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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.

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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.

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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)

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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.

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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.

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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)

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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.

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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)

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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.

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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)

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