Oral microbiota of Brazilian captive snakes

The present work aimed to determine the oral microbiotic composition of snakes from Sao Jose do Rio Preto city, São Paulo State, Brazil. Ten snake species, comprising the families Boidae, Colubridae, Elapidae and Viperidae, were submitted to microbiological examination of their oral cavity, which indicated positivity for all buccal samples. Gram-negative bacilli, gram-negative cocci bacilli, gram-positive bacilli and gram-positive cocci were isolated from the snakes. Among isolated bacterium species, the occurrence of coagulase-negative staphylococci in the buccal cavity of Crotalus durissus (Viperiade), Eunectes murinus (Boidae), Mastigodryas bifossatus (Colubridae) and Bacillus subtilis, common to oral cavity of Bothrops alternatus (Viperidae) and Phalotris mertensi (Colubridae), was detected. It was observed higher diversity of isolated bacteria from the oral cavity of Micrurus frontalis (Elapidae) and Philodryas nattereri (Colubridae), as well as the prevalence of gram-positive baccillus and gram-positive cocci. The composition of the oral microbiota of the studied snakes, with or without inoculating fangs, is diverse and also related to the formation of abscesses at the bite site in the victims of the ophidian accidents, and to pathogenic processes in the snakes that host these microorganisms.

Antibiotic 26-deoxylaidlomycin isolated from Streptomyces sp. Ar386 from Brazilian soil

An actinomycete strain (Ar386) was isolated from the soil of the Araraquara regio, SP, Brazil. The strain, named Streptomyces jacareensis, formed irregular rayed, rugose, grayish-white mycelium with sinuous, branched hyphae carrying rare isolated spores; assimilated glucose, galactose, inositol, ribose, maltose, sucrose, melibiose and starch but not mannitol, rhamnose, arabinose, xylose, lactose and raffinose; and contained LL- diaminopimelic acid in its cell wall. An antibiotic active against Gram- positive bacteria, which was characterized as being 26-deoxylaidlomycin and which may have application against poultry coccidiosis, was isolated from cultures of the strain. This was the first isolation of this antibiotic from a microorganism of the genus Streptomyces and also the first isolation of this antibiotic in Brazil.

In vitro evaluation of the antimicrobial activity of a castor oil-based irrigant

The antimicrobial activity of irrigating solutions - Endoquil (castor oil detergent), 2% chlorhexidine gluconate solution, and 0.5% NaOCI solution - was evaluated against Gram-positive cocci (Micrococcus luteus, Staphylococcus aureus, Enterococcus faecalis, Staphylococcus epidermidis, Streptococcus mutans, and Streptococcus sobrinus), Gramnegative rods (Escherichia coli and Pseudomonas aeruginosa), and the yeast Candida albicans. Activity was evaluated using the two-layer agar diffusion technique. The base layer was obtained by pouring 10.0 ml of Muller Hinton Medium or 10.0 ml of Brain Heart Infusion agar in a Petri dish. After solidification a 5.0 ml seed layer of Muller Hinton Medium or Brain Heart Infusion agar with inoculum (106/ml) was added. Absorbent paper disks (6.0 mm in diameter) immersed in the solutions were placed at equidistant points. Plates were maintained at room temperature for 2 h for prediffusion of the solutions and incubated at 37°C for 24 h. The candle jar system was used for the Brain Heart Infusion agar plates. All tests were performed in duplicate. After incubation the medium was optimized with 0.05 g% triphenyltetrazolium chlorate gel and inhibition halos were measured. All bacterial strains were inhibited by 2.0% chlorhexidine gluconate. Endoquil was effective against Grampositive microorganisms, and 0.5% NaOCI was effective only against S. aureus. Copyright © 2001 by The American Association of Endodontists.

Comparação entre o sistema Autoścan-4 e métodos convencionais na identificação de bacilos Gram-negativos não fermentadores, isolados de materiais clínicos de pacientes em Presidente Prudente - SP

Ninety eight strains of glucose-nonfermenting Gram-negative bacilli were analyzed and isolated from several clinical materials of 95 patients admitted at the Dr. Domingos Leonardo Cerávolo University Hospital and three from outpatients. All of them were assisted in the Laboratory of Clinical Analysis of Unoeste University, Presidente Prudente, SP, from the period of October of 1999 to April of 2001. In this work, the level of agreement between the semi-automated commercial system AutoScan-4 and the conventional system for the identification of those bacteria were studied comparatively. There was agreement in 81 (82.7%), showing that both methodologies are useful for identification; partial agreement in six strains (6.1%) and disagreement in 11 (11.2%). The comercial system did not identify nine (9.2%) of the strains and reported them as very rare biotypes.

Padrão de sensibilidade aos antimicrobianos de bacilos gram-negativos não fermentadores isolados de materiais clínicos de pacientes em Presidente Prudente - SP

The aim of this study was the assessment of isolation frequency and antimicrobial susceptibility pattern of nonfermenting Gram-negative bacilli. Ninety eight strains of nonfermenting Gram-negative bacilli, isolated from several clinical materials of patients admited at the Dr. Domingos Leonardo Cerávolo University Hospital and at Dr. Odilo Antunes Siqueira State Hospital, as well as from every outpatient; assisted at Laboratory of Clinical Analysis of Unoeste University, Presidente Prudente, São Paulo, in the period of October 1999 to April 2001 were analyzed. The most frequent species were Pseudomonas aeruginosa (65.3%) and Acinetobacter baumannii (23.5%). The frequency of the other isolated species was smaller than 2.5%. In the antimicrobial susceptibility tests, the two species more prevalent showed high resistance. The antibiotic most active in vitro was the imipenem, with 79.6% in microdiluition method, and 76.6% in diffusion method, for Pseudomonas aeruginosa strains and 100.0% in both microdiluition and diffusion methods, for Acinetobacter baumannii. The cephalosporins of third generation, the ciprofloxacin and the aminoglycosides, presented percentage of susceptibility varying from 22.4 to 69.7%. These results bring implications to the emergency use of the antimicrobial agents in the treatment of patients with severe infection.

Immune Response to the Rhodococcus equi infection in high and low antibody-producing mice (selection IV-A)

Rhodococcus equi is a Gram-positive, facultative intracellular bacterium which infects macrophages and causes rhodococcal pneumonia and enteritis in foals. Recently, this agent has been recognized as an opportunistic pathogen for immunocompromised humans. Several murine experimental models have been used to study R. equi infection. High (H IV-A) and Low (L IV-A) antibody (Ab)-producers mice were obtained by bi-directional genetic selections for their ability to produce antibodies against sheep and human erythrocytes (Selection IV-A). These lines maintain their phenotypes of high and low responders also for other antigens than those of selection (multispeciflc effect). A higher macrophage activity in L IV-A mice has been described for several intracellular infectious agents, which could be responsible for their intense macrophage antigens (Ag)-handling and low Ab production. Due to these differences, L IV-A mice were found to exhibit a better performance to trigger an effective immune response towards intracellular pathogens. The objective of this work was to characterize the immune response of Selection IV-A against R. equi. H IV-A and L IV-A mice were infected with 2.0 × 10 6 CFU of ATCC 33701 +R. equi by intravenous route. With regards to bacterial clearance and survival assays, L IV-A mice were more resistant than H IV-A mice to virulent R. equi. L IV-A mice presented a higher hydrogen peroxide (H 2O 2) and nitric oxide (NO) endogenous production by splenic macrophages than H IV-A mice. L IV-A expressed the most intense cellular response, available by the Delayed-Type Hypersensitivity (DTH) reaction, which activated macrophages and produced more H 2O 2 and NO. The three times higher specific antibodies titres in H IV-A indicated that Selection IV-A maintained the multispecific effect and the polygenic control of humoral and cellular responses also to R. equi.

Qualitative analysis of preputial and vaginal bacterial microbiota in owl monkeys (Aotus azarai infulatus) raised in captivity

Background: The aim of this study was to identify the aerobic bacteria of the preputial and vaginal microbiota in owl monkeys that have been raised in captivity and to evaluate the antimicrobial susceptibility of these bacteria by gender and social organization. Methods: Thirty clinically healthy Aotus azarai infulatus were used. A total of 134 samples were collected, 60 from the preputial mucosa and 74 from the vaginal mucosa. An automated system of bacterial identification was used. Results and Conclusions: Staphylococcus intermedius and Proteus mirabilis were the microorganisms that were most frequently identified according to gender and social organization. The antimicrobial susceptibility of the isolated gram-positive bacteria was similar in both sexes. However, the gram-negative strains had some differences. The aerobic bacterial population of the vaginal and preputial microbiota is similar in owl monkeys, and there are no differences in the number and bacterial species according to sex and social organization. © 2012 John Wiley & Sons A/S. Published by Blackwell Publishing Ltd.

Determinação do potencial antimicrobiano, antioxidante e da motibilidade intestinal de extrato de Paepalanthus geniculatus

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Purificação e caracterização parcial de proteínas presentes no veneno de Bothrops leucurus

Pós-graduação em Microbiologia - IBILCE

Riboflavin analgs from Streptomyces davawensisas antiinfectives: mode of action, mechanism of resistance of the producer and metabolization by humans

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estudo de genômica comparativa de corynebacterium pseudotuberculosis linhagem 226 (biovar ovis)

Corynebacterium pseudotuberculosis é uma bactéria Gram-positiva, intracelular facultativa, nãoesporulante, não-capsulada e sem mobilidade, contudo possui fímbria, e pode assumir formas cocóides e filamentosas (pleomórfica), além disto, apresenta crescimento ótimo à 37°C. Este patógeno apresenta dois biovares: ovis que geralmente acomete pequenos ruminantes, e causa a doença linfadenite caseosa, e biovar equi, mais comum em equinos, bovinos, camelídeos, e bubalinos causando a Linfangite ulcerativa. A infecção por esta bactéria pode levar a condenação das carcaças e redução de lã (em ovinos e caprinos), leite e carne destes animais, e consequentemente a perdas econômicas para a indústria agropecuária mundial. Atualmente, ainda não existe uma vacina eficaz para estas doenças. A fim de obter um maior entendimento biológico entre as espécies o presente trabalho tem como objetivo principal analisar, por meio da genômica comparativa a linhagem C. pseudotuberculosis 226 biotipo ovis isolada de um caprino na Califórnia com outras linhagens do biovarar ovis e equi. Na análise de sintenia entre as linhagens foi possível identificar que a linhagem 226 apresenta alta conservação da ordem gênica entre as linhagens do biótipo ovis. Através de análises filogenômicas foi possível identificar que as linhagens I19 e 267 apresentaram maior e menor proximidade filogenética com a linhagem 226. A linhagem 1/06-A foi a que apresentou maior proximidade filogenômica entre as linhagens do biovar equi, quando comparadas a linhagem 226. Foram preditas 8 ilhas de patogenicidade, estando presente na ilha 1 os genes relacionados a virulência de C. pseudotuberculosis mais bem descritos na literatura. Não houveram regiões novas relacionadas a genes de virulência entre nenhuma das linhagens. Foram identificados 248 genes ortólogos entre as linhagens I19, 267 e 226 e 282 genes ortólogos entre as linhagens 258,1/06-A e 226. Com base nesse estudo é possível inferir que as linhagens do biovar ovis possuem um repertório gênico pouco variado e que as linhagens do biovar equi apresentam uma quantidade menor de genes compartilhados com a linhagem 226, corroborando com a diversidade gênica entre os biovares.

Atividade in vivo de parasporina extraída de B. thuringiensis sobre o câncer colorretal murino (CT‐26)

Bacillus thuringiensis (Bt), is an environmental Gram-positive spore-forming bacterium that produces crystalline parasporal protein (Cry) during sporulation. The inclusions often exhibit strong and specific insecticidal activity, making Bt an agent for agricultural controlling insects pest, mites, protozoa and nematodes. Recent studies reported that some of these Crys do not show cytotoxicity against insects but they are capable to kill some human and animal cancer cells. These proteins were denominated parasporins (PS). However, antitumor activity of Bt parasporin on the development of murine colorectal cancer (CT-26), are not well studies and these are no reports on the in vivo effect of these proteins. Thus, the present study evaluated the in vitro and in vivo anti-tumoral activity of Bt parasporin against the murine colorectal cancer line CT-26. Therefore, Balb/c mice were s.c. inoculated with CT-26 cells and weekly treated with parasporin (i.p.) pre-activated by enzymatic digestion with trypsin or proteinase K. Our results have shown, for the first time, that despite the anti-tumor activity in vitro, parasporin crystals couldn’t combat tumor growth in vivo. Instead, this protein was highly toxic, affecting the liver and spleen, with possible effect on other organs, decreasing the survival of treated animals. The results indicate the need for studies to better detoxification or manipulation of parasporin for therapeutic use and new studies for analysis of toxicological effects of repetitive exposure of farmers to this toxin

Green tea extract and its major constituent epigallocatechin-3-gallate inhibit growth and halitosis-related properties of Solobacterium moorei

Background: Solobacterium moorei is a volatile sulfide compound (VSC)-producing Gram-positive anaerobic bacterium that has been associated with halitosis. The aim of this study was to investigate the effects of green tea extract and its major constituent epigallocatechin-3-gallate (EGCG) on growth and severalhalitosis-related properties of S. moorei.Methods: A microplate dilution assay was used to determine the antibacterial activity of green tea extract and EGCG against S. moorei. Their effects on bacterial cell membrane integrity were investigated by transmission electron microscopy and a fluorescence-based permeability assay. Biofilm formation was quantified by crystal violet staining. Adhesion of FITC-labeled S. moorei to oral epithelial cells was monitored by fluorometry. The modulation of beta-galactosidase gene expression in S. moorei was evaluated by quantitative RT-PCR.Results: The green tea extract as well as EGCG inhibited the growth of S. moorei, with MIC values of 500 and 250 mu g/ml, respectively. Transmission electron microscopy analysis and a permeabilization assay brought evidence that the bacterial cell membrane was the target of green tea polyphenols. Regarding the effects of green tea polyphenols on the S. moorei colonization properties, it was found that biofilm formation on EGCG-treated surfaces was significantly affected, and that green tea extract and EGCG can cause the eradication of pre-formed S. moorei biofilms. Moreover, both the green tea extract and EGCG were found to reduce the adherence of S. moorei to oral epithelial cells. The beta-galactosidase activity of S. moorei, which plays a key role in VSC production, was dose-dependently inhibited by green tea polyphenols. In addition, EGCG at 1/2 MIC significantly decreased the beta-galactosidase gene expression.Conclusion: Our study brought evidence to support that green tea polyphenols possess a number of properties that may contribute to reduce S. moorei-related halitosis. Therefore, these natural compounds may be of interest to be used to supplement oral healthcare products.

Influence of seasonality and production method on the antibacterial activity of propolis

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