Expression of genes encoding resistance in Staphylococcus spp. isolated from bovine subclinical mastitis in Brazil.

INTRODUCTION: Staphylococci are the most important agents associated with bovine mastitis. This study aimed at characterizing resistance factors to antimicrobials in Staphylococcus spp. isolated from the milk of cows with subclinical mastitis. METHODOLOGY: In vitro resistance of 243 Staphylococcus spp. isolates to antimicrobials commonly used in clinical practice was evaluated. The detection and expression of genes encoding resistance mecA (gene encoding penicillin binding protein 2a) mecALGA251 (mecA homologue), blaZ (gene encoding penicillin resistance), femA and femB (genes encoding essential factors - A and B - for the expression of methicillin resistance) and aacA-aphD (gene encoding for a bifunctional enzyme that confers resistance to gentamicin) using PCR and RT-PCR was investigated. RESULTS: One or more genes encoding resistance to different antimicrobials were detected in 184 Staphylococcus spp. SAMPLES: The femA and femB genes were the most frequent. Regarding the variables' detection (N = number of strains) and expression (% of strains), the following results were obtained: blaZ (N = 40 - 82.5%), femA (N = 147 - 47.6%), aacAaphD (N = 30 - 43.3%), femB (N = 138 - 29.7%), mecA (N = 33 - 27.3%), mecALGA251 (N = 01 - 0.0%). There was a higher occurrence of phenotypic resistant strains for amoxicillin, ampicillin and penicillin in isolates positive for detection and/or expression of blaZ gene when compared with the other genes. CONCLUSIONS: The present study provides new information on genotypic traits of Staphylococcus isolates from bovine subclinical mastitis especially regarding the evaluation of expression of genes associated with antimicrobial resistance in Staphylococcus spp. using molecular tools.

Cryptococcus gattii molecular type VGII infection associated with lung disease in a goat.

BACKGROUND: Cryptococcus gattii-induced cryptococcosis is an emerging infectious disease of humans and animals with worldwide distribution and public health importance due to its significant morbidity and mortality rate. The present study aimed to report a case of pulmonary infection by C. gattii molecular type VGII in State of São Paulo, Brazil. CASE PRESENTATION: A 5-year-old goat showing intermittent dry cough, ruminal tympany, anorexia, fever, tachycardia and tachypnea was presented for necropsy at the Veterinary Hospital of the School of Veterinary Medicine and Animal Sciences, São Paulo University, São Paulo, Brazil. Postmortem examination revealed numerous 2.0-6.0 cm diameter yellow gelatinous pulmonary masses. Tissues were evaluated by a combination of pathological, mycological, and molecular diagnostic techniques. Microscopically, pneumonia granulomatous, multifocal to coalescing, moderate, with many intralesional carminophilic yeasts was observed. The immunohistochemistry and mycological culture confirmed Cryptococcus spp. Internal transcribed spacers and orotidine monophosphate pyrophosphorylase nucleotide differentiation demonstrated that the isolate corresponds to the C. gattii VGII molecular subtype. CONCLUSIONS: To our knowledge, this is the first report of a pulmonary infection in a goat linked to C. gattii molecular type VGII in Southeastern Brazil. Our findings emphasize the need for an active surveillance program for human and animal new infections to improve the current public health policies due to expansion of the epidemiological niche of this important microorganism.

In vitro photoinactivation of bovine mastitis related pathogens.

BACKGROUND: Bovine mastitis is considered the most important disease of worldwide dairy industry. Treatment of this disease is based on the application intramammary antibiotic, which favors an increase in the number of resistant bacteria in the last decade. Photodynamic inactivation (PDI) has been investigated in different areas of Health Sciences, and has shown great potential for inactivating different pathogens, without any selection of resistant microorganisms. The objective of this study was to investigate the efficacy of PDI in the inactivation of pathogens associated with bovine mastitis. METHODS: We tested the effectiveness of PDI against antibiotic resistant strains, isolated from bovine mastitis, from the following species: Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae, Corynebacterium bovis, and the alga Prototheca zopfii. Nine experimental groups were evaluated: control, no treatment; light only, irradiation of a red light-emitting diode (λ=662 (20) nm) for 180 s; exposure to 50 µM methylene blue alone for 5 min; and PDI for 5, 10, 30, 60, 120 and 180 s. RESULTS: S. dysgalactiae, S. aureus, and C. bovis were inactivated after 30s of irradiation, whereas S. agalactiae was inactivated after 120 s and P. zopfii at 180 s of irradiation. CONCLUSION: These results show that PDI can be an interesting tool for inactivating pathogens for bovine mastitis.

Somatic cell counts as an indicator of mammary infection in periparturient cows / Contagem de células somáticas como indicador de infecção mamária em vacas periparturientes

The somatic cell count (SCC) is a screening test for the evaluation of intramammary infection; however, changes in mammary secretion during colostrogenesis can promote a physiological increase in the SCC, potentially reducing its reliability in the diagnosis of mastitis. Therefore, the aim of this study was to evaluate bovine colostrum SCC as an indicative parameter of breast infection in Holstein periparturient cows. A total of 80 samples were harvested from the first milking colostrum of 20 cows and were subjected to manual SCC and bacteriological examination. Bacterial growth was present in 36.62% of the crops; coagulase-negative Staphylococcus (CNS) was the predominant microorganism (76.92%). The median SCC in infected cows (1.8 × 106 cells/mL) was significantly higher than in uninfected cows (0.9 × 106 cells/mL) (p = 0.0451). The sensitivity (10015%), specificity (1002.2%), and false positive (1002.2%) of the SCC decreased gradually when thresholds of 0.210.0 × 106 cells/mL were adopted. In contrast, the proportion of false negatives (084.6%) revealed an opposite trend. The threshold of greatest concordance between SCC and bacteriological examination was 10.0 × 106 cells/mL; however, the sensitivity rates (15.4%), specificity (2.2%), and false positive (2.2%) were very low. Based on these results, we conclude that SCCs increased prior to the infectious processes of the mammary gland, particularly in the CNS group. However, physiological changes caused by colostrogenesis resulted in poor concordance between the SCC and bacteriological examination of the colostrum...

A contagem de células somáticas (CCS) é um teste de triagem para avaliação de infecção intramamária, porém, as alterações na composição do colostro observadas durante a colostrogênese podem promover o aumento fisiológico da CCS e inviabilizar o uso desta prova para diagnóstico da mastite. Assim, o objetivo desta pesquisa foi avaliar a CCS do colostro como parâmetro indicativo de infecção mamária em vacas Holandesas periparturientes. Para tanto, foram colhidas 80 amostras de colostro de primeira ordenha, provenientes de 20 vacas, para a CCS manual e exame bacteriológico (EB). Observou-se crescimento bacteriano em 36,62% dos cultivos, com predomínio de Staphylococcus coagulase-negativa (SCN) (76,92%). O valor mediano da CCS de vacas infectadas (1,8 x106 células/mL) foi maior do que o obtido para as vacas sadias (0,9 x106 células/mL) (P=0,0451). A sensibilidade (100 a 15%), especificidade (100 a 2,2%) e falso-positivo (100 a 2,2%) diminuíram gradativamente quando os limiares de 0,2 a 10,0 x106 células/mL foram adotados. Em contrapartida, a proporção de falso-negativo (0 a 84,6%) apresentou perfil inverso. O limiar de maior concordância entre a CCS e EB foi de 10,0 x106 células/mL, porém os índices de sensibilidade (15,4%), especificidade (2,2%) e falso-positivo (2,2%) foram muito baixos. Com base nos resultados encontrados, conclui-se que a CCS apresentou elevação diante dos processos infecciosos da glândula mamária (GM), causados especialmente por bactérias do grupo SCN. No entanto, as alterações fisiológicas decorrentes da colostrogênese resultaram em baixa concordância entre a CCS e o exame bacteriológico do colostro...

Virulence factors genes of Staphylococcus spp. isolated from caprine subclinical mastitis.

The aim of this study was to investigate genes involved in adhesion expression, biofilm formation, and enterotoxin production in isolates of Staphylococcus spp. from goats with subclinical mastitis and associate these results with the staphylococcal species. One hundred and twenty-four isolates were identified and polymerase chain reaction (PCR) was performed to detect the following genes: cna, ebpS, eno, fib, fnbA, fnbB, bap, sea, seb, sec, sed and see. The most commonly Staphylococcus species included S. epidermidis, S. lugdunensis, S. chromogenes, S. capitis ss capitis and S. intermedius. With the exception of fnbB, the genes were detected in different frequencies of occurrence in 86.3% of the Staphylococcus spp. isolates. Eno (73.2%) and bap (94.8%) were more frequently detected in coagulase-negative staphylococci (CNS); ebpS (76%), fib (90.9%) and fnbA (87%) were the most frequent genes in coagulase-positive staphylococci (CPS). Regarding enterotoxins, genes sed (28.2%) and see (24.2%) had a higher frequency of occurrence; sec gene was more frequently detected in CPS (58.8%). There was no association between the presence of the genes and the Staphylococcus species. Different virulence factors genes can be detected in caprine subclinical mastitis caused by CNS and CPS. The knowledge of the occurrence of these virulence factors is important for the development of effective control and prevention measures of subclinical mastitis caused by CNS and CPS in goats.

The neutrophil function and lymphocyte profile of milk from bovine mammary glands infected with Streptococcus dysgalactiae.

Streptococcus dysgalactiae is a bacterium that accounts for a notable proportion of both clinical and subclinical intramammary infections (IMIs). Thus, the present study explores the function of milk neutrophils and the lymphocyte profile in mammary glands naturally infected with Streptococcus dysgalactiae. Here, we used 32 culture-negative control quarters from eight clinically healthy dairy cows with low somatic cell counts and 13 S. dysgalactiae-infected quarters from six dairy cows. Using flow cytometry, we evaluated the percentage of milk monocytes/macrophages and neutrophils, expression of CD62L, CD11b and CD44 by milk neutrophils, the levels of intracellular reactive oxygen species (ROS) production and phagocytosis of Staphylococcus aureus by milk neutrophils, and neutrophil viability. Furthermore, the percentages of B cell (CD21(+)) and T lymphocyte subsets (CD3(+)/CD4(+)/CD8(-); CD3(+)/CD8(+)/CD4(-); and CD3(+)/CD8(-)/CD4(-)), and the expression of CD25 by T milk lymphocytes (CD3(+)) and T CD4(+) milk cells were also assessed by flow cytometry using monoclonal antibodies. The present study showed a higher SCC and percentage of milk neutrophils, and a decrease in the percentage of milk monocytes/macrophages from S. dysgalactiae-infected quarters when compared to uninfected ones. We also observed a higher expression of CD11b by milk neutrophils and a tendency toward a decrease in neutrophil apoptosis rate in S. dysgalactiae-infected quarters. In addition, the S. dysgalactiae-infected quarters had higher percentages of milk T cells (CD3(+)) and their subset CD3(+)CD8(+)CD4(-) cells. Overall, the present study provided new insights into S. dysgalactiae IMIs, including distinct lymphocyte profiles, and a tendency toward an inhibition of apoptosis in milk neutrophils.

Pilot study: evaluation of homeopathic treatment of Escherichia coli infected swine with identification of virulence factors involved

Aim: To evaluate the efficacy of prophylactic homeopathic treatment of swine diarrhea to achievea satisfactory balance between weight gain and cost. Methods: Escherichia coli (E. coli) wasisolated from 4 piglets with diarrhea, their clinical signs were used to select the adequate homeopathic medicine China officinalis (Chin). Newborn piglets were divided in 4 groups (n=11or 12): control group treated with the routinely used antimicrobial agent (ceftiofur 4 mg 1ml/piglet for 3 days); one group with homeopathic medicine Chin 30 cH; one group treated with biotherapy prepared from locally isolated E. coli; and one group was treated with association ofChin and biotherapy. All isolated samples of E. coli were subjected to polymerase chain reaction(PCR) to identify virulence factors in each group. Results: There was no difference in the numberof animals with diarrhea between groups treated with homeopathic medicine and control; weightgain was greater in groups treated with homeopathic medicine alone and associated withbiotherapy compared to control (p<0.05). Cost of homeopathic treatment was lower compared toceftiofur. Only one E. coli sample used to prepare biotherapy medicine tested positive forvirulence factors (F41); 3 samples from the Chin treated group tested positive for Stbenterotoxins; 1 sample from the homeopathic medicine plus biotherapy treated group and 1 sample from the biotherapy treated group tested positive for F41; 2 samples from the controlgroup tested positive for F41. Conclusion: Strain variability was too large in PCR to allow forany conclusion; despite feces tested positive for E. coli, weight gain of piglets was greater ingroups with homeopathic treatment that thus can be rated more effective in newborn piglets diarrhea.(AU)

Tuberculosis in goats and sheep slaughtered in the semiarid of the Paraíba state, Northeast region of Brazil / Tuberculose em caprinos e ovinos abatidos no semiárido da Paraíba, Brasil

The aim of this work was to isolate and characterize microorganisms in hypertrophied lymph nodes or gross lesions suggestive of tuberculosis collected from 12 goats and 28 sheep slaughtered at the public slaughterhouse of Patos municipality, Paraíba State, in the Northeast region of Brazil. The identification of mycobacteria was performed by the PRA method (PCR-Restriction Enzyme Analysis). Histopathological examination of lesions was also performed. Organs affected were liver, lung, mammary gland, bladder and mediastinal, mesenteric, submandibular, parotid, popliteal, precrural, prescapular and superficial inguinal lymph nodes. Histopathological examination showed the presence of granulomas in 8 (20.00%) animals. Of the 12 goats, 1 (8.33%) was positive in the culture of mycobacteria, and by PRA method the isolate was classified as belonging to the M. tuberculosis complex. Two (7.14%) sheep were positive for the presence of environmental mycobacteria. There was isolation of Corynebacterium pseudotuberculosis in 8 (66.66%) goats and 17 (60.71%) sheep, and simultaneous isolation of mycobacteria and C. pseudotuberculosis in 1 (8.33%) goat and 1 (3.57%) sheep. The isolation of mycobacteria of the M. tuberculosis complex in goats in this study raises concerns of public health, as professionals involved in handling these animals and the meat and milk consumers are exposed to the risk [...]

O objetivo do presente trabalho foi isolar e tipificar micro-organismos presentes em linfonodos hipertrofiados ou lesões macroscópicas sugestivas de tuberculose colhidos de 12 caprinos e 28 ovinos abatidos no matadouro público do município de Patos, Paraíba. A identificação de micobactérias foi feita com o método PRA (PCR-Restriction Enzyme Analysis). Também foi realizado o exame histopatológico das lesões. Os órgãos afetados foram fígado, pulmão, glândula mamária, bexiga e linfonodos mediastínicos, mesentéricos, submandibulares, parotídeos, poplíteos, pré-crural, pré-escapular e inguinal superficial. O exame histopatológico apontou a presença de granulomas em 8 (20,00%) animais. Dos 12 caprinos, 1 (8,33%) foi positivo no cultivo de micobactérias, e pelo método PRA o isolado foi classificado como pertencente ao complexo M. tuberculosis. Dois (7,14%) ovinos foram positivos para a presença de micobactérias ambientais. Houve isolamento de Corynebacterium pseudotuberculosis em 8 (66,66%) caprinos e em 17 (60,71%) ovinos, e isolamento simultâneo de micobactérias e C. pseudotuberculosis em 1 (8,33%) caprino e 1 (3,57%) ovino. O isolamento de micobactéria do complexo M. tuberculosis em caprinos no presente trabalho levanta preocupações do ponto de vista de saúde pública, uma vez que profissionais envolvidos na manipulação destes animais, bem como a população consumidora de carne [...]

Occurrence of Chlamydophila felis in a cattery in Osasco city, São Paulo state / Ocorrência de Chlamydophila felis em um gatil na região de Osasco, São Paulo

Chlamydophila felis is associated with upper respiratory tract infections. In the present study, 31 cats from a noncommercial shelter located in Osasco, SP, Brazil, were examined. The cats presented with clinical manifestations, which were classified from grade 1 to 4, with 4 indicating severe manifestations. In total, 16.13% of the cats presented with grade 1 severity of clinical manifestations, 25.81% presented with grade 2, 38.71% presented with grade 3, and 19.35% presented with grade 4. PCR was used to detect C. felis in samples taken from the oral mucosa and ocular conjunctiva of both eyes using sterile, dry cotton swabs. Overall, 58% of the samples were positive for C. felis. Of these animals, none showed clinical manifestations that were classified as grade 1, whereas 5.56% of cats were classified as grade 2, 61.11% were classified as grade 3, and 33.33% were classified as grade 4. The median clinical manifestation intensity score for the first group was 3 and ranged from 2 to 4. In the second group not positive for C. felis, 38.45% of the animals (5/13) presented with manifestations classified as grade 1, 53.85% (7/13) were classified as grade 2, 7.69% (1/13) were classified as grade 3, and no animals were classified as grade 4. The median clinical manifestation intensity score for the second group was 2 and ranged from 1 to 3. In this study, there was a high occurrence of C. felis in animals with clinical manifestations.

A Chlamydophila felis está associada à infecção de trato respiratório superior. No presente estudo, foram utilizados 31 felinos de um gatil não comercial, localizado em Osasco/SP. Os gatos apresentavam manifestações clínicas, classificadas de 1 a 4, sendo 4 atribuído àqueles que apresentavam pior manifestação clínica. Foi observada a intensidade de manifestação clínica grau 1 em 16,13% dos gatos, a 2 em 25,81%, a 3 em 38,71% e a 4 em 19,35%. A detecção de C. felis foi realizada por técnica de PCR em amostras obtidas com suabes de algodão, seco e estéril, de mucosa oral e de conjuntiva ocular de ambos os olhos. Verificou-se que 58% das amostras para C. felis foram positivas, entre esses animais, nenhum apresentou manifestação clínica classificada como grau 1, o grau 2 foi observado em 5,56% dos gatos, 61,11% para o 3 e 33,33% dos animais apresentava a intensidade 4. Verificou-se que para o primeiro grupo a mediana dos escores de intensidade das manifestações clínicas observadas foi de 3, variando de 2 a 4. No segundo grupo, foi observado 38,45% (5/13) dos animais para a intensidade 1, 53,85% (7/13) para a 2 e 7,69% (1/13) para a 3, nenhum animal deste grupo apresentou o grau 4. Verificou-se para o segundo grupo, a mediana dos escores de intensidade das manifestações clínicas observadas foi de 2, variando de 1 a 3. Neste trabalho foi observada uma elevada ocorrência de C. felis em animais com manifestação clínica.

Genetic similarity between APEC and Escherichia coli strains isolated from Guaruba guarouba in a survey on healthy captive psittacine birds / Similaridade genética entre APEC e cepas de Escherichia coli isoladas de Guaruba guarouba em um estudo com psitacídeos hígidos de cativeiro

The role of psittacine birds as a reservoir of avian pathogenic Escherichia coli (APEC) is not known but would be helpful in understanding the human – animal interface, since the enteric microbiota of these birds consists of Gram positive bacteria. The aim of this study was to identify the presence of APEC in feces of clinically healthy Guaruba guarouba. To do this, we isolated and analyzed E. coli from cloacal fecal samples taken from 87 psittacine birds from six zoologic parks, three commercial breeders and one conservation breeder. Of the 87 birds examined, 46 (52.87%) presented E. coli in feces. The presence of the following eight virulence genes was determined by the polymerase chain reaction (PCR): irp2, iucD, iss, vat, cvi/cva, tsh, astA, and papC, and 29 (63.04%) of 46 E. coli isolates tested were positive at least one of the eight genes studied. The frequency of virulence genes observed in isolates of E. coli were 32.6% (15/46) irp2, 26% (12/46) iucD, 19.5% iss (9/46), 17.4% vat (8/46), 17.4% cvi/cva (8/46), 8.7% tsh (4/46), 4.4% astA (2/46) and 0% papC (0/46). The isolates were grouped in 13 genotypic profiles according to virulence gene combinations, but only 2 isolates were classified as APEC, with the pattern iuc, iss, cvi/cva, irp + and iuc, iss, cvi/cva, irp, tsh, vat +. This study reveals the presence of APEC in clinically healthy captive G. guarouba, suggesting that these psittacine birds may act as reservoir for pathogenic microorganisms. Epidemiological studies are needed to determine the relevance of this species as a reservoir and the implications for conservation of endangered species G. guarouba.

O papel dos psitacídeos como reservatório de Escherichia coli patogênicas para aves (APEC) não é conhecido, mas será útil para a compreensão da interface humano-animal, uma vez que a microbiota entérica destas aves é composta por bactérias Gram-positivas. O objetivo deste estudo foi identificar a presença de APEC em fezes de Guaruba guarouba clinicamente saudáveis. Para isso, foram isoladas e analisadas E. coli presentes em fezes cloacais coletadas de 87 psitacídeos, alojados em seis zoológicos, três criatórios comerciais e um criatório conservacionista. Das 87 aves examinadas, 46 (52,87%) apresentaram E. coli nas fezes. A presença de oito genes de virulência foi determinada pela reação em cadeia pela polimerase (PCR): irp2, iucD, iss, vat, cvi/cva, tsh, astA, e papC, e 29 (63,04%) dos 46 isolados foram positivos para pelo menos um dos oito genes estudados. A frequência dos genes de virulência observada nos isolados de E. coli foi 32.6% (15/46) irp2, 26% (12/46) iucD, 19.5% iss (9/46), 17.4% vat (8/46), 17.4% cvi/cva (8/46), 8.7% tsh (4/46), 4.4% astA (2/46) e 0% papC (0/46). Os isolados foram agrupados em 13 perfis genotípicos de acordo com combinações de genes de virulência, mas apenas duas amostras foram classificadas como APEC, com o perfil iuc, iss, cvi/cva, irp + e iuc, iss, cvi/cva, irp, tsh, iuc, iss, +. Este estudo revela a presença de APEC em aves de cativeiro (G. guarouba) clinicamente saudáveis, sugerindo que estes psitacídeos possam atuar como reservatórios de micro-organismos patogênicos. Estudos epidemiológicos são necessários para determinar a relevância desta espécie como reservatório e as implicações para a conservação da espécie ameaçada G. guarouba.

Molecular detection of enteropathogenic Escherichia coli in asymptomatic captive psittacines / Detecção molecular de Escherichia coli enteropatogênica em psitacídeos assintomáticos em cativeiro

Psittaciformes are one of the most endangered groups of birds, and several Brazilian species are classified between vulnerable and critically endangered. It is thus necessary to identify agents that cause infections in captive wild animals and to assess the risks posed thereof and to design interventions to minimize the possibility of disease outbreaks, leading to the conservation of endangered species. The purpose of this study was to identify enteropathogenic Escherichia coli (EPEC) cloacal isolates from asymptomatic psittacines in captivity and evaluate the distribution of the EPEC pathotype. Cloacal swabs were obtained from 46 asymptomatic birds, and resulting isolates were tested by polymerase chain reaction (PCR) for the presence of the attaching and effacing gene (eae) and bundle-forming pilus structural gene (bfpA) of EPEC. Samples from several species were tested, and three samples were found to be positive for the eae and bfpA genes and characterized as typical EPEC. This is the first report of this pathotype in asymptomatic psittacines. Although certain E. coli strains are more pathogenic than others, various factors should be considered when determining the potential of E. coli isolates to cause disease in captive psittacines. Birds that are positive for the EPEC (typical) strain could be zoonotic sources of infection, and may have acquired these strains through contact with humans or domestic animals. These findings may also be valuable for the long-term management of endangered species ex situ as one EPEC sample was isolated from a Red-tailed Amazon (Amazona brasiliensis).

Os psitacídeos são um dos grupos de aves mais ameaçadas no mundo e diversas espécies brasileiras são classificadas desde vulneráveis à criticamente ameaçadas de extinção. Torna-se, portanto, necessário identificar os agentes que causam infecções em animais selvagens em cativeiro e determinar os riscos relacionados de modo a intervir sobre os fatores envolvidos para diminuir a possibilidade de surtos de doenças e promover a conservação de espécies ameaçadas. O objetivo deste estudo foi identificar Escherichia coli Enteropatogência (EPEC) de isolados cloacais de psitacídeos assintomáticos em cativeiro e avaliar a distribuição do patotipo EPEC. Suabes cloacais foram coletados de 46 psitacídeos assintomáticos e os isolados foram testados pela reação em cadeia pela polimerase (PCR) para a presença do gene attaching and effacing (eae) e bundle forming pilus (bfpA) de EPEC. Amostras oriundas de diversas espécies foram testadas e três amostras resultaram positivas para os genes eae e bfp e caracterizadas como EPEC típicas. Esse é o primeiro relato em psitacídeos assintomáticos para esse patotipo. Apesar de que algumas cepas de E.coli serem mais patogênicas do que outras, diversos fatores devem ser considerados para determinar o potencial de isolados de E.coli de causar doença em psitacídeos em cativeiro. Aves positivas para cepas de EPEC (típicas) poderiam ser fontes de infecção zoonóticas e adquirir essas cepas através do contato com humanos e animais domésticos. Esses achados também podem ser valiosos para o manejo a longo prazo de espécies ameaçadas ex situ já que uma amostra de EPEC foi isolada de um Papagaio-de-cara-roxa (Amazona brasiliensis).

Mycobacterium bovis infection in goats from the Northeast region of Brazil

A total of 8,058 male and female mixed-breed goats and 1-4 years of age were slaughtered over a period of 7 months at the public slaughterhouse of Patos city, Paraíba state, in the Northeast region of Brazil; 822 animals were inspected for gross lesions of tuberculosis, and 12 (1.46 percent) had lesions suggestive of tuberculosis in the mammary gland, lungs, liver and mediastinal, mesenteric, submandibular, parotid and prescapular lymph nodes. Presence of granulomatous lesions was confirmed in the submandibular lymph node of one (8.3 percent) goat at the histopathological examination and at the mycobacterium culture the same sample was confirmed positive. Isolate was confirmed as belonging to the M. tuberculosis complex by PCR restriction enzyme analysis (PRA). Spoligotyping identified the isolate into spoligotype SB0295 on the M. bovis Spoligotype Database website (www.mbovis.org), and it was classified as M. bovis. The occurrence of M. bovis in goats in this study suggests that this species may be a potential source of infection for humans and should be regarded as a possible problem in the advancement of control and eradication program for bovine tuberculosis in Brazil.

Mycobacterium bovis infection in goats from the Northeast region of Brazil.

A total of 8,058 male and female mixed-breed goats and 1-4 years of age were slaughtered over a period of 7 months at the public slaughterhouse of Patos city, Paraíba state, in the Northeast region of Brazil; 822 animals were inspected for gross lesions of tuberculosis, and 12 (1.46%) had lesions suggestive of tuberculosis in the mammary gland, lungs, liver and mediastinal, mesenteric, submandibular, parotid and prescapular lymph nodes. Presence of granulomatous lesions was confirmed in the submandibular lymph node of one (8.3%) goat at the histopathological examination and at the mycobacterium culture the same sample was confirmed positive. Isolate was confirmed as belonging to the M. tuberculosis complex by PCR restriction enzyme analysis (PRA). Spoligotyping identified the isolate into spoligotype SB0295 on the M. bovis Spoligotype Database website (www.mbovis.org), and it was classified as M. bovis. The occurrence of M. bovis in goats in this study suggests that this species may be a potential source of infection for humans and should be regarded as a possible problem in the advancement of control and eradication program for bovine tuberculosis in Brazil.

In vitro algaecide effect of sodium hypochlorite and iodine based antiseptics on Prototheca zopfii strains isolated from bovine milk.

Prototheca zopfii has been considered one of the most important causes of environmental mastitis in Brazil. These algae are refractory to conventional therapy and cause great damage to the mammary gland. The present study evaluated the in vitro algaecide effect of sodium hypochlorite and iodine based antiseptics on 27 P. zopfii strains isolated from the milk of cattle. Low concentrations of sodium hypochlorite (0.0390625-0.15625%) and iodine (0.15625-0.625%) were effective against the isolates. These antiseptics may be recommended for hygiene routines, pre and postdipping and cauterization of bovine mammary glands infected by P. zopfii.

EMJH medium with 5-fluorouracil and nalidixic acid associated with serial dilution technique used to recover Leptospira spp from experimentally contaminated bovine semen / Meio de EMJH com 5-fluorouracil e ácido nalidíxico associado a técnica das diluições seriadas usados para recuperar Leptospira spp do sêmen bovino experimentalmente contaminado

Bovine semen experimentally contaminated with Leptospira santarosai serovar Guaricura was submitted tothe modified EMJH medium with 5-fluorouracil (300mg/L) and nalidixic acid (20mg/L), named as ôselectivemediumõ and using the serial dilution technique, in order to evaluate the percentage of recovery of the addedmicroorganism. The selective EMJH medium was found with higher percentage of recovery of leptospirasand minor losses of samples due to contamination with opportunistic microorganisms than the non-selectiveEMJH medium: 151/376 (40.0%) of positive growth; and 38/376 (10.0%) contamination and 58/376 (15%) and129/376 (34.0%), respectively. These results were statistically significant (p<0. 0001; Fisher). Differences werefound when the frequencies of positive leptospires recovery have been compared in the serial dilution technique (10-1 to 10-4) between the selective and non-selective media at different dilution factors. At 1/10th dilution the percentages found were (0%, 0/80) and (38%, 30/80), at 1/100th dilution, (3%, 2/80) and (49%, 39/ 80) and at 1/1,000th dilution, (25%, 20/80) and (50%, 40/80), respectively. The percentage of recovery of leptospires was found to be directly proportional to the dilution used. The methodology of the serial dilution technique (setting at least three dilutions) and the use of selective EMJH medium have been found to be efficient for the isolation of leptospires from the bovine semen samples.

Sêmen bovino experimentalmente contaminado com a estirpe Leptospira santarosai Sorovar Guaricura foi utilizado para verificar a porcentagem de recuperação de leptospiras, utilizandoo meio de EMJH com 5-fluorouracil (300 mg/L) e ácido nalidíxico (20 mg/L), denominado ômeio seletivoõ, associado à técnica das diluições seriadas. O meio seletivo EMJH foi encontrado comporcentagem de recuperação mais elevada e com menos perda de amostras devido à contaminação com microorganismos oportunistas quando comparado com o meio EMJH não seletivo, que foram encontrados, respectivamente, com 151/376 (40.0%) de crescimento positivo e 38/376 (10.0%) de contaminação, 58/376 (15%) e 129/376 (34.0%). Estes resultados foram estatíticamente significantes (p<0, 0001; Fisher). Diferenças foram encontradas quando as freqüências de leptospiras recuperadas foram comparadas com a técnica de diluição seriada (10-1 a 10-4) e entre os meios seletivo e não-seletivo, em diferentes níveis de diluição. À diluição de 1/10, as porcentagens encontradas foram (0%, 0/80) e (38%, 30/80); à diluição 1/100, (3%, 2/80) e (49%, 39/80); e à diluição 1/1000, (25%, 20/80) e (50%, 40/80), respectivamente. A porcentagem de recuperação de leptospiras foi diretamente proporcional às diluições empregadas. A metodologia das diluições seriadas (estabelecendo pelo menos três diluições) e o uso de meio EMJH seletivo mostraram ser eficientes no isolamento de leptospiras presentes nas amostras de sêmen bovino.

EMJH medium with 5-fluorouracil and nalidixic acid associated with serial dilution technique used to recover Leptospira spp from experimentally contaminated bovine semen.

Bovine semen experimentally contaminated with Leptospira santarosai serovar Guaricura was submitted to the modified EMJH medium with 5-fluorouracil (300mg/L) and nalidixic acid (20mg/L), named as "selective medium" and using the serial dilution technique, in order to evaluate the percentage of recovery of the added microorganism. The selective EMJH medium was found with higher percentage of recovery of leptospiras and minor losses of samples due to contamination with opportunistic microorganisms than the non-selective EMJH medium: 151/376 (40.0%) of positive growth; and 38/376 (10.0%) contamination and 58/376 (15%) and 129/376 (34.0%), respectively. These results were statistically significant (p<0. 0001; Fisher). Differences were found when the frequencies of positive leptospires recovery have been compared in the serial dilution technique (10(-1) to 10(-4)) between the selective and non-selective media at different dilution factors. At 1/10(th) dilution the percentages found were (0%, 0/80) and (38%, 30/80), at 1/100(th) dilution, (3%, 2/80) and (49%, 39/ 80) and at 1/1,000(th) dilution, (25%, 20/80) and (50%, 40/80), respectively. The percentage of recovery of leptospires was found to be directly proportional to the dilution used. The methodology of the serial dilution technique (setting at least three dilutions) and the use of selective EMJH medium have been found to be efficient for the isolation of leptospires from the bovine semen samples.

Evaluation of preventive homeopathic treatment against Colibacillosis in swine production / Tratamento homeopático preventivo para Colibacillosis, na produção de suínos

Escherichia coli is the most important etiologic agent implied in neonatal diarrhea in swine; colibacillosis is the disease with highest impact in production of swine. The demands of consumers for meat without chemical residues and the ban on the use of antibiotics and chemotherapics in production of swine compelled to find alternative therapeutic and preventive treatments. Aims: to assess homeopathic treatment as preventive against colibacillosis in swine. Methods: the study was conducted in a farm in Mato Grosso, Brazil; stools of 4 piglets with diarrhea were collected to establish the presence of E. coli; concomitantly it was performed a clinical exam to identify the symptoms leading to the choice of a homeopathy remedy. Newborn piglets were divided into 4 groups (n= 11 or 12): 1) control, subjected to antibiotic treatment against diarrhea; 2) homeopathic treatment, performed with Phosphorus 30cH according to the symptoms collected; 3) biotherapic treatment, performed with Escherichia coli 30cH prepared from the locally obtained bacteria; 4) homeopathic + biotherapics treatment. Results: all 3 groups treated with homeopathy/isotherapy presented a significant reduction of diarrhea compared to the control group (p=0.02); the group treated with Phosphorus 30cH + Escherichia coli 30cH presented the highest weight gain which was significant by comparison to all other groups (p=0.001). Conclusion: homeopathic and biotherapics treatment were more effective than antibiotics in the control of diarrhea in newborn piglets; combination of homeopathic and isopathic treatment resulted in the highest weight gain. These results suggest that homeopathy and isopathy are effective alternatives for the treatment of diarrhea by E. coli in newborn swine.(AU)

Escherichia coli é o agente etiológico mais importante implicado na diarréia neonatal em suínos; a colibacilose é a enfermidade com o maior impacto na produção suína. As demandas dos consumidores por carne sem resíduos químicos e a proibição do uso de antibióticos e quimioterápicos na produção de suínos têm forçado os produtores a procurar por tratamentos curativos e preventivos alternativos. Objetivos: avaliar o tratamento homeopático como profilático na colibacilose em suínos. Métodos: o estudo foi realizado numa granja em Mato Grosso, Brasil; foram colhidas fezes de 4 leitões para pesquisar a presença de E. coli; concomitantemente foi realizado exame clínico para identificar os sintomas necessários para a escolha do medicamento homeopático. Leitões recém-nascidos foram divididos em 4 grupos (n=11-12): 1) controle: tratado com antibiótico contra a diarréia; 2) tratamento homeopático, realizado com Phosphorus 30cH segundo os sintomas colhidos; 3) bioterápico, realizado com Escherichia coli 30cH preparado da cultura da bactéria localmente isolada; 4) tratamento homeopático + bioterápico. Resultados: os 3 grupos com tratamento homeopático/isoterápico apresentaram uma redução significativa da diarréia por comparação ao grupo controle (p=0,02); o grupo tratado com Phosphorus 30cH + Escherichia coli 30cH apresentou o maior ganho de peso em relação aos outros 3 grupos (p=0,001). Conclusão: os tratamentos homeopático e bioterápico foram mais efetivos que o antibiótico no controle da diarréia em leitões recém-nascidos; a combinação do tratamento homeopático e isopático resultou no maior ganho de peso. Esses resultados sugerem que homeopatia e isopatia são alternativas efetivas no tratamento da diarréia por E. coli em suínos recém-nascidos.(AU)

Evaluation of preventive homeopathic treatment against Colibacillosis in swine production

Escherichia coli is the most important etiologic agent implied in neonatal diarrhea in swine; colibacillosis is the disease with highest impact in production of swine. The demands of consumers for meat without chemical residues and the ban on the use of antibiotics and chemotherapics in production of swine compelled to find alternative therapeutic and preventive treatments. Aims: to assess homeopathic treatment as preventive against colibacillosis in swine. Methods: the study was conducted in a farm in Mato Grosso, Brazil; stools of 4 piglets with diarrhea were collected to establish the presence of E. coli; concomitantly it was performed a clinical exam to identify the symptoms leading to the choice of a homeopathy remedy. Newborn piglets were divided into 4 groups (n= 11 or 12): 1) control, subjected to antibiotic treatment against diarrhea; 2) homeopathic treatment, performed with Phosphorus 30cH according to the symptoms collected; 3) biotherapic treatment, performed with Escherichia coli 30cH prepared from the locally obtained bacteria; 4) homeopathic + biotherapics treatment. Results: all 3 groups treated with homeopathy/isotherapy presented a significant reduction of diarrhea compared to the control group (p=0.02); the group treated with Phosphorus 30cH + Escherichia coli 30cH presented the highest weight gain which was significant by comparison to all other groups (p=0.001). Conclusion: homeopathic and biotherapics treatment were more effective than antibiotics in the control of diarrhea in newborn piglets; combination of homeopathic and isopathic treatment resulted in the highest weight gain. These results suggest that homeopathy and isopathy are effective alternatives for the treatment of diarrhea by E. coli in newborn swine.

Microbiological and histopathological aspects of canine pyometra / Aspectos microbiológicos e histopatológicos da piometria canina

As pyometra is recognized as one of the main causes of disease and death in the bitch the purposes of this study were to evaluate microbiological and histopathological aspects of canine pyometra and to research the virulence factors of the E. coli isolates identifying possible risks to human health. The microbiological isolation from the intrauterine contents of 100 dogs with pyometra was carried out and the virulence factors in the E. coli strains were identified using PCR method. This study also consisted of the counting of microorganisms colonies forming units in samples of intrauterine content, tests of antimicrobial susceptibility of the E. coli isolates and the histological examination of the uterus. E. coli was the most prevalent microorganism isolated (76.6 percent) and 120 strains (79.5 percent) were positive for sfa, 86 (56.9 percent) were positive for cnf, 87 (57.6 percent) were positive for pap, 52 (34.4 percent) were positive for hly, 51 (33.8 percent) were positive for iuc and 5 (3.3 percent) were positive for afa genes. One observed more sensitivity of E. coli to norfloxacin, polimixin B, sulphazotrin, chloranfenicol and enrofloxacin. In 42 percent of the samples of uterine walls where microorganisms were isolated, the sizes of the areas of the inflammatory responses corresponded to 39-56 percent. Virulence factors were identified in 98.0 percent of the strains evaluated, demonstrating a high frequency of potentially pathogenic E. coli. It must be considered that dogs are animals that are living in close proximity to man for thousands of years and have an important role in the transmission of E. coli to other animals and to man.

A piometra é uma enfermidade da cadela adulta, sendo a doença reconhecida como uma das causas mais comuns de morte desta espécie animal. Os objetivos deste trabalho foram a avaliação de aspectos microbiológicos e histopatológicos da piometra canina e pesquisa de fatores de virulência de E. coli, identificando possíveis riscos para a saúde humana. Foi realizado o exame microbiológico de conteúdo intra-uterino de 100 cadelas com piometra bem como a contagem de unidades formadoras de colônias de microrganismos nestas amostras, testes de susceptibilidade "in vitro" aos antimicrobianos de E. coli e pesquisa de fatores de virulência nestas estirpes, e exame histopatológico de amostras de útero. Escherichia coli foi o microrganismo mais freqüentemente isolado (76,6 por cento), sendo que 120 estirpes (79,5 por cento) foram positivas para os genes sfa, 86 (56,9 por cento) foram positivas para cnf, 87 (57,6 por cento) foram positivas para pap, 52 (34,4 por cento) foram positivas para hly, 51 (33,8 por cento) foram positivas para iuc e 5 (3,3 por cento) foram positivas para afa. Observou-se maior sensibilidade de E.coli à norfloxacina, polimixina B, sulfazotrim, cloranfenicol e enrofloxacina. Em 42 por cento das amostras de parede uterina nas quais foram isolados microrganismos, os tamanhos das áreas de processo inflamatório corresponderam a 39-56 por cento. Foram identificados fatores de virulência em 98,0 por cento das estirpes de Escherichia coli avaliadas, demonstrando uma alta freqüência de E. coli potencialmente patogênica. Deve-se considerar que os cães são animais que vivem em proximidade aos homens há milhares de anos e possuem um papel importante na transmissão de E. coli aos outros animais e também ao homem.

Microbiological and histopathological aspects of canine pyometra.

As pyometra is recognized as one of the main causes of disease and death in the bitch the purposes of this study were to evaluate microbiological and histopathological aspects of canine pyometra and to research the virulence factors of the E. coli isolates identifying possible risks to human health. The microbiological isolation from the intrauterine contents of 100 dogs with pyometra was carried out and the virulence factors in the E. coli strains were identified using PCR method. This study also consisted of the counting of microorganisms colonies forming units in samples of intrauterine content, tests of antimicrobial susceptibility of the E. coli isolates and the histological examination of the uterus. E. coli was the most prevalent microorganism isolated (76.6%) and 120 strains (79.5%) were positive for sfa, 86 (56.9%) were positive for cnf, 87 (57.6%) were positive for pap, 52 (34.4%) were positive for hly, 51 (33.8%) were positive for iuc and 5 (3.3%) were positive for afa genes. One observed more sensitivity of E. coli to norfloxacin, polimixin B, sulphazotrin, chloranfenicol and enrofloxacin. In 42% of the samples of uterine walls where microorganisms were isolated, the sizes of the areas of the inflammatory responses corresponded to 39-56%. Virulence factors were identified in 98.0% of the strains evaluated, demonstrating a high frequency of potentially pathogenic E. coli. It must be considered that dogs are animals that are living in close proximity to man for thousands of years and have an important role in the transmission of E. coli to other animals and to man.