Molecular characterization and biofilm-formation analysis of Listeria monocytogenes, Salmonella spp., and Escherichia coli isolated from Brazilian swine slaughterhouses.

This study aimed to verify the presence of Listeria monocytogenes, Salmonella spp., and Escherichia coli in two Brazilian swine slaughterhouses, as well as to perform antibiograms, detect virulence and antimicrobial resistance genes, and evaluate the in vitro biofilm-forming capability of bacterial isolates from these environments. One Salmonella Typhi isolate and 21 E. coli isolates were detected, while L. monocytogenes was not detected. S. Typhi was isolated from the carcass cooling chamber's floor, resistant to several antimicrobials, including nalidixic acid, cefazolin, chloramphenicol, doxycycline, streptomycin, gentamicin, tetracycline, and sulfonamide, and contained resistance genes, such as tet(B), tet(C), tet(M), and ampC. It also showed moderate biofilm-forming capacity at 37°C after incubating for 72 h. The prevalence of the 21 E. coli isolates was also the highest on the carcass cooling chamber floor (three of the four samplings [75%]). The E. coli isolates were resistant to 12 of the 13 tested antimicrobials, and none showed sensitivity to chloramphenicol, an antimicrobial prohibited in animal feed since 2003 in Brazil. The resistance genes MCR-1, MCR-3, sul1, ampC, clmA, cat1, tet(A), tet(B), and blaSHV, as well as the virulence genes stx-1, hlyA, eae, tir α, tir ß, tir γ, and saa were detected in the E. coli isolates. Moreover, 5 (23.8%) and 15 (71.4%) E. coli isolates presented strong and moderate biofilm-forming capacity, respectively. In general, the biofilm-forming capacity increased after incubating for 72 h at 10°C. The biofilm-forming capacity was the lowest after incubating for 24 h at 37°C. Due to the presence of resistance and virulence genes, multi-antimicrobial resistance, and biofilm-forming capacity, the results of this study suggest a risk to the public health as these pathogens are associated with foodborne diseases, which emphasizes the hazard of resistance gene propagation in the environment.

WHO Critical Priority Escherichia coli as One Health Challenge for a Post-Pandemic Scenario: Genomic Surveillance and Analysis of Current Trends in Brazil.

The dissemination of carbapenem-resistant and third generation cephalosporin-resistant pathogens is a critical issue that is no longer restricted to hospital settings. The rapid spread of critical priority pathogens in Brazil is notably worrying, considering its continental dimension, the diversity of international trade, livestock production, and human travel. We conducted a nationwide genomic investigation under a One Health perspective that included Escherichia coli strains isolated from humans and nonhuman sources, over 45 years (1974-2019). One hundred sixty-seven genomes were analyzed extracting clinically relevant information (i.e., resistome, virulome, mobilome, sequence types [STs], and phylogenomic). The endemic status of extended-spectrum ß-lactamase (ESBL)-positive strains carrying a wide diversity of blaCTX-M variants, and the growing number of colistin-resistant isolates carrying mcr-type genes was associated with the successful expansion of international ST10, ST38, ST115, ST131, ST354, ST410, ST648, ST517, and ST711 clones; phylogenetically related and shared between human and nonhuman hosts, and polluted aquatic environments. Otherwise, carbapenem-resistant ST48, ST90, ST155, ST167, ST224, ST349, ST457, ST648, ST707, ST744, ST774, and ST2509 clones from human host harbored blaKPC-2 and blaNDM-1 genes. A broad resistome to other clinically relevant antibiotics, hazardous heavy metals, disinfectants, and pesticides was further predicted. Wide virulome associated with invasion/adherence, exotoxin and siderophore production was related to phylogroup B2. The convergence of wide resistome and virulome has contributed to the persistence and rapid spread of international high-risk clones of critical priority E. coli at the human-animal-environmental interface, which must be considered a One Health challenge for a post-pandemic scenario. IMPORTANCE A One Health approach for antimicrobial resistance must integrate whole-genome sequencing surveillance data of critical priority pathogens from human, animal and environmental sources to track hot spots and routes of transmission and developing effective prevention and control strategies. As part of the Grand Challenges Explorations: New Approaches to Characterize the Global Burden of Antimicrobial Resistance Program, we present genomic data of WHO critical priority carbapenemase-resistant, ESBL-producing, and/or colistin-resistant Escherichia coli strains isolated from humans and nonhuman sources in Brazil, a country with continental proportions and high levels of antimicrobial resistance. The present study provided evidence of epidemiological and clinical interest, highlighting that the convergence of wide virulome and resistome has contributed to the persistence and rapid spread of international high-risk clones of E. coli at the human-animal-environmental interface, which must be considered a One Health threat that requires coordinated actions to reduce its incidence in humans and nonhuman hosts.

Molecular characterization of Salmonella spp. and Listeria monocytogenes strains from biofilms in cattle and poultry slaughterhouses located in the federal District and State of Goiás, Brazil.

Listeria monocytogenes and Salmonella spp. are considered important foodborne pathogens that are commonly associated with foods of animal origin. The aim of this study was to perform molecular characterization of L. monocytogenes and Salmonella spp. isolated from biofilms of cattle and poultry slaughterhouses located in the Federal District and State of Goiás, Brazil. Fourteen L. monocytogenes isolates and one Salmonella sp. were detected in poultry slaughterhouses. No isolates were detected in cattle slaughterhouses. All L. monocytogenes isolates belonged to lineage II, and 11 different pulsotypes were detected. Pulsed-field gel electrophoresis analysis revealed the dissemination of two strains within one plant, in addition to the regional dissemination of one of them. The Salmonella isolate was identified via whole genome sequencing as Salmonella enterica serovar Minnesota ST548. In the sequence analysis, no premature stop codons were detected in the inlA gene of Listeria. All isolates demonstrated the ability to adhere to Caco-2 cells, while 50% were capable of invading them. Antimicrobial resistance was detected in 57.1% of the L. monocytogenes isolates, and resistance to sulfonamide was the most common feature. The tetC, ermB, and tetM genes were detected, and four isolates were classified as multidrug-resistant. Salmonella sp. was resistant to nine antimicrobials and was classified as multidrug-resistant. Resistance genes qnrB19, blaCMY-2, aac(6')-Iaa, sul2, and tetA, and a mutation in the parC gene were detected. The majority (78.5%) of the L. monocytogenes isolates were capable of forming biofilms after incubation at 37°C for 24 h, and 64.3% were capable of forming biofilms after incubation at 12°C for 168 h. There was no statistical difference in the biofilm-forming capacity under the different evaluated conditions. Salmonella sp. was capable of forming biofilms at both tested temperatures. Biofilm characterization was confirmed by collecting the samples consistently, at the same sampling points, and by assessing biofilm formation in vitro. These results highlight the potential risk of cross-contamination in poultry slaughterhouses and the importance of surveillance and pathogen control maintenance programs within the meat production industry.

Assessment of Internalin A Gene Sequences and Cell Adhesion and Invasion Capacity of Listeria monocytogenes Strains Isolated from Foods of Animal and Related Origins.

Listeria monocytogenes is a foodborne pathogen of global relevance that causes outbreaks and sporadic cases of listeriosis, acquired through the consumption of contaminated products, including milk or meat products and ready-to-eat meat products subjected to intensive handling. The objective of the present study was to classify L. monocytogenes isolated from various food-related sources in the Federal District of Brazil and surrounding areas to sequence internalin A (inlA) genes from these isolates and assess their adhesion and invasion capacity using Caco-2 cells. In addition, 15 were classified as group I, 3 as group II, and 7 classified as group IV. Premature stop codons (PMSCs) at the nucleotide position 976 (GAA→TAA) of the inlA gene were identified in 5 of the 25 isolates. Adhesion and invasion tests in Caco-2 cells showed that all the isolates were capable of adhesion and cellular invasion, with isolates containing PMSCs exhibiting on average higher invasion capacity than those without PMSCs (p = 0.041) and a median of adhesion very distinctive from those without stop codons. These results are the first report of PMSCs in the inlA gene of L. monocytogenes from the Federal District of Brazil and Brazil.

Conservation of the internal quality of eggs using a biodegradable coating.

This study aimed to evaluate the effect of a pectin biofilm on the preservation of refrigerated and unrefrigerated eggs during 5 wk of storage based on egg weight loss, albumen height, Haugh unit (HU), and the yolk index (YI). A total of 1,200 nonfertile eggs from GLK Bankiva laying hens (40 wk of age), which were freshly laid and came from a single collection, were obtained from a model poultry rearing system (Planaltina, Federal District, Brazil) that meets all animal welfare criteria. The experimental outline was entirely randomized, with 20 treatments in a factorial scheme of 2 × 2 × 5, with 2 biofilm treatments (with and without) × 2 storage temperatures (refrigeration: 5°C and ambient: 25°C) × 5 storage periods (7, 14, 21, 28, and 35 d), with 12 repetitions per treatment. Starting from the third storage week, increased weight loss (%) was observed in noncoated eggs (4.46 ± 1.06; 5.61 ± 1.37; 6.93 ± 1.66%) compared with biofilm-coated eggs (3.57 ± 1.26; 4.74 ± 1.8; 6.05 ± 2.21%), respectively. The HU variation in the pectin-coated eggs (86.84-78.02) was smaller than that in the noncoated eggs (83.01-64.36) between the beginning (7 d) and the end (35 d) of the experimental period. Eggs with and without biofilm stored in the refrigerator presented average HU values of 91.26 ± 6.27 and 88.35 ± 6.96, respectively. In contrast, when kept at room temperature, eggs with the coating presented higher HU values (71.27 ± 10.78) than eggs without the coating (59.11 ± 15.97). Coated eggs (0.37 ± 0.16) showed higher YI values than noncoated eggs (0.35 ± 0.16). A pectin-based biofilm effectively maintained egg quality during the 35 d of storage.

Effect of transportation distances, seasons and crate microclimate on broiler chicken production losses.

The goal of this research was to evaluate the microclimate (temperature, relative humidity and ECI-enthalpy comfort index) of commercial loads of broiler chickens at different transport distances: Dist15 (15 km on average) and Dist90 (90 km on average) in the summer and winter seasons and their effects on the production parameters body weight difference (BWD), mortality (%) and bruising prevalence (%). Twelve broiler loads were monitored using dataloggers to record temperature and humidity, with a total of 24 target crates per load. The experiment followed a factorial design [2 seasons (rainy and dry) × 2 distances (Dist15 and Dist90)] with a randomized complete block arrangement, 3 sexes (all males, all females, or mixed shipments) and one shipment per combination. BWD had a heterogeneous distribution throughout the load, and this distribution was not significantly correlated with the mean ECI measured during transport at 12 positions along the load. In terms of comfort, summer is the most critical period for broiler transport. In the interaction between rainy season and Dist90, the highest ECI was scored in the lethal zone (where physiological mechanisms are not enough to control body temperature). Mortality during the rainy season was not significantly different between distances. However, during the dry season, mortality was twice as high as broilers that travelled for 15 km. The prevalence of bruising on carcasses was not affected by the interaction between season and distance. As we know, broiler chicken performance, during transport, can be also related to road conditions, being hard to evaluate the real impact of seasons and distances on animal welfare. Load microclimate can compromise broiler chicken welfare during transport and it does not necessary reflect significant losses pre and post-slaughter.

Isolamento de Listeria monocytogenes em salsichas comercializadas no Distrito Federal / Isolation of Listeria monocytogenesin sausages commercialized in the Distrito Federal

A bactéria Listeria monocytogenes é o patógeno causador da listeriose, importante doença transmitida por alimentos que acomete humanos e animais podendo causar morte. Foram coletadas 100 amostras de salsicha de mercados do Distrito Federal. A metodologia usada para o isolamento foi a descrita na Instrução Normativa Nº 40 do Ministério da Agricultura, Pecuária e Abastecimento. As amostras que apresentaram motilidade característica de Listeria sp p. foram submetidas a técnica de PCR para confirmação da espécie L. monocytogenes. Os resultados positivos para Listeria sp p. foram vistos em 14 amostras e em 5 confirmou-se a presença de L. monocytogenes. Através dos resultados desse experimento conclui-se que salsichas podem estar contaminadas com bactérias do gênero Listeria sp p. e L. monocytogenes, oferecendo risco de listeriose aos consumidores.

Pesquisa de Clostridium perfringens em carnes bovinas embaladas a vácuo comercializadas no Distrito Federal e entorno / Clostridium perfringens research on vacuum packed beef commercialized on Distrito Federal and in the surrounding area

O objetivo deste trabalho foi detectar a presença de Clostridium perfringens em 54 amostras de carne bovina embaladas a vácuo comercializadas na região do Distrito Federal, bem como detectar a presença da toxina cpe por PCR, ainda avaliar os meios de cultivo ágar SPS® e ágar TSC®, com e sem etapa de pré-enriquecimento das amostras em caldo infusão de cérebro e coração (BHI) na câmara de anaerobiose, e posterior incubação das placas de SPS® e TSC® tanto em jarra de anaerobiose, como em câmara de anaerobiose. Na análise da incubação das placas em ágar SPS® e TSC®, sem a etapa de pré-enriquecimento em caldo BHI na câmara de anaerobiose, observou-se o crescimento em apenas uma (1,85%) das 54 amostras analisadas, em ambos os meios de cultivo e formas de incubação. Com a etapa de pré-enriquecimento com caldo BHI em câmara de anaerobiose, observou-se crescimento em todas as 54 amostras (100%), em ambos os meios de cultivo e formas de incubação. Na reação em cadeia de polimerase (PCR) nenhuma das cepas oriundas das amostras analisadas apresentaram a amplificação de fragmento do gene da toxina cpe. Os resultados evidenciam a presença de C. perfringens em carnes embaladas a vácuo comercializadas no Distrito Federal e Entorno, porém não foi detectada a toxina cpe em nenhuma cepa isolada analisada. Na comparação estatística aplicando o teste qui-quadrado de McNemar, observou-se que houve diferença significativa (p<0,001) entre as análises sem e com a etapa de pré-enriquecimento em caldo BHI, verificando-se a influencia positiva do meio na recuperação de esporos, destacando desta forma a importância do enriquecimento prévio em meio BHI e a incubação em câmara de anaerobiose, na recuperação de esporos deste microrganismo.

The aim of this work was to detect Clostridium perfringens in 54 samples of vacuum packed beef sold at the Federal District area, and to detect the presence of the cpe toxin gene by Polymerase chain reaction (PCR), also evaluate the culture medium SPS® agar and TSC® agar, with and without pre-enrichment step of the samples in Brain Heart Infusion (BHI) broth in the anaerobic chamber, and to promote the incubation of the plates in anaerobic jar and anaerobic chamber. The results of the incubation on SPS® agar and TSC® agar, without the pre-enrichment step in BHI, growth was observed in only one (1,85%) of the 54 analyzed samples, in both culture media and incubation methods. With the pre-enrichment step with BHI broth in the anaerobic chamber, growth was observed in all 54 samples (100%), in both culture media and incubation methods. At the PCR, In the polymerase chain reaction (PCR), none of the strains from the analyzed samples showed fragment amplification of the toxin cpe gene. The results showed the presence of C. perfringens in vacuum packed beef samples commercialized at the Distrito Federal area and surroundings, however the cpe toxin was not detected in any isolated strain analyzed. In the statistical test using the McNemar chi-square test, a significant difference (p<0,001) was observed between the analysis with and without pre-enrichment step in BHI broth, verifying the positive influence of the medium in spore recovery, therefore to enhance the importance of the pre-enrichment stage in BHI broth and the incubation in anaerobic chamber in spore recovery for this microorganism.

Avaliação da qualidade higienicossanitária de carcaças de bovinos oriundos de abatedouros frigoríficos do Distrito Federal e entorno / Evaluation of the hygiene and sanitary quality of carcasses of cattle from slaughterhouses of the Federal District and reversal

O trabalho realizado teve como objetivo avaliar a qualidade higienicossanitária de carcaças bovinas, oriundas de abatedouros-frigoríficos sob Inspeção Distrital (Dipova) e Federal (SIF), localizados na região do Distrito Federal e Entorno. Foram realizadas análises microbiológicas para Contagem de micro-organismos Mesófilos aeróbios estritos e facultativos, identificação do Número mais Provável de Coliformes a 45°C e Contagem de Staphylococcus aureus em 05 (cinco) diferentes pontos de 07 (sete) meias carcaças, nas quais as coletas foram realizadas antes e depois da lavagem com água clorada, perfazendo assim um total de 70 análises. Os valores médios dos resultados observados, antes da lavagem com água clorada, para Contagem de micro-organismos Mesófilos aeróbios estritos e facultativos, foram de 9,7 UFC/cm²; para Número Mais provável de Coliformes a 45ºC, de 29 germes/cm² e para Staphylococcus coagulase positivo, de 1 UFC/cm². Os valores observados nas meias carcaças, após a lavagem com água clorada na Contagem de micro- -organismos Mesófilos aeróbios estritos e facultativos foram de 1,5 UFC/ cm²; para a análise de Número Mais Provável de Coliformes a 45ºC, foi de 8,2 germes/cm² e não houve crescimento de Staphylococcus coagulase positiva. Neste trabalho verificou-se a presença de Coliformes a 45ºC e a Contagem de micro-organismos Mesófilos aeróbios estritos e facultativos antes e após a lavagem das meias carcaças e a presença de Staphylococcus coagulase positiva apenas antes das lavagem das mesmas. A lavagem das meias carcaças com água clorada diminuiu o grau de contaminação.(AU)

PERDA ELETROLÍTICA DE CÁLCIO, MAGNÉSIO E FERRO NO SUOR DURANTE CORRIDA EM ESTEIRA / ELECTROLYTIC LOSS OF CALCIUM, MAGNESIUM AND IRON IN THE SWEAT DURING RUNNING IN A TREADMILL / PÉRDIDA ELECTROLÍTICA DE CALCIO, MAGNESIO Y HIERRO EN EL SUDOR DURANTE CARRERA EN CINTA RODANTE

RESUMO Introdução: O suor e sua consequente evaporação são fundamentais para manutenção da temperatura corporal durante o exercício. Objetivo: Avaliar a perda de cálcio (Ca++), magnésio (Mg++) e ferro (Fe++) no suor de corredores e de indivíduos ativos. Métodos: Foram avaliados 15 atletas corredores de fundo {VO2máx = 68 ± 5,4 ml(kg.min)-1} e 15 indivíduos ativos não atletas {VO2máx = 50,3 ± 6,3 ml(kg.min)-1}, com média de idade, respectivamente, de 25,3 ± 2,4 e 23,1 ± 4,3 anos. Ambos os grupos se exercitaram por 80 minutos em esteira, com intensidade de 75% a 85% da frequência cardíaca de reserva, e ingeriram 3 ml de água/kg de peso corporal a cada 15 minutos. As condições ambientais da prova foram 21,9 ± 1,5 °C e 89,2 ± 5,6% de umidade relativa para os atletas e 21,8 ± 1,6 °C e 93,2 ± 3,5% de UR para os ativos. As amostras de suor foram coletadas em intervalos regulares de 20 minutos nas regiões do peito, torácica e lombar das costas, para posterior análise dos minerais Ca++, Mg++ e Fe++ por espectrofotômetro de absorção atômica. Resultados: Não foram registradas diferenças significativas para os minerais em função do nível de condicionamento. Observou-se tendência à diminuição na concentração do Mg++ e Fe++ do suor ao longo do exercício. Conclusão: Nas condições ambientais e de exercício estudadas, o condicionamento não interfere na perda de Ca++, Mg++ e Fe++.

ABSTRACT Introduction: Sweat and its consequent evaporation are essential for maintaining body temperature during the exercise. Objective: To evaluate the loss of calcium (Ca++), magnesium (Mg++) and iron (Fe++) in the sweat of runners and active individuals. Methods: Fifteen long-distance-runners {VO2máx = 68 ± 5.4 ml (kg.min)-1} and 15 non- athletes active subjects {VO 2máx = 50.3 ± 6.3 ml (kg.min)-1} with mean ages of 25.3 ± 2.4 and 23.1 ± 4.3 years, respectively. Both groups exercised for 80 minutes on a treadmill, with intensity of 75-85% of reserve heart rate, and ingested 3 ml of water/kg of body weight every 15 minutes. The environmental conditions of the test were 21.9 ± 1.5 °C and 89.2 ± 5.6% relative humidity for athletes and 21.8 ± 1.6 °C and 93.2 ± 3.5% RH for active individuals. The sweat samples were collected at regular intervals of 20 minutes in the chest, thoracic and lower back regions for further analysis of the minerals Ca++, Mg++ and Fe++ by atomic absorption spectrophotometer. Results: No significant differences were recorded for the minerals in relation to fitness level. There was a tendency for a decrease in Mg++ and Fe++ concentrations of sweat over the course of the exercise. Conclusion: In the environmental and exercise conditions studied, the conditioning does not interfere in the loss of Ca++, Mg++ and Fe++.

RESUMEN Introducción: El sudor y su consecuente evaporación son esenciales para mantener la temperatura corporal durante el ejercicio. Objetivo: Evaluar la pérdida de calcio (Ca++), magnesio (Mg++) y hierro (Fe++) en el sudor en corredores y personas activas. Métodos: Se evaluaron 15 corredores de larga distancia {VO2máx = 68 ± 5,4 ml (kg.min)-1} y 15 individuos activos no atletas {VO 2 máx = 50,3 ± 6,3 ml (kg.min)-1}, con promedio de edades de, respectivamente, 25,3 ± 2,4 e 23,1 ± 4,3 años. Ambos grupos hicieron ejercicio durante 80 minutos en una cinta rodante con una intensidad del 75% al 85% de la frecuencia cardíaca de reserva, siendo hidratados con 3 ml de agua/Kg de peso corporal cada 15 minutos. Las condiciones ambientales de la prueba fueron 21,9 ± 1,5 °C y 89,2 ± 5,6% de humedad relativa para los atletas y 21,8 ± 1,6 °C y 93,2 ± 3,5% de HR para los sujetos activos. Las muestras de sudor se recogieron a intervalos regulares de 20 minutos en las regiones del pecho, torácica y lumbar de la espalda para el análisis del Ca++, Mg++y Fe++ por espectrofotometría de absorción atómica. Resultados: No se observaron diferencias significativas para los minerales en función del nivel de acondicionamiento. Hubo una tendencia a la disminución de la concentración de Mg++ y Fe++ en el sudor durante el ejercicio. Conclusión: En el ejercicio estudiado y las condiciones ambientales, el acondicionamiento no interfiere en la pérdida de Ca++, Mg++ y Fe++.

Caracterização molecular de Listeria monocytogenes oriundas de cortes cárneos bovinos e de abatedouros frigoríficos de bovinos localizados no Distrito Federal, Brasil / Molecular characterization of Listeria monocytogenes from beef samples and cattle slaughterhouses located in the Federal District, Brazil

Este trabalho teve como objetivo realizar a detecção de cepas de Listeria monocytogenes de cortes cárneos bovinos bem como no ambiente de abatedouros frigoríficos localizados no Distrito Federal, promover a sorotipificação pela reação em cadeia da polimerase (PCR), realizar antibiograma e submeter às cepas à eletroforese de campo pulsado (Pulsed-field gel electrophoresis - PFGE). Foram analisados um total de 125 cortes cárneos bovinos, 45 amostras de swabs de carcaças e 43 amostras de swabs em que foram detectados 13 cepas de Listeria monocytogenes, sendo 11 em cortes cárneos bovinos e 2 swabs de ambiente em um abatedouro frigorifico. Não foram isoladas cepas de swabs de carcaça. Dentre as 13 cepas de Listeria monocytogenes foram encontradas seis cepas do sorotipo 4b, cinco do sorotipo 1/2c e duas cepas do sorotipo 1/2a. Dentre as 11 cepas de L. monocytogenes encontradas em cortes cárneos bovino, uma (9,1%) cepa apresentou resistência a eritromicina, outra (9,1%) cepa a gentamicina e outra a ciprofloxacina (9,1%) e todas as cepas (100%) apresentaram resistência ao Ác. Nalidíxico. Das duas (2) cepas oriundas de ralos de abatedouro frigorífico, todas (100%) apresentaram resistência ao Ác. Nalidíxico e a sulfonamidas. A análise por eletroforese de campo pulsante (PFGE) demonstrou 13 diferentes pulsotipos, em que foram agrupados em 3 diferentes grupos clonais, que coincidentemente se correlacionavam com os 3 diferentes sorotipos encontrados sugerindo uma ampla disseminação desses perfis no Distrito Federal.(AU)

The aim of the study was the analysis of Listeria monocytogenes strains in beef samples as well as slaughterhouse environment, located in the Federal District, promote serotyping by polymerase chain reaction (PCR), perform antibiotic susceptibility and submit the strains to Pulsed-field gel electrophoresis (PFGE). A total of 125 beef samples were analyzed, 45 samples of carcasses swabs and 43 swab samples. It detected 13 strains of Listeria monocytogenes, 11 in beef samples. and 2 in slaughterhouse environment. No carcass swabs strains were isolated. Among the 13 strains of L. monocytogenes six strains of serotype 4b were found, five serotype 1/2c and two strains of serotype 1/2a. Among the 11 strains of L. monocytogenes found in beef, one (9.1%) strain showed resistance to erythromycin, one (9.1%) strain to gentamicin, one to ciprofloxacin (9.1%) and all strains (100%) were resistant to nalidixic acid. The two strains coming from the slaughterhouse drains, all (100%) were resistant to nalidixic acid and Sulfonamides. The analysis by pulsed field gel electrophoresis (PFGE) showed 13 different pulsotypes; they were grouped into three different clonal groups, coincidentally correlated with the three different serotypes found, what suggests a widespread dissemination of these profiles in the Federal District, Brazil.(AU)

Hydration practices of runners during training vs competition / Prácticas de hidratación en corredores durante el entrenamiento vs la competición

Background: Appropriate hydration ensures that the athlete is in ideal condition during exercise. Inadequate hydration practices are still present among athletes participating in different types of sports. This could be the result of a low level of knowledge concerning the importance of hydration for athletic performance. The purpose of this study was to evaluate the level of knowledge and the hydration practices of the runners in training and competition. Method: This study used an exploratory method involving a questionnaire containing 19 objective questions related to hydration habits and level of knowledge about hydration, as reported in prior studies, which included questions categorized into three parts: demographic characteristics, hydration habits during competition and training, and hydration knowledge. A total of 412 athletes (36.1 ± 12.9 years old) with a high experience in the sport (11.2 ± 10.9 years) were evaluated. Results: A significant difference (p = 0.0012) was observed for reported hydration practices between training and competitions. Moreover, the data revealed that a large number of runners (41 % in competitions and 54.4% in training) had inadequate hydration habits because they did not always consume fluids. Increased consumption of fluids in the competitions was also observed. Only 35.4% of athletes consumed sports drinks, while 3.88 % had never consumed fluids during training, and 1.5% never consumed fluids in competition. Few athletes (23.2%) weighed themselves before and after exercising. Participants answered 66.6% of hydration knowledge questions correctly. Conclusion: Runners were not able to transfer their knowledge about proper hydration during both training and competition practices

Introducción: Una hidratación adecuada asegura que el atleta está en condición ideal durante el ejercicio. Se suelen presentar prácticas de hidratación inadecuadas entre los deportistas de diferentes disciplinas que podrían deberse a un bajo nivel de conocimiento sobre la importancia de una correcta hidratación para el rendimiento deportivo. El objetivo del estudio fue evaluar el nivel de conocimiento y las prácticas de hidratación en corredores durante el entrenamiento frente a la competición. Métodos: Este estudio utilizó un método exploratorio que incluía un cuestionario con 19 preguntas objetivas relacionadas con los hábitos de hidratación y el nivel de conocimiento sobre hidratación, según lo informado en estudios previos, cuyas preguntas estaban divididas en tres partes: características demográficas; hábitos de hidratación durante el entrenamiento y competición, y conocimientos sobre hidratación. Se evaluaron 412 corredores (36,1 ± 12,9 años) con una elevada experiencia en la práctica del atletismo (11,2 ± 10,9 años). Resultados: Hubo diferencia significativa (p = 0,0012) de las prácticas de hidratación durante los entrenamientos y la competición, con mayor consumo de líquidos durante las competiciones. Por otra parte, los datos revelaron que un gran número de corredores (41,0% en competición y 54,4 % en entrenamiento) tenían hábitos de hidratación inadecuados al no consumir siempre líquidos. Sólo el 35,4 % de los atletas declararon consumir bebidas deportivas. La ingesta de líquidos era inexistente en el 3,8% de los atletas durante el entrenamiento y el 1,5% en competiciones. Sólo el 23,2% de los encuestados controlaban el peso corporal antes y después del ejercicio. Los participantes respondieron correctamente al 66,6% de las preguntas sobre conocimiento de hidratación. Conclusiones: Los corredores participantes no fueron capaces de trasladar sus conocimientos sobre el tema de hidratación en prácticas adecuadas tanto en entrenamiento como en competición

Avaliação da qualidade microbiológica de queijos do tipo Minas padrão de produção industrial, artesanal e informal / Microbiological evaluation of Minas type cheeses from industrial, artisanal and informal manufacturing

Os objetivos deste estudo foram de avaliar a qualidade microbiológica de queijos do tipo Minas padrão,produzidos de forma industrial com inspeção federal, artesanal com inspeções estadual e municipal (a partir de leite não pasteurizado) e informal (produção caseira), bem como de analisar os hábitos de consumo desse tipo de queijo no Distrito Federal, Brasil. As amostras (n = 21) foram submetidas a análises para a pesquisa de coliformes a 30 °C e 45 °C, Staphylococcus coagulase positiva, Listeria monocytogenes e Salmonella spp. Os resultados foram avaliados de acordo com a RDC 12/2001 da ANVISA; 57,14% das amostras de queijos industriais, 100% das informais e 100% das artesanais estavam em desacordo quanto às contagens de Staphylococcus coagulase positiva. As contagens de coliformes a 45°C estavam em desacordo em 71,42% das amostras de queijos informais e 14,28% das industrializadas e artesanais. Nenhuma amostra foi positiva para L. monocytogenes ou Salmonella spp. Um questionário simplificado sobre o consumo de queijo Minas foi aplicado a 50 pessoas no momento da compra e houve indicação de 47% de preferência ao sabor de queijos informais. Em virtude destes queijos não serem inspecionados e não seguirem padrões de produção, representam um risco à saúde pública...

Bactérias nativas indutoras de precipitação de minerais de carbonato de cálcio em solos tropicais / Native bacteria which induce calcium carbonate mineral precipitations in tropical soils / Bacterias nativas inductoras de precipitación de minerales de carbonato de calcio en suelos tropicales

As bactérias, consideradas como o grupo de microrganismos de maior abundância e diversidade que habitam os solos, são vitais no processo de bioprecipitação, no qual, estes microrganismos formam precipitados minerais cristalinos ou amorfos, como resultado de suas atividades metabólicas sob certas condições ambientais. Nesta pesquisa foram aplicados os conhecimentos sobre bactérias, para dar solução a alguns problemas da engenharia, estudando a capacidade das bactérias nativas existentes num solo tropical de precipitar minerais de carbonato de cálcio, a partir da adição do meio nutriente. Este nutriente se mostrou ótimo para induzir a bioprecipitação gerando fibras e cristais de carbonato de cálcio, que mudam as propriedades do solo, causando uma maior estabilidade estrutural dos grãos, um melhor desempenho dos agregados e uma menor deformabilidade da massa de solo.

Bacteria, considered as the group of soil microorganisms with the greatest abundance and diversity, are vital for bioprecipitation processes, where these microorganisms form crystalline or amorphous mineral precipitates as result of their metabolic activity under certain environmental conditions. In this investigation, knowledge about bacteria was applied for solving some engineering problems, studying the capacity of native bacteria found in tropical soil for precipitating calcium carbonate after the addition of a nutrient medium. This medium was optimal for inducing bioprecipitation, generating calcium carbonate fibers and crystals which modify soil characteristics, producing a greater structural stability of the grains, better performance of the aggregates, and less deformation of the ground mass.

Las bacterias, consideradas como el grupo de microrganismos de mayor abundancia y diversidad que habitan los suelos, son vitales en el proceso de bioprecipitación, en el cual estos microrganismos forman precipitados minerales cristalinos o amorfos, como resultado de sus actividades metabólicas sobre ciertas condiciones ambientales. En esta investigación, se aplicaron los conocimientos sobre bacterias para dar solución a algunos problemas de ingeniería, estudiando la capacidad de las bacterias nativas existentes en un suelo tropical de precipitar minerales de carbonato de calcio, a partir de la adición de un medio nutriente. Este nutriente resultó óptimo para inducir la bioprecipitación, generando fibras y cristales de carbonato de calcio, que varían las propiedades del suelo causando una mayor estabilidad estructural de los granos, un mejor desempeño de los agregados y una menor deformación en la masa de suelo.

Ocorrência e diferenciação de espécies de Listeria spp. em salsichas tipo hot dog a granel e em amostras de carne moída bovina comercializadas no Distrito Federal / Occurrence and differentiation of Listeria spp. in “hot dog” sausages sold in bulk and ground beef samples marketed in the Federal District, Brazil

Listeria monocytogenes é um patógeno relevante veiculado por alimentos. Sua identificação precisa é importante para a correta determinação do risco associado à ingestão do alimento. O objetivo deste trabalho foi verificar a ocorrência das espécies de Listeria spp. em amostras de salsichas tipo hot dog e carne moída bovina, comercializadas a granel no Distrito Federal. Foram analisadas 162 amostras, sendo 127 de salsichas tipo hot dog e 35 amostras de carne moída bovina. O isolamento e a identificação do gênero foram feitos por metodologia convencional e a distinção das espécies foi verificada por kit bioquímico específico (API-Listeria®) e por análise de restrição de fragmentos da reação em cadeia da polimerase (RFLP-PCR) do gene 23 rRNA. Foram isoladas 26 cepas de Listeria spp. das amostras de salsichas tipo hot dog, sendo identificadas 18 cepas de Listeria innocua e 08 cepas de Listeria monocytogenes. Das 35 amostras de carne moída bovina, foram isoladas 16 cepas de Listeria spp., sendo identificadas 12 cepas de Listeria innocua e 04 de Listeria monocytogenes. Houve concordância total na distinção das espécies de Listeria spp. através dos dois métodos empregados. A presença de Listeria spp. em amostras de salsicha do tipo hot dog a granel e em carne moída bovina a granel, de estabelecimentos comerciais do Distrito Federal, representa risco à saúde do consumidor.

Listeria monocytogenes is a pathogen disclosed by foods. Its accurate identification is important for the correct determination of the risk associated with the ingestion of food. The aim of this research was to determine the occurrence of Listeria spp. in samples of hot dog sausages and ground beef, sold in bulk in the Federal District. A total of 162 samples, 127 hot dog sausages and 35 samples of ground beef cattle, were analyzed. The isolation and identification of the genus were made by conventional methodology and distinction of species was verified by specific biochemical kit (API-Listeria®) and by restriction fragment length polymorphism (PCR-RFLP) of 23S rRNA gene. Twenty-six strains of Listeria spp. were isolated. Samples of hot dog sausages identified 18 strains of Listeria innocua and 08 strains of Listeria monocytogenes. Of the 35 samples of ground beef cattle were isolated, 16 strains of Listeria spp., 12 strains of Listeria innocua and 04 strains of Listeria monocytogenes. There was total agreement in the distinction of Listeria species using the two methods. The presence of Listeria spp. in samples of hot dog sausages and ground beef, sold in bulk in commercial establishment of the Federal District represents a risk to consumer health.

Resistência antimicrobiana em Enterococcus faecalis e Enterococcus faecium isolados de carcaças de frango / Antimicrobial Resistance of Enterococcus faecalis and Enterococcus faecium isolated from poultry carcasses

O objetivo deste trabalho foi realizar o isolamento e analisar o perfil de resistência antimicrobiana de Enterococcus de carcaças de frango resfriadas e congeladas comercializadas no Distrito Federal, detectando genes de resistência antimicrobiana e identificando as espécies Enterococcus faecalis e Enterococcus faecium por reação polimerase em cadeia. Foram analisadas 100 carcaças de frangos, das quais foram isoladas 50 cepas de Enterococcus spp., sendo 42% de E. faecalis e 2% de E. faecium. O teste de susceptibilidade antimicrobiana demonstrou que todas as cepas isoladas apresentaram resistência a pelo menos um antimicrobiano, dos quais 90,47% das cepas de E. faecalis, 100% das cepas de E. Faecium e 82,14% dos Enterococcus spp. apresentaram resistência à Tetraciclina; 80,95% das cepas de E. faecalis e 35,71% das cepas de Enterococcus spp. foram resistentes à Eritromicina; 39,28% dos Enterococcus spp. e 23,80% dos E. faecalis à Ciprofloxacina e 28,57% dos E. faecalis apresentaram resistência ao Cloranfenicol. Foram detectados os genes de resistência antimicrobiana erm(B), vanC-1, aph(3')-llla, ant(6)-la, vanB, vanA, aac(6')-le-aph(2'')-la, erm(A) e tet(M) - este último mais frequente. Estes resultados sugerem sérios problemas para a Saúde Pública, uma vez que esses microrganismos podem possuir a capacidade de transmitir genes de resistência antimicrobiana para outros microrganismos presentes na microbiota intestinal de humanos e animais, podendo inviabilizar o uso destas drogas para tratamentos clínicos.

The aim of this study was to isolate and analyze the antimicrobial profile resistance of Enterococcus from cooled and frozen poultry carcasses commercialized at the Federal District area, detecting the antimicrobial resistance genes and identifying Enterococcus faecalis and Enterococcus faecium by polymerase chain reaction. One hundred poultry carcasses were analyzed and 50 strains of Enterococcus spp. were isolated, of which 42% were E. faecalis and 2% E. faecium. The antimicrobial susceptibility test showed that all isolates were resistant to at least one antibiotic; 90,47% of E. faecalis, 100% of E. faecium and 82,14% of Enterococcus spp. were resistant to Tetracycline; 80,95% of E. faecalis and 35,71% of Enterococcus spp. strains were resistant to Erythromycin; 39,28% of Enterococcus spp. and 23,80% of E. faecalis to Ciprofloxacin, and 28,57% of E. faecalis were resistant to Chloramphenicol. There were detected erm(B), vanC-1, aph(3')-llla, ant(6)-la, vanB, vanA, aac(6')-le-aph(2'')-la, erm(A) and tet(M) resistance genes, the last one most often verified. The results might suggest problems for public health due the high resistance, since these microorganisms have ability to transmit genes for antimicrobial resistance to others in the intestinal tract of humans and animals. Thus, the use of these drugs for clinical treatment could be hindered.

Antimicrobial resistance of Campylobacter jejuni isolated from chicken carcasses in the Federal District, Brazil.

The aim of the present study was to perform microbiological isolation of Campylobacter jejuni from chilled chicken carcasses marketed in the Federal District of Brazil and to subject the strains to an antibiogram. A total of 92 samples from chilled chicken carcasses were acquired, 18 of which (19.56%) tested positive for C. jejuni. A total of 16 strains were tested for susceptibility to eight antimicrobial drugs. All 16 strains were resistant to ciprofloxacin, 15 strains to nalidixic acid, streptomycin, tetracycline, and gentamycin, 14 strains to amoxicillin, 11 strains to erythromycin, and 6 strains to chloramphenicol. The present study is the first to report on the presence of C. jejuni in chilled chicken carcasses marketed in the Federal District region of Brazil. These results may indicate flaws in certain steps of this food processing and highlight a possible public health problem due to the high level of resistance exhibited by the isolated strains.

Caracterização das lesões por Cysticercus bovis, na inspeção post mortem de bovinos, pelos exames macroscópico, histopatológico e pela reação em cadeia da polimerase (PCR) / Characterization of Cysticercus bovis lesions at postmortem inspection of cattle by gross examination, histopathology and polymerase chain reaction (PCR)

Considerando a necessidade do conhecimento da cisticercose bovina e do aperfeiçoamento dos métodos de diagnóstico desta doença, objetivou-se verificar a ocorrência do Cysticercus bovis nos diversos locais anatômicos, tais como: cabeça, coração, esôfago, diafragma, língua, fígado e carcaça, examinados pelo Serviço de Inspeção Federal. O diagnóstico foi feito por macroscopia, microscopia e PCR com extração de DNA por fervura para a identificação do metacestóide. Dos 22043 bovinos abatidos, 713 (3,23%) estavam infectados. O coração foi o sítio anatômico mais afetado, com 1,90% (420/22043), seguido da cabeça, 1,11% (245/22043), do esôfago, 0,08% (18/22043), da carcaça, 0,07% (15/22043), do diafragma, 0,03% (7/22043), do fígado, 0,02% (5/22043) e da língua, 0,01% (3/22043). Dos cistos obtidos, 58,35% (416/713) estavam mortos e 41,65% (297/713), vivos. As diferenças entre os sítios anatômicos e a condição morfológica dos cistos foram significativas (p < 0,05). Dos 416 cistos mortos, 253 foram examinados por apresentarem características de: lesões nodulares firmes, brancacentas, com material amarelado, por vezes com aspecto calcário, no interior. O exame microscópico revelou granulomas comumente representados por centro necrótico e/ou mineralizado, envolto por histiócitos dispostos em paliçada, células gigantes multinucleadas, infiltrado misto, predominantemente de mononucleares, e fibrose. Por vezes, a periferia das lesões tinha características de tecido de granulação e mineralização em forma de lâminas lineares. Os restos parasitários foram identificados como um material hialino acelular, contendo elementos ovais e circulares, basofílicos, acidófilos e incolores, denominados corpúsculos calcários. Em algumas lesões foram observados raros corpúsculos, dispersos na reação inflamatória. Nódulos fibrosos, ricos em infiltrado linfóide ou crônico ativos, foram frequentemente visualizados. Dos cistos vivos examinados, 65% (13/20) foram positivos para C. bovis , confirmando o diagnóstico ambulatorial e a eficácia do método de PCR utilizado. Em virtude da positividade observada para C. bovis nos exames histopatológico e PCR, particularmente em fígado e esôfago, sugere-se que seja reformulado o artigo 176 do Regulamento de Inspeção Industrial e Sanitária de Produtos de Origem Animal, incluindo estes locais na rotina de inspeção nos matadouros.

Considering the importance of improving methods for diagnosis of bovine Cysticercosis, this study aimed to verify Cysticercus bovis occurrence in different anatomical sites, as head, heart, esophagus, diaphragm, tongue, liver and carcass, examined by federal inspection service. Diagnosis was performed by gross examination, histopatholgy and PCR with boiling DNA extraction for metacestode identification. Of 22043 slaughtered cattle, 713 (3.23%) were infected. The heart was mostly affected with 1.90% (420/22043), followed by head, 1.11% (245/22043), esophagus, 0.08% (18/22043), carcass, 0.07% (15/22043), diaphragm, 0.03% (7/22043), liver, 0.02% (5/22043) and tongue, 0.01% (3/22043). Of the cysts obtained, 58.35% (416/713) were dead and 41.65% (297/713) were alive. The differences among anatomical sites and cysts status were significant (p<0.05). Of the 416 dead cysts 253, characterized by nodular firm whitish lesions, containing yellowish material, some times in calcareous aspect were examined for histopathology. The histological exams of these cysts yielded granulomatous lesions, whose centers were characterized by caseous and/or calcareous material, multinucleate giant cells, histiocytes in palisade and infiltrate composed predominantly by lymphoid cells, wrapped up by fibrosis. Some times the lesions peripheries had granulation tissue and mineralized areas, like linear blade. The parasite debris were like a hyaline, non cellular material with spherical and ovoid, basophilic, eosinophilic and colorless corpuscles. These corpuscles were seen rarely, some times, among inflammatory reaction. Fibrous nodules, rich in lymphoid or mixed infiltrates, were frequently seen. Of the live cysts subjected to PCR with boiling DNA extraction, 65% (13/20) were positive for C. bovis, confirming the ambulatory diagnosis and the efficacy of the PCR procedure used. Due to microscopic and PCR diagnostic exams of C. bovis, mainly in the liver and esophagus, it is suggested changes in the 176 article of the regulatory inspection, by including these sites in the bovine routine inspection at the slaughterhouses.