Causes of equine abortion, stillbirth, and perinatal mortality in Brazil / Causas de abortamento, natimortalidade e mortalidade em equinos no Brasil

Abortion and complications in reproduction are important causes of economic loss in horse breeding. Studies of its causal agents can help to identify the primary pathogens or other factors involved and define appropriate measures to reduce its occurrence. This research aimed to investigate the primary causes of equine abortion, stillbirth, and perinatal mortality in regions of Brazil. Tissue from aborted fetuses, stillbirths, neonates and foals submitted to the Biological Institute of São Paulo, Brazil, from January 2010 to July 2013 were processed for viral and bacterial isolation, polymerase chain reaction (PCR), histology, and immunohistochemistry. Bacterial infection was the primary detected cause of abortion, found in 16 of the 53 animals submitted for bacterial analysis followed by viruses analysis in 2 of 105 animals, and noninfectious causes (neonatal isoerythrolysis) in 2 of 105 animals. Fungi were found in a single sample of 53 tested. The most frequent bacteria recovered were Escherichia coli, Enterobacter aerogenes, combined E. coli and Streptococcus spp., Staphylococcus spp., and Bacillus spp. The following agents were each observed in a single sample: Arcanobacterium pyogenes, Streptococcus spp., Corynebacterium spp., Actinobacillus spp., and Rhodococcus equi. The predominant identification of fecal and other opportunistic bacteria as opposed to pathogens commonly associated with equine abortion, such as Leptospira spp. and equine herpesvirus type 1 (EHV-1), suggests the need of improving hygiene management of breeding mares to prevent bacterial infection that may cause fetal loss, stillbirth, and perinatal mortality.(AU)

Abortamento e complicações na reprodução são importantes causas de perda econômica na equideocultura. Estudos dos agentes causais podem ajudar a identificar patógenos ou outros fatores envolvidos e definir medidas apropriadas para reduzir sua ocorrência. Esta pesquisa investigou as causas primárias de aborto, natimortalidade e mortalidade perinatal em equinos de diversas regiões do Brasil. Tecidos de fetos abortados, natimortos e potros submetidos ao Instituto Biológico de São Paulo, Brasil, no período de janeiro de 2010 a julho de 2013, foram processados por meio de técnicas de isolamento viral e bacteriano, PCR, histologia e imuno-histoquímica. Infecção bacteriana foi a causa mais detectada, encontrada em 16 de 53 amostras submetidas à análise bacteriana, seguida de causa viral em 2 de 105 amostras, e causas não infecciosas (isoeritrólise neonatal) em 2 de 105 amostras. Fungo foi encontrado em uma única amostra de 53 testadas. As bactérias isoladas mais frequentemente foram Escherichia coli, Enterobacter aerogenes, E. coli associada a Streptococcus spp., Staphylococcus spp. associado a Bacillus spp. Os seguintes agentes foram observados em uma única amostra cada: Arcanobacterium pyogenes, Streptococcus spp., Corynebacterium spp., Actinobacillus spp. e Rhodococcus equi. A identificação predominante de bactérias fecais e outras bactérias oportunistas, ao invés de outros patógenos comumente associados a quadros de abortamento equino, tais como Leptospira spp. e Herpesvírus equino tipo 1, sugere a necessidade de maior atenção no manejo higiênico das éguas em reprodução, a fim de prevenir infecções bacterianas que possam causar perda fetal, natimortalidade e mortalidade perinatal.(AU)

Toxic shock syndrome caused by Streptococcus dysgalactiae subsp. equisimilis in a Mexican preschool patient / Síndrome de choque tóxico por Streptococcus dysgalactiae subsp. equisimilis en un paciente preescolar mexicano

Abstract Background: Severe infections due to Streptococcus dysgalactiae subsp. equisimilis (SDSE) have been identified in adults and may cause toxic shock syndrome, although with a low frequency. Case report: A preschool-age female patient, who started with an upper respiratory tract infection developing a gradual deterioration in the following three days, is described. She was admitted to the hospital in severe conditions, with tachypnea, tachycardia (200/min), hypotension (blood pressure 68/40 mmHg), capillary refill of 7 s, and erythematous maculopapular rash in thorax, abdomen and lower extremities. She received intensive management with an inadequate response. Furthermore, she developed multiple organ failure and died 8 h after admission. The blood culture was positive for S. dysgalactiae subsp. equisimilis. Conclusions: SDSE is a rare pathogen in children. In Mexico, cases of SDSE have not been reported probably due to an inaccurate identification. Mexican pediatricians should be alert to this situation.

Resumen Introducción: En adultos, se han identificado infecciones graves por Streptococcus dysgalactiae subsp. equisimilis (SDSE), que pueden causar el síndrome de choque tóxico causado por SDSE, aunque es de baja frecuencia. Caso clínico: Paciente de sexo femenino en edad preescolar. Comenzó con una infección del tracto respiratorio superior, y desarrolló un deterioro gradual en los siguientes tres días. Ingresó en el hospital en condiciones graves, con taquipnea, taquicardia (200/min), hipotensión (tensión arterial, TA 68/40 mmHg), llenado capilar de 7 s y erupción maculopapular eritematosa en el tórax, abdomen y extremidades inferiores. Recibió manejo intensivo, sin una buena respuesta. Posteriormente, desarrolló datos de falla orgánica múltiple y murió 8 h después de su ingreso. El hemocultivo fue positivo para S. dysgalactiae subsp. equisimilis. Conclusiones: El SDSE es un patógeno raro en los niños. En México, no se han reportado casos de SDSE probablemente debido a una identificación errónea. Los pediatras mexicanos deben estar atentos a esta situación.

Frequency of microorganisms isolated at different stages of lactation and milk production loss associated with somatic cell count and to mastitis-causing pathogens / Frequência de microrganismos isolados em diferentes fases da lactação e perda de produção de leite associada à contagem de células somáticas e aos patógenos causadores de mastite

The aim of this study was to evaluate the frequency of isolation of agents causing subclinical mastitis in a herd and to estimate production losses associated with SCC> 200,000cells/mL. Three 7-day interval microbiological cultures were performed in all lactating animals from the same farm that was evaluated from June to July. To evaluate the negative and positive isolation frequencies between the lactation phases, a Chi-square test was performed. Simple linear regressions were performed to evaluate the lactation curve of animals grouped by pathogens isolated from negative cows in the microbiological culture and with SCC <200,000cells/mL. To estimate the production losses between the groups, regression coefficients were used. Results found in this experiment were: Culture-negative cows with SCC ≥ 200,000cells/mL, cows testing positive in milk culture, with SCC <200,000cells/mL and cows testing positive in milk culture, with SCC ≥ 200,000cells/mL. Milk production was -3.5; -0.5 and -4.27kg, respectively, when compared to culture-negative cows with SCC <200,000cells/mL. Cows infected with yeast cells, Coagulase-negative staphylococci (CNS), Staphylococcus aureus and environmental streptococci produced -3.42; -0.5; -0.168 and -2.5kg of milk when compared to culture-negative cows with SCC <200,000cells/mL. SCC indicates an inflammatory reaction in the mammary gland and it is directly associated with milk production losses and with presence of microorganisms in the mammary gland.(AU)

O objetivo deste estudo foi avaliar a frequência de isolamento de agentes causadores de mastite subclínica em um rebanho e estimar as perdas de produção associadas com CCS>200.000 cél./mL. Três cultivos microbiológicos intervalados por sete dias foram realizados em todos os animais em lactação da propriedade avaliada, no período de junho a julho. Para avaliar as frequências de isolamento negativo e positivo entre as fases da lactação, foi realizado um teste de qui-quadrado. Foram realizadas regressões lineares simples para avaliar a curva de lactação dos animais agrupados por patógenos isolados em relação a vacas negativas na cultura microbiológica e com CCS < 200.000 cél./mL. Os coeficientes das regressões foram utilizados para estimar as perdas de produção entre os grupos. Vacas com resultado negativo na microbiologia, mas com CCS ≥ 200.000 cél./mL, positivas na microbiologia com CCS < 200.000 cél./mL e positivas com CCS ≥ 200.000 cél./mL, produziram por dia, respectivamente, -3,5; -0,5 e -4,27kg de leite em relação às vacas negativas com CCS < 200.000 cél./mL. Vacas infectadas com células leveduriformes, Staphylococcus coagulase negativa, Staphylococcus aureus e Streptococcus ambientais produziram, respectivamente, -3,42; -0,5; -0,168 e -2,5kg de leite, comparadas a vacas negativas com CCS < 200.000 cél./mL. A CCS, indicativa de reação inflamatória, encontra-se diretamente associada às perdas de produção de leite, assim como a presença do microrganismo na glândula mamária.(AU)

Comparison of rRNA-based reverse transcription PCR and rDNA-based PCR for the detection of streptococci in root canal infections

Abstract Objective The rDNA-based method is unable to distinguish between alive and dead cells. Alternatively, bacterial viability can be assessed by molecular methods based on ribosomal RNA (rRNA). Therefore, this study aimed to detect viable streptococci in root canal samples using rRNA-based reverse transcription polymerase chain reaction (RT-PCR), compared to an rDNA-based PCR assay. Methodology Microbiological root canal samples were obtained from 32 teeth with primary endodontic infections before (S1) and after chemomechanical preparation (S2), and after removal of intracanal medication (S3). RNA and DNA were extracted, and complementary DNA (cDNA) was synthesized from RNA using RT reaction. cDNA and genomic DNA were subjected to PCR with primers complementary to the 16S rRNA sequences of Streptococcus spp. McNemar's test was used to compare the detection rate of both assays (P<0.05). Results Streptococci were detected in 28.12% (9/32) and 37.5% (12/32) of S1 samples using rRNA- and rDNA-based PCR assays, respectively. In contrast, they were detected in only 6.25% (2/32) of S2 samples using rRNA-based RT-PCR, compared to 15.62% (5/32) using rDNA-based PCR. Finally, in S3 samples, streptococci were not detected by rRNA, whereas rDNA-based PCR still detected the bacteria in 12.5% (4/32) of the samples. The total number of PCR-positive reactions in the rDNA-based PCR was higher than in the rRNA-based assay (P<0.05). Conclusions The rRNA-based RT-PCR showed a lower detection rate of streptococci when compared to the rDNA-based PCR, suggesting that the latter may have detected dead cells of streptococci in root canal samples.

Direct identification of bovine mastitis pathogens by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in pre-incubated milk

ABSTRACT The present study aimed to compare two MALDI-TOF identification methods [(a) direct sample identification after pre-incubation; or (b) use of bacteria isolated on pre-culture)] to standard, traditional bench microbiology. A total of 120 quarter milk samples from 40 Holstein lactating cows were screened based on culture-positive results obtained by microbiological culture (reference method) with the following numbers of quarters positive per cow: 4 cows with 1, 8 cows with 2, 12 cows with 3 and 16 cows with 4 infected quarters per cow. For direct identification method, quarter milk samples (n = 120) were skimmed by centrifugation (10,000 × g/10 min) and pre-incubated at 37 ºC for 12 h. After pre-incubation, quarter milk samples were submitted to total bacterial count by flow cytometry and for a preparation protocol for bacterial ribosomal protein extraction followed by MALDI-TOF MS analysis. The direct MALDI-TOF MS identification method compared to microbiological culture correctly identified isolates of coagulase-negative Staphylococci (27.2%), Streptococcus agalactiae (21.8%), Staphylococcus aureus (14.2%), and Streptococcus uberis (5.2%). The pre-incubation protocol of milk samples, associated to the direct identification method by MALDI-TOF MS, did not increase the identification at species level (score >2.0) of pathogens causing subclinical mastitis in comparison to the method without previous incubation.

Microbial load after selective and complete caries removal in permanent molars: a randomized clinical trial

Abstract The aim of this randomized clinical trial was to compare the remaining microbial load after treatments based on complete and selective caries removal and sealing. Patients with active carious lesions in a permanent molar were randomly allocated into 2 groups: a test group (selective caries removal-SCR; n=18) and a control group (complete caries removal - CCR; n=18). Dentin samples were collected following the excavation and three months after sealing. Streptococcus species, Streptococcus mutans, Lactobacillus species, and total viable microorganisms were cultured to count the viable cells and frequency of species isolation. CCR resulted in significant lower total viable microorganisms counts (p≤0.001), Streptococcus species (p≤0.001) and Lactobacillus species (p≤0.001) initially. However, after sealing, a decrease in total viable microorganisms, Streptococcus species, and Lactobacillus species in the SCR resulted in no difference between the groups after 3 months. In conclusion, selective caries removal is as effective as complete caries removal in reducing dentin bacterial load 3 months after sealing.

Resumo O objetivo deste ensaio clínico randomizado foi comparar os microrganismos remanescentes após tratamentos baseados em remoção total de tecido cariado e selamento e a remoção seletiva de tecido cariado e selamento. Pacientes com lesões de cárie ativas em molares permanentes foram divididos aleatoriamente em dois grupos: grupo teste (remoção seletiva de tecido cariado-SCR; n=18), e grupo de controle (remoção total de tecido cariado-CCR; n=18). Amostras de dentina foram obtidas após a remoção da tecido cariado e após 3 meses de selamento das cavidades. Streptococcus spp., Streptococcus mutans, Lactobacillus spp. e microrganismos viáveis totais foram cultivados para contagem de células e frequência de isolamento de espécies. CCR resultou em menores contagens totais de microorganismos viáveis (p≤0,001), Streptococcus spp. (p≤0,001) e Lactobacillus spp. (p≤0,001) inicialmente. Entretanto, após o selamento, uma redução significativa nas contagens totais de microrganismos viáveis, Streptococcus spp. e Lactobacillus spp. resultou em nenhuma diferença entre os grupos após 3 meses. Conclui-se que a remoção seletiva de cárie é tão seletiva quanto a remoção completa de cárie na redução da infecção dentinária após três meses com selamento da lesão.

Dental caries and bacterial load in saliva and dental biofilm of type 1 diabetics on continuous subcutaneous insulin infusion

Abstract Objectives Since most of the studies evaluates diabetics on multiple daily injections therapy and continuous subcutaneous insulin infusion may help gain better metabolic control and prevent complications, the objective of this study was to evaluate the prevalence of dental caries, the unstimulated salivary flow rate and the total bacteria load, Streptococcus spp. levels and Lactobacillus spp. levels in saliva and supragingival dental biofilm of type 1 diabetics on insulin pump. Material and Methods Sixty patients with type 1 diabetes on insulin pump and 60 nondiabetic individuals were included. The dental caries evaluation was performed using ICDAS and the oral hygiene was assessed according to Greene and Vermillion Simplified Oral Hygiene Index. Unstimulated saliva and supragingival dental biofilm were collected. Total bacteria, Streptococcus spp. and Lactobacillus spp. was quantified by qPCR. Results Patients with type 1 diabetes had a higher prevalence of dental caries and filled and missing teeth when compared with the control group. These patients were associated with more risk factors for the development of dental caries, namely a lower unstimulated salivary flow rate and a higher bacterial load in saliva and dental biofilm. Conclusion Some risk factors related to dental caries were associated with type 1 diabetics. An early diagnosis combined with the evaluation of the risk profile of the diabetic patient is imperative, allowing the dental caries to be analyzed through a perspective of prevention and the patient to be integrated into an individualized oral health program.

Caracterización molecular de cepas invasivas de Streptococcus dysgalactiae subsp. equisimilis: estudio multicéntrico Argentina 2011-2012 / Molecular characterization of invasive Streptococcus dysgalactiae subsp. equisimilis. Multicenter study: Argentina 2011-2012

Streptococcus dysgalactiae subsp. equisimilis (SDSE) has virulence factors similar to those of Streptococcus pyogenes. Therefore, it causes pharyngitis and severe infections indistinguishable from those caused by the classic pathogen. The objectives of this study were: to know the prevalence of SDSE invasive infections in Argentina, to study the genetic diversity, to determine the presence of virulence genes, to study antibiotic susceptibility and to detect antibiotic resistance genes. Conventional methods of identification were used. Antibiotic susceptibility was determined by the disk diffusion and the agar dilution methods and the E-test. Twenty eight centers from 16 Argentinean cities participated in the study. Twenty three isolates (16 group G and 7 group C) were obtained between July 1 2011 and June 30 2012. Two adult patients died (8.7%). Most of the isolates were recovered from blood (60.9%). All isolates carried speJ and ssa genes. stG62647, stG653 and stG840 were the most frequent emm types. Nineteen different PFGE patterns were detected. All isolates were susceptible to penicillin and levofloxacin, 6 (26.1%) showed resistance or reduced susceptibility to erythromycin --#91;1 mef(A), 3 erm(TR), 1 mef(A) + erm(TR) and 1 erm(TR) + erm(B)--#93; and 7 (30.4%) were resistant or exhibited reduced susceptibility to tetracycline --#91;2 tet(M), 5 tet(M) + tet(O)--#93;. The prevalence in Argentina was of at least 23 invasive infections by SDSE. A wide genetic diversity was observed. All isolates carried speJ and ssa genes. Similarly to other studies, macrolide resistance (26.1%) was mainly associated to the MLS B phenotype.

Streptococcus dysgalactiae subsp. equisimilis (SDSE) posee factores de virulencia similares a Streptococcus pyogenes y, en consecuencia, produce faringitis e infecciones graves indistinguibles de las generadas por este patógeno clásico. Los objetivos del estudio fueron conocer la prevalencia de SDSE en infecciones invasivas en Argentina, estudiar su diversidad genética, determinar la presencia de genes de virulencia, ensayar su sensibilidad a los antibióticos y conocer los genes de resistencia. Se emplearon métodos convencionales de identificación. La sensibilidad se determinó por difusión, Etest y dilución en agar. Participaron 28 centros de 16 ciudades argentinas. Se obtuvieron 23 aislamientos (16 del grupo G y 7 del grupo C) desde el 1-7-2011 hasta el 30-6-2012. Se registraron 2 muertes en adultos (8,7%). La mayoría de los aislamientos fueron obtenidos de sangre (60,9%). Todos eran portadores de los genes speJ y ssa. Los genotipos más frecuentes fueron stG62647, stG653 y stG840. Se detectaron 19 pulsotipos distintos. Todos los aislamientos fueron sensibles a penicilina y levofloxacina, 6 (26,1%) presentaron resistencia o sensibilidad disminuida a eritromicina (1 mef--#91;A--#93;, 3 erm--#91;TR--#93;, 1 mef--#91;A--#93; + erm--#91;TR--#93; y 1 erm--#91;TR--#93; + erm--#91;B--#93;) y 7 (30,4%) fueron resistentes o tuvieron sensibilidad disminuida a tetraciclina (2 tet--#91;M--#93;, 5 tet--#91;M--#93; + tet--#91;O--#93;). La prevalencia anual en la Argentina fue de al menos 23 infecciones invasivas por SDSE y se observó una amplia diversidad genética. Todos los aislamientos presentaron los genes ssa y speJ. Como en otros estudios, la resistencia a macrólidos (26,1%) estuvo asociada, principalmente, al fenotipo MLS B.

Evaluación de la eficacia antimicrobiana del aceite esencial de Minthostachys verticillata y limoneno contra cepas de Streptococcus uberis aislados de mastitis bovina / Evaluation of the antimicrobial efficacy of Minthostachys verticillata essential oil and limonene against Streptococcus uberis strains isolated from bovine mastitis

Bovine mastitis is a disease that causes great economic losses per year, being Streptococcus uberis the main environmental pathogen involved. The aim of the present study was to determine the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of Minthostachys verticillata essential oil and limonene for S. uberis strains isolated from bovine mastitis. In addition, the effect of MIC on biofilm formation was analyzed. MIC values for the essential oil ranged from 14.3 to 114.5 mg/ml (1.56-12.5% v/v) and MBC between 114.5 and 229 mg/ml (12.5-25% v/v). MICs for limonene ranged from 3.3 to 52.5 mg/ml (0.39-6.25% v/v) and MBC was 210 mg/ml (25% v/v). Both compounds showed antibacterial activity and affected the biofilm formation of most of the strains tested. In conclusion, these compounds could be used as an alternative and/or complementary therapy for bovine mastitis caused by S. uberis.

La mastitis bovina es una enfermedad que causa grandes pérdidas económicas por año, Streptococcus uberis es el principal patógeno ambiental involucrado. El objetivo del presente estudio fue determinar la concentración inhibitoria mínima (CIM) y la concentración bactericida mínima (CBM) del aceite esencial de Minthostachys verticillata y del limoneno sobre cepas de S. uberis aisladas de mastitis bovina. Además, se analizó el efecto del aceite esencial y el limoneno en la CIM determinada en caso sobre la formación de biofilm de estas cepas. Los valores de CIM del aceite esencial oscilaron entre 14,3 y 114,5 mg/ml (1,56%-12,5% v/v) y los de CBM entre 114,5 y 229 mg/ml (12,5%-25% v/v). Las CIM del limoneno oscilaron entre 3,3 y 52,5 mg/ml (0,39% - 6,25% v/v) y la CBM fue de 210 mg/ml (25% v/v). Ambos compuestos mostraron actividad antibacteriana y afectaron la formación de biofilm de la mayoría de las cepas. En conclusión, estos compuestos podrían ser utilizados como terapia alternativa o complementaria para la mastitis bovina causada por S. uberis.

Microbiological analysis of bile in patients with benign and malignant biliopancreatic diseases and its consequences / Análise microbiológica da bile em pacientes com doenças biliopancreáticas benignas e malignas e suas consequências

ABSTRACT Background Bactibilia has several consequences to human health. Objetive Assessing the bile microbiology of patients with biliopancreatic diseases in order to identify bacteria and their possible infectious complications. Methods Retrospective study of 30 bile culture samples from patients with benign and malignant biliopancreatic diseases. The samples were assessed to set the bile microbiological flora and to search for its possible link with comorbidity, carcinogenesis and postoperative infectious complications. Results Thirty bile samples from patients at mean age ≈57.7 years, mostly female (n=18), were assessed. Bactibilia was found in 12 cases, mostly in patients with benign diseases (n=8), older than 50 years (n=23) and female (n=10). Adenocarcinoma of the duodenal papilla (n=9) and cholelithiasis (n=8) were the most common diseases. Escherichia coli (n=5) and Klebsiella sp (n=3) were predominantly found in patients with benign diseases; and Klebsiella sp (n=2) and Streptococcus sp (n=2) were prevalent in cancer patients. There were postoperative infectious complications in seven cases, five of them in bactibilia-associated patients (P=0.084). Conclusion Bactibilia was found in 12 samples and Escherichia coli and Klebsiella sp were most often identified in patients with benign diseases, as well as Streptococcus sp and Klebsiella sp in cancer patients. There was a trend of higher postoperative infectious complication incidence in patients with bactibilia.

RESUMO Contexto Bacteriobilia pode produzir várias consequências para a saúde humana. Objetivo Avaliar a microbiologia da bile de pacientes com doenças biliopancreáticas para identificar bactérias e possíveis consequências. Métodos Estudo retrospectivo microbiológico. Trinta amostras de bile de pacientes com doenças biliopancreáticas benignas e malignas foram avaliadas para determinar a flora microbiológica da bile e procurar alguma possível relação dessa com comorbidades, carcinogênese e complicações infecciosas pós-operatórias. Resultados As amostras de bile foram avaliadas em pacientes, com idade média ≈57,7 anos, a maioria mulheres (n=18). Evidenciou-se bacteriobilia em 12 casos, a maioria em pacientes com doenças benignas (n=8); pacientes com mais de 50 anos (n=23) e mulheres (n=10). As doenças mais comuns foram o adenocarcinoma de papila duodenal (n=9) e a colelitíase (n=8). Escherichia coli (n=5) e Klebsiella sp (n=5) foram as bactérias mais identificadas em pacientes com doenças benignas; sendo a Klebsiella sp (n=2) e o Streptococcus sp (n=2) as que predominaram nos pacientes com cânceres. As complicações pós-operatórias exclusivamente infecciosas ocorreram em sete casos, sendo em cinco desses associados à bacteriobilia (P=0,084). Conclusão Bacteriobilia foi evidenciada em 12 amostras, sendo as bactérias mais identificadas Escherichia coli e Klebsiella sp em pacientes com doenças benignas; e Streptococcus sp e Klebsiella sp nos pacientes com câncer. Existiu uma tendência a maior incidência de complicações infecciosas pós-operatórias em pacientes com bacteriobilia.

Isolamento microbiológico do canal auditivo de cães saudáveis e com otite externa na região metropolitana de Recife, Pernambuco / Microbiological isolation from the ear canal of healthy dogs and with otitis externa in the metropolitan region of Recife, Pernambuco

Otite externa (OE) é o termo utilizado para definir a inflamação do conduto auditivo externo; esta doença possui diversas etiologias, ocorre em várias espécies e é particularmente frequente em cães. Os microrganismos da microbiota residente comumente estão envolvidos na etiopatogenia da OE, sendo apontados como agentes perpetuadores da doença. O objetivo deste estudo foi investigar o perfil microbiológico de cães com conduto auditivo saudável e com otite na região metropolitana do Recife. Com o auxílio de suabes estéreis foram coletadas amostras das orelhas direita e esquerda de 41 cães, sendo 11 com OE e 30 sem OE. Foi realizado o isolamento bacteriano e fúngico das amostras cultivadas; observou-se positividade em 80% dos cães com orelhas saudáveis e presença de mais de um microrganismo em 38 amostras (63,3%); já nos cães com OE, a positividade foi 95,3%, com infecção polimicrobiana em 77,3% das amostras. No que se refere aos gêneros bacterianos, o perfil de isolamento microbiológico foi idêntico entre os cães otopatas e sadios. Os microrganismos isolados foram Staphylococcus sp., Micrococcus, Bacillus sp., Streptococcus sp. e Malassezia sp.

Otitis externa (OE) is the term used to describe inflammation of the external ear canal. This disease has many etiologies, occurs in several species and is particularly common in dogs. The resident microbiota microorganisms are commonly involved in the OE etiopathogenesis, being frequently appointed as perpetuator agents. The aim of this study was to investigate the microbiological profile of dogs with healthy ears and of others with otitis in the metropolitan region of Recife, Pernambuco, Brazil. With the aid of sterile swabs, samples of right and left ear of 41 dogs, 11 with and 30 without OE, were collected. Bacterial and fungal isolation was performed with cultured samples; positivity was observed in 80% of animals with healthy ears, with the presence of more than one microrganism in 38 samples (63.3%), whereas in dogs with OE, the positivity was 95.3% with polymicrobial infection in 77.3% samples. With regard to the genus, the microbiological profile was identical between healthy and diseased dogs. The microrganisms isolated were Staphylococcus sp., Micrococcus, Bacillus sp., Streptococcus sp. and Malassezia sp.

Genotipificación y estudio de los genes pauA y sua de aislamientos de Streptococcus uberis de mastitis bovina / Genotyping and study of the pauA and sua genes of Streptococcus uberis isolates from bovine mastitis

This study aimed to determine the clonal relationship among 137 Streptococcus uberis isolates from bovine milk with subclinical or clinical mastitis in Argentina and to assess the prevalence and conservation of pauA and sua genes. This information is critical for the rational design of a vaccine for the prevention of bovine mastitis caused by S. uberis. The isolates were typed by random amplified polymorphic DNA (RAPD) analysis and by pulsed-field gel electrophoresis (PFGE). The 137 isolates exhibited 61 different PFGE types and 25 distinct RAPD profiles. Simpson's diversity index was calculated both for PFGE (0.983) and for RAPD (0.941), showing a high discriminatory power in both techniques. The analysis of the relationship between pairs of isolates showed 92.6 % concordance between both techniques indicating that any given pair of isolates distinguished by one method tended to be distinguished by the other. The prevalence of the sua and pauA genes was 97.8 % (134/137) and 94.9 % (130/137), respectively. Nucleotide and amino acid sequences of the sua and pauA genes from 20 S. uberis selected isolates, based on their PFGE and RAPD types and geographical origin, showed an identity between 95 % and 100 % with respect to all reference sequences registered in GenBank. These results demonstrate that, in spite of S. uberis clonal diversity, the sua and pauA genes are prevalent and highly conserved, showing their importance to be included in future vaccine studies to prevent S. uberis bovine mastitis.

Este estudio pretendió determinar la relación clonal entre 137 aislamientos de S. uberis obtenidos de leche de bovinos con mastitis clínica o subclínica en la Argentina, como así también la prevalencia y la conservación de los genes sua y PauA entre dichos aislamientos. Esta información es crítica para el diseño racional de una vacuna que prevenga la mastitis bovina por S. uberis. Los aislamientos se tipificaron molecularmente por amplificación al azar del ADN polimórfico (RAPD) y mediante electroforesis de campos pulsados (PFGE). Los 137 aislamientos mostraron 61 pulsotipos mediante PFGE y 25 tipos de RAPD diferentes. Los índices de Simpson calculados fueron 0,983 por PFGE y 0,941 por RAPD; esto evidencia el elevado poder discriminatorio de ambas técnicas. El análisis de la relación entre pares de aislamientos mostró un 92,6 % de concordancia entre ambas técnicas, lo que indica que cualquier par de aislamientos que fue distinguido por un método tendió a ser distinguido por el otro. La prevalencia de los genes sua y puaA fue del 97,8 % (134/137) y 94,9 % (130/137), respectivamente. Las secuencias de nucleótidos y de aminoácidos codificados por los genes sua y pauA de los 20 aislamientos de S. uberis seleccionados sobre la base de su tipo de PFGE y RAPD y origen geográfico tuvieron un porcentaje de identidad de entre 95 % y 100 % con respecto a todas las secuencias de referencia registradas en GenBank. Estos resultados demuestran que, a pesar de la diversidad clonal de S. uberis, los genes sua y pauA son prevalentes y están altamente conservados y deberían ser incluidos en futuros estudios de vacunas para prevenir mastitis bovina causada por S. uberis.

Sepsis neonatal temprana, incidencia y factores de riesgo asociados en un hospital público del occidente de México / Early neonatal sepsis, incidence and associated risk factors in a public hospital in western Mexico

Background: Early neonatal sepsis (EOS) is the second leading cause of death in the first week of life. Epidemiology differs in developed and developing countries. Aim: To describe the epidemiology of EOS among newborn patients in a public hospital in western Mexico. Methods: A prospective cohort study was performed in newborns of Nuevo Hospital Civil de Guadalajara "Dr. Juan I Menchaca". EOS was diagnosed with blood cultures or cultures of cerebrospinal fluid within the first 72 h of life. We analyzed risk factors (RF) by multivariate analysis with logistic regression. Results: We identified an EOS incidence of 4.7 events per 1,000 live births. Seventy two percent of the isolated bacteria were gram negative bacilli. Factors associated with EOS were maternal age ≤ 15 years (OR 3.50; 95% CI 1.56-7.85), rupture of membranes > 18 h (OR 2.65; 95% CI 1.18-5.92), maternal fever (OR 6.04; 95% CI 1.54-23.6), birth weight ≤ 2,500 g (OR 4.82; 95% CI 2.38-9.75) and gestational age < 37 weeks (OR 3.14; 95% CI 1.58-6.22). Conclusions: In addition to the RF known for EOS an independent association was observed with maternal age ≤ 15 years.

Introducción: La sepsis neonatal temprana (SNT) es la segunda causa de muerte en la primer semana de vida; la epidemiología difiere en países desarrollados y en vías de desarrollo. Objetivo: Describir la epidemiología de SNT en recién nacidos (RN) de un hospital público del occidente de México. Material y Métodos: Estudio de cohorte prospectivo en RN del Nuevo Hospital Civil de Guadalajara "Dr. Juan I Menchaca". Se diagnosticó SNT con cultivos de sangre o líquido cefalorraquídeo en las primeras 72 h de vida. Se indagaron factores de riesgo (FR) mediante análisis multivariado con regresión logística. Resultados: La incidencia de SNT fue de 4,7 eventos por 1.000 RN vivos. El 72% de las bacterias aisladas correspondió a bacilos gramnegativos. Los factores asociados a SNT fueron la edad materna ≤ 15 años (OR 3,50; IC 95% 1,56-7,85), ruptura de membranas > 18 h (OR 2,65; IC 95% 1,18-5,92), fiebre materna (OR 6,04; IC 95%1,54-23,6), peso al nacimiento ≤ 2.500 g (OR 4,82; IC 95% 2,38-9,75) y edad gestacional < 37 semanas (OR 3,14; IC 95% 1,58-6,22). Conclusiones: Además de los FR ya conocidos para SNT se observó asociación independiente con edad materna ≤ 15 años.

Perfiles genéticos de Streptococcus uberis aislados de mastitis bovina / Genetic patterns of Streptococcus uberis isolated from bovine mastitis

El objetivo del presente estudio fue evaluar las relaciones genotípicas entre 40 Streptococcus uberis aislados de mastitis bovina mediante la técnica de electroforesis de campos pulsantes (pulsed-field gel electrophoresis [PFGE]). Además, se investigó la asociación entre los patrones de PFGE y los perfiles de virulencia. Los aislamientos mostraron 17 patrones de PFGE. Se encontraron diferentes cepas dentro de los tambos y en los distintos tambos, y un bajo número de aislamientos dentro del mismo tambo compartieron un perfil idéntico de PFGE. No se encontró ninguna asociación entre los patrones de PFGE y los perfiles de virulencia. Sin embargo, la detección de cepas particulares en algunos tambos podría indicar que algunas de ellas son más virulentas que otras. Sería importante avanzar en las investigaciones para identificar nuevos genes relacionados con la virulencia que podrían contribuir a la capacidad infecciosa de estas cepas

The aim of this study was to evaluate the genotypic relationships among 40 Streptococcus uberis isolated from bovine mastitis by using pulsed-field gel electrophoresis (PFGE). Additionally, the association between PFGE patterns and virulence profiles was investigated. The isolates exhibited 17 PFGE patterns. Different strains were found within and among herds; however, a low number of isolates within the same herd shared an identical PFGE type. No association between PFGE patterns and virulence profiles was found. However, the detection of specific strains in some herds could indicate that some strains are more virulent than others. Further research needs to be undertaken to elucidate new virulence-associated genes that might contribute to the capability of these strains to produce infection

Piometra em uma leoa (Panthera leo): relato de caso / Pyometra in a lion (Panthera leo): case report

A piometra é uma infecção aguda ou crônica do útero que ocorre frequentemente em cadelas não castradas, podendo também ocorrer em gatas domésticas e selvagens, sendo poucos os estudos relacionados à piometra em grandes felídeos. O objetivo deste relato foi descrever um caso de piometra em uma leoa (Panthera leo) de cativeiro, as lesões de necropsia e histológicas, bem como os resultados da análise microbiológica. Uma leoa com aproximadamente 23 anos, pertencente a um criadouro conservacionista de Santa Maria-RS, foi encontrada morta pela manhã em seu recinto. Após coleta de dados, procedeu-se à necropsia e à coleta de material para análise histopatológica e bacteriológica. A análise microbiológica revelou predomínio das bactérias Streptococcus sp. e Escherichia coli no conteúdo purulento do útero, caracterizando como piometra, e a bactéria predominante em plasma, fígado e medula óssea foi E. coli. De acordo com o laudo histopatológico, as alterações observadas nessa leoa sugerem um quadro de septicemia grave, sendo a origem do foco infeccioso bacteriano, provavelmente, a piometra. Considera-se importante chamar a atenção dos médicos veterinários de animais selvagens para um diagnóstico precoce dessa doença, que é comum em cadelas, mas que pode acometer também felídeos selvagens e levá-los à morte.

Pyometra is an acute or chronic uterus infection that occurs often in not spayed dogs, but may also occur in wild and domestic cats, and there are few studies related to pyometra in big cats. The aim of this report was to describe a case of pyometra in a lion (Panthera leo) in captivity, the gross and microscopic lesions found at necropsy, and the results of the microbiological analysis. A female with approximately 23 years of age, belonging to a Conservationist Breeding Center located in Santa Maria-RS-Brazil was found dead in her enclosure in the morning. After data collection, we proceeded to the autopsy and collection of material for histopathological and bacteriological analyzes. Microbiological analysis revealed a predominance of the bacteria Streptococcus sp and Escherichia coli in the uterus content, characterized as pyometra and the predominant bacterium in the plasma, liver and bone marrow was Escherichia coli. According to histopathology, the changes observed in this lioness suggest a framework of severe septicemia, being the source of the bacterial infection, probably, pyometra. It is considered important to draw the attention of zoo and wildlife veterinarians for an early diagnosis of this common disease in dogs, which can also affect large wild cats and lead them to death.

Is Streptococcus pseudopneumoniae a neglected respiratory pathogen.

Streptococcus pseudopneumoniae is a member of Viridans streptococci, associated with chronic obstructive lung disease and lower respiratory infection. It is non-capsulated, bile insoluble and optochin susceptible in ambient air but intermediate or resistant in 5%CO2. But, its role as a pathogen is yet to be established. The objective of the study was to detect the prevalence and to determine the clinical significance of S.pseudopneumoniae in sputum samples. The study period was 5 months. Good quality sputum samples (>25 neutrophils and <10 epithelial cells/LPF) of patients suspected of LRTIs, were inoculated onto chocolate agar. Alpha haemolytic colonies were identified by testing for capsule by India ink preparation, bile solubility and optochin susceptibility in ambient air and 5%CO2. Antibiotic sensitivity testing was done by Kirby-Bauer's disc diffusion method. Out of 66 alpha haemolytic colonies, 4 were S.pseudopneumoniae, 17 were S.pneumoniae and 45 were other Viridans streptococci. Antibiotic sensitivity patterns of S.pseudopneumoniae and S.pneumoniae were analyzed. S.pseudopneumoniae is misinterpreted as S.pneumoniae unless specifically looked for. As there is scant information available, studies have to be encouraged for better determination of clinical importance of S.pseudopneumoniae. This will help in right diagnosis and successful treatment.

Survey of canine tick-borne diseases in Lábrea, Brazilian Amazon: ‘accidental’ findings of Dirofilaria immitis infection / Pesquisa de agentes transmitidos por carrapatos em cães de Lábrea, Amazonas: achados “acidentais” de Dirofilaria immitis

Blood samples were collected from 99 domestic dogs from the urban and rural areas of the Lábrea municipality, state of Amazonas, Brazil. Canine serum samples were tested by immunofluorescence assay against Rickettsia spp., which revealed that only 3.0% (1/33) and 7.6% (5/66) of the dogs from urban and rural areas, respectively, reacted positively to at least one Rickettsia species. DNA was extracted from canine blood and tested by a battery of PCR assays targeting protozoa of the genera Babesia and Hepatozoon, and bacteria of the genera Rickettsia and Ehrlichia and family Anaplasmataceae. All samples were negative in the PCR assays targeting the genera Babesia, Hepatozoon, Ehrlichia and Rickettsia. For Anaplasmataceae, 3% (1/33) and 39.4% (26/66) of the urban and rural dogs, respectively, yielded amplicons that generated DNA sequences 100% identical to the corresponding sequence of Wolbachia endosymbiont of Dirofilaria immitis. Because of these results, all canine DNA samples were further tested in a PCR assay targeting filarial nematodes, which was positive for 18.2% (6/33) and 57.6% (38/66) urban and rural dogs, respectively. Filarial-PCR products generated DNA sequences 100% identical to D. immitis. While tick-borne infections were rare in Lábrea, D. immitis infection rates were among the highest reported in South America.

Amostras de sangue foram coletadas de 99 cães domésticos de áreas urbana e rural do município de Lábrea, estado do Amazonas. Soros caninos foram testados pela técnica de imunofluorescência indireta contra Rickettsia spp., resultando em apenas 3,0% (1/33) e 7,6% (5/66) de cães soropositivos nas áreas urbana e rural, respectivamente. DNA foi extraído do sangue canino e testado por diferentes protocolos da PCR para detecção de protozoários dos gêneros Babesia e Hepatozoon, e bactérias dos gêneros Rickettsia e Ehrlichia e da família Anaplasmataceae. Todas as amostras foram negativas nos protocolos de PCR para os gêneros Babesia, Hepatozoon, Ehrlichia e Rickettsia. Para Anaplasmataceae, 3% (1/33) e 39,4% (26/66) dos cães de áreas urbana e rural, respectivamente, geraram sequências de DNA 100% idênticas ao endosimbionte Wolbachia de Dirofilaria immitis. Posteriormente, as amostras foram testadas pela PCR para nematódeos filarídeos, resultando em 18,2% (6/33) e 57,6% (38/66) de amostras positivas nas áreas urbana e rural, respectivamente. Os produtos geraram sequências de DNA 100% idênticas a D. immitis. Em contraste com várias outras regiões do Brasil, infecções transmitidas por carrapatos foram raras em Lábrea. Por outro lado, as frequências de infecção por D. immitis estiveram entre as mais altas relatadas na América do Sul.

Caracterización clínico - epidemiológica de pacientes con artritis séptica: año 2013: Instituto Traumatológico de Chile / Clinical - epidemiological characteristics of patients with septic arthritis: year 2013 Instituto Traumatologico of Chile

Artritis séptica (AS) es la reacción inflamatoria de una articulación desencadenada por microorganismos, provocando destrucción osteocondral y pérdida funcional irreversible si no es tratada a tiempo. El cuadro clínico de esta patología debe despertar sospecha del médico, obligando a realizar diagnóstico y manejo adecuado. OBJETIVOS: Caracterizar clínica y epidemiológicamente a pacientes con AS del InstitutoTraumatológico de Santiago, Chile (IT) y describir su manejo realizado. MATERIALES Y MÉTODOS: Estudio descriptivo retrospectivo de corte transversal, analizando fichas clínicas electrónicas de pacientes mayores de 15 años atendidos en elServicio de Urgencia del IT con diagnóstico de AS, durante el año 2013 (n=28). Los datos fueron tabulados en una planilla Microsoft® Excel® 2010. RESULTADOS: 68 por ciento fueron pacientes masculinos. Edad promedio de 54 años. Se realizaron 2 aseos quirúrgicos en promedio por paciente. 17 días fue el promedio de estadía hospitalaria. Principales comorbilidades fueron hipertensión arterial (29 por ciento) y diabetes mellitus 2 (21 por ciento). En síntomas predominó dolor (93 por ciento) y aumento de volumen (82 por ciento).La localización más frecuente fue rodilla (53,6 por ciento). Etiología más habitual fue Staphyloccocus aureus (60 por ciento). Las alteraciones de laboratorio fueron VHS elevada (90 por ciento), PCR elevada (80 por ciento) y leucocitosis(45 por ciento). Tratamiento antibiótico empírico más utilizado fue esquema tri asociado de cloxacilina, gentamicina y penicilinasódica (60 por ciento). Antibioticoterapia ambulatoria más indicada fuecloxacilina (60 por ciento). DISCUSIÓN: Recalcamos la importancia del diagnóstico y tratamiento oportuno de la AS, cuyo pronóstico y complicaciones dependen directamente de su pesquisa temprana. Las características clínico - epidemiológicas del presente estudio son consistentes con lo descrito en la literatura nacional e internacional...

Septic arthritis (SA) is the inflammatory reaction of an articular surface by the presence of microorganisms. It can cause joint destruction and rapid functional loss. Clinical manifestations of septic arthritis should be considered a medical emergency, therefore proper diagnosis and management is mandatory. OBJECTIVES: To characterize clinically and epidemiologically adult patients with SA from Instituto Traumatologico de Santiago de Chile (IT) and to describe its treatment. MATERIALS AND METHODS: A retrospective, cross-sectional, descriptive study analyzing electronic health records of patients over 15 years old, with diagnosis of SA, attended in the ER of the IT during 2013 (n=28). Data were tabulated in an Excel2010spreadsheet. RESULTS: 68 percent of patients were males. Average age was 54 years. An average of 2 surgical toilettes was performed per patient. The mean impatient stay was 17 days. Major comorbidities were hypertension (29 percent) and diabetes mellitus type 2(21 percent). The main symptoms were pain (93 percent) and swelling (82 percent).The most common location was the knee (53,6 percent). Most common etiology was Staphyloccocus aureus (60 percent). The laboratory abnormalities were high ESR (90 percent), elevated CRP (80 percent) and leukocytosis (45 percent). The most used empiric antibiotic the rap was the triassociated scheme of cloxacillin, gentamicin and sodium penicillin (60 percent). Most indicated outpatient antibiotic was cloxacillin (60 percent). DISCUSSION: We emphasize the importance of early diagnosis and treatment of SA, having direct consequences in complications and prognosis if not made quickly. Both clinical and epidemiological findings of our study were comparable to those found in national and international literature...

Evaluación inmediata y a distancia de tres técnicas de sellado canalicular en dientes apicectomizados: experiencia en conductos de corte transversal achatado / Immediate and long-term evaluation of three root canal filling techniques in apicectomized teeth: an experience in flat cross-cut canals

Con el propósito de estudiar in vitro la capacidad de sellado en sentido corono-apical de tres técnicas de obturación canalicular (condensación lateral con conos de gutapercha + sellador de Grossman, cono único de gutapercha + sellador de Grossman y cono único de gutapercha + cemento de ionómero vítreo), en dientes apicectomizados de conductos de corte transversal achatado, se utilizó un método bacteriológico que permitió comprobar el grado de permeabilidad de bacterias conocidas (Proteus mirabilis y Streptococcus salivarius) a través de las obturaciones realizadas. Los resultados permitieron establecer que ninguna de las técnicas estudiadas resultó totalmente efectiva, tanto a la valoración inmediata como a distancia, apreciándose una marcada tendencia al aumento de la permeabilidad a la microfiltración bacteriana en las muestras conservadas (p<0,02). Resulta evidente, según esta experiencia, que la anatomía del conducto radicular condiciona la calidad de la obturación en aquellos dientes que deben ser apicectomizados

Infecciones invasivas por estreptococos de los grupos A, C y G en la Argentina / Invasive infectios due to group A, C and G streptococci in Argentina

Streptococcus pyogenes (estreptococo beta-hemolítico del grupo A) (SGA) y Streptococcus dysgalactiae subsp. equisimilis, (estreptococos beta-hemolíticos grupos C y G) (SDSE) son capaces de provocar enfermedades graves como la fascitis necrotizante y el síndrome de shock tóxico estreptocócico (SSTE) y de causar complicaciones posinfecciosas. El objetivo de este trabajo fue presentar resultados de un estudio multicéntrico y compararlo con diferentes estudios descriptivos previos sobre infecciones invasivas por estreptococos beta-hemolíticos de los grupos A, C y G, también realizados en la Argentina. Se incluyeron 54 pacientes de 0 a 15 años con infecciones invasivas por SGA (N=50) o SDSE (N=4) en forma prospectiva entre julio de 2011 y junio de 2012 en 28 centros de 17 ciudades argentinas. Se aisló S. pyogenes en 28 pacientes que presentaron bacteriemia, 6 de ellas sin foco. Cuatro pacientes (7,4%) presentaron SSTE, en todos los casos por S. pyogenes. La mortalidad fue del 2,0% para SGA. La evolución de los pacientes fue peor en los tres estudios anteriores respecto del actual: mayor porcentaje de casos de SSTE (diferencias no significativas) y mayor mortalidad (diferencia significativa respecto de dos estudios previos). Es probable que la morbimortalidad haya decrecido en esta última década en la Argentina posiblemente debido al uso temprano de clindamicina en las infecciones invasivas por S. pyogenes y SDSE, aunque no se puede descartar la diferente circulación de cepas virulentas. Esta apreciación además está sesgada por la inclusión de pacientes de distintos centros con diferentes formas de presentación inicial (AU)

Streptococcus pyogenes (group A ß-hemolytic streptococcus (GAS)) and Streptococcus dysgalactiae subsp. equisimilis (group C and G ß-hemolytic streptococcus (GCGS)) may cause severe diseases such as necrotizing fasciitis and streptococcal toxic shock syndrome (STSS) as well as postinfectious complications. The aim of this report was to present the results of a multicenter study and compare them with the results of different previous descriptive studies on invasive infections due to beta-hemolytic streptococcus groups A, C, and G that were also conducted in Argentina. Forty-five patients between 0 and 15 years of age with invasive infections due to GAS (N=50) or GCGS (N=4) were prospectively included in the study between July 2011 and June 2012 from 28 centers in 17 Argentine cities. S. pyogenes was isolated in 28 patients who presented with bacteremia, without a focus in six. Four patients (7.4%) had STSS, due to S. pyogenes in all of them. In patients with GAS, mortality rate was 2.0%. Outcome of the patients was worse in previous studies than in the present one: Percentages of cases with STSS (no significant difference) and mortality (significant difference) were higher. It is probable that over the last decade morbidity and mortality have decreased in Argentina, possibly due to the early use of clindamycin in invasive infections due to S. pyogenes and GCGS, although a different circulation of virulent strains cannot be ruled out. Additionally, this observation is biased by the inclusion of patients from different centers with different presentations at onset (AU)