Detecção e identificação de genes de β-lactamases de espectro estendido em Salmonella spp. e Escherichia coli isoladas de carnes de frango, suína e bovina destinadas ao consumo humano

Pós-graduação em Microbiologia - IBILCE

Estudo da atividade antimicrobiana de algumas árvores medicinais nativas com potencial de conservação/recuperação de florestas tropicais

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ocorrência de mosca-branca em Annona squamosa no estado de São Paulo

In Brazil the knowledge about the geographical distribution and host plants of whiteflies (Hemiptera: Aleyrodidae) is limited, mainly on crops. In this communication, the occurrence of Aleurodicus mirabilis (Cockerell, 1898) in high infestations in custard apple (Annona squamosa Linnaeus, 1753) is recorded in the state of São Paulo, Brazil.

Clinical and Microbiologic Evaluation of Chronic Leg Ulcers: A Cross-sectional Study

OBJECTIVE: This study aimed at evaluating the flora and bacterial load of chronic leg ulcers (CLUs) according to the clinical judgment of colonization or infection.DESIGN: This was an analytical and cross-sectional study.SETTING: This study was conducted in an outpatient wound care unit in the Dermatology Department of the Botucatu School of Medicine-UNESP, Brazil.PARTICIPANTS: The participants were patients with CLUs who did not use systemic antibiotics.METHODS: The ulcers were clinically divided into 3 groups: ulcers with good granulation tissue (GGT), critical colonization (CC), and infection. Secretion was collected from a 1-cm(2) area using a swab and seeded by the semiquantitative method.OUTCOME MEASURES: The main outcome measures were genus and species of the bacteria found in the cultures and result of the semiquantitative culture correlating with the clinical diagnosis of GGT, CC, and infection.MAIN RESULTS: Seventy-seven ulcers were evaluated: 27 with GGT, 29 with CC, and 21 with infection. Gram-negative bacteria were most often found in all groups (81%): Pseudomonas aeruginosa, in granulation and colonized ulcers, and Proteus mirabilis, in infected ulcers. Ulcers from the infected group showed higher bacterial load.CONCLUSIONS: The flora of CLUs was predominantly constituted by gram-negative bacteria, and P aeruginosa was the most prevalent. The bacterial load of infected ulcers was higher as compared with the others, although some ulcers with GGT also presented a high load. The interpretation of microbiologic tests based on the swab techniques and even on semiquantitative analysis requires close clinical correlation.

Etiologia e perfil de sensibilidade aos antimicrobianos de pacientes ambulatoriais de Botucatu e região

As infecções do trato urinário (ITU) são definidas como a invasão e posterior multiplicação de microorganismos nos tecidos das vias urinárias, sendo a bactéria gram-negativa E.coli o principal uropatógeno envolvido nessa infecção. A avaliação para a presença de ITU é feita através do quadro clínico característico desse tipo de paciente e o exame de cultura de urina. As amostras positivas para as culturas são avaliadas quanto ao perfil de sensibilidade aos antimicrobianos dos agentes etiológicos encontrados. Descrever a porcentagem de ocorrência dos agentes etiológicos e o perfil de sensibilidade aos antimicrobianos das Infecções do Trato Urinário em pacientes ambulatoriais de Botucatu e região. Foram incluídas neste estudo culturas de urinas de pacientes atendidos em UBS de Botucatu e região Pólo Cuesta, no período de Janeiro de 2008 a Maio de 2010. As culturas positivas foram avaliadas quanto a porcentagem de ocorrência dos uropatógenos segundo sexo, faixa etária e origem. Foi utilizado o teste do qui-quadrado para verificar associação entre cada antimicrobiano e as porcentagens de sensibilidade e resistência encontradas. Foram avaliadas 11.768 culturas de urina provenientes de 11 municípios da região, sendo consideradas as culturas positivas (28%). Quanto à ocorrência dos agentes etiológicos verificou-se que a E.coli é responsável 58,76% dos casos. A distribuição dos uropatógenos segundo sexo revelou maior ocorrência de E.coli nos dois sexos. O segundo agente etiológico de maior ocorrência foi, nos homens o Proteus mirabilis e nas mulheres, o S. saprophyticus. Dentre as faixas etárias, a E.coli foi o patógeno de maior ocorrência. O perfil de sensibilidade aos antimicrobianos revelou que a E.coli é mais sensível a Ertapenem, Imipenem, Meropenem, Fosfomicina, Amicacina, Ceftriaxona, Nitrofurantoína e Norfloxacina; e mais resistente... (Resumo completo, clicar acesso eletrônico abaixo)

Avaliação de métodos de triagem e de detecção fenotípica de ß-lactamase de espectro ampliado (ESBL)

The catalytic function of extended-spectrum β-lactamases can result in high degrees of bacterial resistance to β-lactamic antimicrobials and in the emergence of ESBL among the members of Enterobacteriaceae family, especially Klebsiella pneumoniae and Escherichia coli. This occurs due to the dissemination and emergence of new variants of these enzymes caused by the high utilization of antibiotics like broad-spectrum cephalosporins. The ESBL are β-lactamases capable of conferring bacterial resistance to the penicillins, 1st, 2nd and 3rd generation cephalosporins, and aztreonam (but not cephamycins and carbapenems) through the hydrolysis of these antibiotics. In view of this phenomenon, the exact screening and detection of the producers of ESBL are essential for the appropriate selection of the antimicrobial therapy. The purposes of this study were to evaluate the best antimicrobial for the selection of ESBL producers and to determine the best method for the detection of such microorganisms. We evaluated 200 sequential bacterial samples including the species Klebsiella pneumoniae (56.5%), Escherichia coli (34%), Proteus mirabilis (8.5%) and Klebsiella oxytoca (1%), previously characterized as ESBL producers between February and September 2008 in the Laboratory of Microbiology, Botucatu Medical School - UNESP, Botucatu, São Paulo State, Brazil. To select the ESBL-producer bacteria, we used the disks recommended by CLSI 2008, aztreonam (ATM), cefpodoxime (CPD), ceftriaxone (CRO), cefotaxime (CTX) and ceftazidime (CAZ), besides cefepime (FEP). ESBL production was confirmed by three methods: double disk screening, ESBL Etest®, and Vitek® automated system. The disks employed in the double disk screening were: penicillin associated with β-lactamase inhibitor, amoxicillin-clavulanic acid, and two β-lactamic antibiotics, ceftazidime and cefotaxime...(Complete abstract click electronic access below)

Detection of bla(CTX-M)-type genes in complex class 1 integrons carried by Enterobacteriaceae isolated from retail chicken meat in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Meningite supurativa secundária a otite média e interna em um gato: relato de caso

Suppurative meningitis are rarely reported in dogs and cats. The present study aims to report the occurrence of suppurative meningitis secondary to otitis media and interna in a five-year-old female Persian cat examined at the Veterinary School of Unesp-Botucatu with acute progressive multifocal neurological alterations. Analysis of the cerebrospinal fluid (CSF) demonstrated predominance of neutrophils (90%) and the presence of bacteria. Therapy with antibiotics was initiated, but the animal came to die. beta-hemolytic Staphylococcus, beta-hemolytic Streptococcus and Proteus mirabilis were detected in fragments sent to culture. The present report demonstrates the importance of performing more specific exams such as complete CSF analysis in cases of otitis media and interna. This way, central nervous infections can be diagnosed and treated in a timely manner, as those are acute diseases with poor prognosis.

Ocorrência de fungos negros em ambientes domésticos

Black fungi are able to adapt to extreme environmental conditions, such as: high temperatures, the presence of toxic chemical substances and lack of nutrients. Besides, they are also potential pathogens to humans. The natural environment of many black fungi is still unknown and some studies are being conducted to evaluate the biodiversity of this group and their different habitats. This study aimed to isolate black fungi in domestic environments and facilities, such as toothbrushes, fridge sealing rubbers, bathroom strainers and divisions, windows, wall tiles and bath sponge. For the collection, material surfaces were scratched with a scalpel and the resulting fragments were sewed in Mycosel agar (DifcoTM), supplemented with actidione to inhibit the growth of highly-sporulating fungi. Plates were incubated at 25ºC for three weeks. The 46 isolated fungi were maintained on MA2% slants at 8ºC and cryopreserved at -80ºC. Fungal identification was performed through the analysis of macro and microscopic features and ITS rDNA sequencing. The following black fungi taxa were found: Ascomycota sp., Cladosporium spp., Dothideomycete sp., Exophiala alcalophila, Ochroconis mirabilis and Rhinocladiella atrovirens. Non-melanized fungi were also found, such as Geosmithia sp., Penicillium sp. and Rhodotorula mucilaginosa. The temperature tests showed that isolated black fungi were not able to grow at 37°C, however, this temperature proved to be fungistatic to 43% of them. According to literature, all black fungi isolated in this study are opportunistic pathogens and additional studies are necessary to evaluate the risk that these micro-organisms offer to health, once they were isolated from domestic environments

Ocorrência de fungos negros em ambientes domésticos

Black fungi are able to adapt to extreme environmental conditions, such as: high temperatures, the presence of toxic chemical substances and lack of nutrients. Besides, they are also potential pathogens to humans. The natural environment of many black fungi is still unknown and some studies are being conducted to evaluate the biodiversity of this group and their different habitats. This study aimed to isolate black fungi in domestic environments and facilities, such as toothbrushes, fridge sealing rubbers, bathroom strainers and divisions, windows, wall tiles and bath sponge. For the collection, material surfaces were scratched with a scalpel and the resulting fragments were sewed in Mycosel agar (DifcoTM), supplemented with actidione to inhibit the growth of highly-sporulating fungi. Plates were incubated at 25ºC for three weeks. The 46 isolated fungi were maintained on MA2% slants at 8ºC and cryopreserved at -80ºC. Fungal identification was performed through the analysis of macro and microscopic features and ITS rDNA sequencing. The following black fungi taxa were found: Ascomycota sp., Cladosporium spp., Dothideomycete sp., Exophiala alcalophila, Ochroconis mirabilis and Rhinocladiella atrovirens. Non-melanized fungi were also found, such as Geosmithia sp., Penicillium sp. and Rhodotorula mucilaginosa. The temperature tests showed that isolated black fungi were not able to grow at 37°C, however, this temperature proved to be fungistatic to 43% of them. According to literature, all black fungi isolated in this study are opportunistic pathogens and additional studies are necessary to evaluate the risk that these micro-organisms offer to health, once they were isolated from domestic environments

The Pst system of Streptococcus mutans is important for phosphate transport and adhesion to abiotic surfaces

The Pst system is a high-affinity inorganic phosphate transporter found in many bacterial species. Streptococcus mutans, the etiological agent of tooth decay, carries a single copy of the pst operon composed of six cistrons (pstS, pstC1, pstC, pstB, smu.1134 and phoU). Here, we show that deletion of pstS, encoding the phosphate-binding protein, reduces phosphate uptake and impairs cell growth, which can be restored upon enrichment of the medium with high concentrations of inorganic phosphate. The relevance of Pst for growth was also demonstrated in the wild-type strain treated with an anti-PstS antibody. Nevertheless, a reduced ability to bind to saliva-coated surfaces was observed, along with the reduction of extracellular polysaccharide production, although no difference on pH acidification was observed between mutant and wild-type strains. Taken together, the present data indicate that the S.similar to mutans Pst system participates in phosphate uptake, cell growth and expression of virulence-associated traits.

Community-acquired urinary tract infection: age and gender-dependent etiology

LO, Denise Swei et al. Community-acquired urinary tract infection: age and gender-dependent etiology. J. Bras. Nefrol. [online]. 2013, vol.35, n.2, pp. 93-98. ISSN 0101-2800. http://dx.doi.org/10.5935/0101-2800.20130016. INTRODUCTION: Choosing the antimicrobial agent for initial therapy of urinary tract infection (UTI) is usually empirical and should consider the prevalence of uropathogens in different age groups and gender. OBJECTIVE: To establish prevalence rates of uropathogens in community-acquired UTI in relation to age and gender. METHODS: Crosssectional study conducted in the emergency department (ED) of a general hospital, from January to December, 2010, in patients younger than 15 years old who had clinical suspicion of UTI and collected quantitative urine culture. UTI was defined as urine culture with growth of a single agent > 100.000 colony forming units (cfu)/mL in a midstream collection or > 50.000 cfu/mL in urethral catheterization. RESULTS: There were 63.464 visits to ED. 2577 urine cultures were obtained, of whom 291 were positive for UTI (prevalence = 11.3% of clinical suspicion and 0.46% of visits), 212 cases (72.8%) in females, median age = 2.6 years. The predominant uropathogen was E. coli (76.6%), followed by Proteus mirabilis (10.3%) and Staphylococcus saprophyticus (4.1%). Among infants < 3 months, prevalence rates of E. coli were significantly lower (50% vs 78.4%; OR = 0.276; p = 0.006). Higher prevalences of Staphylococcus saprophyticus occurred among patients > 10 years (24.4% vs 0.4%; OR = 79.265; p < 0.0001). Proteus mirabilis was significantly more prevalent in boys than girls (24.0% vs 5.2%; OR = 5.786; p < 0.001). CONCLUSIONS: E. coli was the most prevalent community-acquired uropathogen. Nevertheless, initial empiric antimicrobial treatment of UTI should consider the significant prevalence of other agents different from E. coli in infants < 3 months, the high prevalence of Staphylococcus saprophyticus in patients > 10 years and Proteus mirabilis in males.

Cerebral revascularization model in a swine

The purpose of this study was to analyze the suitability of the cerebral vasculature of the pig regarding a revascularization procedure. In two 60 kg pigs the femoral artery was exposed and canulated for selective angiography and interventional procedures. After the angiography, the pigs were brought to the animal OR for craniotomy and analysis of the intracranial cerebral arteries and the surgical exposure of the carotid arteries under the microscope. Angiography demonstrated the presence of a true internal-, external carotid artery and vertebral arteries. Both the vertebral and internal carotid arteries are feeding a rete mirabilis both at the cranial base and the cranio-cervical junction. At these sites further advancement of the angiography catheter was not possible. Out of these rete mirabilis, an intracranial carotid artery and an intracranial vertebral artery were formed, respectively. The intracranial cerebral vessels were of the dimension of 1 mm and less. The extracranial portion of the internal carotid artery was 2.5 mm of diameter. From these findings, we conclude that a direct cerebral revascularization procedure of the intracranial vessels is not possible in the swine. However, a global revascularization procedure on the extracranial portion of the internal carotid artery is thus feasible, both using a low- and high-flow anastamosis technique.

A multiplex real-time PCR assay for rapid detection and differentiation of 25 bacterial and fungal pathogens from whole blood samples

Early detection of bloodstream infections (BSI) is crucial in the clinical setting. Blood culture remains the gold standard for diagnosing BSI. Molecular diagnostic tools can contribute to a more rapid diagnosis in septic patients. Here, a multiplex real-time PCR-based assay for rapid detection of 25 clinically important pathogens directly from whole blood in <6 h is presented. Minimal analytical sensitivity was determined by hit rate analysis from 20 independent experiments. At a concentration of 3 CFU/ml a hit rate of 50% was obtained for E. aerogenes and 100% for S. marcescens, E. coli, P. mirabilis, P. aeruginosa, and A. fumigatus. The hit rate for C. glabrata was 75% at 30 CFU/ml. Comparing PCR identification results with conventional microbiology for 1,548 clinical isolates yielded an overall specificity of 98.8%. The analytical specificity in 102 healthy blood donors was 100%. Although further evaluation is warranted, our assay holds promise for more rapid pathogen identification in clinical sepsis.

Eemian and Holocene sea-surface conditions in the southern Black Sea: organic-walled dinoflagellate cyst record from core 22-GC3

In order to compare the sea-surface conditions in the Black Sea during the Holocene and Eemian, sapropelic parts of marine core 22-GC3 (42°13.53′N/36°29.55′E, 838 m water depth) were studied for organic-walled dinoflagellate cyst content. The record shows a change from freshwater/brackish assemblages (Pyxidinopsis psilata, Spiniferites cruciformis, and Caspidinium rugosum) to more marine assemblages (Lingulodinium machaerophorum and Spiniferites ramosus complex) during each interglacial, due to the inflow of saline Mediterranean water. The lacustrine–marine transitions in 22-GC3 occurred at ~ 8.3 cal kyr BP during the early Holocene and ~ 128 kyr BP during the early Eemian, slightly later compared to the onset of interglacial conditions on the adjacent land. Dinoflagellate cyst assemblages reveal higher sea-surface salinity (~ 28–30) (e.g. Spiniferites pachydermus, Bitectatodinium tepikiense, and Spiniferites mirabilis) around ~ 126.5–121 kyr BP in comparison to the Holocene (~ 15–20) as well as relatively high sea-surface temperature (e.g. Tuberculodinium vancampoae, S. pachydermus, and S. mirabilis) especially at ~ 127.6–125.3 kyr BP. Establishment of high sea-surface salinity during the Eemian correlates very well with reconstructed relatively high global sea-level and is explained as a combined effect of increased Mediterranean supply and high temperatures at the beginning of the last interglacial. The observed changes in the dinocyst record highlight the importance of nutrients for the composition of the Eemian and Holocene dinocyst assemblages.