Whole-genome sequencing analysis of multidrug-resistant Serratia marcescens isolates in an intensive care unit in Brazil.

INTRODUCTION: Serratia marcescens is an opportunistic pathogen found ubiquitously in the environment and associated with a wide range of nosocomial infections. This multidrug-resistant bacterium has been a cause of concern for hospitals and healthcare facilities due to its ability to spread rapidly and cause outbreaks. Next generation sequencing genotyping of bacterial isolates has proven to be a valuable tool for tracking the spread and transmission of nosocomial infections. This has allowed for the identification of outbreaks and transmission chains, as well as determining whether cases are due to endogenous or exogenous sources. Evidence of nosocomial transmission has been gathered through genotyping methods. The aim of this study was to investigate the genetic diversity of carbapenemase-producing S. marcescens in an outbreak at a public hospital in Cuiaba, MT, Brazil. METHODOLOGY: Ten isolates of S. marcenses were sequenced and antibiotic resistance profiles analyzed over 12 days. RESULTS: The isolates were clonal and multidrug resistant. Gentamycin and tigecycline had sensitivity in 90% and 80% isolates, respectively. Genomic analysis identified several genes that encode ß-lactamases, aminoglycoside-modifying enzymes, efflux pumps, and other virulence factors. CONCLUSIONS: Systematic surveillance is crucial in monitoring the evolution of S. marcescens genotypes, as it can lead to early detection and prevention of outbreaks.

Pan-genomic analysis of Corynebacterium amycolatum gives insights into molecular mechanisms underpinning the transition to a pathogenic phenotype.

Corynebacterium amycolatum is a nonlipophilic coryneform which is increasingly being recognized as a relevant human and animal pathogen showing multidrug resistance to commonly used antibiotics. However, little is known about the molecular mechanisms involved in transition from colonization to the MDR invasive phenotype in clinical isolates. In this study, we performed a comprehensive pan-genomic analysis of C. amycolatum, including 26 isolates from different countries. We obtained the novel genome sequences of 8 of them, which are multidrug resistant clinical isolates from Spain and Tunisia. They were analyzed together with other 18 complete or draft C. amycolatum genomes retrieved from GenBank. The species C. amycolatum presented an open pan-genome (α = 0.854905), with 3,280 gene families, being 1,690 (51.52%) in the core genome, 1,121 related to accessory genes (34.17%), and 469 related to unique genes (14.29%). Although some classic corynebacterial virulence factors are absent in the species C. amycolatum, we did identify genes associated with immune evasion, toxin, and antiphagocytosis among the predicted putative virulence factors. Additionally, we found genomic evidence for extensive acquisition of antimicrobial resistance genes through genomic islands.

Genome Report of Emergent Fish Pathogen Edwardsiella piscicida Recovered from Pseudoplatystoma corruscans in Brazil.

Edwardsiella piscicida is a Gram-negative bacteria belonging to the Hafniaceae family which affects several species of marine and freshwater fish. We present the complete genome of E. piscicida strain BEP80 recovered from the Brazilian catfish named Surubim (Pseudoplatystoma corruscans), consisting a chromosome of 3,883,256 bp and no plasmids.

Characterization of Multidrug-Resistant Staphylococcus aureus Isolates and Comparison of Methods of Susceptibility to Vancomycin.

S. aureus are among the main bacteria causing problems related to multidrug resistance in nosocomial infections. Therefore, it is necessary to carry out a reliable and rapid diagnosis for the identification of the bacteria and characterization of its susceptibility profile, especially vancomycin, which is an alternative treatment against multidrug-resistant (MDR) S. aureus. Thus, the goal of this study was to characterize isolates of S. aureus regarding the resistance and virulence and to check the susceptibility to vancomycin, through different methods, for comparative purposes. Seventeen antimicrobials were tested to assess the susceptibility profile. It was evaluated the presence of identification (nuc), resistance (mecA and blaZ), biofilm (icaA and icaD) and siderophore (sfaD and sbnD) genes. The susceptibility to vancomycin was evaluated by Minimum Inhibitory Concentration (MIC) by broth microdilution (BMD), E-test, commercial panel (Kit), and Phoenix equipment. Most S. aureus (93,33%) was classified as MDR. These isolates were 100% positive for nuc, mecA, icaA, icaD, and sfaD genes; 96.67% for sbnD and 33.33% for blaZ. In relation to BMD, all methods correctly classified the susceptibility of the isolates; however, regarding the exact MIC value for vancomycin, Phoenix showed agreement of 63.33%, E-test (33.33%) and Kit (26.66%). In conclusion, most of S. aureus was considered MDR. Also, they presented resistance, biofilm production, and siderophores genes, showing the pathogenic potential of these bacteria. Besides, the Phoenix test was considered the most effective, as it presents advantages, such as identification of the microorganism and a greater number of antimicrobials tested at a time.

Antibacterial and antibiofilm activities and synergism with florfenicol from the essential oils of Lippia sidoides and Cymbopogon citratus against Aeromonas hydrophila.

AIMS: Aeromonas hydrophila is an opportunistic bacterium, with a high capacity for biofilm production, which can cause severe damage in aquaculture. The objective of this study was to identify the chemical compounds of the essential oils of Lippia sidoides (EOLS) and Cymbopogon citratus (EOCC), and to evaluate the biocidal, antibiofilm and synergistic action with the antimicrobial florfenicol of these essential oils (EOs) against A. hydrophila. METHODS AND RESULTS: The antibacterial activity of EOLS and EOCC was verified by the minimum bactericidal concentration and by the action of these EOs against both forming and consolidated biofilms. The synergistic activity of EOs with florfenicol was performed using the checkerboard technique. The main component of EOLS and EOCC was carvacrol (44.50%) and α-citral (73.56%), respectively. Both EOs showed weak inhibitory activity (≥3125.00 µg ml-1 ). Two bacterial isolates were able to produce biofilm, and EOLS and EOCC acted upon the bacterial isolates to prevent biofilm formation. A bactericidal effect was verified for EOLS in the previously consolidated biofilm for both isolates and for EOCC in only one of the isolates. In general, EOLS had a synergistic effect with florfenicol, while EOCF had an additive effect. CONCLUSIONS: Both EOs were able to interfere with biofilm formation and did not have an antagonistic effect in combination with florfenicol. The best results were found for EOLS, which showed a synergistic effect with florfenicol and the ability to interfere in the formation of consolidated biofilm. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights the potential of EOLS and EOCC to interfere in biofilm and act in synergy with florfenicol to reduce the occurrence of A. hydrophila. Development of these compounds may contribute to the development of herbal medicines in aquaculture.

Phytochemical identification and quantification of quercetin in Triplaris gardneriana wedd. leaves by HPLC-DAD with evaluation of antibacterial activity.

Triplaris gardneriana is used by traditional medicine. The objective of this work was the leaves chemical study with isolate, identify and quantify the chemical constituent, validate the analytical method and evaluate the antibacterial activity. The ethyl acetate and chloroform fractions were subjected to column chromatography for isolation of the compounds quercetin and lupeol, respectively. For the identification, quantification of quercetin in the samples and validation of method were performed using HPLC-DAD. The antibacterial activity was evaluated by the microdilution method. The isolated phytochemicals are being reported for the first time in the species. The ethyl acetate fraction showed a higher content of quercetin with 9.967 ± 1.01 mg.g-1. The method was validated. The samples showed good antibacterial activity. In this study, quercetin was isolated and quantified in the species being a great alternative as a producer of this secondary metabolite, which can be safely applied in the quality control analysis.

Cymbopogon flexuosus essential oil as an additive improves growth, biochemical and physiological responses and survival against Aeromonas hydrophila infection in Nile tilapia.

The objective of the present study was to evaluate growth, biochemical, hematological and intestinal enzymes responses and survival of Nile tilapia juveniles fed a diet containing the essential oil of lemongrass Cymbopogum flexuosus (EOCF) and infected by Aeromonas hydrophila. Five diets were evaluated (in quadruplicate) with increasing levels of EOCF (0.0 - control; 0.25; 0.50; 1.0 and 2.0 mL kg diet-1). On day 45, eight fish per treatment were sampled and blood, liver and intestine samples were taken. Others eight fish per treatment were infected with A. hydrophila followed by a 15-day period of observation. Citral is the main constituent of EOCF. The inclusion of 2.0 mL EOCF kg diet-1 increased specific growth rate and survival after A. hydrophila infection and decreased feed conversion ratio of Nile tilapia. In general, higher concentrations of EOCF in the diet reduced plasma glucose and triglycerides levels, and increased plasma amino acids, alanine aminotransferase (ALT) and hepatic ALT levels, hematological parameters, and the activity of intestinal enzymes. It was concluded that the inclusion of 2.0 mL EOCF kg diet-1 improved growth performance, biochemical and physiological responses and decreased mortality of Nile tilapia after A. hydrophila infection.

Low frequency of antibodies anti-Neospora caninum in rodents in Fernando de Noronha Island, Brazil.

This study aimed to evaluate the frequency of Neospora caninum antibodies in rodents from the Island of Fernando de Noronha, Brazil, and to contribute to epidemiological data on neosporosis in this region. Blood samples were collected from 154 black-rats (Rattus rattus) and 12 rock-cavies (Kerodon rupestris). The search for anti-N. caninum antibodies was made using the NAT technique, with a cut-off of 1:20. The frequency of positive mice was 5.19% (8/154 - 1:20 titer) and all rock-cavies were negative. The low frequency of antibodies against N. caninum in rodents found in the Island shows the low contamination of the environment where these animals live.

Antimicrobial activity of propolis extract fractions against Staphylococcus spp. isolated from goat mastitis / Atividade antimicrobiana de frações do extrato de própolis frente à Staphylococcus spp. isolados de mastite caprina

The indiscriminate use of antibiotics in the treatment of caprine mastitis causes the appearance of resistant microorganisms, besides leaving residues in milk, putting at risk to human health. In this way, propolis is an alternative in the treatment of diseases because it has antimicrobial activity, mainly because of the presence of flavonoids in its composition. The aim of this study was to evaluate the antimicrobial potential of propolis to Staphylococcus spp. Isolated from cases of goat mastitis and qualify the crude ethanoic extract by high performance liquid chromatography (HPLC). In this study, the minimum bactericidal concentration values of propolis extracts in ethanol, ethyl acetate and hexane showed that the best concentrations capable of promoting the highest mortality of the isolates of Staphylococcus spp. from mastitis in goats, were 6250, 3125 and 1562.5µg/mL, respectively. By the microplate adherence test, it was found that 20.78% isolates were not able to form biofilm, 14.70% were classified as moderate and 64.70% were weak and none as a strong biofilm producer. Propolis in its different diluents was able to affect the formation of biofilm and showed a pronounced marked antimicrobial activity against Staphylococcus spp. strains and may be indicated for use in in vivo studies.(AU)

O uso indiscriminado de antibióticos no tratamento de mastite caprina leva ao desenvolvimento de micro-organismos resistentes que poderão estar presentes em alimentos, colocando em risco a saúde humana. Dessa forma, a própolis surge como uma alternativa para o tratamento de doenças por possuir uma ação antimicrobiana, principalmente pela presença de flavonoides em sua composição. O objetivo desse estudo foi avaliar o potencial antimicrobiano da própolis frente à Staphylococcus spp. isolados de casos de mastite caprina e qualificar o extrato etanoico bruto por cromatografia líquida de alta eficiência (CLAE-DAD). Neste estudo, os valores de concentração bactericida mínima (CBM) dos extratos de própolis em álcool etílico, acetato de etila e hexano nos isolados foram de 6250, 3125 e 1562,5µg/mL, respectivamente. Pelo teste de aderência à microplacas, observou-se que 20,78% dos microorganismos, não foram capazes de formar biofilme, 14,70% foram classificados como moderados, 64,70% em fracos e nenhum como forte produtor de biofilme. A própolis em seus diferentes diluentes foi capaz de afetar a formação de biofilme e apresentou significativa atividade antimicrobiana frente a cepas de Staphylococcus spp., podendo ser indicada para utilização em estudos "in vivo".(AU)

Antimicrobial activity of propolis extract fractions against Staphylococcus spp. isolated from goat mastitis / Atividade antimicrobiana de frações do extrato de própolis frente à Staphylococcus spp. isolados de mastite caprina

The indiscriminate use of antibiotics in the treatment of caprine mastitis causes the appearance of resistant microorganisms, besides leaving residues in milk, putting at risk to human health. In this way, propolis is an alternative in the treatment of diseases because it has antimicrobial activity, mainly because of the presence of flavonoids in its composition. The aim of this study was to evaluate the antimicrobial potential of propolis to Staphylococcus spp. Isolated from cases of goat mastitis and qualify the crude ethanoic extract by high performance liquid chromatography (HPLC). In this study, the minimum bactericidal concentration values of propolis extracts in ethanol, ethyl acetate and hexane showed that the best concentrations capable of promoting the highest mortality of the isolates of Staphylococcus spp. from mastitis in goats, were 6250, 3125 and 1562.5µg/mL, respectively. By the microplate adherence test, it was found that 20.78% isolates were not able to form biofilm, 14.70% were classified as moderate and 64.70% were weak and none as a strong biofilm producer. Propolis in its different diluents was able to affect the formation of biofilm and showed a pronounced marked antimicrobial activity against Staphylococcus spp. strains and may be indicated for use in in vivo studies.(AU)

O uso indiscriminado de antibióticos no tratamento de mastite caprina leva ao desenvolvimento de micro-organismos resistentes que poderão estar presentes em alimentos, colocando em risco a saúde humana. Dessa forma, a própolis surge como uma alternativa para o tratamento de doenças por possuir uma ação antimicrobiana, principalmente pela presença de flavonoides em sua composição. O objetivo desse estudo foi avaliar o potencial antimicrobiano da própolis frente à Staphylococcus spp. isolados de casos de mastite caprina e qualificar o extrato etanoico bruto por cromatografia líquida de alta eficiência (CLAE-DAD). Neste estudo, os valores de concentração bactericida mínima (CBM) dos extratos de própolis em álcool etílico, acetato de etila e hexano nos isolados foram de 6250, 3125 e 1562,5µg/mL, respectivamente. Pelo teste de aderência à microplacas, observou-se que 20,78% dos microorganismos, não foram capazes de formar biofilme, 14,70% foram classificados como moderados, 64,70% em fracos e nenhum como forte produtor de biofilme. A própolis em seus diferentes diluentes foi capaz de afetar a formação de biofilme e apresentou significativa atividade antimicrobiana frente a cepas de Staphylococcus spp., podendo ser indicada para utilização em estudos "in vivo".(AU)

Improvement of Baker's yeast-based fuel cell power output by electrodes and proton exchange membrane modification.

The production of more efficient yeast-based fuel cells (YFCs) depends on a combination of effective proton exchange membranes, electron mediators and current collectors. The adhesion of organisms on electrode surface plays a key role in the electron transfer process optimizing the generated power density. In this work, it is reported the preparation of a new YFC prototype using membranes of polyvinyl alcohol/ phosphoric acid and anodes of carbon nanotubes/polyurethane. The high surface area for yeast adhesion and the strong interaction established between cells/carbon nanotubes favor the energy generation in fuel cell. To evaluate the influence of external mediators and the consumption of feed solution (glucose) on performance of YFC, the kinetics of current generation of resulting fuel cells was analyzed. Results reveal that increases in the impedance of electrodes on generated power can be minimized by periodical infusion of feed fuel, preserving 70% of maximum power, representing an important condition for prolonged activity of fuel cell.

Chemical composition and antimicrobial activity of two extract of propolis against isolates of Staphylococcus spp. and multiresistant bacterials / Composição química e atividade antimicrobiana de dois extratos de própolis contra isolados de Staphylococcus spp. e bactérias multirresistentes

There is a growing need to discover and develop alternative therapies for the treatment of mastitis caused by Staphylococcus spp. and multidrug-resistant bacterial infections. This study examined the chemical composition and antimicrobial potential of two propolis extracts (EPA and EPB) against seventy-seven isolates of Staphylococcus spp. obtained from subclinical bovine mastitis; three clinical strains of MRSA and two from clinical strains of S. aureus ATCC, identified as S. aureus ATCC 6538 and S. aureus ATCC 25923. The total phenolic content was determined by the Folin-Ciocalteau method, the total flavonoid content by the Dowd method and the phenolic profile was quantified by HPLC-DAD. The MBC values of the extracts were evaluated by broth microdilution method. The amount of total phenolic and flavonoid compounds was higher in EPA than EPB. Both extracts revealed the presence of caffeic, coumaric, cinnamic, ferulic and 3,4-dihydroxybenzoic acids, with higher concentrations of coumaric and cinnamic acids. Staphylococcus spp. isolates were susceptible to EPA (90.9%), EPB (83.1%) and oxacillin (80.5%). The oxacillin susceptible isolates were also susceptible to EPA (70.1%) and EPB (80.6%), whereas those oxacillin-resistant strains were also susceptible to EPA (40.0%) and to EPB (26.7%). MBC ranged from 34.3 to 68.7µm/mL for EPA and from 68.7 to 137.5µg/mL for EPB. Both extracts inhibited significantly (100%) the clinical strains of MRSA, S. aureus ATCC 6538 and S. aureus ATCC 25923 at the concentration of 68.7µg/mL. It is concluded that both extracts of propolis, whose main constituents are coumaric and cinnamic acids, have high antimicrobial activity against the microorganisms studied, and EPA also against oxacillin-resistant strains. These findings reinforce its potential use for the treatment of bovine mastitis.(AU)

É cada vez mais oportuna a necessidade de descobrir e desenvolver terapias alternativas para tratamento da mastite causada por Staphylococcus spp. e de infecções bacterianas multirresistentes. Este estudo examinou a composição química e o potencial antimicrobiano de dois extratos etanólicos de própolis (EPA e EPB) contra setenta e sete isolados de Staphylococcus spp. obtidos a partir de mastite bovina subclínica; três estirpes clínicas de MRSA e duas de linhagens clínicas de S. aureus ATCC, identificadas como, S. aureus ATCC 6538 e S. aureus ATCC 25923, ambas metacilina resistentes. O teor total de fenólicos foi determinado pelo método de Folin-Ciocalteau, o teor de flavonoides totais pelo método Dowd e o perfil fenólico foi quantificado por HPLC-DAD. CBM dos extratos foi avaliada pelo método de microdiluição em caldo. A quantidade total de compostos fenólicos e flavonoides foi maior no EPA do que no EPB. Ambos os extratos revelaram a presença dos ácidos cafeico, cumárico, cinâmico, ferúlico e 3,4-di-hidroxibenzóico, com maiores concentrações de ácidos cumárico e cinâmico. Os isolados de Staphylococcus spp. foram sensíveis a EPA (90,9%), EPB (83,1%) e oxacilina (80,5%). Os isolados suscetíveis à oxacilina também foram suscetíveis ao EPA (70,1%) e ao EPB (80,6%), enquanto os do resistente à oxacilina foram suscetíveis ao EPA (40,0%) e ao EPB (26,7%). MBC variou de 34,3 a 68,7µm/mL para EPA e de 68,7 a 137,5µg/mL para EPB. Ambos os extratos inibiram significativamente (100%) as linhagens clínicas de MRSA, S. aureus ATCC 6538 e S. aureus ATCC 25923 na concentração de 68,7µg/mL. Conclui-se que os extratos etanólicos da própolis, cujos principais constituintes são os ácidos cumário e cinâmico, possuem atividade antimicrobiana contra os micro-organismos estudados, e o EPA também contra as cepas resistentes à oxacilina. Estes achados reforçam seu potencial uso para o tratamento da mastite bovina.(AU)

Chemical composition and antimicrobial activity of two extract of propolis against isolates of Staphylococcus spp. and multiresistant bacterials / Composição química e atividade antimicrobiana de dois extratos de própolis contra isolados de Staphylococcus spp. e bactérias multirresistentes

There is a growing need to discover and develop alternative therapies for the treatment of mastitis caused by Staphylococcus spp. and multidrug-resistant bacterial infections. This study examined the chemical composition and antimicrobial potential of two propolis extracts (EPA and EPB) against seventy-seven isolates of Staphylococcus spp. obtained from subclinical bovine mastitis; three clinical strains of MRSA and two from clinical strains of S. aureus ATCC, identified as S. aureus ATCC 6538 and S. aureus ATCC 25923. The total phenolic content was determined by the Folin-Ciocalteau method, the total flavonoid content by the Dowd method and the phenolic profile was quantified by HPLC-DAD. The MBC values of the extracts were evaluated by broth microdilution method. The amount of total phenolic and flavonoid compounds was higher in EPA than EPB. Both extracts revealed the presence of caffeic, coumaric, cinnamic, ferulic and 3,4-dihydroxybenzoic acids, with higher concentrations of coumaric and cinnamic acids. Staphylococcus spp. isolates were susceptible to EPA (90.9%), EPB (83.1%) and oxacillin (80.5%). The oxacillin susceptible isolates were also susceptible to EPA (70.1%) and EPB (80.6%), whereas those oxacillin-resistant strains were also susceptible to EPA (40.0%) and to EPB (26.7%). MBC ranged from 34.3 to 68.7µm/mL for EPA and from 68.7 to 137.5µg/mL for EPB. Both extracts inhibited significantly (100%) the clinical strains of MRSA, S. aureus ATCC 6538 and S. aureus ATCC 25923 at the concentration of 68.7µg/mL. It is concluded that both extracts of propolis, whose main constituents are coumaric and cinnamic acids, have high antimicrobial activity against the microorganisms studied, and EPA also against oxacillin-resistant strains. These findings reinforce its potential use for the treatment of bovine mastitis.(AU)

É cada vez mais oportuna a necessidade de descobrir e desenvolver terapias alternativas para tratamento da mastite causada por Staphylococcus spp. e de infecções bacterianas multirresistentes. Este estudo examinou a composição química e o potencial antimicrobiano de dois extratos etanólicos de própolis (EPA e EPB) contra setenta e sete isolados de Staphylococcus spp. obtidos a partir de mastite bovina subclínica; três estirpes clínicas de MRSA e duas de linhagens clínicas de S. aureus ATCC, identificadas como, S. aureus ATCC 6538 e S. aureus ATCC 25923, ambas metacilina resistentes. O teor total de fenólicos foi determinado pelo método de Folin-Ciocalteau, o teor de flavonoides totais pelo método Dowd e o perfil fenólico foi quantificado por HPLC-DAD. CBM dos extratos foi avaliada pelo método de microdiluição em caldo. A quantidade total de compostos fenólicos e flavonoides foi maior no EPA do que no EPB. Ambos os extratos revelaram a presença dos ácidos cafeico, cumárico, cinâmico, ferúlico e 3,4-di-hidroxibenzóico, com maiores concentrações de ácidos cumárico e cinâmico. Os isolados de Staphylococcus spp. foram sensíveis a EPA (90,9%), EPB (83,1%) e oxacilina (80,5%). Os isolados suscetíveis à oxacilina também foram suscetíveis ao EPA (70,1%) e ao EPB (80,6%), enquanto os do resistente à oxacilina foram suscetíveis ao EPA (40,0%) e ao EPB (26,7%). MBC variou de 34,3 a 68,7µm/mL para EPA e de 68,7 a 137,5µg/mL para EPB. Ambos os extratos inibiram significativamente (100%) as linhagens clínicas de MRSA, S. aureus ATCC 6538 e S. aureus ATCC 25923 na concentração de 68,7µg/mL. Conclui-se que os extratos etanólicos da própolis, cujos principais constituintes são os ácidos cumário e cinâmico, possuem atividade antimicrobiana contra os micro-organismos estudados, e o EPA também contra as cepas resistentes à oxacilina. Estes achados reforçam seu potencial uso para o tratamento da mastite bovina.(AU)

Morphology and biometry of the reproductive organs of adult males of Trachemys scripta elegans reared in São Paulo state, Brazil / Morfologia e biometria dos órgãos reprodutivos de machos adultos de Trachemys scripta elegans criados em São Paulo, Brasil

Trachemys scripta elegans is an American underwater chelonian illegally marketed in Brazilian pet shops. When abandoned in nature, it compromises native species, threatening local biodiversity. However, little is known about the body development and structure of its reproductive tract. The objective of the present study was to investigate the morphology and biometry of testis, epididymis and penis, as well as the biometry of the body and secondary sexual characters in this species. Twenty-seven adult males were used aiming to contribute to preservation actions in captivity, population control, and scientific research, as well as to interspecific comparisons. Sex identification by the third claw length was effective, and the specimens presented harmonious and positive body development between mass, carapace, plastron, and height, with unimodal tendency and higher frequency of maximum carapace length at 15cm. The testes and epididymides presented biometric similarity between the antimeres and anatomical and histological structure similar to that of other species of chelonians and mammals, except for the type of epithelium. The findings suggest that there is conserved morphology between slider turtles and homology in relation to mammals. Histological similarity to the reproductive organs of other amniotes, including humans, may give rise to scientific and comparative studies, essential for the establishment of conservation strategies in reptiles.(AU)

Trachemys scripta elegans é um quelônio subaquático americano ilegalmente comercializado em pet shops brasileiros. Ao ser abandonado na natureza, compromete as espécies nativas, ameaçando à biodiversidade local. No entanto, pouco se conhece sobre o desenvolvimento corporal e a estrutura do seu aparelho reprodutor. O objetivo do presente trabalho foi investigar a morfologia e a biometria dos testículos, epidídimos e pênis, a biometria corporal e dos caracteres sexuais secundários. Foram utilizados 27 machos adultos desta espécie, visando contribuir com ações de preservação em cativeiro, controle populacional e pesquisas científicas, além de comparações interespecíficas. A identificação sexual pelo comprimento da terceira garra foi efetiva e os espécimes apresentaram desenvolvimento corporal harmônico e positivo entre massa, carapaça, plastrão e altura, com tendência unimodal e maior frequência de comprimento máximo de carapaça em 15,0cm. Testículos e epidídimos apresentaram semelhança biométrica entre os antímeros e estrutura anatômica e histológica semelhantes à de outras espécies de quelônios e mamíferos, excetuando-se pelo tipo de epitélio. Os achados sugerem haver morfologia conservada entre os cágados e homologia em relação aos mamíferos. A semelhança histológica com os órgãos reprodutivos de outros amniotas, incluindo os humanos, pode dar ensejo a estudos científicos e comparativos, essenciais para estabelecimento de estratégias de conservação em répteis.(AU)

Morphology and biometry of the reproductive organs of adult males of Trachemys scripta elegans reared in São Paulo state, Brazil / Morfologia e biometria dos órgãos reprodutivos de machos adultos de Trachemys scripta elegans criados em São Paulo, Brasil

Trachemys scripta elegans is an American underwater chelonian illegally marketed in Brazilian pet shops. When abandoned in nature, it compromises native species, threatening local biodiversity. However, little is known about the body development and structure of its reproductive tract. The objective of the present study was to investigate the morphology and biometry of testis, epididymis and penis, as well as the biometry of the body and secondary sexual characters in this species. Twenty-seven adult males were used aiming to contribute to preservation actions in captivity, population control, and scientific research, as well as to interspecific comparisons. Sex identification by the third claw length was effective, and the specimens presented harmonious and positive body development between mass, carapace, plastron, and height, with unimodal tendency and higher frequency of maximum carapace length at 15cm. The testes and epididymides presented biometric similarity between the antimeres and anatomical and histological structure similar to that of other species of chelonians and mammals, except for the type of epithelium. The findings suggest that there is conserved morphology between slider turtles and homology in relation to mammals. Histological similarity to the reproductive organs of other amniotes, including humans, may give rise to scientific and comparative studies, essential for the establishment of conservation strategies in reptiles.(AU)

Trachemys scripta elegans é um quelônio subaquático americano ilegalmente comercializado em pet shops brasileiros. Ao ser abandonado na natureza, compromete as espécies nativas, ameaçando à biodiversidade local. No entanto, pouco se conhece sobre o desenvolvimento corporal e a estrutura do seu aparelho reprodutor. O objetivo do presente trabalho foi investigar a morfologia e a biometria dos testículos, epidídimos e pênis, a biometria corporal e dos caracteres sexuais secundários. Foram utilizados 27 machos adultos desta espécie, visando contribuir com ações de preservação em cativeiro, controle populacional e pesquisas científicas, além de comparações interespecíficas. A identificação sexual pelo comprimento da terceira garra foi efetiva e os espécimes apresentaram desenvolvimento corporal harmônico e positivo entre massa, carapaça, plastrão e altura, com tendência unimodal e maior frequência de comprimento máximo de carapaça em 15,0cm. Testículos e epidídimos apresentaram semelhança biométrica entre os antímeros e estrutura anatômica e histológica semelhantes à de outras espécies de quelônios e mamíferos, excetuando-se pelo tipo de epitélio. Os achados sugerem haver morfologia conservada entre os cágados e homologia em relação aos mamíferos. A semelhança histológica com os órgãos reprodutivos de outros amniotas, incluindo os humanos, pode dar ensejo a estudos científicos e comparativos, essenciais para estabelecimento de estratégias de conservação em répteis.(AU)

Low intensity electric field inactivation of Gram-positive and Gram-negative bacteria via metal-free polymeric composite.

The adhesion of pathogenic bacteria in medical implants and surfaces is a health-related problem that requires strong inhibition against bacterial growth and attachment. In this work, we have explored the enhancement in the antibacterial activity of metal free-based composites under external electric field. It affects the oxidation degree of polypyrrole-based electrodes and consequently the antibacterial activity of the material. A conductive layer of carbon nanotubes (graphite) was deposited on porous substrate of polyurethane (sandpaper) and covered by polypyrrole, providing highly conductive electrodes characterized by intrinsic antibacterial activity and reinforced by electro-enhanced effect due to the external electric field. The bacterial inhibition of composites was monitored from counting of viable cells at different voltage/time of treatment and determination of biofilm inhibition on electrodes and reactors. The external voltage on electrodes reduces the threshold time for complete bacterial inactivation of PPy-based composites to values in order of 30 min for Staphylococcus aureus and 60 min for Escherichia coli.

Inclusion of discarded banana in sugarcane silage decreases dry matter losses and improves its nutritional value / La inclusión de banano de descarte en el ensilaje de caña de azúcar reduce las pérdidas de materia seca y mejora su valor nutricional / A inclusão de banana de descarte em silagem de cana de açúcar decresce as perdas de matéria seca e melhora o seu valor nutricional

Abstract Background: Sugarcane silage (SCS) has often been characterized by large dry matter (DM) losses associated with reduction in soluble nutrients and increase in indigestible fiber. Objective: To evaluate the effect of including discarded fruits on the losses and nutritional value in SCS. Methods: A completely randomized design was used with five treatments and five replications. Silages were added with 15% (as-fed basis) papaya (Carica papaya) (SP), acerola (Malpighia glabra) (SA), banana (Musa sp.) (SB) or tomato (Solanum lycopersicum) (ST). The control treatment was composed only of SCS. Twenty-five (25-L) plastic buckets were used to make the experimental silos, which were opened on the 90th day after their closure. Data were analyzed by analysis of variance. Results: Gas losses and total DM losses were lower and DM recovery was higher in SB and SA than in the control (p<0.05). The SB had lower content of neutral detergent fiber and acid detergent fiber compared to SP, SA, ST and the control group (p<0.05). There was no effect of treatment for ethanol content (p>0.05). However, there was treatment effect for lactic and acetic acids, ammonia nitrogen contents, lactic acid bacteria, and yeasts and mold counts (p<0.05). Conclusion: The inclusion of 15% banana or acerola in SCS decreases losses and increases DM recovery. Furthermore, the inclusion of 15% banana improves the nutritional value of SCS.

Resumen Antecedentes: El ensilaje de caña de azúcar (SCS) es frecuentemente caracterizado por grandes pérdidas de materia seca (DM) asociadas con reducciones en los nutrientes solubles y aumentos en la fibra indigestible. Objetivo: Evaluar el efecto de la inclusión de frutas de descarte sobre las perdidas y el valor nutricional del SCS. Métodos: Se utilizó un diseño completamente aleatorizado con cinco tratamientos y cinco repeticiones. Fueron adicionados a los ensilajes, 15 % (materia natural) de papaya (Carica papaya) (SP), acerola (Malpighia glabra) (SA), banano (Musa sp.) (SB) o tomate (Solanum lycopersicum) (ST). El control estuvo compuesto apenas por SCS. Veinticinco baldes de 25 L fueron usados para hacer los silos, los cuales se abrieron al día 90 posterior a su cierre. Los datos fueron analizados por análisis de varianza. Resultados: Las pérdidas de gases y de DM total fueron menores y la recuperación de DM fue mayor en SB y SA que en el grupo control (p<0,05). Los contenidos de fibra detergente neutra y ácida fueron menores en SB comparados con SP, SA, ST y el grupo control (p<0,05). No hubo efecto para el contenido de etanol (p>0,05). Sin embargo, hubo efecto de tratamiento para los contenidos de ácidos láctico y acético, y nitrógeno amoniacal y recuento de bacterias ácido lácticas, levaduras y hongos (p<0,05). Conclusiones: La inclusión de 15% de banano y acerola en SCS decreció las perdidas, y aumentó la recuperación de DM. Además, la inclusión de un 15% de banano mejoró el valor nutricional de SCS.

Resumo Antecedentes: A silagem de cana de açúcar (SCS) é frequentemente caracterizada por grandes perdas de matéria seca (DM) associadas com reduções nos nutrientes solúveis e aumento na fibra indigestível. Objetivo: Avaliar o efeito da inclusão de frutas de descarte sobre as perdas e o valor nutricional de SCS. Métodos: Um delineamento completamente casualizado foi utilizado, com cindo tratamentos e cinco repetições. As silagens foram adicionadas com 15% (matéria natural) de mamão (Carica papaya) (SP), acerola (Malpighia glabra) (SA), banana (Musa sp.) (SB) ou tomate (Solanum lycopersicum) (ST). O controle foi composto apenas por SCS. Vinte e cinco baldes de 25 L foram usados para fazer os silos, que foram abertos no 90º dia após o fechamento deles. Os dados foram analisados por meio de análise de variância. Resultados: As perdas por gases e de DM total foram menores e a recuperação de DM foi maior em SB e SA que no controle (p<0,05). Os conteúdos de fibra em detergente neutro e ácido foram menores em SB comparado a SP, SA, ST e o grupo de controle (p<0,05). Não houve efeito para conteúdo de etanol (p>0,05). Contudo, houve efeito de tratamento para os conteúdos de ácidos lático e acético, e nitrogênio amoniacal e contagens de bactérias láticas, e leveduras e fungos (p<0,05). Conclusões: A inclusão de 15% de banana e acerola em SCS decresceu as perdas, e aumentou a recuperação de DM. Adicionalmente, a inclusão de 15% de banana melhorou o valor nutricional da SCS.

Antimicrobial activity of propolis extract fractions against Staphylococcus spp. isolated from goat mastitis

ABSTRACT: The indiscriminate use of antibiotics in the treatment of caprine mastitis causes the appearance of resistant microorganisms, besides leaving residues in milk, putting at risk to human health. In this way, propolis is an alternative in the treatment of diseases because it has antimicrobial activity, mainly because of the presence of flavonoids in its composition. The aim of this study was to evaluate the antimicrobial potential of propolis to Staphylococcus spp. Isolated from cases of goat mastitis and qualify the crude ethanoic extract by high performance liquid chromatography (HPLC). In this study, the minimum bactericidal concentration values of propolis extracts in ethanol, ethyl acetate and hexane showed that the best concentrations capable of promoting the highest mortality of the isolates of Staphylococcus spp. from mastitis in goats, were 6250, 3125 and 1562.5g/mL, respectively. By the microplate adherence test, it was found that 20.78% isolates were not able to form biofilm, 14.70% were classified as moderate and 64.70% were weak and none as a strong biofilm producer. Propolis in its different diluents was able to affect the formation of biofilm and showed a pronounced marked antimicrobial activity against Staphylococcus spp. strains and may be indicated for use in in vivo studies.

RESUMO: O uso indiscriminado de antibióticos no tratamento de mastite caprina leva ao desenvolvimento de micro-organismos resistentes que poderão estar presentes em alimentos, colocando em risco a saúde humana. Dessa forma, a própolis surge como uma alternativa para o tratamento de doenças por possuir uma ação antimicrobiana, principalmente pela presença de flavonoides em sua composição. O objetivo desse estudo foi avaliar o potencial antimicrobiano da própolis frente à Staphylococcus spp. isolados de casos de mastite caprina e qualificar o extrato etanoico bruto por cromatografia líquida de alta eficiência (CLAE-DAD). Neste estudo, os valores de concentração bactericida mínima (CBM) dos extratos de própolis em álcool etílico, acetato de etila e hexano nos isolados foram de 6250, 3125 e 1562,5g/mL, respectivamente. Pelo teste de aderência à microplacas, observou-se que 20,78% dos microorganismos, não foram capazes de formar biofilme, 14,70% foram classificados como moderados, 64,70% em fracos e nenhum como forte produtor de biofilme. A própolis em seus diferentes diluentes foi capaz de afetar a formação de biofilme e apresentou significativa atividade antimicrobiana frente a cepas de Staphylococcus spp., podendo ser indicada para utilização em estudos in vivo.

Chemical composition and antimicrobial activity of two extract of propolis against isolates of Staphylococcus spp. and multiresistant bacterials

ABSTRACT: There is a growing need to discover and develop alternative therapies for the treatment of mastitis caused by Staphylococcus spp. and multidrug-resistant bacterial infections. This study examined the chemical composition and antimicrobial potential of two propolis extracts (EPA and EPB) against seventy-seven isolates of Staphylococcus spp. obtained from subclinical bovine mastitis; three clinical strains of MRSA and two from clinical strains of S. aureus ATCC, identified as S. aureus ATCC 6538 and S. aureus ATCC 25923. The total phenolic content was determined by the Folin-Ciocalteau method, the total flavonoid content by the Dowd method and the phenolic profile was quantified by HPLC-DAD. The MBC values of the extracts were evaluated by broth microdilution method. The amount of total phenolic and flavonoid compounds was higher in EPA than EPB. Both extracts revealed the presence of caffeic, coumaric, cinnamic, ferulic and 3,4-dihydroxybenzoic acids, with higher concentrations of coumaric and cinnamic acids. Staphylococcus spp. isolates were susceptible to EPA (90.9%), EPB (83.1%) and oxacillin (80.5%). The oxacillin susceptible isolates were also susceptible to EPA (70.1%) and EPB (80.6%), whereas those oxacillin-resistant strains were also susceptible to EPA (40.0%) and to EPB (26.7%). MBC ranged from 34.3 to 68.7m/mL for EPA and from 68.7 to 137.5g/mL for EPB. Both extracts inhibited significantly (100%) the clinical strains of MRSA, S. aureus ATCC 6538 and S. aureus ATCC 25923 at the concentration of 68.7g/mL. It is concluded that both extracts of propolis, whose main constituents are coumaric and cinnamic acids, have high antimicrobial activity against the microorganisms studied, and EPA also against oxacillin-resistant strains. These findings reinforce its potential use for the treatment of bovine mastitis.

RESUMO: É cada vez mais oportuna a necessidade de descobrir e desenvolver terapias alternativas para tratamento da mastite causada por Staphylococcus spp. e de infecções bacterianas multirresistentes. Este estudo examinou a composição química e o potencial antimicrobiano de dois extratos etanólicos de própolis (EPA e EPB) contra setenta e sete isolados de Staphylococcus spp. obtidos a partir de mastite bovina subclínica; três estirpes clínicas de MRSA e duas de linhagens clínicas de S. aureus ATCC, identificadas como, S. aureus ATCC 6538 e S. aureus ATCC 25923, ambas metacilina resistentes. O teor total de fenólicos foi determinado pelo método de Folin-Ciocalteau, o teor de flavonoides totais pelo método Dowd e o perfil fenólico foi quantificado por HPLC-DAD. CBM dos extratos foi avaliada pelo método de microdiluição em caldo. A quantidade total de compostos fenólicos e flavonoides foi maior no EPA do que no EPB. Ambos os extratos revelaram a presença dos ácidos cafeico, cumárico, cinâmico, ferúlico e 3,4-di-hidroxibenzóico, com maiores concentrações de ácidos cumárico e cinâmico. Os isolados de Staphylococcus spp. foram sensíveis a EPA (90,9%), EPB (83,1%) e oxacilina (80,5%). Os isolados suscetíveis à oxacilina também foram suscetíveis ao EPA (70,1%) e ao EPB (80,6%), enquanto os do resistente à oxacilina foram suscetíveis ao EPA (40,0%) e ao EPB (26,7%). MBC variou de 34,3 a 68,7m/mL para EPA e de 68,7 a 137,5g/mL para EPB. Ambos os extratos inibiram significativamente (100%) as linhagens clínicas de MRSA, S. aureus ATCC 6538 e S. aureus ATCC 25923 na concentração de 68,7g/mL. Conclui-se que os extratos etanólicos da própolis, cujos principais constituintes são os ácidos cumário e cinâmico, possuem atividade antimicrobiana contra os micro-organismos estudados, e o EPA também contra as cepas resistentes à oxacilina. Estes achados reforçam seu potencial uso para o tratamento da mastite bovina.

Morphology and biometry of the reproductive organs of adult males of Trachemys scripta elegans reared in São Paulo state, Brazil

ABSTRACT: Trachemys scripta elegans is an American underwater chelonian illegally marketed in Brazilian pet shops. When abandoned in nature, it compromises native species, threatening local biodiversity. However, little is known about the body development and structure of its reproductive tract. The objective of the present study was to investigate the morphology and biometry of testis, epididymis and penis, as well as the biometry of the body and secondary sexual characters in this species. Twenty-seven adult males were used aiming to contribute to preservation actions in captivity, population control, and scientific research, as well as to interspecific comparisons. Sex identification by the third claw length was effective, and the specimens presented harmonious and positive body development between mass, carapace, plastron, and height, with unimodal tendency and higher frequency of maximum carapace length at 15cm. The testes and epididymides presented biometric similarity between the antimeres and anatomical and histological structure similar to that of other species of chelonians and mammals, except for the type of epithelium. The findings suggest that there is conserved morphology between slider turtles and homology in relation to mammals. Histological similarity to the reproductive organs of other amniotes, including humans, may give rise to scientific and comparative studies, essential for the establishment of conservation strategies in reptiles.

RESUMO: Trachemys scripta elegans é um quelônio subaquático americano ilegalmente comercializado em pet shops brasileiros. Ao ser abandonado na natureza, compromete as espécies nativas, ameaçando à biodiversidade local. No entanto, pouco se conhece sobre o desenvolvimento corporal e a estrutura do seu aparelho reprodutor. O objetivo do presente trabalho foi investigar a morfologia e a biometria dos testículos, epidídimos e pênis, a biometria corporal e dos caracteres sexuais secundários. Foram utilizados 27 machos adultos desta espécie, visando contribuir com ações de preservação em cativeiro, controle populacional e pesquisas científicas, além de comparações interespecíficas. A identificação sexual pelo comprimento da terceira garra foi efetiva e os espécimes apresentaram desenvolvimento corporal harmônico e positivo entre massa, carapaça, plastrão e altura, com tendência unimodal e maior frequência de comprimento máximo de carapaça em 15,0cm. Testículos e epidídimos apresentaram semelhança biométrica entre os antímeros e estrutura anatômica e histológica semelhantes à de outras espécies de quelônios e mamíferos, excetuando-se pelo tipo de epitélio. Os achados sugerem haver morfologia conservada entre os cágados e homologia em relação aos mamíferos. A semelhança histológica com os órgãos reprodutivos de outros amniotas, incluindo os humanos, pode dar ensejo a estudos científicos e comparativos, essenciais para estabelecimento de estratégias de conservação em répteis.