Detection of Cryptosporidium spp and other intestinal parasites in children with acute diarrhea and severe dehydration in Rio de Janeiro

The objective of the present study was to estimate the frequency of infection by Cryptosporidium spp and other intestinal parasites in dehydrated children with gastroenteritis who were admitted to a pediatric hospital. Stool examinations from 218 children were performed. Cryptosporidium spp was identified in eighteen out of 193 stool samples (9.3%) subjected to safranin-methylene blue staining. Giardia lamblia was detected in ten out of 213 (4.7%) samples examined via the direct or Ritchie methods. Other parasites identified were Ascaris lumbricoides (4.2%), Blastocystis hominis (1.4%), Entamoeba coli (0.9%), Entamoeba histolytica/Entamoeba dispar (0.5%), Endolimax nana (0.5%), Trichuris trichiura (0.5%) and Enterobius vermicularis (0.5%).

Endocardite infecciosa causada por Eikenella corrodens

Os microorganismos do grupo HACEK (Haemophilus spp, Actinobacillus actinomycetemcomitans, Cardiobacterium hominis, Eikenella corrodens e Kingella kingae) são responsáveis por 3% dos casos de endocardites. Eles apresentam propriedades clínicas e microbiológicas semelhantes entre si: são bacilos gram-negativos, isolados mais facilmente em meios aeróbicos, suas culturas necessitam de tempo prolongado de incubação para crescimento (média 3,3 dias) e podem ser considerados como parte da flora normal do trato respiratório superior e da orofaringe1,2. Algumas características foram identificadas nas endocardites por esses agentes, como o quadro clínico insidioso¹, diagnóstico difícil pela natureza fastidiosa e culturas negativas3,4. A endocardite por Eikenella corrodens foi descrita pela primeira vez em 1972(5) e continua sendo um agente etiológico raro. Relatamos o caso de uma paciente com valva nativa que apresentou endocardite infecciosa causada por Eikenella corrodens.

Comparison of methods for the identification of coagulase-negative staphylococci

Coagulase-negative staphylococci (CNS) species identification is still difficult for most clinical laboratories. The scheme proposed by Kloos and Schleifer and modified by Bannerman is the reference method used for the identification of staphylococcal species and subspecies; however, this method is relatively laborious for routine use since it requires the utilization of a large number of biochemical tests. The objective of the present study was to compare four methods, i.e., the reference method, the API Staph system (bioMérieux) and two methods modified from the reference method in our laboratory (simplified method and disk method), in the identification of 100 CNS strains. Compared to the reference method, the simplified method and disk method correctly identified 100 and 99% of the CNS species, respectively, while this rate was 84% for the API Staph system. Inaccurate identification by the API Staph method was observed for Staphylococcus epidermidis (2.2%), S. hominis (25%), S. haemolyticus (37.5%), and S. warneri (47.1%). The simplified method using the simple identification scheme proposed in the present study was found to be efficient for all strains tested, with 100% sensitivity and specificity and proved to be available alternative for the identification of staphylococci, offering, higher reliability and lower cost than the currently available commercial systems. This method would be very useful in clinical microbiology laboratory, especially in places with limited resources.

Automated systems in the identification and determination of methicillin resistance among coagulase negative staphylococci

Coagulase-negative staphylococci (CoNS) are an important cause of nosocomial bacteremia, specially in patients with indwelling devices or those submitted to invasive medical procedures. The identification of species and the accurate and rapid detection of methicillin resistance are directly dependent on the quality of the identification and susceptibility tests used, either manual or automated. The objective of this study was to evaluate the accuracy of two automated systems MicroScan and Vitek - in the identification of CoNS species and determination of susceptibility to methicillin, considering as gold standard the biochemical tests and the characterization of the mecA gene by polymerase chain reaction, respectively. MicroScan presented better results in the identification of CoNS species (accuracy of 96.8 vs 78.8%, respectively); isolates from the following species had no precise identification: Staphylococcus haemolyticus, S. simulans, and S. capitis. Both systems were similar in the characterization of methicillin resistance. The higher discrepancies for gene mec detection were observed among species other than S. epidermidis (S. hominis, S. saprophyticus, S. sciuri, S. haemolyticus, S. warneri, S. cohnii), and those with borderline MICs.

Blastocystis subtypes in irritable bowel syndrome and inflammatory bowel disease in Ankara, Turkey

Blastocystis infection has been reported to be associated with irritable bowel syndrome (IBS), inflammatory bowel disease (IBD) and chronic diarrhoea. The availability of data on the subtypes of Blastocystis found in these patient groups would be of interest in understanding the significance of Blastocystis infection in chronic illness. In this study, we identify Blastocystis subtypes found in patients presenting with IBS, IBD, chronic diarrhoea and asymptomatic patients in Ankara, Turkey. Blastocystis was detected in 11 symptomatic patients by microscopy and 19 by stool culture. Stool culture was more sensitive than microscopy in identifying Blastocystis. Using standard nomenclature adopted in 2007, Blastocystis sp. subtype 3 was the most common in all groups, followed by Blastocystis sp. subtype 2. Identical subtypes of Blastocystis are found in patients with IBS, IBD and chronic diarrhoea. These particular subtypes show low host specificity and are carried by humans and some farm animals. The subtypes of Blastocystis that are commonly found in rodents and certain wild birds were not found in these patients. We suggest a model in which the severity of enteric protozoan infection may be mediated by host factors.

Detection and differentiation of Cryptosporidium by real-time polymerase chain reaction in stool samples from patients in Rio de Janeiro, Brazil

This study reports the first genetic characterisation of Cryptosporidium isolates in Brazil using real-time polymerase chain reaction (RT-PCR). A total of 1,197 faecal specimens from children and 10 specimens from human immunodeficiency virus-infected patients were collected between 1999-2010 and screened using microscopy. Forty-eight Cryptosporidium oocyst-positive isolates were identified and analysed using a generic TaqMan assay targeting the 18S rRNA to detect Cryptosporidium species and two other TaqMan assays to identify Cryptosporidium hominis and Cryptosporidium parvum. The 18S rRNA assay detected Cryptosporidium species in all 48 of the stool specimens. The C. parvum TaqMan assay correctly identified five/48 stool samples, while 37/48 stool specimens were correctly amplified in the C. hominis TaqMan assay. The results obtained in this study support previous findings showing that C. hominis infections are more prevalent than C. parvum infections in Brazil and they demonstrate that the TaqMan RT-PCR procedure is a simple, fast and valuable tool for the detection and differentiation of Cryptosporidium species.

Coagulase-negative staphylococci strains resistant to oxacillin isolated from neonatal blood cultures

Coagulase-negative staphylococci (CoNS) are the microorganisms most frequently isolated from clinical samples and are commonly found in neonatal blood cultures. Oxacillin is an alternative treatment of choice for CoNS infections; however, resistance to oxacillin can have a substantial impact on healthcare by adversely affecting morbidity and mortality. The objective of this study was to detect and characterise oxacillin-resistant CoNS strains in blood cultures of newborns hospitalised at the neonatal ward of the University Hospital of the Faculty of Medicine of Botucatu. One hundred CoNS strains were isolated and the mecA gene was detected in 69 of the CoNS strains, including 73.2% of Staphylococcus epidermidis strains, 85.7% of Staphylococcus haemolyticus strains, 28.6% of Staphylococcus hominis strains and 50% of Staphylococcus lugdunensis strains. Among these oxacillin-resistant CoNS strains, staphylococcal cassette chromosome mec (SCCmec) type I was identified in 24.6%, type II in 4.3%, type III in 56.5% and type IV in 14.5% of the strains. The data revealed an increase in the percentage of CoNS strains isolated from blood cultures from 1991-2009. Furthermore, a predominant SCCmec profile of the oxacillin-resistant CoNS strains isolated from neonatal intensive care units was identified with a prevalence of SCCmec types found in hospital-acquired strains.

Genetic diversity of Cryptosporidium identified in clinical samples from cities in Brazil and Argentina

The identification and characterisation of Cryptosporidiumgenotypes and subtypes are fundamental to the study of cryptosporidiosis epidemiology, aiding in prevention and control strategies. The objective was to determine the genetic diversity ofCryptosporidium in samples obtained from hospitals of Rio de Janeiro, Brazil, and Buenos Aires, Argentina. Samples were analysed by microscopy and TaqMan polymerase chain reaction (PCR) assays forCryptosporidium detection, genotyped by nested-PCR-restriction fragment length polymorphism (RFLP) analysis of the 18S rRNA gene and subtyped by DNA sequencing of the gp60 gene. Among the 89 samples from Rio de Janeiro, Cryptosporidium spp were detected in 26 by microscopy/TaqMan PCR. In samples from Buenos Aires,Cryptosporidium was diagnosed in 15 patients of the 132 studied. The TaqMan PCR and the nested-PCR-RFLP detected Cryptosporidium parvum, Cryptosporidium hominis, and co-infections of both species. In Brazilian samples, the subtypes IbA10G2 and IIcA5G3 were observed. The subtypes found in Argentinean samples were IbA10G2, IaA10G1R4, IaA11G1R4, and IeA11G3T3, and mixed subtypes of Ia and IIa families were detected in the co-infections. C. hominis was the species more frequently detected, and subtype family Ib was reported in both countries. Subtype diversity was higher in Buenos Aires than in Rio de Janeiro and two new subtypes were described for the first time.

Fatores Associados ao Aborto Espontâneo Recorrente

Objetivo: identificar fatores mais freqüentemente associados ao aborto espontâneo recorrente. Casuística: no período de março de 1993 a março de 1997, 175 pacientes foram avaliadas no Ambulatório de Aborto Recorrente do CAISM/UNICAMP. Os critérios de seleção foram: história de três ou mais abortos espontâneos consecutivos em pacientes com idade inferior a 35 anos e/ou dois abortos e idade superior a 35 anos. Métodos: o protocolo de investigação incluiu: cariótipo; histerossalpingografia; dosagem seriada de progesterona e/ou biópsia de endométrio; pesquisa sorológica de infecções: toxoplasmose, listeriose, brucelose, sífilis e citomegalovírus e pesquisa para Mycoplasma hominis e Chlamydia trachomatis na secreção cérvico-uterina. A investigação também incluiu dosagem de hormônios tiroideanos e da glicemia de jejum; pesquisa de fator auto-imune por meio de painel de auto-anticorpos; pesquisa de fator aloimune mediante anticorpos contra antígenos leucocitários humanos (anti-HLA), prova cruzada por microlinfocitotoxicidade e cultura mista unidirecional de linfócitos, em que se comparam as respostas maternas diante das células paternas e de um doador não-relacionado para pesquisa de fator inibidor destas respostas no soro materno. O exame dos parceiros incluiu: exame físico geral e especial, sorologias para sífilis, doença de Chagas, hepatite B e C e síndrome da imunodeficiência adquirida (AIDS), além da prova cruzada por microlinfocitotoxicidade e da cultura mista de linfócitos. Resultados: o fator mais freqüentemente encontrado foi o aloimune (86,3% dos casos), representado por prova cruzada negativa e inibição na cultura mista de linfócitos inferior a 50%. O segundo fator mais freqüentemente encontrado foi a incompetência istmo-cervical (22,8%), seguido pelo fator hormonal (21,2%), representado principalmente pela insuficiência lútea. Algumas pacientes apresentavam mais de um fator concomitantemente. Conclusão: para uma investigação completa dos fatores associados ao aborto espontâneo recorrente faz-se necessária a inclusão do fator aloimune, sem a qual a maior parte dos casos não terá etiologia esclarecida.

Infecção Assintomática do Líquido Amniótico

Objetivo: determinar a presença de infecção assintomática do líquido amniótico em gestantes, identificar os agentes bacterianos envolvidos na infecção e determinar o perfil de suscetibilidade a antimicrobianos in vitro. Métodos: foram obtidas 81 amostras de líquido amniótico, colhidas por amniocentese, em gestantes sem sinais de trabalho de parto e sem suspeita clínica de infecção, atendidas na Maternidade Escola Assis Chateaubriand, entre agosto/97 e janeiro/99. Pesquisou-se a presença de bactérias aeróbias, anaeróbias estritas/facultativas e micoplasmas genitais. As bactérias anaeróbias foram identificadas pelo sistema ATBÒ (Bio-Mérieux) e os micoplasmas pelo kit Micoplasmas ISTÒ (Bio-Mérieux). Resultados: entre as amostras obtidas, oito (9,8%) apresentaram culturas positivas, sendo que em duas foram identificadas duas espécies bacterianas. Os patógenos isolados foram: Ureaplasma urealyticum (7 casos, 8,6%), Mycoplasma hominis (1 caso, 1,2%) e Peptostreptococcus sp (2 casos 2,4%). O padrão de resistência aos antimicrobianos caracterizou-se pela maior resistência dos micoplasmas à eritromicina (37,5%) e nenhuma resistência às ciclinas. Conclusões: o percentual de infecções assintomáticas foi muito elevado, havendo necessidade de serem realizadas novas pesquisas para avaliar as conseqüências da infecção subclínica nas grávidas e em seus conceptos, que envolvam métodos que identifiquem micoplasmas genitais, já que foram as bactérias mais freqüentemente isoladas.

Antimicrobial resistance and detection of mecA and blaZ genes in coagulase-negative Staphylococcus isolated from bovine mastitis

The present study evaluated the pheno- and genotypical antimicrobial resistance profile of coagulase-negative Staphylococcus (CNS) species isolated from dairy cows milk, specially concerning to oxacillin. Of 100 CNS isolates, the S. xylosus was the prevalent species, followed by S. cohnii, S. hominis, S. capitis and S. haemolyticus. Only 6% were phenotypically susceptible to the antimicrobial agents tested in disk diffusion assay. Penicillin and ampicillin resistance rates were significantly higher than others antimicrobials. Four isolates were positive to mecA gene (4%), all represented by the S. xylosus species. The blaZ gene was detected in 16% of the isolates (16/100). It was noticed that all mecA + were also positive to this gene and the presence of both genes was correlated to phenotypic beta-lactamic resistance. We conclude that CNS species from bovine milk presented significantly distinct antimicrobial resistance profiles, evaluated by phenotypic and genotypic tests, which has implications for treatment and management decisions.