Cutaneous phaeohyphomycosis in a greater bulldog bat (Noctilio leporinus) in northeastern Brazil.

An adult male greater bulldog bat (Noctilio leporinus) was found dead in a suburban area in the municipality of Patos, Paraiba, northeastern Brazil. At post-mortem examination, the bat was emaciated and had multifocal to coalescent grey, crusted, dry, scaly cutaneous lesions, irregularly distributed over the dorsal thoracoabdominal region, muzzle, labial commissures, ears and dorsoventral surfaces of the patagia. Histopathology revealed numerous longitudinal and transverse sections of fungal organisms, with weakly basophilic walls, associated with multifocal areas of ulceration of the epidermis, necrosis, rupture and discontinuity of collagen fibres in the dermis without any inflammatory response. Molecular identification matched the organism to Cladosporium spp, Curvularia spp, Exserohilum spp, Bipolaris spp (100%) and Alternaria spp (97%), all of which have been associated with phaeohyphomycosis. Phaeohyphomycosis should be included as a differential diagnosis of cutaneous lesions in chiropterans.

Atividade antibacteriana de nanopartículas de polipirrol diante de Staphylococcus aureus isolados de amostras de leite de vacas e cabras com mastite / Antibacterial behavior of polypyrrole nanoparticles against Staphylococcus aureus isolated from cows and goats with mastitis

The aim of the present study was to evaluate the antibacterial behavior of polypyrrole nanoparticles (PPy-NPs) in water against biofilm producer or not S. aureus isolated from cows and goats with mastitis. One hundred and thirty-eight isolates of S. aureus were initially evaluated for biofilm formation by spectrophotometry in microplates. In addition, the minimum inhibition concentration (MIC) and minimum bactericidal concentration (MBC) of PPy-NPs in water for planktonic S. aureus were determined. From the bovine samples analyzed, 5 (4.46%) S. aureus isolates showed a strong biofilm production, 17 (15.18%) moderate production, 36 (32.14%) with weak production and 54 (48.21%) did not produce biofilms. Strains from goats (26) showed no biofilm production in 18 (69.23%) strains and weak biofilm production in 8 (30.76%) strains. The MIC and MBC of S. aureus to PPy-NPs were found in the same concentration (125搭/mL) in all strains tested, regardless of biofilm production or not. This finding provides a new insight into the interaction between PPy-NPs and S. aureus, and will offer potential benefits for the control of mastitis.(AU)

Avaliação da contaminação por Staphylococcus aureus em queijo coalho artesanal elaborado com leite de cabra produzido no estado de Pernambuco / Evaluation of contamination by Staphylococcus aureus in artisanal curd cheese made from goat's milk produced in the state of Pernambuco

A fabricação de queijo coalho artesanal elaborado com leite de cabra é composta pelas etapas de obtenção do leite, refrigeração, manipulação e armazenamento, que aumentam o risco de contaminação do produto. Objetivou-se neste estudo avaliar o nível de contaminação por Staphylococcus aureus em amostras de queijo coalho artesanal produzido com leite de cabra cru no estado de Pernambuco, Brasil, bem como avaliar a concordância entre a técnica oficial da Instrução Normativa nº62/2003 (Mapa) e a técnica molecular (gene nuc) para identificar S. aureus no queijo. Houve crescimento de colônias típicas de Staphylococcus aureus em 100% das amostras, e a contagem variou de 7,0×103 a 8,6×106 UFC/g. Das 30 amostras analisadas, 18 (60,0%) apresentaram valores superiores ou iguais a 105UFC/g, e 21 (70,0%) estavam contaminadas por S. aureus. A concordância entre os métodos de diagnóstico de S. aureus em queijo coalho caprino foi moderada. O nível de contaminação dos queijos revela a necessidade de ações de melhoria das condições de elaboração do produto, a fim de garantir um produto seguro aos consumidores.(AU)

The manufacture of artisanal Coalho cheese made from goat's milk is composed of the steps of obtaining milk, refrigeration, handling and storage that increase the risk of product contamination. The objective of this study was to evaluate the level of contamination by Staphylococcus aureus in samples of artisanal Coalho cheese produced with raw goat's milk in the state of Pernambuco, Brazil. In addition to evaluating the agreement between the official technique of Normative Instruction nº62/2003 (MAPA) and the molecular technique (nuc gene) to identify S. aureus in cheese. There was growth of typical Staphylococcus aureus colonies in 100% of the samples and the count ranged from 7.0×103 to 8.6×106 CFU/g. Of the 30 analyzed samples, 18 (60.0%) presented values greater than or equal to 105CFU/g and 21 (70.0%) were contaminated by S. aureus. The agreement between the diagnostic methods of S. aureus in goat cheese was moderate. The level of contamination of cheeses reveals the need for actions to improve the preparation conditions of the product in order to guarantee a safe product to consumers.(AU)

Occurrence of Mycobacterium avium subsp. paratuberculosis in coalho cheese in the State of Pernambuco, Brazil / Ocorrência de Mycobaterium avium subsp. paratuberculosis em queijo coalho do Estado de Pernambuco, Brasil

Paratuberculosis is a chronic and incurable disease that affects ruminants and other domestic animals. It is caused by Mycobacterium avium subsp. paratuberculosis (MAP) that may also be involved in some human diseases such as Crohn's disease, type 1 diabetes, sarcoidosis, multiple sclerosis, and Hashimoto's thyroiditis. The objective of this study was to investigate the occurrence of MAP DNA in samples of artisanal coalho cheese purchased in the State of Pernambuco. Forty samples of coalho cheese submitted to the Real Time Polymerase Chain Reaction (qPCR) technique were analyzed for the detection of the MAP region IS900. 11 (27.5%) were positive with a mean of 195.9 MAP colony forming unit (CFU) per gram of each sample, with a minimum of 30.3 CFU/g and a maximum of 324.2 CFU/g. Thus, this type of cheese that is one of the most consumed in this region of Brazil constitutes a source of human exposure to MAP. Further research in this area should be performed to evaluate the viability of the bacteria in this cheese type.(AU)

Paratuberculose é uma enfermidade crônica e incurável que acomete ruminantes e outras espécies de animais domésticos. É causada pelo Mycobacterium avium subsp. paratuberculosis (MAP) e ainda há a suspeita do seu envolvimento em enfermidades nos humanos como a doença de Crohn, diabetes tipo 1, sarcoidose, esclerose múltipla e tireoidite de Hashimoto. Objetivou-se com esta pesquisa investigar a ocorrência do DNA de MAP em amostras de queijo coalho artesanal adquiridas em estabelecimentos comerciais do Estado de Pernambuco. 40 amostras de queijo coalho artesanal foram submetidas a técnica de Reação em Cadeia da Polimerase em Tempo Real (qPCR) para detecção da região IS900 do MAP. 11 (27,5%) foram positivas com uma média de 195,9 unidades formadoras de colônia (UFC) de MAP por grama de queijo, com detecção mínima de 30,3UFC/g e máxima de 324,2UFC/g. Sendo assim, esse tipo de queijo que é um dos mais consumidos nesta região do Brasil constitui uma fonte de exposição humana ao MAP. Mais pesquisas nessa área devem ser realizadas para avaliar a viabilidade dessa bactéria no queijo coalho.(AU)

Elevada frequência de Aeromonas spp. e genes de virulência em cultivos de tilápia-do-nilo (Oreochromis niloticus) em tanques-rede, na região semiárida de Pernambuco, Brasil / High frequency of Aeromonas spp. and virulence genes in Nile Tilapia farms (Oreochromis niloticus) in semi-arid regions of Pernambuco, Brazil

Objetivou-se avaliar a ocorrência de Aeromonas spp. em peixes e amostras de água na região semiárida de Pernambuco e avaliar a frequência de aerolissina (aerA), enterotoxina citotóxica (act), enterotoxina citotônica (alt) e serina protease (ahp) nesses isolados. Foram analisados 70 peixes vivos e oito mortos com sinais clínicos de aeromoniose e 16 amostras de água. Aeromonas spp. foram identificadas por análises microbiológicas (provas bioquímicas) e molecular, usando-se primers específicos para a região 16S rRNA, e a distribuição dos quatro fatores de virulência (aerA, alt, act e ahp) foi investigada por ensaio de PCR. Cento e cinquenta e cinco (84,7%) isolados foram confirmados como Aeromonas spp. na análise molecular. Os genes de virulência mais frequentes foram act (53,55%) e aerA (51,61%). De acordo com o tipo de amostra, observou-se maior frequência do gene aerA (87,5% P=0,0474) em isolados de peixes mortos e a menor frequência do gene act (47,73% P=0,0002) em peixes vivos. Este estudo demonstrou a presença de aeromoniose no cultivo de tilápias em tanques-rede, nos municípios de Jatobá e Petrolândia, na região semiárida de Pernambuco. A detecção de aerA, act e alt pode ser utilizada na tipagem de virulência de Aeromonas spp.(AU)

The purpose of this study was to evaluate the occurrence of Aeromonas spp. from fishes and tilapia net-cage farm water in semi-arid regions of Pernambuco and to evaluate the frequency of the aerolysin (aerA), cytotoxic enterotoxin (act), cytotonic enterotoxin (alt) and serine protease (ahp) genes in Aeromonas isolates. 70 live and eight dead fish with aeromoniosis clinical signs and 16 water samples were analyzed. Aeromonas spp. isolated were identified by microbiological (biochemical evidence) and molecular analysis using specific primers for 16SrRNA region, while the distribution of four virulence factors, including aerA, alt, act and ahp, was investigated by PCR assay. One hundred fifty-five (84.7%) isolates were confirmed as Aeromonas spp. by molecular analysis. The most frequent virulence genes in isolates were act (53.55%) and aerA (51,61%). According to the kind of sample, the higher frequency of aerA gene (87.5% P= 0.0474) was observed in isolates from dead fish and the lowest frequency of act gene (47.73% P= 0.0002) from live fish. This study found the presence of aeromoniosis on tilapia farming in net-cages on Jatobá and Petrolândia counties in the semiarid Pernambuco region. The detection of aerA, act and alt can be used for virulence typing of Aeromonas spp. isolates.(AU)

Padronização de uma PCR para diagnóstico molecular de Microsporum canis em amostras de pelos e crostas de cães e gatos / Standardization of a pcr for molecular diagnosis of microsporum canis in samples of fur and crusts of dogs and cats

Objetivou-se neste estudo padronizar um protocolo de reação em cadeia da polimerase (PCR) para detecção de Microsporum canis em amostras de pelos e/ou crostas de cães e gatos. Foram selecionadas 48 amostras previamente identificadas por meio de cultura. Destas, 23 foram positivas para dermatófitos no cultivo. Padronizou-se a PCR a partir de primers desenhados para o alvo M. canis. Sessenta e um por cento (14/23) das amostras positivas para dermatófitos foram identificadas como M. canis em cultura. Desse total, 71,4% (10/14) apresentaram um fragmento de 218pb compatível com o esperado para a espécie fúngica alvo dessa reação. Observou-se uma sensibilidade de 71,4% e especificidade de 100% na PCR, além de uma boa concordância entre essas técnicas de diagnóstico (Kappa: 0,78; P<0,0001). O protocolo utilizado neste estudo apresentou alta especificidade na detecção de M. canis diretamente de amostras de pelos e/ou crostas de cães e gatos, viabilizando um diagnóstico mais rápido e específico, podendo esse protocolo ser empregado como um método confirmatório para agilizar a detecção de M. canis.(AU)

The aim of this study was to standardize a Polymerase Chain Reaction protocol (PCR) for the detection of Microsporum canis in fur and/or crusts of dogs and cats. 48 samples previously identified by culture were selected. Of these, 23 were positive for dermatophytes in culture. PCR was standardized from drawn primers whose target is M. canis. A total of 61% (14/23) of the dermatophyte positive samples were identified as M. canis in culture. Of this total, 71.4% (10/14) presented a fragment of 218bp compatible with that expected for the fungal species target of the reaction. A sensitivity of 71.4% and specificity of 100% in the PCR were observed, in addition to a good agreement between the techniques (Kappa: 0.78; P<0.0001). The protocol used in this study showed high specificity in the detection of M. canis directly from fur and/or crusts of dogs and cats, making possible a faster and more specific diagnosis. This protocol could be used as a confirmatory method, speeding the detection of M. canis.(AU)

Short communication: High frequency of ß-lactam-resistant Staphylococcus aureus in artisanal coalho cheese made from goat milk produced in northeastern Brazil.

Reports of ß-lactam-resistant Staphylococcus aureus in artisanal goat cheese are increasing, and this phenomenon is relevant to public health. The objective of the present study was to determine the prevalence of S. aureus strains carrying the blaZ and mecA resistance genes, as well as the genes encoding the staphylococcal enterotoxins SEA, SEB, SEC, SED, SEE, and TSST-1 in artisanal coalho cheese made from goat milk produced in northeastern Brazil. We used biochemical and molecular tests to characterize 54 S. aureus isolates found in artisanal coalho cheese collected from commercial establishments producing animal products in 11 municipalities of Pernambuco State, Brazil. A PCR analysis revealed that 42.6% (23/54) of the isolates were positive for the blaZ gene, and 7.4% (4/54) were resistant to methicillin by phenotypic testing. We did not detect mecA or any genes encoding enterotoxins. The presence of S. aureus carriers of the blaZ gene and the identification of methicillin-resistant S. aureus strains are of concern for the health of consumers of this type of cheese.

Imunodetecção de Toxoplasma gondii em tecido placentário de cabras naturalmente infectadas / Immunodetection of Toxoplasma gondii in placenta tissue of naturally infected goats

A imuno-histoquímica (IHQ) é considerada uma ferramenta rápida e precisa para a identificação de protozoários, como Toxoplasma gondii, em tecidos fetais e placentários. Neste estudo foi avaliada a imunodetecção de Toxoplasma gondii em tecido placentário de cabras naturalmente infectadas. Foram coletadas e analisadas 80 amostras de placentas de cabras procedentes de único rebanho com sorologia positiva para T. gondii na técnica de ELISA. Na histopatologia, 27/80 amostras apresentaram lesões sugestivas de infecção por protozoários. Após a avaliação histopatológica, procedeu-se à realização da técnica de imuno-histoquímica, obtendo-se 85,2% (23/27) de amostras com marcação positiva. A imunodetecção ocorreu no epitélio de revestimento das vilosidades coriônicas e foi classificada de acordo com o grau de intensidade da imunomarcação. Também foi evidenciada imunomarcação no interior dos vasos sanguíneos fetais em 8,69% (2/23) das amostras. Este estudo demonstrou que a técnica de IHQ se comportou como uma ferramenta valiosa no diagnóstico da infeção por T. gondii em tecido placentário de cabras naturalmente infectadas e complementou, de forma decisiva, o diagnóstico, além de agregar maior valor aos resultados obtidos nas análises histopatológica e sorológica.(AU)

Immunohistochemistry (IHC) is considered to be a rapid and accurate tool for the identification of protozoa such as Toxoplasma gondii in fetal and placental tissues. In this study, we evaluated the immunodetection of Toxoplasma gondii in placental tissue from naturally infected goats. A total of 80 samples of goat placentas from a single herd with positive ELISA serology for T. gondii were collected and analyzed. In the histopathology, 27/80 samples presented lesions suggestive of protozoal infection. After the histopathological evaluation, the immunohistochemistry technique was performed, obtaining 85.2% (23/27) of samples with positive marking. Immunodetection occurred in the lining epithelium of the chorionic villi and was classified according to the degree of intensity of the immunostaining. Immunostaining within the fetal blood vessels was also evidenced in 8.69% (2/23) of the samples. This study demonstrated that the IHQ technique behaved as a valuable tool in the diagnosis of T. gondii infection in placental tissue of naturally infected goats completing the diagnosis in a decisive way and adding greater value to the results obtained in the histopathological and serological analysis.(AU)

Development and standardization of a western blotting test for detection of antibodies against B. abortus / Desenvolvimento e padronização do teste Western blotting para detecção de anticorpos contra B. abortus

Brucellosis is an infectious disease caused by bacteria of the genus Brucella spp. with diagnosis based on use of serological techniques. The present study aimed to develop and standardize a western blotting (WB) test for detection of antibodies against B. abortus. Samples from two groups of cattle were analyzed: group I: 60 serum samples from true positive and true negative vaccinated animals (30 positive samples from infected animals according to rose bengal test (RBT), 2-mercaptoethanol, serum agglutination test (SAT) and complement fixation test (CFT) and 30 RBT negatives samples); group II: 383 field samples (90 positive and 293 CFT negative sera). The most reactive band in the western blotting, which properly identified and separated infected from non - infected had a molecular weight of ≤ 20kDa. The sensitivity, specificity and accuracy of the WB compared to RBT was 93%, 99%, 98%, respectively and k= 0.938. When compared to CFT, the sensitivity, specificity and accuracy of the WB was 97%, 98% and 97%, respectively and k= 0.929. The WB developed and standardized in the present study is a serological test with potential use as a confirmatory test for the diagnosis of bovine brucellosis.(AU)

A brucelose é uma doença infectocontagiosa, causada por bactérias do gênero Brucella spp., com diagnóstico baseado no emprego de técnicas sorológicas. Objetivou-se neste estudo desenvolver e padronizar um teste Western blotting (WB) para detecção de anticorpos contra B. abortus. Foram analisados dois grupos de amostras bovinas: grupo I, com 60 amostras de animais verdadeiros positivos e verdadeiros negativos vacinados (30 amostras positivas de animais infectados e positivos nos testes de antígeno acidificado tamponado (AAT), 2 - mercaptoetanol (2 - ME), soroaglutinação lenta em tubos (SAT) e fixação do complemento e de 30 amostras negativas no AAT); grupo II, com 383 amostras de campo, sendo 90 soropositivas e 293 soronegativas no TFC. O resultado da análise do WB revelou peso molecular ≤20kDa como sendo a área mais reativa e característica para identificação e separação dos animais infectados dos não infectados. A sensibilidade, a especificidade e a acurácia do WB, quando este foi comparado com o AAT, foram, respectivamente, 93%, 99% e 98%, e k= 0,938. Quando comparadas com a TFC, a sensibilidade, a especificidade e a acurácia foram 97%, 98% e 97%, respectivamente, e k= 0,929. O WB padronizado neste estudo mostrou-se um teste sorológico com potencial uso como teste confirmatório no diagnóstico da brucelose bovina.(AU)

Biofilm formation on different pH conditions by Streptococcus agalactiae isolated from bovine mastitic milk.

Streptococcus agalactiae is among the most relevant aetiologic agent of bovine clinical and subclinical mastitis, a major problem for the dairy industry. In Brazil, clonal diversity, capsular typing and multidrug resistance profiles of S. agalactiae related to human and bovine infections need further investigation. Presently, S. agalactiae isolates of bovine subclinical mastitis, from Brazilian Northeastern region, were submitted to capsular and pulsed-field gel electrophoresis (PFGE)-typing, antimicrobial susceptibility and assays of biofilm formation at different time incubation and pH levels. Sixteen bovine isolates were characterized by polymerase chain reaction assay as S. agalactiae capsular type II (CTII) and classified by PFGE in A1/A2 (n = 06), B1/B2 (n = 06), C (n = 03) and D (n = 01) patterns. Bovine S. agalactiae CTII strains were classified as 25% multidrug-resistant (MDR) with susceptibility to penicillin, linezolid and vancomycin. Biofilm formation on abiotic surface was strain- and time-dependent with significantly higher rates at pH 6·5. In conclusion, S. agalactiae capsular type II isolates recovered from bovine subclinical mastitis produced different pH-dependent biofilm levels. Our findings suggest that biofilm production is modulated by environmental factors and provides S. agalactiae advantageous in colonizing mammary gland during mastitis development, including MDR strains. SIGNIFICANCE AND IMPACT OF THE STUDY: Streptococcus agalactiae is among the most relevant aetiologic agent of bovine clinical and subclinical mastitis, a major problem for the dairy industry. The disease may cause significant economic loss due to decreased production and milk quality and increased use of medicaments. Presently, data demonstrated that biofilm formation favours the establishment of infectious process in health mammary tissue by S. agalactiae and emphasizes that an acidic pH promotes adhesion by biofilm-forming bacterial strains. S. agalactiae strains (25%) showed resistance to tetracycline, azithromycin, erythromycin and clindamycin, and consequently were classified as multidrug-resistant strains.

Seroprevalence of selected flaviviruses in free-living and captive capuchin monkeys in the state of Pernambuco, Brazil.

Mosquito-borne diseases such as dengue, yellow fever and, more recently, Chikungunya virus (CHIKV) and Zika virus (ZIKV) have a great impact in the public health. In addition, the presence of such viruses might have an impact on wild animal conservation as well as their possible role as animal reservoir. Here, we performed a serological survey searching for antibodies against a panel of flaviviruses [ZIKV, Dengue virus (DENV), Yellow Fever virus (YFV), West Nile virus (WNV), Saint Louis Encephalitis virus (SLEV), Ilheus virus (ILHV) and Rocio virus (ROCV)] using plaque reduction neutralization test (PRNT90 ) in both free-ranging and captive capuchin monkeys (Sapajus flavius and Sapajus libidinosus). Captive and free-living monkeys were sampled between June 2015 and January 2016 in the state of Pernambuco, including in the border with State of Paraíba, the epicentre of the ZIKV epidemics in Brazil. We have found neutralizing antibodies for ZIKV, DENV-1, DENV-2, DENV-3, DENV-4, YFV, ILHV and SLEV in both S. flavius and S. libidinosus samples. No positives samples were found for ROCV and WNV. Our results suggest that these flaviviruses might be circulating in capuchin monkey in the studied region. The possible presence of these viruses represents a risk for public health, as well as for animal conservation, especially for S. flavius which is a critically endangered species, facing high risk of extinction.

Anticorpos anti-Toxoplasma gondii em carcarás (Caracara plancus) procedentes da região aeroportuária do Recife, Pernambuco, Brasil / Antibodies to Toxoplasma gondii in Crested Caracara (Caracara plancus) from Recife Airport, Pernambuco, Brazil

Objetivou-se com este estudo pesquisar a ocorrência de anticorpos anti-Toxoplasma gondii em carcarás (Caracara plancus) capturados no Aeroporto Internacional do Recife/Guararapes Gilberto Freyre, Pernambuco, Brasil. Foram analisadas 115 amostras de soros sanguíneos pelo teste de aglutinação modificada (IgG, MAT≥25) utilizando taquizoítos inativados em formalina. Do total de amostras analisadas, 5,21% (6/115) foram positivas para presença de anticorpos anti-T. gondii, 16,67% com título 1:25 (1/06) e 83,33% (5/06) com título 1:50. A ocorrência de anticorpos em carcarás procedentes de região aeroportuária fomenta a preocupação em relação a aspectos ainda pouco elucidados, relacionados principalmente à inserção de aves silvestres na cadeia epidemiológica da toxoplasmose conectada à ação antrópica, tornando próxima a interação entre animais silvestres, domésticos e o homem. Dessa forma, é notória a necessidade de estudos relacionados à dinâmica de transmissão entre os diferentes genótipos existentes nessa tríade e sua relação com o meio ambiente, a fim de determinar a influência dessa espécie animal na cadeia epidemiológica da toxoplasmose.(AU)

The objective of this study was to investigate the occurrence of antibodies against Toxoplasma gondii in carcarás (Caracara plancus) captured in the Recife/Guararapes Gilberto Freyre International Airport, in the State of Pernambuco, Brazil. 115 samples of blood sera were tested by the Modified Agglutination Test technique (IgG, MAT > 25) using tachyzoites inactivated in formalin. Of the total of the analyzed samples, 5,21% (6/115) were positive for the presence of antibodies against T. gondii, 16,67% with a titer of 1:25 (1/06) and 83,33% (5/06) with a titration of 1:50. The occurrence of antibodies in caracaras coming from airport region generate concern about aspects still poorly understood, mainly related to the inclusion of wild birds in the epidemiological chain of toxoplasmosis connected to human action, making close interaction between wild animals, domestic and man. Thus, the need for studies related to the dynamics of transmission between the different existing genotypes in this triad is evident as is its relationship with the environment to determine the influence of this animal species in the epidemiological chain of toxoplasmosis.(AU)

Phylogenetic analysis, biofilm production, and antimicrobial resistance profile of Escherichia coli isolated from slaughtered pigs / Análise filogenética, produção de biofilme e perfil de resistência antimicrobiana de Escherichia coli isoladas de suínos abatidos

Enteric diseases of bacterial origin are frequent in the pig industry, of particular notoriety are the colibacillosis that mainly affect piglets and cause great damage to the swine industry worldwide. The aim of the study was to analyze phylogenetics, to detect biofilm production, and to determine antimicrobial resistance profile in 126 strains of Escherichia coli isolated from swabs obtained from fragments of the small intestines of 235 healthy pigs killed in slaughterhouses in Pernambuco (Brazil) using polymerase chain reaction (PCR), adherence to microplates test and disc diffusion technique. Of the analyzed samples, 88.10% (111/126) were classified in phylogenetic group B1; 4.76% (6/126) in group D; 3.97% (5/126) in group B2 and, 3.17% (4/126) in group A. Antimicrobial resistance rates observed were: lincomycin 100% (126/126), erythromycin 100% (126/126), chlortetracycline 94.44% (119/126), cephalothin 51.59% (65/126), ampicillin 38.89% (49/126), sulfamethoxazole + trimethoprim 37.3% (47/126), ciprofloxacin 19.84% (25/126), norfloxacin 14.29% (18/126), gentamicin 8.73% (11/126) and, chloramphenicol 5.55% (7/126). Multiple antibiotic resistance (MAR) ranged from 0.2 to 0.9. Of the strains tested 46.03% (58/126) produced biofilm, and 99.21% (125/126) of the strains exhibited multi-resistance. Further studies are required to elucidate the importance of each phylogenetic group in pigs and to prevent the propagation of multi-resistant E. coli strains.(AU)

Doenças entéricas de origem bacteriana são frequentes na indústria de suínos, destacando-se a colibacilose, que afeta principalmente leitões e causa grandes danos à indústria suína em todo o mundo. Cento e vinte e seis cepas de Escherichia coli foram isoladas de swabs obtidos de fragmentos de intestino delgado de 235 suínos saudáveis abatidos em matadouros de Pernambuco (Brasil). O objetivo do presente estudo foi analisar filogeneticamente essas cepas, bem como detectar a produção de biofilme e determinar o perfil de resistência antimicrobiana delas, utilizando-se a reação em cadeia da polimerase (PCR), o teste de adesão em microplacas e a técnica de disco-difusão. 88,10% (111/126) das amostras foram classificadas no grupo filogenético B1; 4,76% (6/126) no grupo D; 3,97% (5/126) no grupo B2; e 3,17% (4/126) no grupo A. As taxas de resistência antimicrobiana observadas foram: lincomicina 100% (126/126), eritromicina 100% (126/126), clortetraciclina 94,44% (119/126), cefalotina 51,59% (65/126), ampicilina 38,89% (49/126), sulfametoxazol + trimetoprima 37,3% (47/126), ciprofloxacina 19,84% (25/126), norfloxacina 14,29% (18/126), gentamicina 8,73% (11/126) e cloranfenicol 5,55% (7/126). O índice de resistência múltipla (IRMA) variou de 0,2 a 0,9. Entre as amostras, 46,03% (58/126) produziram biofilme e 99,21% (125/126) foram multirresistentes. São necessários mais estudos para elucidar a importância de cada grupo filogenético em suínos e evitar a propagação de estirpes de E. coli multirresistentes.(AU)

Isolation of Clostridium perfringens and C. difficile in crab-eating fox ( Cerdocyon thous - Linnaeus 1776) from Northeastern Brazil / Isolamento de Clostridium perfringens e C. difficile em cachorro-do-mato (Cerdocyon thous - Linnaeus 1776) da região Nordeste do Brasil

The aim of the present study was to isolate Clostridium perfringens and C. difficile in crab-eating fox (Cerdocyon thous) from Northeastern Brazil. Stool samples of 18 captive crab-eating foxes from four states of Northeastern Brazil (Alagoas, Bahia, Paraíba e Pernambuco) were collected and subjected to C. perfringens and C. difficile isolation. Suggestive colonies of C. perfringens were then analyzed for genes encoding the major C. perfringens toxins (alpha, beta, epsilon and iota), beta-2 toxin (cpb2), enterotoxin (cpe), and NetB- (netB) and NetF- (netF) encoding genes. C. difficile strains were analyzed by multiplex-PCR for a housekeeping gene (tpi), toxins A (tcdA) and B (tcdB) and a binary toxin gene (cdtB). Unthawed aliquots of stool samples positive for toxigenic C. difficile were subjected to a commercial ELISA to evaluate the presence of A/B toxins. Clostridium perfringens (type A) was isolated from five (27%) samples, and only one sample was positive for beta-2 enconding gene (cpb2). Two (11%) stool samples were positive for C. difficile, but negative for A/B toxins. These two wild canids were also positive for C. perfringens type A. This is the first report of C. difficile in crab-eating fox.(AU)

O objetivo deste estudo foi isolar Clostridium perfringens e C. difficile em cachorro-do-mato (Cerdocyon thous) da região Nordeste do Brasil. Amostras de fezes de 18 cachorros-do-mato mantidos em cativeiro e oriundos de quatro estados da região Nordeste do Brasil (Alagoas, Bahia, Paraíba e Pernambuco) foram coletadas e submetidas a isolamento de C. perfringens e C. difficile. As colônias sugestivas de C. perfringens foram analisadas para os genes que codificam as principais toxinas de C. perfringens (alfa, beta, épsilon e iota), toxina beta-2 (cpb2), enterotoxina (cpe) e NetB- (netB) e NetF- (netF). As cepas de C. difficile foram analisadas por PCR-multiplex para o gene tpi, toxinas A (tcdA) e B (tcdB) e um gene de toxina binária (cdtB). Alíquotas de amostras de fezes positivas para C. difficile toxigênico foram submetidas a um ELISA comercial para avaliar a presença de toxinas A/B. Clostridium perfringens (tipo A) foi isolado de cinco (27%) amostras, e apenas uma amostra foi positiva para o gene da toxina beta-2 (cpb2). Duas (11%) amostras de fezes foram positivas para C. difficile, mas negativas para toxinas A/B. Estes dois canídeos silvestres também foram positivos para C. perfringens tipo A. Este é o primeiro relato de C. difficile em cachorro-do-mato.(AU)

Occurrence of Mycoplasma bovigenitalium and Ureaplasma diversum in dairy cattle from to Pernambuco state, Brazil / Ocorrência de Mycoplasma bovigenitalium e Ureaplasma diversum em bovinos leiteiros do estado de Pernambuco, Brasil

The objective of this study was to conduct an investigation of Mycoplasma bovigenitalium and Ureaplasma diversum infections in cattle in the microregion of the Ipanema Valley, state of Pernambuco, Brazil. Vaginal swabs were collected from 355 breeding cows in reproductive age and were analyzed by multiplex PCR (mPCR) and culture. An epidemiological investigation of risk factors was performed for Mollicutes. mPCR analysis showed that, 9.29% (33/355) of the cows were positive for M. bovigenitalium and 21.69% (77/355) for U. diversum; coinfection was observed in 2.81% (10/355) of the cows. The microbiological isolation showed, 81.81% (27/33) of Mycoplasma spp. and 24.67% (19/77) of Ureaplasma spp.. The risk factors related to Mollicutes infection identified were semi-intensive breeding system (OR= 4.6), pasture rent (OR= 3.6), non-isolation of animals with reproductive disorders (OR= 3.2), and natural mounting and artificial insemination (OR= 3.5). There was a significant association between Mollicutes infection and abortions in the first gestational third (P= 0.001). This is the first record of M. bovigenitalium and U. diversum infection in cows in the semiarid region of the state of Pernambuco, Brazil. Preventive measures directed to the identified risk factors can decrease the occurrence of Mollicutes in these herds.(AU)

O objetivo deste estudo foi realizar uma investigação de Mycoplasma bovigenitalium e Ureaplasma diversum em bovinos leiteiros da microrregião do Vale do Ipanema, estado de Pernambuco, Brasil. Foram coletados suabes vaginais de 355 vacas em idade reprodutiva. As amostras foram analisadas por multiplex PCR (mPCR) e cultura. Foi realizada uma investigação dos fatores de risco para Mollicutes. Na mPCR, 9,29% (33/355) das vacas foram positivas para M. bovigenitalium e 21,69% (77/355) para U. diversum; coinfecção foi observada em 2,81% (10/355) das vacas. O isolamento microbiológico mostrou crescimento de Mycoplasma spp. em 81,81% (27/33) das amostras e em 24,67% (19/77) para Ureaplasma spp. Os fatores de risco relacionados à infecção por Mollicutes identificados foram sistema de produção semi-intensivo (OR= 4,6), aluguel de pastagem (OR= 3,6), não isolamento de animais com desordens reprodutivas (OR= 3,2) e monta natural e inseminação artificial (OR= 3,5). Houve uma associação significativa entre a infecção por Mollicutes e abortos no primeiro terço gestacional (P=0,001). Este é o primeiro relato da infecção por M. bovigenitalium e U. diversum em vacas na região semiárida do estado de Pernambuco, Brasil. As medidas preventivas direcionadas aos fatores de risco identificados podem diminuir a ocorrência de Mollicutes nesses rebanhos.(AU)

Atypical Toxoplasma gondii genotype in feral cats from the Fernando de Noronha Island, northeastern Brazil.

Toxoplasma gondii isolates from Brazil have a different phenotypic and genotypic pattern, with predominance of virulent isolates and recombinant genotypes, compared to the North Hemisphere. Considering that a new T. gondii genotype, non-pathogenic to mice, was previously identified from free-range chickens from the Fernando de Noronha Island, Brazil, this study aimed to identify genotypes of this parasite in tissue samples of feral cats (Felis catus) from this Brazilian Island. Anti-T. gondii IgG antibodies were detected in 18/31 (58%) feral cats. Two non-virulent T. gondii isolates were obtained by mouse bioassay. Genotyping was performed by PCR-RFLP using 10 genetic markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, PK1, L358 and Apico) and an atypical strain of T. gondii (ToxoDB #146) was identified. This is the first report of this genotype in feral cats.

Detecção molecular e isolamento de Mycoplasma spp. em psitacídeos no estado de Pernambuco, Brasil / Molecular detection and isolation of Mycoplasma spp. in psittacines in Pernambuco state, Brazil

Objetivou-se com este estudo investigar a ocorrência de Mycoplasma spp., Mycoplasma galissepticum (MG) e Mycoplasma synoviae (MS) em psitacídeos de cativeiro localizado no estado de Pernambuco, Brasil. Foram estudadas 85 aves provenientes do Parque Estadual Dois Irmãos, localizado no estado do Pernambuco, Brasil. De cada psitacídeo analisado foram obtidas três amostras por meio de swabs da cloaca, palato e conjuntiva totalizando 255 amostras. As amostras coletadas foram submetidas à extração de DNA e à reação em cadeia da polimerase (PCR), sendo as positivas submetidas ao isolamento em ágar Frey. O DNA de Mycoplasma spp. foi detectado em 16,47% (14/85) dos psitacídeos estudados. Das 255 amostras analisadas, 6,66% (17/255) foram positivas para a presença de Mycoplasma spp., sendo 41,18% (7/17) provenientes da conjuntiva, 35,29% (6/17) do palato e 23,53% (4/17) da cloaca. Nenhuma amostra foi positiva para MG ou MS na PCR. Os resultados obtidos permitem confirmar a presença do DNA de Mycoplasma spp. em conjuntiva, palato e cloaca nas aves estudadas. Foram detectadas colônias semelhantes a membros da classe Mollicutes em 17,64% das amostras (3/17). Esse é o primeiro relato da presença de Mycoplasma spp. em psitacídeos de cativeiro no Nordeste do Brasil.

The aim of this study was to investigate the occurrence of Mycoplasma spp., Mycoplasma galissepticum (MG) and Mycoplasma synoviae (MS) in captive psittacines. Eighty-five wild birds from Parque Estadual Dois Irmãos, Pernambuco state, northeastern Brazil, were used. From each psittacid analyzed three samples were obtained through cloaca, palate and conjunctiva swabs, totaling 255 samples. Samples collected were submitted to DNA extraction and Polimerase Chain Reaction (PCR). Mycoplasma spp. DNA was detected in 16.47% (14/85) of psittacines studied. From 255 samples, 6.66% (17/255) were positive for Mycoplasma spp.: 41.18% (7/17) of positivity in conjunctiva, 35.29% (6/17) in palate and 23.53% (4/17) in cloaca. There was no positive sample for MG or MS in PCR. Similar colonies were found for members of the Mollicutes Class in 17.64% of the samples (3/17). The results confirmed Mycoplasma spp. DNA in conjunctiva, palate and cloaca from the wild birds analyzed. This is the first record of Mycoplasma spp. in captive psittacines from northeastern Brazil.

Occurrence of Neospora caninum and Toxoplasma gondii DNA in brain tissue from hoary foxes (Pseudalopex vetulus) in Brazil.

The hoary fox (Pseudalopex vetulus) is a wild canid native to Brazil and is commonly found in the semiarid northeastern area living in contact with cattle. The main purpose of this study was to investigate the presence of Neospora caninum and Toxoplasma gondii DNA in hoary foxes, in the state of Paraíba, Brazil. Brain tissue samples were collected from 49 hoary foxes. From the samples, DNA extraction and the polymerase chain reaction (PCR) were performed using specific primers for N. caninum and T. gondii. The prevalences found were 14.3% (7/49) for T. gondii and 12.2% (6/49) for N. caninum. The molecular identities of the amplified products were confirmed by means of the sequencing reaction. This study demonstrated the presence of N. caninum and T. gondii DNA in free-ranging hoary foxes in Brazil for the first time, thus confirming that this species is an intermediate host.

Risk factors associated with Toxoplasma gondii infection in captive Sapajus spp.

The aim of this study was to identify risk factors associated with prevalence of Toxoplasma gondii infection in captive capuchin monkeys at a facility in the northeastern Brazil. Serum samples from 116 bearded capuchin (Sapajus libidinosus), nine blonde capuchin (Sapajus flavius), five black-capped capuchin (Sapajus apella), and four capuchin monkeys (Sapajus spp.) were tested for T. gondii antibodies using the modified agglutination test (MAT, cut-off ≥25); antibodies were found in 85.3% (99/116) of S. libidinosus, 55.6% (5/9) of S. flavius, 80.0% (4/5) of S. apella, and 75.0% (3/4) of S. spp. The risk factors associated with T. gondii seropositivity were ingestion of raw meat [OR = 4.13 (1.26; 13.50)] and old age [OR = 4.90 (1.70; 14.13)]. Results indicate a very high T. gondii seropositivity in these primate populations. To minimize exposure to T. gondii raw meat should not be fed to these animals.

Detection of Toxoplasma gondii in the milk of naturally infected goats in the Northeast of Brazil.

The aim of the present study was to detect the genomic DNA of Toxoplasma gondii in milk samples from naturally infected goats in the state of Pernambuco, (Brazil). In total, 248 blood serum samples were collected and processed from lactating goats and then submitted to a search for antibodies to T. gondii through the indirect immunofluorescence reaction. Samples with a score of 64 or more were considered positive. In total, 248 milk samples were collected and processed from the same group of goats in order to study the DNA of T. gondii using the polymerase chain reaction (PCR) technique. In the serum samples, 56/248 (22.58%) of the animals were positive, whereas the DNA of the parasite was detected in 15/248 (6.05%) of the milk samples. Five of these 15 samples were animals who were also positive in the serology. This study reports the first occurrence of the elimination of T. gondii from the milk of naturally infected goats in the north-east of Brazil. It is suggested that the consumption of in natura goat milk may constitute a potential risk to the health of milk consumers in this region.