In Vitro Antimicrobial Susceptibility of Mycobacterium massiliense Recovered from Wound Samples of Patients Submitted to Arthroscopic and Laparoscopic Surgeries

Testing of rapidly growing species of mycobacteria (RGM) against antibacterial agents has been shown to have some clinical utility. This work establishes the MICs of seven antimicrobial agents following the guidelines set forth by the Clinical and Laboratory Standards Institute (CLSI) against eighteen isolates of Mycobacterium massiliense recovered from wound samples of patients submitted to minimally invasive surgery such as arthroscopy and laparoscopy. The isolates showed susceptibility to amikacin (MIC90 = 4 μg/mL) and clarithromycin (MIC90 < 1 μg/mL) but resistance to ciprofloxacin (MIC90 > 16 μg/mL), doxycycline (MIC90 > 32 μg/mL), sulfamethoxazole (MIC90 > 128 μg/mL), and tobramycin (MIC90 = 32 μg/mL), and intermediate profile to cefoxitin (MIC90 = 64 μg/mL). Therefore, we suggest that the antimicrobial susceptibilities of any clinically significant RGM isolate should be performed

Paracoccidioides brasiliensis pancreatic destruction in Calomys callosus experimentally infected

<p>Abstract</p> <p>Background</p> <p>The wild rodent <it>Calomys callosus </it>is notably resistant to <it>Trypanosoma cruzi </it>infection. In order to better characterize this animal model for experimental infections, we inoculated <it>C</it>. <it>callosus </it>intraperitoneally with <it>Paracoccidioides brasiliensis</it>, a thermally dimorphic fungus that causes a chronic disease with severe granuloma formation in the mouse and humans. The dissemination of <it>P. brasiliensis </it>cells through the lungs, liver, pancreas, and spleen was assessed by histological analysis.</p> <p>Results</p> <p>The animals were susceptible to infection and showed a granulomatous reaction. <it>C. callosus </it>presented peritonitis characterized by the presence of exudates containing a large number of yeast cells. Extensive accumulation of yeast cells with intense destruction of the parenchyma was observed in the pancreas, which reduced the glucose levels of infected animals. These lesions were regressive in the liver, spleen, and lungs until complete recovery. The role of estrogen during <it>C. callosus </it>infection with <it>P. brasiliensis </it>was addressed by infecting ovariectomized animals. It was observed a reduced inflammatory response as well as reduced extension of tissue damage. Removal of ovaries reestablished the normal glucose levels during infection.</p> <p>Conclusion</p> <p>Taken together, the results presented here reveal the pancreas as being an important organ for the persistence of <it>P. brasiliensis </it>during infection of <it>C. callosus </it>and that estrogen plays an important role in the susceptibility of the animals to this pathogen.</p

The activity of a Hexameric M17 Metallo-Aminopeptidase is associated with survival of Mycobacterium tuberculosis

Mycobacterium tuberculosis is one of the most prevalent human pathogens causing millions of deaths in the last years. Moreover, tuberculosis (TB) treatment has become increasingly challenging owing to the emergence of multidrug resistant M. tuberculosis strains. Thus, there is an immediate need for the development of new anti-TB drugs. Proteases appear to be a promising approach and may lead to shortened and effective treatments for drug-resistant TB. Although the M. tuberculosis genome predicts more than 100 genes encoding proteases, only a few of them have been studied. Aminopeptidases constitute a set of proteases that selectively remove amino acids from the N-terminus of proteins and peptides and may act as virulence factors, essential for survival and maintenance of many microbial pathogens. Here, we characterized a leucine aminopeptidase of M. tuberculosis (MtLAP) as a cytosolic oligomeric metalloaminopeptidase. Molecular and enzymatic properties lead us to classify MtLAP as a typical member of the peptidase family M17. Furthermore, the aminopeptidase inhibitor bestatin strongly inhibited MtLAP activity, in vitro M. tuberculosis growth and macrophage infection. In murine model of TB, bestatin treatment reduced bacterial burden and lesion in the lungs of infected mice. Thus, our data suggest that MtLAP participates in important metabolic pathways of M. tuberculosis necessary for its survival and virulence and consequently may be a promising target for new anti-TB drugs

New vectors derived from pUC18 for cloning and thermal-induced expression in Escherichia coli

We report the construction of two vectors for Escherichia coli: pUC72, for molecular cloning, and pPLT7, for thermal-induced expression. The main feature of pUC72 is a novel polylinker region that includes restriction sites for Nde I and Nco I which provide an ATG codon for proper translation initiation of expressed genes. Vector pPLT7 is ideal for thermo-inducible expression in host cells that carry the cI857 repressor gene. The use of pPLT7 was validated by the successful expression of the genes encoding carp and porcine growth hormones. These vectors provide novel cloning possibilities in addition to simple, non-expensive, high level expression of recombinant proteins in E. coli

Non-disulfide-Bridge peptide 5.5 from the Scorpion Hadrurus gertschi Inhibits the growth of mycobacterium abscessus subsp. massiliense

Multi-drug resistant microorganisms have been a growing concern during the last decades due to their contribution in mortality rates worldwide. Antimicrobial peptides (AMPs) are broad spectrum antimicrobial agents that display potent microbicidal activity against a wide range of microorganisms. AMPs generally have a rapid mode of action that reduces the risk of resistance developing among pathogens. In this study, an AMP derived from scorpion venom, NDBP-5.5, was evaluated against Mycobacterium abscessus subsp. massiliense, a rapidly growing and emerging pathogen associated with healthcare infections. The minimal bactericidal concentration of NDBP-5.5, AMP quantity necessary to stop bacteria visible growth, against M. abscessus subsp. massiliense was 200 μM, a concentration that did not induce hemolysis of human red blood cells. The therapeutic index was 3.05 indicating a drug with low toxicity and therefore good clinical potential. Treatment of infected macrophages with NDBP-5.5 or clarithromycin presented similar results, reducing the bacterial load. M. abscessus subsp. massiliense-infected animals showed a decrease in the bacterial load of up to 70% when treated with NDBP-5.5. These results revealed the effective microbicidal activity of NDBP-5.5 against Mycobacterium, indicating its potential as an antimycobacterial agent

Microbiological quality of human milk from a Brazilian milk bank

OBJECTIVE: The objectives of the present study were to determine the prevalence of potentially pathogenic microorganisms that indicate the hygienic and sanitary conditions of human milk samples collected at a Human Milk Bank. METHODS: Three hundred and thirty eight (338) samples of human milk collected from a milk bank in a maternity in the municipality of Goiânia, in the state of Goias, Brazil were submitted to microbiological analysis. The latter were plated on McConkey agar according to the type of bacteria. Among the total number of samples collected, 194 consisted of raw milk and the remaining 144 were pasteurized milk. RESULTS: The presence of Staphylococcus spp., Streptococcus spp., yeasts and molds, and Enterobacteriaceae was verified in the raw milk samples. Staphylococcus aureus were isolated in 10 (5.2%) samples, Staphylococcus epidermidis in 28 (14.4%) samples, Streptococcus spp. in three (1.6%) samples, yeasts and molds in 43 (22.2%) and Enterobacteriaceae in 49 (25.3%) samples. In a hundred and forty four (144) samples which underwent thermal treatment Staphylococcus aureus was detected in five (3.5%) samples, Staphylococcus epidermidis in 15 (10.4%), Staphylococcus lugdenensis in two (1.4%), Streptococcus spp. in four (2.8%), yeasts and molds in 37 (25.7%), and Enterobacteriaceae in nine (6.3%). CONCLUSIONS: Analysis indicated a high degree of contamination in raw human milk, and as for the pasteurized milk, despite elimination of the great majority of potentially pathogenic microorganisms, the percentage of yeasts and molds was higher than in raw milk, demonstrating that a lower degree of initial contamination would be necessary for pasteurization to be an efficient means of microbiological control.OBJETIVO: Determinar a prevalência de microrganismos indicadores e potencialmente patogênicos que indicam as condições higiênico-sanitárias das amostras de leite humano ordenhado coletadas em banco de leite. MÉTODOS: Foram realizadas análises microbiológicas de 338 amostras de leite humano ordenhado, sendo 194 de leite cru e 144, pasteurizado, coletadas em banco de leite humano de um hospital materno infantil de Goiânia, GO. As análises microbiológicas foram realizadas com semeadura em ágar Mc Conkey, de acordo com o tipo de bactéria. RESULTADOS: No leite cru, verificou-se a presença de Staphylococcus spp. Streptococcus spp., bolores e leveduras e Enterobacteriaceae. Observou-se que Staphylococcus aureus esteve presente em 10 (5,2%) amostras, Staphylococcus epidermidis em 28 (14,4%), Streptococcus spp. em três (1,6%), bolores e leveduras em 43 (22,2%) e Enterobacteriaceae em 49 (25,3%). Das 144 amostras de leite humano ordenhado pasteurizado, detectaram-se Staphylococcus aureus em cinco (3,5%), Staphylococcus epidermidis em 15 (10,4%), Staphylococcus lugdenensis em duas (1,4%), Streptococcus spp. em quatro (2,8%), bolores e leveduras em 37 (25,7%) e Enterobacteriaceae em nove (6,3%). CONCLUSÕES: Os resultados mostraram um alto grau de contaminação no leite cru. No leite pasteurizado, apesar da eliminação da grande maioria de microrganismos potencialmente patogênicos, a percentagem de bolores e leveduras excedeu a de leite cru, mostrando a necessidade de obtenção de um leite com carga microbiana inicial mais baixa para que a pasteurização seja eficiente no controle microbiológico

AVALIAÇÃO DE RESISTÊNCIA ASSOCIADA AO BIOFILME DE ISOLADOS DE Candida E IDENTIFICAÇÃO DE ESPÉCIES POR PCR EM TEMPO REAL

Introdu&ccedil;&atilde;o e objetivos: Esp&eacute;cies do g&ecirc;nero Candida s&atilde;o comensais end&oacute;genos em indiv&iacute;duos saud&aacute;veis e infec&ccedil;&atilde;ocausada por esses agentes &eacute; denominada candid&iacute;ase. Pacientes hospitalizados tempredisposi&ccedil;&atilde;o para adquirir candid&iacute;ase invasiva apresentando isoladosresistentes aos antif&uacute;ngicos em decorr&ecirc;ncia da forma&ccedil;&atilde;o de biofilme. Dessa forma, o objetivo dotrabalho foi avaliar o biofilme de isolados de Candida de pacientes hospitalizados diferenciando as esp&eacute;cies porPCR em tempo real. Metodologia: O ensaio de redu&ccedil;&atilde;o do sal tetraz&oacute;lio foi utilizadopara determinar a suscetibilidade das c&eacute;lulas s&eacute;sseis de 21 isolados de Candida frente &agrave; caspofungina,anfotericina B, fluconazol e voriconazol. Diferencia&ccedil;&atilde;o das esp&eacute;cies foirealizada por compara&ccedil;&atilde;o da temperatura de melting de amplicons da regi&atilde;o doDNA ribossomal 18S. Resultadose discuss&otilde;es: Foramisolados 10 C. albicans, 10 C. parapsilosis e 01 C. tropicalis. As c&eacute;lulas s&eacute;sseis foramresistentes em 71,4% dos isolados, apresentando sensibilidade apenas &agrave;anfotericina B. Foi poss&iacute;vel diferenciar a Tm das esp&eacute;cies por an&aacute;liseestat&iacute;stica (p&lt;0,05). Conclus&otilde;es: Os dados evidenciam a mudan&ccedil;a no perfil epidemiol&oacute;gico com aumento non&uacute;mero de esp&eacute;cies n&atilde;o albicans. Amaioria dos isolados apresentou resist&ecirc;ncia aos antif&uacute;ngicos. Apesar dadiferencia&ccedil;&atilde;o da Tm dos isolados pela PCR em tempo real, a regi&atilde;o amplificadan&atilde;o apresentou varia&ccedil;&atilde;o para distin&ccedil;&atilde;o das esp&eacute;cies

Early Holocene ritual complexity in South America: the archaeological record of Lapa do Santo (east-central Brazil)

Early Archaic human skeletal remains found in a burial context in Lapa do Santo in eastcentral Brazil provide a rare glimpse into the lives of hunter-gatherer communities in South America, including their rituals for dealing with the dead. These included the reduction of the body by means of mutilation, defleshing, tooth removal, exposure to fire and possibly cannibalism, followed by the secondary burial of the remains according to strict rules. In a later period, pits were filled with disarticulated bones of a single individual without signs of body manipulation, demonstrating that the region was inhabited by dynamic groups in constant transformation over a period of centuries

The Oldest Case of Decapitation in the New World (Lapa do Santo, East-Central Brazil)

We present here evidence for an early Holocene case of decapitation in the New World (Burial 26), found in the rock shelter of Lapa do Santo in 2007. Lapa do Santo is an archaeological site located in the Lagoa Santa karst in east-central Brazil with evidence of human occupation dating as far back as 11.7-12.7 cal kyBP (95.4% interval). An ultra-filtered AMS age determination on a fragment of the sphenoid provided an age range of 9.1-9.4 cal kyBP (95.4% interval) for Burial 26. The interment was composed of an articulated cranium, mandible and first six cervical vertebrae. Cut marks with a v-shaped profile were observed in the mandible and sixth cervical vertebra. The right hand was amputated and laid over the left side of the face with distal phalanges pointing to the chin and the left hand was amputated and laid over the right side of the face with distal phalanges pointing to the forehead. Strontium analysis comparing Burial 26's isotopic signature to other specimens from Lapa do Santo suggests this was a local member of the group. Therefore, we suggest a ritualized decapitation instead of trophy-taking, testifying for the sophistication of mortuary rituals among hunter-gatherers in the Americas during the early Archaic period. In the apparent absence of wealth goods or elaborated architecture, Lapa do Santo's inhabitants seemed to use the human body to express their cosmological principles regarding death