Caracterização da microbiota de queijos artesanais provenientes da Serra da Canastra - MG e da cultura iniciadora natural utilizada em sua produção / Microbiota characterization of artisanal cheeses from Serra da Canastra - MG and their endogenous starter culture

O queijo Canastra possui grande importância na cultura e economia local, é parte do Patrimônio Imaterial do Brasil (IPHAN, 2014) e recebeu o selo de produto com designação de origem em 2012 (INPI, 2016). Sua produção utiliza leite, sal, coalho e uma cultura iniciadora natural, chamada popularmente de pingo. Esse estudo visou a caracterização da microbiota presente no queijo maturado da Serra da Canastra e no pingo utilizado em sua produção utilizando técnicas avançadas de sequenciamento em larga escala para identificação das bactérias e fungos ali presentes. Nossos dados da microbiota bacteriana foram comparados com dados da microbiota de outros queijos brasileiros e do mundo disponíveis na literatura. As principais bactérias encontradas em amostras de pingo pertencem aos gêneros Lactococcus (45.6%), Streptococcus (30.3%), Staphylococcus (5.1%), e em amostras de queijo aos gêneros Lactococcus (22.5%), Streptococcus (27.2%), Corynebacterium (18.8%), Staphylococcus (13.6%), Leuconostoc (6.3%) e Weissella (6%). Os principais gêneros de fungos encontrados nos queijos foram Debaryomycesa (78.6%), Trichosporona (7.8%). Nosso estudo foi capaz de separar a microbiota dos queijos produzidos na Serra da Canastra de outros queijos na Europa e América do Norte, sendo o pH um possível fator de segregação. Também foi observada uma diferença entre a microbiota do queijo Canastra com outros queijos Brasileiros. Além disso, visualizamos que a distância geográfica entre produtores e a sazonalidade possuem um efeito sobre a microbiota dos pingos e queijos. A partir da análise de todos os microrganismos encontrados na microbiota bacteriana, foram detectados táxons que discriminam produtores por suas aplicações de boas práticas de fabricação e por sua infraestrutura. Observamos proporções menores de um táxon de Kocuria Kristinae nos pingos e um de Streptococcus nos queijos e proporções maiores de um táxon de Staphylococcus nos queijos. Também pudemos observar uma diminuição nas proporções de táxons de Debaryomycesa e aumento na proporção de táxons de Trichosporona na composição fúngica dos queijos, possivelmente devido a transição sazonal do período seco para o chuvoso. Usando técnicas moleculares de sequenciamento em larga escala, demonstramos que há uma diferença na microbiota presente em diferentes áreas da Serra da Canastra, um possível efeito da sazonalidade na composição fúngica e bacteriana. E evidenciamos que táxons de Streptococcus, Staphylococcus e Kocuria estão correlacionados às boas práticas de produção e elucidamos a conexão existente entre a microbiota do pingo e a do queijo. Estes resultados podem influenciar o desenvolvimento de métodos de rastreamento de sub-regiões específicas da Canastra e auxiliar os produtores na produção de queijos de boa qualidade, mantendo as características específicas de sua região

The Canastra cheese has great importance for the local culture and economy, being part of the Intangible Heritage of Brazil (IPHAN, 2014). It has received the protected designation of origin certification in 2012 (INPI, 2016). It's made using milk, salt, rennet and a endogenous starter culture, popularly called as "pingo". This study aimed to characterize the microbiota present in the Serra da Canastra's cheese and the pingo used in its production. In order to conduct this research we used next generation sequencing to identify the bacteria and fungi present there. Our bacterial microbiota dataset was compared with microbiota datasets from other Brazilian and world cheeses available in the literature. The main bacteria found were Lactococcus (45.6%), Streptococcus (30.3%) and Staphylococcus (5.1%) in the endogenous starter samples and Lactococcus (22.5%), Streptococcus (27.2%), Corynebacterium (18.8 %), Staphylococcus (13.6%), Leuconostoc (6.3%) and Weissella (6%) in cheese samples. The main fungi found in the cheeses were Debaryomycesa (78.6%) and Trichosporona (7.8%). We were able to separate the microbiota from Serra da Canastra cheeses and other cheeses in Europe and North America, being the pH a possible segregation factor. Furthermore, a difference was also observed between the microbiota of Canastra and other Brazilian cheeses. In addition, we observed that the geographical distance between producers and the seasonality could be affecting the pingos and cheeses microbiota. We found bacterial taxa that could discriminate producers by their good manufacturing practices and their local infrastructure. Low levels of good manufacturing practices (GMPs) were assigned to bigger proportions of a Kocuria Kristinae taxon in the pingos and a Staphylococcus taxon in the cheeses. Also, higher levels of GMPs were assigned to smaller proportions of Streptococcus taxons in the cheeses. Furthermore We could observe a decrease of Debaryomycesa and an increase of Trichosporona proportions in the fungal composition of cheeses. This could be due to a climate transition: from the dry season to the rainy season. Using large-scale sampling coupled with molecular sequencing techniques, we observe a connection between pingo and cheeses microbiota. We show that the microbiota of different areas in Serra da Canastra is different, also, there is a possible effect of seasonality on fungal and bacterial composition. Furthermore, we could see that Streptococcus, Staphylococcus and Kocuria taxons are correlated with good practices. These results may influence the development of tracking methods for specific Canastra subregions and assist producers to manufacture good quality cheeses while maintaining the specific characteristics of their region

Encrusted cystitis caused by corynebacterium urealyticum: a case report with novel treatment strategy of intravesical dimethyl sulfoxide

ABSTRACT Encrusted cystitis (EC) was first described as chronic cystitis with mucosal calcification in 1914 (1). It is a very rare chronic inflammatory disease presenting with dysuria, pelvic pain and gross hematuria. Voided urine contains mucus or calcified mucopurulent stone like particles. Urinalysis always reveals alkaline pH. It may be present in healthy individuals with no predisposing etiological factors (2-4). Etiologically, previous urological diseases, immunosuppression, urinary infection with urea splitting bacteria, or urological interventions resulting in bladder mucosa trauma may also be present (5, 6). In the present case report, we describe a novel treatment for EC with intravesical dimethyl sulfoxide.

Central venous catheter-related infections caused by Corynebacterium amycolatum and other multiresistant non-diphtherial corynebacteria in paediatric oncology patients

ABSTRACT Bloodstream and venous catheter-related corynebacterial infections in paediatric patients with haematological cancer were investigated from January 2003 to December 2014 at the Brazilian National Cancer Institute in Rio de Janeiro, Brazil. We observed that during cancer treatment, invasive corynebacterial infections occurred independent of certain factors, such as age and gender, underlying diseases and neutropenia. These infections were ssscaused by Corynebacterium amycolatum and other non-diphtherial corynebacteria. All cases presented a variable profile of susceptibility to antimicrobial agents, except to vancomycin. Targeted antibiotic therapy may contribute to catheters maintenance and support quality of treatment. Non-diphtherial corynebacteria must be recognized as agents associated with venous access infections. Our data highlight the need for the accurate identification of corynebacteria species, as well as antimicrobial susceptibility testing.

Retrospective evaluation of the clinical characteristics associated with Corynebacterium species bacteremia

ABSTRACT Objectives: Corynebacterium spp. are becoming recognized as pathogens that potentially cause various infections. We aimed to evaluate the clinical characteristics associated with Corynebacterium spp. bacteremia. Patients and methods: We retrospectively reviewed the medical records of all adult patients who had positive blood cultures for Corynebacterium spp. in a single university hospital between January 2014 and December 2016. Patients were divided into a bacteremia group and a contamination group based on microbiological test results and clinical characteristics. Patients' characteristics, antimicrobial susceptibility of isolated species, antimicrobials administered, and patient outcomes were evaluated. Results: Corynebacterium spp. were isolated from blood samples of 63 patients; Corynebacterium striatum was the predominant isolate. Twenty-eight patients were determined to have bacteremia. Younger age (p = 0.023), shorter time to positivity (p = 0.006), longer hospital stay (p = 0.009), and presence of an indwelling vascular catheter (p = 0.002) were observed more often in the bacteremia group compared to the contamination group. The source of infection in most patients with bacteremia was an intravenous catheter. All tested strains were susceptible to vancomycin. Four of the 27 patients with bacteremia died, despite administration of appropriate antimicrobial therapy. Conclusions: We found that younger age, shorter time to positivity, and presence of an indwelling catheter were related to bacteremia caused by Corynebacterium spp. Appropriate antimicrobials should be administered once Corynebacterium spp. are isolated from the blood and bacteremia is suspected.

Tricomicosis (tricobacteriosis) en un infante: aspectos microbiológicos, dermatoscópicos y ultraestructurales / Trichomycosis (trichobacteriosis) capitis in an infant: microbiological, dermoscopic and ultrastructural features

La tricomicosis es una infección superficial causada por Corynebacterium flavescens, que afecta por lo regular pelos axilares, en menor grado los púbicos, los escrotales e interglúteos y excepcionalmente los de la cabeza o tricomicosis capitis (TC). Esta infección se caracteriza por formación de nódulos pilosos. Clínicamente se confunde con infecciones como piedra blanca y pediculosis. El diagnóstico se realiza por microscopia y dermatoscopia de masas bacterianas y confirmado por cultivo. OBJETIVO: Presentar un caso de TC en un infante, y mostrar las características microscópicas, dermatoscópicas y ultraestructurales. CASO CLÍNICO: Niño sano de 6 meses de edad, con dermatosis que afectó los pelos de la cabeza en forma de múltiples nódulos-pilosos amarillentos. Se comprobó TC mediante fluorescencia amarilla a la luz de Wood; a la dermatoscopia se observaron cadenas blanco-amarillentas, como "rosarios de piedras cristalinas"; al examen directo se distinguieron masas bacterianas y al cultivo se identificó Corynebacterium flavescens. A la microscopia electrónica se observó infección superficial, sin perforación de los pelos. Se realizó tratamiento con aplicación de ácido fusídico por 3 semanas y se obtuvo curación clínica y microbiológica. CONCLUSIÓN: La TC es una entidad rara que se presenta en niños, y que suele confundirse con otros padecimientos del pelo como la pediculosis e infecciones micóticas.

Trichomycosis is a superficial infection caused by Corynebacterium flavescens, which regularly affects axillary, and to a a lesser extent, pubic, scrotal and intergluteal, and exceptionally, head hairs or trichomycosis capitis (TC). This condition is characterised by the formation of bacterial nodules. Clinically, it can be confused with white piedra or pediculosis. The diagnosis is made by microscopic and dermoscopic observation and confirmed by culture. OBJECTIVE: To present a case of TC in an infant and illustrate the microscopic, dermoscopic, and ultrastructural characteristics. CLINICAL CASE: A 6 month-old boy, otherwise healthy, with multiple yellowish concretions on the hairs of the head. TC was confirmed by yellow fluorescence with Wood’s light; white-yellowish beads, like "rosaries of crystalline stones" were observed on dermoscopy, direct examination showed bacterial masses, and Corynebacterium flavescens was identified by culture. A superficial infection, without perforation of the hairs, was confirmed by electron microscopy. Treatment with fusidic acid for 3 weeks achieved a clinical and microbiological cure. CONCLUSION: TC is a rare condition that affects children, and tends to be mistaken for other diseases of the hair, such as pediculosis and mycotic infections.

Osteomielitis crónica por Corynebacterium striatum en una adolescente / Chronic osteomyelitis due to Corynebacterium striatum in a female adolescent

A case of a girl with chronic posttraumatic osteomyelitis of the right tibia with microbiological isolation of Bacillus spp. by conventional methods and confirmation by mass spectrometry MALDI-TOF (matrix-assisted laser desorption/ionization time-of-flight mass) of Corynebacterium striatum is presented. Diagnostic methods, clinical manifestations, and resistance pattern of these infections are described.

Se presenta el caso de una adolescente con osteomielitis crónica postraumática de la tibia derecha con aislamiento microbiológico por métodos convencionales de Bacillus spp. y confirmación por espectrometría de masas MALDI-TOF de Corynebacterium striatum, microorganismo considerado generalmente saprófito. Se describen las presentaciones clínicas de estas infecciones descritas en la literatura científica, la necesidad del diagnóstico etiológico por técnicas no convencionales y el patrón de resistencia comúnmente expresado.

Neumonía por Corynebacterium pseudodiphtheriticum / Pneumonia caused by Corynebacterium pseudodiphtheriticum

Corynebacterium pseudodiphtheriticum forma parte de la microbiota normal de orofaringe y de piel. Sin embargo, en las últimas décadas está emergiendo como oportunista causante de infecciones clínicamente significativas en pacientes con algún compromiso previo. Se refiere el caso clínico de una paciente de 76 años con antecedentes de hipertensión arterial, hipotiroidismo, diabetes tipo 2 e insuficiencia renal crónica, que presentó neumonía durante su estadía en terapia intensiva. El examen directo del esputo inducido (coloración de Gram) reveló una muestra representativa con abundante microbiota monomicrobiana constituida por bacilos gram positivos pleomórficos corineformes y el cultivo presencia de C. pseudodiphtheriticum. La paciente recibió medicación empírica con cefalosporina de tercera generación con evolución favorable.

Microorganisms of the genera Corynebacterium, specie pseudodiphtheriticum are a part of the indigenous microbiota of human skin and oropharinx. Nevertheless in recent decades these bacilli are emerging as opportunistic pathogens causing clinically significant infections in patients with previous compromise. We report the case of a 76 years old female patient, with a history of hypertension, hypothyroidism, type 2 diabetes and chronic renal failure, who presented pneumonia during their stay at the intensive care unit. The induced sputum revealed a representative sample with monomicrobial gram positive pleomorphic coryneform rods (Gram stain) and cultures demonstrated the presence of C. pseudodiphtheriticum as the only bacteria recovered. The pacient received an empirical third generation cephalosporin medication with a succesfull recovery.

Endocarditis infecciosa fatal por Corynebacterium pseudodiphtheriticum / Letal endocarditis due to Corynebacterium pseudodiphtheriticum

We present a case report of a infective endocarditis by Corynebacterium pseudodiphtheriticum in a young patient eight years old with bicuspid aortic valve that led to a severe neurological embolism and death. Corynebacterium pseudodiphtheriticum is part of the nasopharyngeal and skin flora. However, there are cases reported of endocarditis usually associated with immunocompromised patients that mostly presented a poor clinical outcome.

Se presenta el caso clínico de una endocarditis infecciosa por Corynebacterium pseudodiphtheriticum en un niño de 8 años, portador de una valvulopatía aórtica bicúspide, que falleció por una embolia cerebral grave. Corynebacterium pseudodiphtheriticum forma parte de la microbiota de la piel y nasofaringe. Sin embargo, se han descrito casos de endocarditis en pacientes inmunocomprometidos, asociados, en su mayoría, a una mala evolución clínica.

Expression of CD14 and toll-like receptors 2 and 4 by milk neutrophils in bovine mammary glands infected with Corynebacterium bovis / Expressão de CD14 e dos receptores do tipo toll 2 e 4 por neutrófilos lácteos provenientes de glândulas infectadas por Corynebacterium bovis

This study evaluated the expression of CD14, toll-like receptor (TLR) 2 and TLR4 on the surface of milk neutrophils in bovine mammary glands infected with Corynebacterium bovis. Here, we used 23 culture-negative control quarters with no abnormal secretion on the strip cup test and milk somatic cell count lower than 1x105 cells/mL, and 14 C. bovis infected quarters. The identification of neutrophils, as well as, the percentage of neutrophils that expressed CD14, TLR2 and TLR4 were analyzed by flow cytometry using monoclonal antibodies. The present study encountered no significant difference in the percentages of milk neutrophils that expressed TLR2 and TLR4 or in the expression of TLR4 by milk neutrophils. Conversely, a lower median fluorescence intensity of TLR2 in milk neutrophils was observed in C. bovis-infected quarters. The percentage of neutrophils that expressed CD14 and the median fluorescence intensity of CD14 in milk neutrophils was also lower in C. bovis-infected quarters...

O presente estudo objetivou avaliar alterações na expressão de CD14, e dos receptores do tipo toll (TLR) 2 e 4 na superfície de neutrófilos lácteos provenientes de glândulas mamárias infectadas por Corynebacterium bovis. O presente estudo utilizou 23 quartos negativos no exame bacteriológico, sem alterações na prova de fundo escuro e com contagem automática de células somáticas menor que 1 x105 células/mL, e 14 quartos mamários infectados por C. bovis A identificação de neutrófilos, assim como a porcentagem de neutrófilos lácteos que expressaram CD14, TLR2 e 4 foram avaliadas por citometria de fluxo utilizando anticorpos monoclonais. A porcentagem de neutrófilos que expressaram TLR2 e TLR4 nos quartos mamários infectados por C. bovis não diferiu dos quartos mamários sadios, assim como na expressão de TLR4. No entanto, a intensidade de fluorescência do TLR2 na superfície dos neutrófilos foi menor nos quartos mamários infectados por C. bovis. A porcentagem de neutrófilos que expressaram CD14 e a intensidade de fluorescência da molécula de CD14 foi menor na superfície dos neutrófilos lácteos dos quartos infectados por C. bovis...

Interdigital foot infections: Corynebacterium minutissimum and agents of superficial mycoses

Interdigital foot infections are mostly caused initially by dermatophytes, yeasts and less frequently by bacteria. Erythrasma caused by Corynebacterium minutissimum can be confused with superficial mycoses. The aim of the study was to determine the prevalence of the etiologic agents of superficial mycoses and the frequency of Corynebacterium minutissimum in interdigital foot infections. All the samples obtained from the 121 patients with interdigital foot infections were examined directly with the use of 20% potassium hydroxide mounts and Gram stain under the microscope and cultured on Sabouraud's dextrose agar plates. In identification of superficial mycoses, the rate was found to be 14% with the cultural method and 14% with direct microscopic examination. Using a combination of direct microscopic examination and culture, a 33.8% ratio was achieved. In the culture of these samples, the most isolated factor was Trichophyton rubrum (33.7%). In 24 of the patients (19.8%) Corynebacterium minutissimum was detected by Gram staining, in 6 of these patients Trichophyton rubrum was found, Trichophyton mentagrophytes was found in 2 and Trichosporon spp. was found in 1. The examination of interdigital foot lesions in the laboratory, the coexistence of erythrasma with dermatophytes and yeast should be considered.

Aspectos clínico-epidemiológicos e microbiológicos de processos infecciosos causados por Corynebacterium spp em pacientes de centro de referência oncológico – Rio de Janeiro, Brasil / Clinical-epidemiological and microbiological aspects of infectious processes caused by Corynebacterium spp in pacients at the National Cancer Institute in Rio de Janeiro, Brazil

Os resultados permitiram a redação de quatro artigos. Aspectos microbiológicos e clínicos de corinebacterioses em pacientes com câncer observados durante cinco anos foram descritos no Artigo 1. No Artigo 2 foram apresentados casos de bacteremia causados por corinebactérias invasivas não toxigênicas em dois períodos com intervalo de sete anos. As infecções em pacientes com câncer por C. diphtheriae, causando casos clínicos atípicos foram descritas no Artigo 3, além do estudo dos principais fatores de virulência de uma cepa de C. diphtheriae isolada de infecção associada ao cateter de nefrostomia foi descrita no Artigo 4. Resumidamente no Artigo 1, além dos aspectos clínico-epidemiológicos foram avaliados os perfis de resistência aos antimicrobianos e o potencial de virulência dos micro-organismos. Em cinco anos, 932 amostras de corinebactérias, com perfis de resistência aos antimicrobianos testados, foram isoladas de pacientes com câncer. As espécies predominantes foram Corynebacterium amycolatum (44,7%), Corynebacterium minutissimum (18,3%) e Corynebacterium pseudodiphtheriticum (8,5%). O uso de catéteres de longa permanência e a neutropenia, foram às condições importantes para infecção por corinebactérias. As doenças de base mais comuns foram os tumores sólidos. Pacientes hospitalizados apresentaram risco seis vezes maior de morrer, quando relacionadas às taxas de mortalidade com 30 dias (RC= 5,5; IC 95%= 1,15-26,30; p= 0,033). As bacteremias (Artigo 2) causadas por corinebactérias foram observadas em dois períodos: 2003-2004 (n=38) e de 2012-2013 (n=24). As espécies multirresistentes C. amycolatum e Corynebacterium jeikeium foram os principais responsáveis pelos quadros de bacteremia. Havia 34 pacientes com tumores sólidos e 28 pacientes com doenças linfoproliferativas, sendo que 21 deles apresentavam neutropenia e 54 utilizavam cateter venoso central. Em 41 pacientes havia infecção relacionada ou associada aos dispositivos intravasculares...

A retrospective study at the InstitutoNacional doCâncer-INCA (National Cancer Institute) in Rio de Janeiro, Brazil examined infections of Corynebacterium sp. in cancer patients. The results were presented in four papers. Article 1 describes microbiological and clinical aspects of corynebacteriosis in cancer patients observed over five years. Article 2 presents cases of bacteremia caused by invasive non-toxigenic corynebacteria observed in 2003-2004 and seven years later in 2012-2013. Article 3 presents atypical clinical cases of cancer patients infected by Corynebacterium diphtheriae, while Article 4 is a study of the major bacterial virulence factors of an isolated strain of C. diphtheriae in infections associated with nephrostomy catheters.In addition to clinical and epidemiological aspects, Article 1 evaluates the antimicrobial resistance profiles and potential virulence factor of microorganisms. Over a period of five years, 932 samples of corynebacteria with antimicrobial resistance profiles were isolated from patients with cancer. The predominant species were Corynebacterium amycolatum (44.7%), Corynebacterium minutissimum (18.3%) and Corynebacterium pseudodiphtheriticum (8.5%).Long-term catheter use and neutropenia were the major conditions for infection by corynebacteria. Solid tumors were the most common underlying illness. The 30-day mortality rate for patients with corynebacteria infections was six times greater in hospitalized patients than for non-hospitalized patients (OR = 5.5, 95% CI = 1.15 to 26.30, p = 0.033).In Article 2, bacteremia caused by corynebacteria were observed in two time frames: 2003 to 2004 (n=38) and 2012 to 2103 (n=24). The multidrug-resistant species C. amycolatum and Corynebacterium jeikeium were responsible for invasive diseases.There were 34 patients with solid tumors and 28 patients with lymphoproliferative diseases, of which 21 had neutropenia and 54 used central venous catheter...

[Molecular identification of corynebacterium minutissimum in skin lesions suspicious for erythrasma by the PCR method]

Erythrasma is a chronic superficial infection of the intertriginous areas. Most laboratories use methylene blue stain and 10% KOH smear to identify Corynebacterium minutissimum [C. minutissimum] by direct observation of filamentous bacilli. Occasionally atypical forms can be seen that create problems in diagnosis. This study aims to use the polymerase chain reaction [PCR] method in order to definitively identify C. minutissimum as an agent of erythrasma. This research was performed during 2013 on 100 skin scrapings suspicious for erythrasma which were obtained from various medical mycology laboratories in Tehran. Samples were tested by three methods - direct examination, culture and PCR. DNA was extracted by the modified phenol-chloroform method after which PCR was performed using designed primers. We sequenced some of the PCR products. The sensitivity and specificity of the PCR method was compared to the direct and culture examinations. Of the 100 samples, there were 25 positive samples according to PCR analysis, 13 positive by direct examination and 23 that cultured positive. DNA sequencing results showed the presence of C. minutissimum. The PCR method in comparison with direct examination had a sensitivity of 100% and a specificity of 86.2%. The study also showed that the PCR method in comparison with culture had a sensitivity of 100% and a specificity of 97.4%. This study showed that the PCR method in comparison with the direct method and culture had a higher sensitivity in the detection of C. minutissimum. The present PCR method confirmed all typical and some of the atypical forms of C. minutissimum which indicated the importance of this method in the definitive diagnosis of erythrasma

Detection of Corynebacterium bovis infection in athymic nude mice from a research animal facility in Korea

Corynebacterium (C.) bovis infection in nude mice causes hyperkeratosis and weight loss and has been reported worldwide but not in Korea. In 2011, nude mice from an animal facility in Korea were found to have white flakes on their dorsal skin. Histopathological testing revealed that the mice had hyperkeratosis and Gram-positive bacteria were found in the skin. We identified isolated bacteria from the skin lesions as C. bovis using PCR and 16S rRNA sequencing. To the best of our knowledge, this is the first report of C. bovis infection in nude mice from Korea.

Etiology, antimicrobial susceptibility profile of Staphylococcus spp. and risk factors associated with bovine mastitis in the states of Bahia and Pernambuco / Etiologia, perfil de sensibilidade dos Staphylococcus spp. aos antimicrobianos e fatores de risco associados à mastite bovina nos estados da Bahia e Pernambuco

The purpose of this paper was to study the etiology of mastitis, determine the antimicrobial susceptibility profile of Staphylococcus spp. and to identify the risk factors associated with infection in dairy cows in the states of Bahia and Pernambuco, Brazil. From the 2,064 milk samples analyzed, 2.6% were associated with cases of clinical mastitis and 28.2% with subclinical mastitis. In the microbiological culture, Staphylococcus spp. (49.1%) and Corynebacterium spp. (35.3%) were the main agents found, followed by Prototheca spp. (4.6%) and Gram negative bacilli (3.6%). In the antimicrobial susceptibility testing, all 218 Staphylococcus spp. were susceptible to rifampicin and the least effective drug was amoxicillin (32.6%). Multidrug resistance to three or more drugs was observed in 65.6% of Staphylococcus spp. The risk factors identified for mastitis were the extensive production system, not providing feed supplements, teat drying process, not disinfecting the teats before and after milking, and inadequate hygiene habits of the milking workers. The presence of multiresistant isolates in bovine milk demonstrates the importance of the choice and appropriate use of antimicrobial agents. Prophylactic and control measures, including teat antisepsis and best practices for achieving hygienic milking should be established in order to prevent new cases of the disease in herds.

Objetivou-se estudar a etiologia da mastite, determinar o perfil de sensibilidade dos Staphylococcus spp. aos antimicrobianos e identificar os fatores de risco associados à infecção em vacas leiteiras nos estados da Bahia e Pernambuco. Das 2.064 amostras de leite analisadas, 2,6% estavam associadas a casos de mastite clínica e 28,2% à mastite subclínica. No exame microbiológico, Staphylococcus spp. (49,1%) e Corynebacterium spp. (35,3%) foram os principais agentes isolados, seguidos de Prototheca spp. (4,6%) e bacilos Gram negativos (3,6%). No teste de sensibilidade aos antimicrobianos, todos os 218 Staphylococcus spp. apresentaram-se sensíveis à rifampicina e a droga menos eficaz foi a amoxicilina (32,6%). A resistência simultânea a três ou mais drogas foi observada em 65,6% dos Staphylococcus spp. Os fatores de risco identificados para a mastite foram o sistema de criação extensivo, não realização de suplementação alimentar, processo de secagem dos tetos, não realização de desinfecção dos tetos antes e após a ordenha e hábitos higiênicos inadequados dos ordenhadores. A presença de isolados multirresistentes no leite bovino demonstra a importância da escolha e da utilização adequada de antimicrobianos. Medidas de controle e profilaxia, incluindo a antissepsia dos tetos e boas práticas para a obtenção de ordenha higiênica devem ser estabelecidas, com o intuito de prevenir novos casos da doença nos rebanhos.

Caracterização da microbiota auricular de cutias (Dasyprocta aguti) criadas em cativeiro / Characterization of auricular natural microbiota from captive agoutis (Dasyprocta aguti)

The aim of this study was to identify the aerobic bacteria of the auricular natural microbiota from healthy agoutis (Dasyprocta aguti Linnaeus, 1758). In the total, 48 agoutis were used in this experiment, being 32 adults and 16 puppies (both groups divided into equal parts between males and females). The animals were raised under captive conditions, in the Brazilian Semiarid. From each animal, a sample of auricular secretion was collected from each auricular pinna and processed for microbiological analyses. A total of 96 samples were collected and analyzed by colony macroscopic format, cytology and by biochemistry proofs with the objective of isolate and identify the microorganisms. The main bacteria found were Staphylococcus spp. (47.26%), Streptococcus spp. (12.80%), Bacillus spp. (22.73%) and Corynebacterium spp. (17.30%). As conclusion, the most frequent bacteria in auricular pinna of healthy agoutis are Gram-positive cocci and rods, similarly to found in some pets.

O presente trabalho teve por objetivo identificar as principais bactérias aeróbias que compõem a microbiota natural do pavilhão auricular de cutias hígidas. Para tanto, foram utilizadas 48 cutias, criadas em cativeiro sob as condições Semiáridas do Nordeste Brasileiro. Esses animais foram distribuídos nas categorias de adultos (N=32) e filhotes (N=16), e, em ambas, distribuídos igualmente entre machos e fêmeas. Através de um swab, em cada animal coletou-se de cada orelha a secreção presente na superfície do pavilhão auricular dos animais, totalizando 96 amostras. Este material foi refrigerado, e encaminhado ao laboratório para a realização das análises microbiológicas (macroscopia das colônias, citologia e provas bioquímicas), com o intuito de isolar e identificar os microrganismos. Os principais microrganismos isolados foram Staphylococcus spp. (47,26%), Streptococcus spp. (12,80%), Bacillus spp. (22,73%) e Corynebacterium spp. (17,30%). Verificou-se também que não houve diferença entre adultos e filhotes em relação aos microrganismos retrocitados. Assim, as bactérias residentes do pavilhão auricular de cutias hígidas são essencialmente cocos e bacilos gram-positivos, similarmente ao encontrado em pequenos animais domésticos.

Isolation, speciation, and antibiogram of clinically relevant non-diphtherial Corynebacteria (Diphtheroids).

Purpose: Coryneform or the non-diphtherial Corynebacterium species largely remains a neglected group with the traditional consideration of these organisms as contaminants. This concept, however, is slowly changing in the light of recent observations. This study has been done to find out the species distribution and antibiogram of various members of the clinically relevant Coryneform group, isolated from various clinical materials. Materials and Methods: One hundred and fourteen non-duplicate isolates of diphtheroids from various clinical isolates were selected for the study. The isolates were identified to the species level by using a battery of tests; and antimicrobial susceptibility was tested by using a combination of Clinical and Laboratory Standards Institute (CLSI) and the British Society for Antimicrobial Chemotherapy (BSAC) guidelines, in the absence of definitive CLSI guidelines. Results: Corynebacterium amycolatum was the predominant species (35.9%) in our series followed by the CDC Group G organisms (15.7%). Each of the remaining 19 species comprised of less than 10% of the isolates. More than half the total isolates were resistant to the penicillins, erythromycin, and clindamycin; while excellent activity (all the strains being susceptible) was shown by vancomycin, linezolid, and tigecycline. Chloramphenicol and tetracycline also had good activity in inhibiting more than 80% of the isolates. Multiply drug resistance was exhibited by all the species. Conclusion: This study was an attempt to establish the clinical significance of coryneform organisms. The high level of resistance shown by this group to some of the common antibacterial agents highlights the importance of processing these isolates in select conditions to guide the clinicians towards an appropriate therapy.

The efficacy and stability of five sanitizing agents challenged with reference microorganisms and clean area isolates / Eficácia e estabilidade de cinco agentes sanitizantes avaliados frente a microrganismos de referência e isolados de áreas limpas

The antimicrobial activity of five sanitizing agents employed in clean areas designated for the pharmaceutical manufacturing of sterile products was tested against nine microorganisms, including four microorganisms from the clean area microbiota. The method consisted of challenging 5 mL of each sanitizing agent - 70% isopropyl alcohol, 0.4% LPH®, 1.16% hydrogen peroxide, 4% hydrogen peroxide, 1% Bioper® and 5% phenol - with 0.1mL each of concentrated suspensions (105 ? 106 CFU/mL) of Staphylococcus aureus, Candida albicans, Corynebacterium sp., Micrococcus luteus, Escherichia coli, Aspergillus niger, Bacillus subtilis, Staphylococcus sp. and Bacillus sp. for 10 minutes, followed by serial dilutions and plating. The results demonstrated that the five agents were effective against S. aureus, C. albicans, Corynebacterium sp., and M. luteus. The same was true of E. coli, except that isopropyl alcohol showed low levels of inactivation. With A. niger, isopropyl alcohol, 0.4% LPH® and hydrogen peroxide were more effective and 5% phenol and 1% Bioper® less effective. 1% Bioper® and 4% hydrogen peroxide showed greater inactivation of Staphylococcus sp., Bacillus sp. and B. subtilis than the other agents. Against S. aureus, C. albicans, Corynebacterium sp. and M. luteus, 5% phenol showed similar activity to other agents, while with A. niger, B. subtilis, Staphylococcus sp. and Bacillus sp., it was similar to or less active than the other agents. It was demonstrated that two microorganisms from the clean area microbiota, Staphylococcus sp. and Bacillus sp., were the most difficult to eradicate, requiring more frequent application of hydrogen peroxide and 1% Bioper® than the other strains.

O objetivo deste estudo é avaliar a atividade antimicrobiana de cinco agentes sanitizantes empregados em áreas limpas construídas para a fabricação de produtos farmacêuticos estéreis contra nove microrganismos, incluindo quatro microrganismos oriundos da área limpa. A metodologia constituiu em desafiar 5 mL de cada agente sanitizante, álcool isopropílico 70%, LPH® 0,400%, peróxido de hidrogênio 1,160% e 4%, Bioper® 1% e fenol 5% com 0,1 mL de suspensão concentrada (105 ? 106 UFC/mL) de Staphylococcus aureus, Candida albicans, Corynebacterium sp., Micrococcus luteus, Escherichia coli, Aspergillus niger, Bacillus subtilis, Staphylococcus sp. e Bacillus sp. individualmente por 10 minutos, seguido de diluições seriadas e plaqueamento. Os resultados demonstraram que os cinco agentes sanitizantes foram efetivos contra S. aureus, C. albicans, Corynebacterium sp., e M. luteus. Os mesmos resultados foram observados com E. coli, exceto para o álcool isopropílico, que demonstrou baixos níveis de inativação. Contra A. niger, álcool isopropílico, 0.4% LPH® e peróxido de hidrogênio foram mais efetivos e fenol e Bioper® menos efetivos. Bioper® e peróxido de hidrogênio 4% demonstraram altos níveis de inativação de Staphylococcus sp., Bacillus sp. e B. subtilis quando comparados com outros agentes. Fenol demonstrou atividade antimicrobiana similar aos outros agentes contra S. aureus, C. albicans, Corynebacterium sp. e M. luteus. Contra A. niger, B. subtilis, Staphylococcus sp. e Bacillus sp., a atividade antimicrobiana do fenol foi similar ou inferior a dos outros agentes. Foi demonstrado que os microrganismos isolados da área limpa, Staphylococcus sp. e Bacillus sp., foram os que apresentaram maior dificuldade para inativar, sendo necessária a aplicação de peróxido de hidrogênio e Bioper® , com maior frequência.

Microbiological and host features associated with corynebacteriosis in cancer patients: a five-year study

During a five-year period, 932 clinical isolates from cancer patients treated in a Brazilian reference centre were identified as corynebacteria; 86 percent of the cultures came from patients who had been clinically and microbiologically classified as infected and 77.1 percent of these patients had been hospitalised (71.1 percent from surgical wards). The adult solid tumour was the most common underlying malignant disease (66.7 percent). The univariate and multivariate analyses showed that hospitalised patients had a six-fold greater risk (OR = 5.5, 95 percent CI = 1.15-26.30 p = 0.033) related to 30-day mortality. The predominant species were Corynebacterium amycolatum (44.7 percent), Corynebacterium minutissimum (18.3 percent) and Corynebacterium pseudodiphtheriticum (8.5 percent). The upper urinary tracts, surgical wounds, lower respiratory tracts, ulcerated tumours and indwelling venous catheters were the most frequent sources of C. amycolatum strains. Corynebacterium jeikeium infection occurred primarily in neutropenic patients who have used venous catheters, while infection caused by C. amycolatum and other species emerged mainly in patients with solid tumours.

Corynebacterium striatum infecting a malignant cutaneous lesion: the emergence of an opportunistic pathogen / Corynebacterium striatum infectando lesão cutânea maligna: a emergência de um patógeno oportunista

We described a case of a 27-year old male patient with skin and soft tissue infection of a neoplastic lesion caused by Corynebacterium striatum, an organism which has been rarely described as a human pathogen. Identification was confirmed by DNA sequencing. Successful treatment with penicillin was achieved. The role of the C. striatum as an emerging opportunistic pathogen is discussed.

Descrevemos infecção de lesão neoplásica em paciente masculino de 27 anos, envolvendo pele e partes moles, causada por Corynebacterium striatum, um microrganismo raramente descrito como patógeno humano. A identificação foi confirmada por seqüenciamento de DNA. O paciente foi tratado com penicilina, com sucesso. O papel do C. striatum como patógeno oportunista é discutido.

Genetic fingerprinting and relationships of some corynebacterium isolates based on DNA polymorphism

Genetic differences and relationships among six Corynebacterium isolates of different geographic origin from Saudi Arabia were studied based on RAPD-PCR polymorphism. In this analysis many random 10-mer primers were screened, but only five primers [A3, A11, B10, B11 and Z6] were able to generate repeatable and informative products among the DNA samples of the studied bacterial isolates. A total of 47 polymorphic bands were generated by the five primers [100% polymorphism]. Of these bands 23 were scored as unique polymorphic bands. The two primers A11 and B10 were found to be the most effective ones in generating unique bands. The former primer generated 8 unique bands in six isolates while the latter primer produced six unique bands in four isolates. In addition, the most pronounced non unique bands categorized the studied isolates into many class patterns. Combination of all data provides a considerable potentiality for discriminating each Corynebacterium isolate by one or more unique bands or a group of combined class patterns. RAPD data were combined together and used to estimate similarity coefficients and to construct a dendrogram that estimates the relationships among the studied Corynebacterium isolates. The similarity coefficient values varied from zero to 0.44% indicating considerable variability within the studied isolates Based on this dendrogram the studied isolates were separated into two main groups. Such clustering in most cases was well correlated to the geographic collection sites