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Purulent vulvar discharges, primarily caused by genito-urinary tract infections, are an important source of economic loss for swine producers due to sow culling and mortality. However, the agents that compose the vaginal microbiota of sows and their changes during infections are not well understood. The first goal of this study was to characterize and compare the vaginal bacterial content of healthy (HE, n = 40) and purulent vulvar discharge sows (VD, n = 270) by a culture-dependent method and MALDI-TOF MS identification. Secondly, we performed 16S rRNA targeted metagenomic approach (n = 72) to compare the vaginal microbiota between these groups. We found a wide variety of bacteria, with Proteobacteria, Firmicutes, and Bacteroidota being the most abundant phyla in both groups, as well as Escherichia-Shigella, Streptococcus, and Bacteroides at the genus level. Most agents identified in the sequencing method also grew in the culture-dependent method, showing the viability of these bacteria. Alpha diversity did not differ between HE and VD sows, regarding sample richness and diversity, but a beta-diversity index showed a different microbiota composition between these groups in two tested herds. ANCOM analysis revealed that Bacteroides pyogenes were more abundant in VD females and can be a marker for this group. Other agents also require attention, such as the Streptococcus dysgalactiae and Staphylococcus hyicus found in remarkably greater relative abundance in VD sows. Network analysis revealed important positive correlations between some potentially pathogenic genera, such as between Escherichia-Shigella, Trueperella, Streptococcus, Corynebacterium, and Prevotella, which did not occur in healthy sows. We conclude that the alteration of the vaginal microbiota between healthy and purulent vulvar discharge sows, although not extreme, could be due to the increase in the relative abundance of specific agents and to associations between potentially pathogenic bacteria.

The grey-breasted parakeet (Pyrrhura griseipectus) is an endangered psittacine species that have been affected by illegal trade and deforestation. Currently, this endemic species is only found in three areas in Ceará state, in Brazil. This study aimed to investigate the frequency and diversity of Enterobacteriaceae in wild adult grey-breasted parakeets and determine their susceptibility to antimicrobial agents. Cloacal swab samples were collected from 27 individuals and environmental swabs (drag swabs) from five nests used by these birds. Twenty-seven strains from nine species of Enterobacteriaceae were recovered from cloacal swabs, and the most prevalent bacteria strains were Hafnia alvei (22%) and Pantoea agglomerans (22%). From environmental nest samples, seven strains from three bacterial species were isolated, being the P. agglomerans the most frequent species (100%). Twenty-two of the 27 isolates (81.4%) exhibited antibiotic resistance, varying from one to eight of the 12 antimicrobials commonly used. Resistance to amoxicillin was the most prevalent (70.4%), followed by azithromycin (22.2%) and ceftriaxone (18.5%). None of the strains were resistant to gentamicin, tobramycin, ciprofloxacin or tetracycline. The H. alvei was the main species presenting multidrug resistance, including resistance against meropenem, which is an important finding. These results could provide interesting information on the health of these endangered wild grey-breasted parakeets. They could also indicate that the obtained isolates are part of a group of bacteria that are typical components of the enteric microbiota of birds, which present elevated rates of resistance to amoxicillin. RESUMO: O periquito-de-cara-suja (Pyrrhura griseipectus) é uma espécie de psitacídeo considerado pela IUCN como ameaçado de extinção, resultado do comércio ilegal e do desmatamento. Atualmente, essa espécie endêmica é encontrada apenas em três áreas no estado do Ceará, Brasil. O objetivo deste estudo foi investigar a frequência e a diversidade de Enterobacteriaceae em periquitos de peito cinza adultos selvagens e determinar sua suscetibilidade a agentes antimicrobianos. Amostras de suabes cloacais foram coletadas de 27 indivíduos e de suabes ambientais (suabes de arrasto) de cinco ninhos utilizados por essas aves. Vinte e sete cepas de nove espécies de Enterobacteriaceae foram isoladas a partir de suabes cloacais, sendo as cepas bacterianas mais prevalentes Hafnia alvei (22%) e Pantoea agglomerans (22%). Das amostras ambientais de ninhos foram isoladas sete linhagens de três espécies bacterianas, sendo P. agglomerans a espécie mais frequente (100%). Vinte e dois dos 27 isolados (81,4%) exibiram resistência a antibióticos, variando de um a oito dos 12 antimicrobianos comumente usados. A resistência a amoxicilina foi a mais prevalente (70,4%), seguida por azitromicina (22,2%) e ceftriaxona (18,5%). Nenhuma das cepas era resistente à gentamicina, tobramicina, ciprofloxacina ou tetraciclina. H. alvei foi a principal espécie que apresentou resistência a múltiplas drogas e que também esteve associada a um outro achado relevante desta pesquisa, que foi a detecção de um caso de resistência ao meropenem. Esses dados fornecem informações relevantes sobre a saúde desses periquitos selvagens ameaçados e permite concluir que os isolados obtidos fazem parte de um grupo de bactérias que normalmente compõe a microbiota entérica das aves, sendo a amoxicilina envolvida em elevadas taxas de resistência.

The objective was to compare standard versus on-plate sample preparation protocols for identification of mastitis bacteria by MALDI-TOF MS. A total of 186 bacterial isolates from cows with subclinical mastitis were identified by MALDI-TOF MS after preparation using two extraction protocols. On-plate protocol was performed by applying the bacterial colony directly from the culture plate onto the plate spot. For the standard protocol, lysis of bacterial colonies using reagents was performed in a cryotube, and the resulting extract was applied onto the plate spot for analysis. The on-plate protocol showed a similar bacteria identification rate (91.4%, n = 170/186) in comparison to the standard (94.6%, n = 176/186). Identification was higher for both protocols when scores used for species-level identification (≥ 2.0) was reduced to genus-level (≥ 1.7); genus-level identification score rate increased from 94.6 to 100% when using the standard protocol, and from 91.4 to 94.6% when using the on-plate protocol. However, when compared standard (as gold standard) versus on-plate protocol, genus-level identification score rate ranged from 87.1 to 89.8%. Therefore, when the on-plate protocol fails to identify any specie, the standard extraction may be more suitable as a reference protocol for use. Strategy for increasing identification with the on-plate protocol may include upgrading the reference database library. Choice of protocol for preparation may be influenced by the bacterial type to be identified. Standard and on-plate extraction protocols of bacterial ribosomal proteins associated with MALDI-TOF MS might be alternatives to conventional microbiology methods for identification of subclinical mastitis pathogens.

Bovine genital campylobacteriosis (BGC) is a venereal and subclinical disease that affects the fertility of cattle herds, and it is caused by Campylobacter fetus subsp. venerealis . This study selected peptides mimetic to the BGC-causing agent from a phage library. Phage display is a technique that applies bacteriophage libraries that reveal peptides fused to the viral capsid in biological selections against target proteins. Biopannings were performed for biological selection in the phage library using rabbit hyperimmune serum and C. fetus subsp. venerealis protein extract. Five selected heptapeptides were considered mimetic to Cfv-NCTC 10354 based on the results of bioinformatics analysis and assays with hyperimmune serum and cervicovaginal mucus obtained from heifers. ALASLPL and LSYLFPP were the most reactive peptides and considered promising as possible mimetic immunogens for C. fetus subsp. venerealis. RESUMO: Campilobacteriose Genital Bovina (CGB) é uma doença venérea e subclínica que causa problemas reprodutivos em rebanhos, causada por Campylobacter fetus subsp. venerealis. Este trabalho teve como objetivo selecionar peptídeos miméticos ao agente da CGB de uma biblioteca de fagos. Phage display é uma técnica que aplica bibliotecas de bacteriófagos que expõem peptídeos fusionados ao capsídeo viral em seleções biológicas contra proteínas alvo. Biopannings foram realizados para seleção biológica na biblioteca de fagos por meio de soro hiperimune de coelho e extrato proteico de C. fetus subsp. venerealis. Cinco heptapeptídeos selecionados foram considerados miméticos para Cfv-NCTC 10354 a partir de análises de bioinformática e ensaios com soro hiperimune e muco cérvico-vaginal de novilhas. ALASLPL e LSYLFPP foram os peptídeos mais reativos e considerados promissores como possíveis imunógenos miméticos para C. fetus subsp. venerealis.

Bacterial diseases are common in ornamental fish, more frequently associated with ubiquitous bacteria from the aquarium environment. The disease can lead to fish mortality and cause high economic losses if not rapidly controlled. The aim of this study was to identify the main causative bacterial agents of infection in ornamental fish with different clinical signs. A total of 126 freshwater fish, from 12 families and 38 species, with clinical signs were collected in a wholesaler in Sao Paulo, SP, Brazil. Samples were taken from the eye, skin ulcers, kidneys, and gills, plated on MacConkey, CHROMagar Orientation, and blood agar and incubated under aerobic and anaerobic conditions. Bacterial identification was performed by MALDI-TOF mass spectrometry. From the 126 studied animals, 112 were positive for bacterial isolation. Among the positive animals, 32.1% presented infection caused by a single bacterial species, while in the remaining 67.9%, two to six different bacterial species were identified. A total of 259 bacterial strains were obtained and classified among 46 bacterial species. The species of higher frequency were Aeromonas veronii (26.3%), Aeromonas hydrophilla (16.2%), Shewanella putrefaciens (7.3%), Citrobacter freundii (8.1%), Vibrio albensis (5.8%), and Klebsiella pneumoniae (4.2%). MALDI-TOF MS showed to be a rapid method for diagnosis of bacterial disease outbreaks in ornamental fish establishments.

Campylobacter species are a significant cause of sheep abortion in most sheep-raising countries. The relationship between the presence of Campylobacter spp. in fecal samples and reproductive disorders was investigated in 274 sheep from 28 properties in the state of São Paulo, Brazil. Biological samples from 16 aborted fetuses, one uterus, six placentas, five uterine secretion samples, five vaginal swabs, 17 semen samples, and three preputial swabs were also subjected to bacterial isolation. The bacteria were isolated from fecal samples of 14.9% (5/28) of the properties, affecting 3.65% (10/274) of the sheep, 3.5% (9/255) of females and 5.3% (1/19) of males. Campylobacter jejuni was the most prevalent species, present in 66.67% (7) of the positive samples, followed by Campylobacter coli, present in 22.22% (2), and one strain was identified as Campylobacter spp. The birth of “weak” lambs (p=0.06, OR=6.83 and CI=1.73 to 27.05) and neonatal death (p=0.087, OR=3.5 and CI=0.83 to 14.72) were associated with the fecal isolation of Campylobacter spp. Diarrhea was also associated with the bacteria (p=0.003, OR=9.83 and CI=2.19 to 44.18). The dissemination of Campylobacter spp. in Brazilian sheep is low and that, at present, the existing strains are not responsible for significant economic losses in sheep production, especially in adult animals.

Erliquiose é uma doença causada por rickettsias pertencentes ao gênero Ehrlichia. No Brasil, estudos sorológicos e moleculares têm avaliado a ocorrência de espécies de Ehrlichia em cães, gatos, animais selvagens e seres humanos. Ehrlichia canis é a principal espécie em cães no Brasil, embora a infecção por E. ewingii tenha, recentemente, despertado suspeita em cinco cães. O DNA de E. chaffeensis foi detectado e caracterizado em cervo-do-pantanal, enquanto que E. muris e E. ruminantium ainda não foram identificadas no Brasil. A erliquiose monocítica canina causada pela E. canis parece ser altamente endêmica em muitas regiões do Brasil, embora dados de prevalência não estejam disponíveis em muitas delas. O DNA de E. canis também foi detectado e caracterizado em três gatos domésticos, enquanto anticorpos contra E. canis foram detectados em felídeos neotropicais de vida livre. Evidências sorológicas sugerem a ocorrência de erliquiose humana no Brasil, entretanto, o agente etiológico ainda não foi identificado. A melhoria do diagnóstico molecular promoverá a identificação e caracterização de espécies associadas à erliquiose humana no Brasil. Ehrlichiosis is a disease caused by rickettsial organisms belonging to the genus Ehrlichia. In Brazil, molecular and serological studies have evaluated the occurrence of Ehrlichia species in dogs, cats, wild animals and humans. Ehrlichia canis is the main species found in dogs in Brazil, although E. ewingii infection has been recently suspected in five dogs. Ehrlichia chaffeensis DNA has been detected and characterized in mash deer, whereas E. muris and E. ruminantium have not yet been identified in Brazil. Canine monocytic ehrlichiosis caused by E. canis appears to be highly endemic in several regions of Brazil, however prevalence data are not available for several regions. Ehrlichia canis DNA also has been detected and molecularly characterized in three domestic cats, and antibodies against E. canis were detected in free-ranging Neotropical felids. There is serological evidence suggesting the occurrence of human ehrlichiosis in Brazil but its etiologic agent has not yet been established. Improved molecular diagnostic resources for laboratory testing will allow better identification and characterization of ehrlichial organisms associated with human ehrlichiosis in Brazil.

Objetivando a an?lise comparativa da ocorrencia da leptospirose canina na Regi?o Urbana do Anhanduizinho, entre estudos de 2002 e 2006, avaliaram-se os ambientes dom?sticos como de risco, os efeitos de interven??o p?blica e de urbaniza??o. Obteve-se uma redu??o da positividade de 15,7% para 5,5%, onde a chance de um c?o adquirir a doen?a em 2002 (OR: 2,4) era maior do que em 2006. A diversidade de sorovares identificados demonstrou a persistencia e o potencial do agente no ambiente, podendo sugerir sorovares aut?ctones. Dentre os animais, os jovens foram os mais acometidos. Quanto ao sexo, os machos com maior positividade nos periodos, tiveram em 2006 um risco maior de contrair a doen?a (OR=2,7). Considerando o aspecto sanit?rio dos domicilios, a queda no padr?o de manuten??o predial e sanit?rio observado em 2002, na maioria com condi??es satisfat?rias de higiene e habitabilidade, inverte a chance de infec??o para 2006 (OR:2,8). A an?lise espacial dos casos positivos demonstrou em ambos estudos, os bairros Aero Rancho, o maior, e o Guanandi, o mais populoso, com persistencia de casos. Apesar das interven??es urbanisticas e melhorias na paisagem urbana empreendidas pelo poder p?blico terem favorecido a redu??o da incidencia da doen?a, ainda persistem condi??es inadequadas de habita??o, propiciadas por fatores educacionais e sociais intrinsecos ao crescimento urbano e suas aglomera??es. Medidas de controle e de educa??o em sa?de devem ser implementadas como estimulo para mudan?a do padr?o comportamental local, com a inclus? de pr?ticas de manejo do ambiente dom?stico, de higiene pessoal e dos animais de estima??o. This work aimed to evaluate the domestic environments and the effects of public intervention and of urbanization, through of the comparative analysis of the occurrence of the canine leptospirosis in the Anhanduizinho's urban area, among studies was held in 2002 and 2006 years. The results indicated the reduction of the positivity of 15,7% for 5,5%, where the chance of the dog to acquire the disease in 2002 (OR:2,4) it was larger than in 2006. The persistence and the agent's potential infection in the environment were showed through of the diversity of identified serovars and could suggest autochthonous serovars. Among the animals, the youths were the more attacked and the males with larger positivity in the periods, had in 2006 a larger risk of contracting the disease (OR=2,7) The household's pattern sanitary performed in 2002, it inverts the infection chance for 2006 (OR:2,8). The space analysis of the positive cases demonstrated in studies, the Aero Rancho, the largest uptown, and Guanandi, the most populous, with persistence of cases. Despite the interventions and improvements in the urban landscape undertaken by the government had supported the reduction of the disease incidence, still persist in household inappropriate sanitary conditions provide by educational and social factors intrinsic to the urban growth and your gatherings. Control measures, health's educational must to be taken for change of the pattern sanitary local, with the inclusion of practices of handling of the domestic atmosphere, personal hygiene and of the pet animals.

O objetivo deste estudo foi estimar a prevalencia da brucelose bovina nos 22 municípios que compõem a região denominada estrato 1 do Estado de Mato Grosso do Sul, e identificar os fatores de risco associados à infecção. A região amostrada constitui uma área de 70.214,1 km2, que representa 19,7% do Estado. O rebanho de região estudada é de, aproximadamente, 5,7 milhões de cabeças, correspondente a 23% do efetivo de 24,9 milhões de bovinos de Mato Grosso do Sul. Nas 210 propriedades amostradas, no período de dezembro de 2003 a março de 2004, foram colhidas 2.376 amostras de sangue de femeas com idade igual ou superior a 24 meses, submetidas a testes diagnósticos em série. A triagem, realizada por meio do teste do antígeno acidificado tamponado, foi seguida pelo teste confirmatório 2- meracaptoetanol. Na mesma ocasião da colheita das amostras, foi preenchido um questionário com informações de identificação, tipo de criação e práticas de manejo. Em animais, a prevalência real foi estimada em 5,6% e em rebanhos, 37,3%. As variáveis que apresentaram associação, por meio da análise univariada odds ratio (OR), com a soropositividade à brucelose foram: o tipo de exploração corte (OR 2,82), a raça Zebu (OR 2,62), aborto (OR 1,83), a existência de áreas alagadiças na propriedade (OR 1,04) e o ingresso de animais nos rebanhos (OR 1,53).Os resultados demonstram que, além da brucelose ser prevalente no estrato estudado em Mato Grosso do Sul, o controle da doença pode consistir na adoção de programa com especial atenção à exploração do tipo corte, à raça Zebu e à presença do aborto. This study aimed to estimate the prevalence of bovine brucellosis in the 22 counties which make up the region called extract 1 of the State of Mato Grosso do Sul and identify the risk factors associated with that infection. The sample region encompasses an area of 70.214,1 km2, which represents 19,7% of the State. The cattle herd in the studied region has approximately 5,7 million individuals, corresponding to 23 % of the whole of 24,9 million of bovines in the State of Mato Grosso do Sul. In the 210 sample estates in the period between December 2003 and March 2004, 2.376 blood samples were collected from females aged 24 months or older, submitted to serial diagnostic tests. The sorting, carried on through the buffered acidified antigen test, was followed by the confirmatory 2-mercaptoetanol test. On the same occasion of the sample collection, a questionnaire with information related to the identification, kind of cattle and management practices was filled out. In animals the real prevalence was estimated in 5,6% and in cattle herd, 37,3%. The variables which presented association through odds ratio univariate analysis (OR) with serum positivity to brucellosis were: the exploration of beef cattle (OR 2,82), Zebu breed (OR 2,62), abortion (OR 1,83), the existence of flooding areas in the estate (OR 1,04) and the joining of animals to the herd (OR 1,53). The results demonstrate that, despite the prevalence of brucellosis in the extract studied in the State of Mato Grosso do Sul, the control of the disease may depend on the adoption of a program focusing on the exploration of beef cattle, the Zebu breed and the presence of abortion.