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.

Seasonal occurrence and variation of ectoparasites in Nile tilapia (Oreochromis niloticus) in the Federal District, Brazil

The present study assesses the diversity and seasonal variation of parasites in Oreochromis niloticus (Nile tilapia) cultivated in excavated tanks in the Federal District, Brazil. A total of 120 specimens of O. niloticus were collected in 12 monthly collections. Water quality parameters were checked at all tanks. The animals, sacrificed by immersion into anesthetic solution, had the gills scraped, in which the mucus was analyzed under an optical microscope. Parasitological examination showed Trichodina sp., Epistylis sp., Ichthyophthirius multifiliis, and Monogenea helminths in the fish gills. We found statistically significant differences between Monogenea parasites collected in the fall and winter and between I. multifiliis protozoans collected in fall-winter and winter-summer periods. Except for Epistylis sp., all parasites showed abundance peaks in O. niloticus specimens collected during the winter, which may characterize the seasonality of these parasites in the Federal District region. Monogenea helminths were the most prevalent among the parasites found, with the highest prevalence during the fall. The mean abundance of parasites was similar between the fall and winter. However, the abundance of monogenetic trematodes was higher between the winter and summer. I. multifiliis showed significant variation between fall-winter and winter-summer periods.(AU)

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.

Incidence of Mycoplasma gallisepticum and Mycoplasma synoviae in broiler flocks at the Federal District of Brazil and its surrounding areas / Incidência de lotes de frangos afetados por Mycoplasma gallisepticum e Mycoplasma synoviae no Distrito Federal e Entorno, Brasil

Mycoplasma is an important avian pathogen that can cause both respiratory disease and synovitis in birds, resulting in considerable economic losses to the poultry industry worldwide. This study aimed to determine the incidence of Mycoplasma gallisepticum and M. synoviae in broiler flocks at the Federal District and its surrounding regions using polymerase chain reaction (PCR). All slaughtered lots (57 flocks) were analyzed from July to November in one of the two slaughterhouses at the Federal District with Federal inspection services. Approximately 10 samples of broiler tracheae per slaughtered batch were collected from the evisceration line. The results obtained from the accumulated incidence over the study period were 7.02% and 35.09% for M. gallisepticum and M. synoviae, respectively. A greater concentration of flocks affected by M. synoviae was observed during October. The sample design as well as the PCR technique assisted in detecting both agents in the broiler batches in the first epidemiological study of these two agents in the region.(AU)

Os micoplasmas são importantes patógenos aviários, que podem causar doenças respiratórias e sinovites em aves, resultando em consideráveis perdas econômicas para a indústria avícola em todo o mundo. O objetivo deste estudo foi determinar a incidência de Mycoplasma gallisepticum e Mycoplasma synoviae em lotes de frangos na região do Distrito Federal e Entorno, por meio da reação em cadeia de polimerase (PCR). Todos os lotes abatidos (57 lotes) foram analisados durante os meses de julho a novembro, em um dos dois abatedouros frigoríficos do Distrito Federal com Serviço de Inspeção Federal. Na linha de evisceração foram coletadas cerca de 10 amostras de traqueia de frangos de corte por lote abatido. Os resultados obtidos da incidência acumulada no período de estudo foram de 7.02% para M. gallisepticum e 35,09% M. synoviae. Uma maior concentração do número de lotes afetados por M. synoviae foi observada durante o mês de outubro. O desenho amostral, assim como a técnica de PCR, permitiu a detecção de ambos os agentes nos lotes de frangos de corte analisados, sendo este o primeiro estudo epidemiológico desses dois agentes na região de estudo.(AU)

Detection of enterotoxin and adhesin genes of Escherichia coli strains isolated from feces of healthy dogs / Detecção de genes de enterotoxinas e adesinas em cepas de Escherichia coliisoladas de fezes de cães saudáveis

This study aimed to investigate the presence of genes encoding the enterotoxins STa and Stx1 and the adhesins K99 and Intimin in E. coli strains isolated from feces of dogs who appeared to be healthy. Rectal swab samples were collected from 50 dogs who visited the Veterinary Hospital of the University of Brasilia and 48 E. coli isolates were obtained. No positive isolates were found for STa and K99. However, positive results were found in 21 isolates (43.7%) for Stx1 and 14 isolates (29%) for the Intimin gene (eae). The antimicrobial sensitivity profile was also assessed for the following antibiotics: sulfazothrim, azithromycin, enrofloxacin, ceftiofur, amoxicillin + clavulanate, doxycycline, ampicillin, and cephalexin. The antibiotics on which the isolates showed the highest resistance were ampicillin (25%), doxycycline (22.9%) and cephalexin (20.8%). As for sensitivity, the isolates were most sensitive to sulfazothrim (87.5%), azithromycin (85.41%) and enrofloxacin (77%). Healthy dogs can carry multidrug-resistant E. coli strains that can also carry enterotoxin and adhesin genes, thus indicating that, the proximity between dogs and humans may contribute to possible zoonotic transmission of these microorganisms.

Este estudo teve como objetivo investigar a presença de genes que codificam as enterotoxinas STa e Stx1 e as adesinas K99 e Intiminaem cepas de E. coliisoladas de fezes de cãesaparentementesaudáveis. Amostras de swab retal foram coletadas de 50 cães que visitaram o Hospital Veterinário da Universidade de Brasília e 48 isolados de E. coliforam obtidos. Não foram encontrados isolados positivos para STa e K99. No entanto, resultados positivos foram encontrados em 21 isolados (43,7%) para Stx1 e 14 isolados (29%) para o gene Intimina(eae). O perfil de sensibilidade antimicrobiana também foi avaliado para os seguintes antibióticos: sulfazotrim, azitromicina, enrofloxacina, ceftiofur, amoxicilina + clavulanato, doxiciclina, ampicilina,e cefalexina. Os antibióticos nos quais os isolados apresentaram maior resistência foramaampicilina (25%), doxiciclina (22,9%) e cefalexina (20,8%). Quanto à sensibilidade, os isolados foram mais sensíveis ao sulfazotrim (87,5%), azitromicina (85,41%) e enrofloxacina (77%). Cães saudáveis podem carrearcepas de E. colimultirresistentes que por sua vez também podem carreargenes codificadores de enterotoxina e adesina, indicando assim que a proximidade entre cães e humanos pode contribuir para a possível transmissão zoonótica desses microrganismos.

Detection of enterotoxin and adhesin genes of Escherichia coli strains isolated from feces of healthy dogs / Detecção de genes de enterotoxinas e adesinas em cepas de Escherichia coliisoladas de fezes de cães saudáveis

This study aimed to investigate the presence of genes encoding the enterotoxins STa and Stx1 and the adhesins K99 and Intimin in E. coli strains isolated from feces of dogs who appeared to be healthy. Rectal swab samples were collected from 50 dogs who visited the Veterinary Hospital of the University of Brasilia and 48 E. coli isolates were obtained. No positive isolates were found for STa and K99. However, positive results were found in 21 isolates (43.7%) for Stx1 and 14 isolates (29%) for the Intimin gene (eae). The antimicrobial sensitivity profile was also assessed for the following antibiotics: sulfazothrim, azithromycin, enrofloxacin, ceftiofur, amoxicillin + clavulanate, doxycycline, ampicillin, and cephalexin. The antibiotics on which the isolates showed the highest resistance were ampicillin (25%), doxycycline (22.9%) and cephalexin (20.8%). As for sensitivity, the isolates were most sensitive to sulfazothrim (87.5%), azithromycin (85.41%) and enrofloxacin (77%). Healthy dogs can carry multidrug-resistant E. coli strains that can also carry enterotoxin and adhesin genes, thus indicating that, the proximity between dogs and humans may contribute to possible zoonotic transmission of these microorganisms.(AU)

Este estudo teve como objetivo investigar a presença de genes que codificam as enterotoxinas STa e Stx1 e as adesinas K99 e Intiminaem cepas de E. coliisoladas de fezes de cãesaparentementesaudáveis. Amostras de swab retal foram coletadas de 50 cães que visitaram o Hospital Veterinário da Universidade de Brasília e 48 isolados de E. coliforam obtidos. Não foram encontrados isolados positivos para STa e K99. No entanto, resultados positivos foram encontrados em 21 isolados (43,7%) para Stx1 e 14 isolados (29%) para o gene Intimina(eae). O perfil de sensibilidade antimicrobiana também foi avaliado para os seguintes antibióticos: sulfazotrim, azitromicina, enrofloxacina, ceftiofur, amoxicilina + clavulanato, doxiciclina, ampicilina,e cefalexina. Os antibióticos nos quais os isolados apresentaram maior resistência foramaampicilina (25%), doxiciclina (22,9%) e cefalexina (20,8%). Quanto à sensibilidade, os isolados foram mais sensíveis ao sulfazotrim (87,5%), azitromicina (85,41%) e enrofloxacina (77%). Cães saudáveis podem carrearcepas de E. colimultirresistentes que por sua vez também podem carreargenes codificadores de enterotoxina e adesina, indicando assim que a proximidade entre cães e humanos pode contribuir para a possível transmissão zoonótica desses microrganismos.(AU)

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.(AU)

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.

Isolation and antimicrobial resistance of Campylobacter jejuni and Campylobacter coli found in chilled chicken carcasses in the Federal District Region and surrounding areas / Isolamento e resistência antimicrobiana de Campylobacter jejuni e Campylobacter coli encontradas em carcaças de frango resfriadas na região do Distrito Federal e entorno

This study aimed to isolate Campylobacter jejuni and Campylobacter coli from chilled chicken carcasses marketed in the Federal District Region and surrounding areas, as well as to detect the occurrence of antimicrobial resistance and genes responsible for the same. A total of 105 chilled chicken carcasses were collected, of which 7 (6.67%) were positive for C. jejuni and 4 (3.81%) were positive for C. coli. These results were obtained using both the conventional microbiological isolation method and polymerase chain reaction assays. All of the positive strains were subjected to antimicrobial susceptibility testing for seven antimicrobials. The resistance incidences found in the C. jejuni strains were as follows: 71.43% for tetracycline and nalidixic acid, 42.86% for streptomycin and gentamicin, 57.14% for ciprofloxacin and erythromycin, and 28.57% for chloramphenicol. Among the C. coli strains, 100% were resistant to tetracycline and streptomycin, 75% were resistant to erythromycin, 50% were resistant to ciprofloxacin, gentamicin, and nalidixic acid, and no strains were resistant to chloramphenicol. While analyzing the presence of antimicrobial resistance genes in the isolated C. jejuni strains, the aph3-1 (resistance to aminoglycosides), aadE (resistance to streptomycin), and tet(O) (resistance to tetracycline) genes were identified, with occurrence rates of 57.14%, 28.57%, and 42.86%, respectively, whereas in the C. coli strains, there was a 25% occurrence rate for both the aph3-1 and tet(O) genes. The aadE gene was not found in the C. coli isolates. The results of this study demonstrated the presence of C. jejuni and C. coli in chilled chicken carcasses marketed in the Federal District Region and surrounding areas, as well as the antimicrobial resistance and the presence of resistance genes in these bacteria, which may pose threats to public health.(AU)

O objetivo deste trabalho foi isolar Campylobacter jejuni e Campylobacter coli de carcaças de frango resfriadas comercializadas no Distrito Federal e entorno, bem como detectar a ocorrência de resistência antimicrobiana e genes responsáveis pela resistência antimicrobiana. Foram coletadas um total de 105 carcaças de frango resfriadas, das quais 7 (6,67%) foram positivas para C. jejuni e 4 (3,81%) para C. coli. Estes resultados foram obtidos usando tanto o método convencional de isolamento microbiológico quanto os ensaios de reação em cadeia da polymerase (PCR). Todas as cepas positivas foram submetidas ao teste de susceptibilidade antimicrobiana para sete antimicrobianos. As incidências de resistência encontradas nas cepas de C. jejuni foram as seguintes: 71,43% para tetraciclina e ácido nalidíxico, 42,86% para estreptomicina e gentamicina, 57,14% para ciprofloxacina e eritromicina e 28,57% para cloranfenicol. Entre as cepas de C. coli, 100% foram resistentes à tetraciclina e à estreptomicina, 75% eram resistentes à eritromicina, 50% eram resistentes à ciprofloxacina, gentamicina e ácido nalidíxico, e nenhuma cepa eram resistentes ao cloranfenicol. Ao analisar a presença de genes de resistência antimicrobiana nas cepas isoladas de C. jejuni, foram identificados os genes aph3-1 (resistência a aminoglicosídeos), aadE (resistência à estreptomicina) e tet (O) (resistência à tetraciclina) genes foram identificados, com taxas de ocorrência de 57,14%, 28,57% e 42,86%, respectivamente, enquanto que nas cepas de C. coli, houve uma taxa de ocorrência de 25% para os genes aph3-1 e tet (O). O gene aadE não foi encontrado nos isolados de C. coli. Os resultados deste estudo demonstraram a presença de C. jejuni e C. coli em carcaças de frango resfriadas comercializadas na Região do Distrito Federal e entorno, além da resistência antimicrobiana e a presença de genes de resistência nestas cepas, que podem se tornar uma possível ameaça a saúde pública.(AU)

Isolation and antimicrobial resistance of Campylobacter jejuni and Campylobacter coli found in chilled chicken carcasses in the Federal District Region and surrounding areas / Isolamento e resistência antimicrobiana de Campylobacter jejuni e Campylobacter coli encontradas em carcaças de frango resfriadas na região do Distrito Federal e entorno

This study aimed to isolate Campylobacter jejuni and Campylobacter coli from chilled chicken carcasses marketed in the Federal District Region and surrounding areas, as well as to detect the occurrence of antimicrobial resistance and genes responsible for the same. A total of 105 chilled chicken carcasses were collected, of which 7 (6.67%) were positive for C. jejuni and 4 (3.81%) were positive for C. coli. These results were obtained using both the conventional microbiological isolation method and polymerase chain reaction assays. All of the positive strains were subjected to antimicrobial susceptibility testing for seven antimicrobials. The resistance incidences found in the C. jejuni strains were as follows: 71.43% for tetracycline and nalidixic acid, 42.86% for streptomycin and gentamicin, 57.14% for ciprofloxacin and erythromycin, and 28.57% for chloramphenicol. Among the C. coli strains, 100% were resistant to tetracycline and streptomycin, 75% were resistant to erythromycin, 50% were resistant to ciprofloxacin, gentamicin, and nalidixic acid, and no strains were resistant to chloramphenicol. While analyzing the presence of antimicrobial resistance genes in the isolated C. jejuni strains, the aph3-1 (resistance to aminoglycosides), aadE (resistance to streptomycin), and tet(O) (resistance to tetracycline) genes were identified, with occurrence rates of 57.14%, 28.57%, and 42.86%, respectively, whereas in the C. coli strains, there was a 25% occurrence rate for both the aph3-1 and tet(O) genes. The aadE gene was not found in the C. coli isolates. The results of this study demonstrated the presence of C. jejuni and C. coli in chilled chicken carcasses marketed in the Federal District Region and surrounding areas, as well as the antimicrobial resistance and the presence of resistance genes in these bacteria, which may pose threats to public health.

O objetivo deste trabalho foi isolar Campylobacter jejuni e Campylobacter coli de carcaças de frango resfriadas comercializadas no Distrito Federal e entorno, bem como detectar a ocorrência de resistência antimicrobiana e genes responsáveis pela resistência antimicrobiana. Foram coletadas um total de 105 carcaças de frango resfriadas, das quais 7 (6,67%) foram positivas para C. jejuni e 4 (3,81%) para C. coli. Estes resultados foram obtidos usando tanto o método convencional de isolamento microbiológico quanto os ensaios de reação em cadeia da polymerase (PCR). Todas as cepas positivas foram submetidas ao teste de susceptibilidade antimicrobiana para sete antimicrobianos. As incidências de resistência encontradas nas cepas de C. jejuni foram as seguintes: 71,43% para tetraciclina e ácido nalidíxico, 42,86% para estreptomicina e gentamicina, 57,14% para ciprofloxacina e eritromicina e 28,57% para cloranfenicol. Entre as cepas de C. coli, 100% foram resistentes à tetraciclina e à estreptomicina, 75% eram resistentes à eritromicina, 50% eram resistentes à ciprofloxacina, gentamicina e ácido nalidíxico, e nenhuma cepa eram resistentes ao cloranfenicol. Ao analisar a presença de genes de resistência antimicrobiana nas cepas isoladas de C. jejuni, foram identificados os genes aph3-1 (resistência a aminoglicosídeos), aadE (resistência à estreptomicina) e tet (O) (resistência à tetraciclina) genes foram identificados, com taxas de ocorrência de 57,14%, 28,57% e 42,86%, respectivamente, enquanto que nas cepas de C. coli, houve uma taxa de ocorrência de 25% para os genes aph3-1 e tet (O). O gene aadE não foi encontrado nos isolados de C. coli. Os resultados deste estudo demonstraram a presença de C. jejuni e C. coli em carcaças de frango resfriadas comercializadas na Região do Distrito Federal e entorno, além da resistência antimicrobiana e a presença de genes de resistência nestas cepas, que podem se tornar uma possível ameaça a saúde pública.

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

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, foide 8,2 germes/cm² e não houve crescimento de Staphylococcus coagulase positiva. Neste trabalho verificou-sea 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)

The aim of this work was to verify the quality hygienic of bovine carcasses from slaughterhouses and cold storage, under the district inspection (Dipova) and federal inspection (SIF), located in Distrito Federal area. It were realized microbiological analysis in 05 (five)different points of 07 (seven) half-carcasses to identify Coliforms at 45°C, Mesophilic bacteria and enumerate Staphylococcus aureus. The samples were collected before and after washing them with chlorinated water, thus making a total of 70 tests. The averages observed before the wash with chlorinated water, for the count of Strict and Facultative microorganisms Aerobic Mesophilic was 9,7 CFU/cm², the Most Probable Number of Coliforms at 45°C was 29 cells/cm² and e de 1 CFU/cm² de Staphylococcus coagulase positive. The values observed in the half-carcasses,after the wash with chlorinated water for the count of Strict and Facultative microorganisms Aerobic Mesophilic were 1,5 CFU/cm², for the analysis of the Most Probable Number of Coliforms at 45º C were 8,2 cells/cm². There was no growth of Staphylococcus coagulase positive,after washing. In this work, we verified the presence of Coliforms at 45ºC and the presence of Strict and Facultative microorganisms Aerobic Mesophilic before and after the washing and the growth of Staphylococcus aureus only before the washing. It was observed a decreased in the degree of contamination, in the washed half-carcasses with chlorinated water.(AU)

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)

Atividade antimicrobiana da erva-mate (Ilex paraguariensis) contra microrganismos isolados da carne de frango / Antimicrobial activity of yerba mate (Ilex paraguariensis) extract against microorganisms isolated from chicken meat

A erva-mate vem sendo estudada devido às diversas atividades biológicas já evidenciadas. Este estudo teve como objetivo avaliar o efeito antimicrobiano, pelo método de difusão em disco, da adição de extratos de erva-mate frente à Escherichia coli e ao Proteus mirabilispreviamente isolados da carne de peito de frango. Foram utilizadas diferentes concentrações de extrato (125 mg/ml, 250 mg/mL, 550 mg/mLl de extrato de erva-mate, controle negativo com solução salina 0,85% e erva-mate pura) em triplicata e os resultados analisados pelo SAS® 5.1. Quanto maior a concentração de extrato aplicada, maior (P<0,0001) foi o halo de inibição formado para as duas bactérias avaliadas, sendo que para a E. coli os halos foram menores quando comparados com os halos para o P. Mirabilis. Foi possível estimar a concentração de 400 mg/mLl como sendo a mais eficiente frente ambas as bactérias avaliadas.

Yerba mate has been studied due to the biological activities already evidenced. This study aimed to evaluate the antimicrobial effect of the addition of yerba mate extracts against Escherichia coli and Proteus mirabilis previously isolated from chicken breast meat using the disk diffusion method. Different extract concentrations were applied (125 mg/ml, 250 mg/ml, 550 mg/ml of yerba mate extract, negative control with saline 0.85% and pure yerba mate) in triplicate and the results were analyzed by the SAS® 5.1. The increase in the concentration of the extract applied resulted in an increase (P<0.0001) in the inhibition zone formed for both bacteria, and it was evident that the inhibition zone for E. coli was smaller compared with the halos for P. mirabilis. It was possible to estimate the concentration of 400 mg/ml as the most efficient for both bacteria.

Atividade antimicrobiana da erva-mate (Ilex paraguariensis) contra microrganismos isolados da carne de frango / Antimicrobial activity of yerba mate (Ilex paraguariensis) extract against microorganisms isolated from chicken meat

A erva-mate vem sendo estudada devido às diversas atividades biológicas já evidenciadas. Este estudo teve como objetivo avaliar o efeito antimicrobiano, pelo método de difusão em disco, da adição de extratos de erva-mate frente à Escherichia coli e ao Proteus mirabilispreviamente isolados da carne de peito de frango. Foram utilizadas diferentes concentrações de extrato (125 mg/ml, 250 mg/mL, 550 mg/mLl de extrato de erva-mate, controle negativo com solução salina 0,85% e erva-mate pura) em triplicata e os resultados analisados pelo SAS® 5.1. Quanto maior a concentração de extrato aplicada, maior (P<0,0001) foi o halo de inibição formado para as duas bactérias avaliadas, sendo que para a E. coli os halos foram menores quando comparados com os halos para o P. Mirabilis. Foi possível estimar a concentração de 400 mg/mLl como sendo a mais eficiente frente ambas as bactérias avaliadas.(AU)

Yerba mate has been studied due to the biological activities already evidenced. This study aimed to evaluate the antimicrobial effect of the addition of yerba mate extracts against Escherichia coli and Proteus mirabilis previously isolated from chicken breast meat using the disk diffusion method. Different extract concentrations were applied (125 mg/ml, 250 mg/ml, 550 mg/ml of yerba mate extract, negative control with saline 0.85% and pure yerba mate) in triplicate and the results were analyzed by the SAS® 5.1. The increase in the concentration of the extract applied resulted in an increase (P<0.0001) in the inhibition zone formed for both bacteria, and it was evident that the inhibition zone for E. coli was smaller compared with the halos for P. mirabilis. It was possible to estimate the concentration of 400 mg/ml as the most efficient for both bacteria.(AU)

ATIVIDADE ANTIMICROBIANA DA ERVA-MATE (Ilex paraguariensis) CONTRA MICRORGANISMOS ISOLADOS DA CARNE DE FRANGO

Abstract Yerba mate has been studied due to the biological activities already evidenced. This study aimed to evaluate the antimicrobial effect of the addition of yerba mate extracts against Escherichia coli and Proteus mirabilis previously isolated from chicken breast meat using the disk diffusion method. Different extract concentrations were applied (125 mg/mL, 250 mg/mL, 550 mg/mL of yerba mate extract, negative control with saline 0.85% and pure yerba mate) in triplicate and the results were analyzed by the SAS® 5.1. The increase in the concentration of the extract applied resulted in an increase (P 0.0001) in the inhibition zone formed for both bacteria, and it was evident that the inhibition zone for E. coli was smaller compared with the halos for P. mirabilis. It was possible to estimate the concentration of 400 mg/mL as the most efficient for both bacteria.

Resumo A erva-mate vem sendo estudada devido às diversas atividades biológicas já evidenciadas. Este estudo teve como objetivo avaliar o efeito antimicrobiano, pelo método de difusão em disco, da adição de extratos de erva-mate frente à Escherichia coli e ao Proteus mirabilis previamente isolados da carne de peito de frango. Foram utilizadas diferentes concentrações de extrato (125 mg/mL, 250 mg/mL, 550 mg/mL de extrato de erva-mate, controle negativo com solução salina 0,85% e erva-mate pura) em triplicata e os resultados analisados pelo SAS® 5.1. Quanto maior a concentração de extrato aplicada, maior (P 0,0001) foi o halo de inibição formado para as duas bactérias avaliadas, sendo que para a E. coli os halos foram menores quando comparados com os halos para o P. Mirabilis. Foi possível estimar a concentração de 400 mg/mL como sendo a mais eficiente frente ambas as bactérias avaliadas.

Principais lesões em carcaças e órgãos de bovinos oriundos de frigoríficos localizados em Goiás e no Distrito Federal / Principals lesions in carcasses and beef organs coming from slaughterhouses located in Goias and the Federal District

o presente trabalho teve como objetivo verificar as principais lesões que ocorrem em órgãos e carcaças de bovinos oriundos de frigoríficos localizados no Distrito Federal e no estado de Goiás. A pesquisa foi realizada a partir do acompanhamento das atividades da Inspeção Federal do Serviço de inspeção do Ministério da Agricultura, Pecuária e Abastecimento (MAPA) e do Distrito federal, Diretoria de Inspeção de Produtos de Origem Vegetal e Animal (DIPOV A), durante o período de março a maio de 2012, junto às linhas de inspeções, totalizando 4012 carcaças de animais abatidos, sendo 3419 no estado do Goiás e animais no Distrito Federal. Foram condenados 1411 órgãos na inspeção post mortem, e as principais lesões observadas nos mesmos, do total de 4012 animais abatidos, foram renais, devido à nefrite com 160 (11,34%), pulmonares devido a enfisema com 114 (8,08%) e aspiração de sangue com 97 (6,87%) e de fígado devido a telangiectasia com 51 (3,61%). Não houve nenhuma condenação total ou parcial de carcaça no período analisado. Os resultados observados sugerem que as principais causas de condenação do pulmão e fígado são devido a falhas na tecnologia de abate, atentando assim para necessidade de treinamento dos funcionários da indústria e também a possíveis falhas no manejo no caso das condenações devido à nefrite e outras detectadas nesse estudo. (AU)

The aim of this work was to verify the lesions that occur in organs and carcasses of cattle from slaughterhouses localized in Federal District and Goiás. This study was performed by monitoring the activities of the Federal inspection Service of the Ministry of Agriculture, Livestock and Supply (MAPA), of the District Board of inspection Products of Plant and Animal Origin from Federal District area (D/- POVA), and along the lines of inspections of four different cattle slaughterhouses. Three slaughterhouses are located in the state of Goiás and the other one in the Federal District. All activities of this study were done between March and May 2012. Were analysed a total of 4012 carcasses of slaughtered animals. From these, 3,419 carcasses were from the state of Goiás and 593 from the Federal District area. It was condemned a total of 1411 organs in the post mortem inspec- I tion procedures, 3419 from the Goiás slaughterhouses and 593 from DF. The major lesions observed in organs of all the 4012 animals slaughtered, were nephritis with 160 (11,34%), in the lung was emphysema with 114 (8,08%) and blood aspiration with 97 (6,87%) and in the liver condemnation was due telangiectasia with 51 (3,61%). No carcass was condemned. The results indicate that the major cause of lung and liver condemnation were caused by failure in the slaughter technology. 1n the case of contamination and blood aspirations, is important paying attention to an improvement in training of the employees of the slaughterhouse, and also possible failures in the cattle ranching management due to the presence of the lesions detected.(AU)