da polimerase em tempo real (qPCR). Dos 24 casos de cães suspeitos de leptospirose, seis (25%) foram positivos no MAT, e três (12,5%) no qPCR, sendo 1 (um) (4,17%) positivo nos dois testes diagnósticos avaliados. Quando se associou MAT + qPCR a positividade foi para 8/24 (33,3%). Os sorogrupos reativos predominantes foram Icterohaemorrhagiae (33,33%) e Djasiman (33,33%), seguido de Ballum (16,60%). A sensibilidade e especificidade da qPCR em relação teste padrão ouro (MAT) foi 16,7 % e 88,9%, respectivamente. Houve alterações nos níveis séricos de creatinina, ureia e FA no grupo positivo aos testes diagnósticos (MAT/qPCR), porém somente ureia e FA apresentaram valores elevados no grupo negativo. Quando se comparou os parâmetros bioquímicos no grupo positivo aos testes diagnósticos (MAT/qPCR), e negativo, não se observou nenhuma diferença estatística significativa entre os elementos avaliados (p>0,05). Conclui-se para o diagnóstico de leptospirose canina aguda, o médico veterinário deve associar os achados clínicos-patológicos aos resultados do MAT de forma pareada e o qPCR de urina.(AU)

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Bovine brucellosis is a disease that significantly impacts animal production and human health. Although many sensitive diagnostic tests are used, there is still no ideal fast serological test for all epidemiological situations. In this context, we developed peptides that mimic regions of antigenic proteins of Brucella abortus and can be used in serological diagnosis. RESULTS: From phage display technology, we randomly selected nine clones of phage displaying peptide binders to B. abortus. These clones were sequenced and translated. After molecular docking analysis, two peptides (Ba4 and Ba9) were selected, chemically synthesized, and verified for their potential diagnostic value. By enzyme-linked immunoassay (ELISA), Ba9 showed a sensitivity of up to 97.5% to detect antibodies circulating in animals with brucellosis. We incorporated the peptide Ba9 onto a bioelectrode (graphite modified with poly-3-hydroxyphenylacetic acid). Then, direct serum detection was demonstrated by differential pulse voltammetry, micrographs, and topographic analyses in addition to the average roughness coefficient (Ra) and the value of the mean squared deviation of the roughness (Rms). CONCLUSION: This work shows that the mimetic epitope of B. abortus can be useful for developing new platforms for diagnosing brucellosis. In addition, we propose a fast test based on an electrochemical sensor using graphite modified with poly-3-hydroxyphenylacetic acid.

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Genotyping methods have led to a better understanding of the epidemiology of Mycobacterium bovis (M. bovis) infection, and its transmission dynamics, as well as the possible phylogenetic relationships between Mycobacterium strains, thus making bovine tuberculosis control programs more efficient. The goal of this study was to characterize the main spoligotypes of M. bovis isolated from cattle in the State of Minas Gerais, Brazil. It was carried out in 28 municipalities of "Triângulo Mineiro" and "Alto Paranaíba" regions of the state. Viscera samples were obtained from 58 bovines positive for tuberculosis according to comparative cervical tests, and from another 100 bovines with lesions suggestive of tuberculosis, which were donated by the National Agricultural Laboratory of Pedro Leopoldo, Minas Gerais. Microbiological isolation was performed in Stonebrink medium, and molecular identification of mycobacteria was performed by PCR. Genotyping was performed using the spoligotyping method at the Agrobiotechnology and Molecular Biology Institute of National Agricultural Technology Institute-National Scientific and Technical Research Council, Buenos Aires, Argentina. Among the 158 viscera samples, we obtained 40 (25%) isolates of M. bovis, and detected 11 spoligotype patterns, with a predominance of SB1142 (37.5%), SB0121 (25.0%), and SB1145 (10.0%). Other standards, SB0295, SB1050, SB0881, SB1144, SB1802, SB0140, SB0120, and SB0849, varied from 2.5 to 7.5%, heterogeneously distributed among the municipalities. The presence of spoligotypes shared with other Brazilian states and different countries indicates their possible exchange through epidemiological relationships, such as the transit of live animals and/or genetic similarity between strains that share a common ancestor.

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Background: Biofilms have been reported as important virulent markers associated with drug resistance in urinary tractinfections (UTIs) in humans and dogs. However, in veterinary medicine, researches involving biofilm formation, treatments and preventions have been limited; yet, it is still possible to find few studies demonstrating biofilm-forming bacteriaassociated with different comorbidities such as otitis, wound infections, UTIs, and endometritis. These studies generallyselect dogs with chronic and recurrent infections, which could be an important factor in antibiotic resistance. We aimed toevaluate biofilms in sporadic cystitis regarding prevalence and drug resistance.Materials, Methods & Results: Urine samples were collected by cystocentesis from 36 client-owned dogs under clinicaland laboratory suspicion of non-recurrent urinary bladder infection (cystitis). Urine was aseptically plated onto bloodagar, MacConkey, and CLED, followed by incubation for 24 to 48 h. Definitive identification of a potential pathogen wasmade by subculture collected from an isolated colony to obtain a pure culture. The gram staining method and specificbiochemical tests (phenol red fermentation, lysine, phenylalanine, citrate, sulfide-indole-motility, and urease) were usedto distinguish and classify the bacteria. After identification, the bacteria were tested for antimicrobial susceptibility by astandard disk diffusion method, using the following antimicrobials: amoxicillin with clavulanic acid, ampicillin, ceftriaxone, ciprofloxacin, clindamycin, cefazolin, cephalothin, erythromycin, gentamicin, norfloxacin, and sulfamethoxazoletrimethoprim. The biofilm-forming ability was determined based on a culture...

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Background: Pseudomonas aeruginosa is a bacterium that belongs to the microbiota of snakes, but it may also be anopportunistic pathogen and contaminate humans through fecal contact, bites, and injuries. In snakes, this microorganismmay present high pathogenicity at certain conditions and have been associated with high morbidity and mortality. Reportsof infection of Boa constrictor by this pathogen are rare. Thus, this study aimed to describe the P. aeruginosa oral infection in a snake specimen (Boa constrictor amarali), approaching the isolation and identification of the infectious agentsinvolved, the antimicrobial sensitivity and resistance, and the therapeutic protocol adopted.Case: A free-living adult female specimen of Boa constrictor amarali (Amarals boa), with no described previous history was rescued in an urban area by the Environmental Police. Clinical evaluations showed structures of caseous aspectin the oral cavity, with hyperemia spots in the mucosa. Samples of these lesions were sent for mycological examination,and fungal forms were not found. Samples were collected for isolation and culture. The antimicrobial susceptibility of theisolated microorganisms was determined by the modified Kirby-Bauer disk diffusion method. P. aeruginosa was isolatedand showed susceptibility to amikacin, gentamicin, and polymyxin-B; intermediate susceptibility to azithromycin, and ciprofloxacin; and resistance to cephalexin, ceftiofur, chloramphenicol, and enrofloxacin. The treatment consisted of cleaningof the oral cavity, local infiltration of lidocaine for debridement of the caseous area that were later cauterized with iodine.Systemic antibiotic therapy was used, with intramuscular administration of amikacin (5 mg/kg) for the first dose and (2.5mg/kg) for the other doses with intervals of 72 h, and oral administration of metronidazole...(AU)

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Background: Pseudomonas aeruginosa is a bacterium that belongs to the microbiota of snakes, but it may also be anopportunistic pathogen and contaminate humans through fecal contact, bites, and injuries. In snakes, this microorganismmay present high pathogenicity at certain conditions and have been associated with high morbidity and mortality. Reportsof infection of Boa constrictor by this pathogen are rare. Thus, this study aimed to describe the P. aeruginosa oral infection in a snake specimen (Boa constrictor amarali), approaching the isolation and identification of the infectious agentsinvolved, the antimicrobial sensitivity and resistance, and the therapeutic protocol adopted.Case: A free-living adult female specimen of Boa constrictor amarali (Amaral’s boa), with no described previous history was rescued in an urban area by the Environmental Police. Clinical evaluations showed structures of caseous aspectin the oral cavity, with hyperemia spots in the mucosa. Samples of these lesions were sent for mycological examination,and fungal forms were not found. Samples were collected for isolation and culture. The antimicrobial susceptibility of theisolated microorganisms was determined by the modified Kirby-Bauer disk diffusion method. P. aeruginosa was isolatedand showed susceptibility to amikacin, gentamicin, and polymyxin-B; intermediate susceptibility to azithromycin, and ciprofloxacin; and resistance to cephalexin, ceftiofur, chloramphenicol, and enrofloxacin. The treatment consisted of cleaningof the oral cavity, local infiltration of lidocaine for debridement of the caseous area that were later cauterized with iodine.Systemic antibiotic therapy was used, with intramuscular administration of amikacin (5 mg/kg) for the first dose and (2.5mg/kg) for the other doses with intervals of 72 h, and oral administration of metronidazole...

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Leptospirosis in sheep is often underestimated, and leads to great economic losses for the sheep farming industry. The aim of this study was to evaluate the humoral immune response in pregnant ewes, after the injection of a commercial polyvalent vaccine for leptospirosis, and to observe the transmission of anti-Leptospira antibodies through the colostrum to the offspring. For this, 24 pregnant ewes were vaccinated for leptospirosis. Blood samples were collected prior to vaccination and then 7, 14, 21, 28, 35, 42 and 49 days after vaccination. In order to evaluate passive immunity transfer, blood samples of 32 lambs were collected during the first 48 hours after birth, and another collection was performed 10 to 21 days after birth. The lambs were placed into 2 groups: Group A (n=16): singleton lambs; and group B (n=16): twins. The sera samples were submitted to the Microscopic Agglutination Test (MAT), in which 21 Leptospira serovars were tested. The results were analyzed in a descriptive form. The number of sheep reactive to MAT gradually increased until 21 days after vaccination, and decreased right after. Of all the serovars contained in the vaccine, the largest proportion of animals were seroconverted to Hardjoprajtino serovar, Serjoe serogroup. Anti-Leptospira antibodies transferred through colostrum to lambs were detected by MAT in the serum collected 24-48 hours after birth. It was observed that 65.6% (21 out of 32) of the lambs were reactive. In the subsequent collections that occurred from 10 to 21 days after birth, a decrease in the number of animals reactive to the MAT was detected. There was no significant statistical difference for the passive transfer of antibodies between single or twin lambs.

A leptospirose é uma doença frequentemente subestimada em rebanhos ovinos e leva a grandes prejuízos à ovinocultura. O objetivo deste estudo foi avaliar a resposta imune humoral em ovelhas gestantes, após imunização com uma vacina polivalente comercial contra leptospirose, bem como verificar a transmissão dos anticorpos adquiridos às crias, pelo colostro. Para isto, 24 ovelhas gestantes foram vacinadas. Amostras de sangue foram coletadas pré-imunização, bem como 7, 14, 21, 28, 35, 42 e 49 dias após. Para avaliar a transferência passiva de anticorpos, os 32 cordeiros que nasceram dessas ovelhas foram amostrados nas primeiras 48 horas após o nascimento, bem como com 10 e 21 dias de vida. Os cordeiros foram divididos em dois grupos: partos simples (Grupo A; n=16) e partos gemelares (Grupo B, n= 16). O soro sanguíneo foi submetido à prova de Soroaglutinação Microscópica (MAT), na qual 21 sorovares de Leptospira foram testados. O número de ovelhas reativas à MAT aumentou gradualmente até os 21 dias após imunização, com posterior decréscimo. De todos os sorovares presentes na vacina, a maior parte dos animais soroconverteu para o sorovar Hardjoprajtino, do sorogrupo Serjoe. Anticorpos anti-Leptospira passaram pelo colostro e puderam ser detectados no soro dos cordeiros entre 24 e 48 horas após o nascimento. Foi observado que 65,5% (21 de 32) cordeiros foram reativos. Em coletas subsequentes, realizadas 10 e 21 dias após o nascimento, houve um decréscimo no número de animais reativos à MAT. Não houve diferença significativa na transferência de anticorpos entre cordeiros de partos simples e gemelares.

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Leptospirosis is a zoonosis that affects several species of domestic and wild animals and is an important cause of economic losses in cattle in Brazil. In this study, we determined the prevalence of bovine leptospirosis in the Triângulo Mineiro region, Minas Gerais, Brazil, identified the most frequent serovars of Leptospira interrogans, and examine the renal pathological changes associated with the disease. Samples of blood serum and kidney fragments from 100 bovine females were collected in cattle abattoir. In the serological investigation 48% of the cows were positive. The serovars for which there were more reactive animals were Wolffi (24%), Hardjo (21%) and Hebdomadis (18%). Among the positive samples, 14/48 showedantibody titers greater than 1:100, and 70.83% of the seropositive animals responded to more than one Leptospira interrogans serovar. Only one farm did not have seropositive cows and in nine farms studied, six (66.66%) presented seropositive animals to the Hebdomadis serovar. At the histopathological examination, the most frequent microscopic lesions in positive animals were hyalinization (81.25%), congestion (81.25%) and hydropic degeneration (70.83%). However, these histopathological alterations were also found in kidneys of animals negative to serology, such as hyalinization (80.77%), congestion (48.07%) and hydropic degeneration (55.77%) and these findings are unrelated to positivity. Histopathological examination of the kidneys is not indicated to replace the serological diagnosis of leptospirosis, and may be used only as a complementary examination. Despite the low frequency of seropositive animals in the Triângulo Mineiro region, the disease is present in a large number of farms. Noteworthy is the high frequency of serovar Hebdomadis and it can be considered an emerging serovar in the region. The evaluation of the frequency of this serovar in other regions becomes important, and once verified should result in the recommendation of the inclusion of this serovar in the leptospirosis control.

A leptospirose é uma zoonose que afeta várias espécies de animais domésticos e silvestres e importante causa de prejuízos à bovinocultura nacional. Objetivou-se determinar a prevalência da leptospirose e identificar os sorovares de Leptospira interrogans mais freqüentes na região do Triângulo Mineiro, além de avaliar microscopicamente as lesões renais e correlacionar estas lesões com sorovares específicos. Foram coletadas amostras de soro sanguíneo e fragmentos de rim de 100 fêmeas bovinas em abatedouro e, destas 100 amostras, 48% foram positivas. Os sorovares para os quais houve mais animais reagentes foram Wolffi (24%), hardjo (21%) e Hebdomadis (18%). Dentre as amostras positivas, 14/48 apresentaram títulos de anticorpos aglutinantes superiores a 1:100 e notou-se que 70,83% dos animais soropositivos reagiram a mais de um sorovar de Leptospira interrogans. Somente uma propriedade não possuía vacas sororreagentes para leptospirose e das nove propriedades estudadas, seis (66,66%) apresentaram animais soropositivos ao sorovar Hebdomadis. Ao exame histopatológico, as alterações microscópicas mais encontradas em animais positivos foram hialinização (81,25%), congestão (81,25%) e degeneração hidrópica (70,83%). Porém, essas alterações histopatológicas também foram encontradas em rins de animais negativos à sorologia, como hialinização (80,77%), congestão (48,07%) e degeneração hidrópica (55,77%) e estes achados não apresentaram correlação com positividade. O exame histopatológico dos rins não é indicado para substituir o diagnóstico sorológico da leptospirose, podendo ser utilizado somente como um exame complementar. Apesar da frequência de animais sororreagentes ser baixa na região do Triângulo Mineiro, a doença está presente em um grande número depropriedades. Chama a atenção a alta frequência do sorovar Hebdomadis, podendo ser considerado um sorovar emergente na região. A avaliação da frequência deste sorovar em outras regiões se torna importante, pois uma vez verificada, deve resultar na recomendação da inclusão deste sorovar no controle da leptospirose.

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Background: Bovine tuberculosis control programs are based on a standard diagnostic method, the intradermal test with purified protein derivatives, which is used to identify and eliminate diseased animals. Currently none of the tests available allow complete differentiation between infected and uninfected animals. The main limitations of the tests available are related to diagnostic sensitivity and specificity, which results in false positive reactions due to the existence of cross infections, and also false negative, inherent to the state of energy of some animals. The aim of this work was to study the intercurrence of paratuberculosis in tuberculosis reactive cattle by the comparative cervical test. Materials, Methods & Results: Three hundred and thirty four cattle were evaluated using the comparative cervical test (CCT) and serology for tuberculosis (TB) and paratuberculosis (PTB) ELISA IDEXX®. All of the animals testing positive by CCT were euthanized and necropsied. Fragments of lymph node, lung and intestine were collected and analyzed using histopathological techniques, with staining by Hematoxylin-Eosin (HE). Samples of lung and lymph nodes (retropharyngeal, submandibular, cervical and mediastinal) of the animals testing positive by CCT were evaluated using qPRC for M. bovis, and intestinal and mesenteric lymph nodes using PCR for PTB. Of the 334 cattle evaluated using the comparative cervical test, 16 were considered positive. No lesions suggestive of tuberculosis were found in the macroscopic inspection of the carcasses. The most evident anatomical and pathological finding was a thickening of intestinal mucosa, found in 12 of the 16 cattle submitted to necropsy. No microscopic lesions suggestive of TB were identified nor was the presence of M. bovis detected by qPCR. The main histopathological findings were observed in the small intestine and mesenteric lymph nodes and identified as enteritis, lymphangitis...

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