Aloysio de Mello Figueiredo Cerqueira

The Tricycle Protocol is a One Health initiative that involves monitoring extended-spectrum β-lactamase-producing Escherichia coli (ESBL-Ec), an opportunistic intestinal and extraintestinal pathogen that is recognized as critically important for the development of new antimicrobial agents and as a key indicator of water, food, and environmental contamination. This study aimed to detect and characterize, phenotypically and genotypically, the presence of ESBL-Ec in coastal waters collected from three urban marine ecosystems and mussels from a mariculture area in the metropolitan region of Rio de Janeiro City, Brazil. Eighteen samples from each beach and from mussels were collected between June 2022 and December 2023. A total of 299 E. coli strains were identified over the study period, of which 33 (11%) were initially identified as potential ESBL producers after selective pressure cultivation. Antimicrobial susceptibility analysis revealed a high frequency of resistance to ampicillin and third-generation cephalosporins, exceeding 80%. Eighteen (54.4%) of the 33 E. coli strains were classified as multidrug resistant (MDR). Genotypic confirmation revealed at least one ESBL gene ( bla _CTX-M-1,2 , bla _CTX-M-8 , bla _CTX-M-14 , bla _TEM , or bla _SHV ) in 30 of the 33 strains, with bla _CTX-M-1,2 being the most prevalent in samples from one estuarine beach and from the mariculture site. Additionally, the genes qnrS , intl1 , and intl2 were detected in 21.5%, 7.6%, and 1.3% of the confirmed ESBL-Ec strains, respectively. Since we could not associate the presence of MDR ESBL-Ec strains with unsuitable water classification, these findings underscore a grave threat to both human and animal health in densely inhabited coastal regions.

Multidrug-resistant (MDR) Staphylococcus aureus has been increasingly isolated from pigs and people in close contact with them, especially livestock-associated methicillin-resistant S. aureus (LA-MRSA). In this cross-sectional study, we investigated S. aureus colonization in pigs and farm workers, their resistance profile, and genetic background to estimate interspecies transmission potential within farms from Rio de Janeiro state, Brazil, between 2014 and 2019. We collected nasal swabs from 230 pigs and 27 workers from 16 and 10 farms, respectively. Five MDR strains were subjected to whole genome sequencing. Fourteen (6.1%) pigs and seven (25.9%) humans were colonized with S. aureus, mostly (64–71%) MDR strains. Resistance to clindamycin, erythromycin, penicillin, and tetracycline was the most common among the pig and human strains investigated. MDR strains shared several resistance genes [blaZ, dfrG, fexA, lsa(E), and tet(M)]. Pig and human strains recovered from the same farm shared the same genetic background and antimicrobial resistance profile. LA-MRSA ST398-SCCmecV-t011 was isolated from pigs in two farms and from a farm worker in one of them, suggesting interspecies transmission. The association between pig management practices and MDR S. aureus colonization might be investigated in additional studies.

Access to genetic resources (GR) and/or traditional knowledge associated with genetic resources (ATK) has been regulated in Brazil since 2001. The law 13,123 / 2015 determined a significant change in the theme, mainly on the rules of distribution of benefits obtained for conservation and sustainable use of biodiversity, the access to technology and technology transfer, the exploitation of products or reproductive material from the GR or ATK and consignment to the outside of part or all the living or dead organism shipped for GR. The implementation of international treaties on GR and ATK for research, biotechnological development and bioprospecting have been causing difficulties for Brazilian researchers, mainly due to the lack of information and dissemination available for compliance with the legislation. In this work, the members of the Committee for Access to Genetic Resources and Associated Traditional Knowledge of the Federal Fluminense University (UFFGEN) - Brazil, and collaborators performed a critical reflection on the new law, helping Brazilian researchers with information necessary to understand the changes made by the new legislation, especially in the field of Biotechnology associated with Brazilian Biodiversity.

In animal husbandry, antimicrobial agents have been administered as supplements to increase production over the last 60 years. Large-scale animal production has increased the importance of antibiotic management because it may favor the evolution of antimicrobial resistance and select resistant strains. Brazil is a significant producer and exporter of animal-derived food. Although Brazil is still preparing a national surveillance plan, several changes in legislation and timely programs have been implemented. Thus, Brazilian data on antimicrobial resistance in bacteria associated with animals come from official programs and the scientific community. This review aims to update and discuss the available Brazilian data on this topic, emphasizing legal aspects, incidence, and genetics of the resistance reported by studies published since 2009, focusing on farm animals and derived foods with the most global public health impact. Studies are related to poultry, cattle, and pigs, and mainly concentrate on non-typhoid Salmonella, Escherichia coli, and Staphylococcus aureus. We also describe legal aspects of antimicrobial use in this context; and the current occurrence of genetic elements associated with resistance to beta-lactams, colistin, and fluoroquinolones, among other antimicrobial agents. Data here presented may be useful to provide a better understanding of the Brazilian status on antimicrobial resistance related to farm animals and animal-derived food products.

Respiratory diseases in birds generate sanitary and economic impacts and may be related to the environment and climate. Mycoplasma gallisepticum, Ornithobacterium rhinotracheale (ORT), Pasteurella multocida, Avibacterium paragallinarum, Escherichia coli, Riemerella anatipestifer, and Bordetella avium are among the most important avian respiratory pathogens. ORT is responsible for causing ornitobacteriosis, a disease characterized by clinical signs ranging from mild to severe respiratory conditions, with high mortality rates, mainly affecting turkeys and chickens. The first report of ornitobacteriosis was in 1981 in Germany. Despite its importance, few studies on ORT have been published. In addition, the presence of this pathogen has been neglected in poultry farms, mainly due to the lack of appropriate diagnostic protocols. The lack of correct isolation and diagnostic protocols along with inappropriate use of antimicrobial agents have been contributing to treatment failure. Due to its economic importance to the poultry industry, ornitobacteriosis should be monitored and included in national programs for the prevention and control of avian respiratory diseases. This review aimed to update and discuss important issues related to ORT since this pathogen has great economic and sanitary implications for the chicken production chain.

Shiga toxin‐producing Escherichia coli (STEC) is a zoonotic pathotype associated with human gastrointestinal disease that may progress to severe complications. Ruminants, especially cattle, are the main reservoirs of STEC from which they can contaminate the environment and foods of animal or vegetable origin. Besides Shiga toxin, other virulence factors are involved in STEC virulence. O157:H7 remains the most frequent serotype associated with disease. In Brazil, the prevalence of STEC reaches values as high as 90% in cattle and 20%inmeat productswhich may impact the Brazilian food export trade.However, only few reports are related to human disease. The stx1 gene prevails in cattle whereas the stx2 gene is more frequent in food. Several STEC serotypes have been isolated from cattle and food in Brazil, including the O157:H7, O111:NT, NT:H19 as well as O26 and O103 serogroups. O113: H21 STEC strains are frequent in ruminants and foods but with no report in human disease. The virulence profile of Brazilian STEC strains from cattle and food suggests a pathogenic potential to humans, although some differences with clinical strains have been detected. Further studies, employing recent and more discriminative techniques are in need to better clarify their virulence potential.

Respiratory diseases are common in young horses but little is known about such infections in mule foals. This study aimed to characterize Escherichia coli and Klebsiella sp. isolated from tracheal wash (TW) and fecal samples (FS) of mule foals, with or without cytological evidence of respiratory disease. Strains were analyzed against 13 antimicrobials, for presence of Extended spectrum beta-lactamase (ESBL), and virulence genes. Phylogrouping and Randomic (RAPD)-PCR profiles were used to evaluate their genetic relatedness. E. coli strains from TW and FS showed greatest resistance to tetracycline, while Klebsiella strains were mainly resistant to ampicillin; multidrug resistance and ESBL production were also detected. The blaCTX gene prevailed among the E. coli isolates, while the blaSHV gene was more frequently found in K. pneumoniae. The fimH gene was detected in most of the isolates and multiple virulence factors were identified in three E. coli isolates. Most of the E. coli isolates belonged to the B1 phylogroup, but B2 strains displayed more virulence genes. The RAPD assay revealed genetic diversity among strains and was able to distinguish FS isolates from TW isolates. Knowledge of the bacteria associated with the respiratory tract of mule foals is important in the treatment of sick animals.

The occurrence of virulence markers, serotypes and invasive ability were investigated in Shiga toxin-producing Escherichia coli (STEC) isolated from fecal samples of healthy dairy cattle at Rio de Janeiro State, Brazil. From 1,562 stx-positive fecal samples, 105 STEC strains were isolated by immuno-magnetic separation (IMS) or plating onto MacConkey agar (MC) followed by colony hybridisation. Fifty (47.6%) strains belonged to 9 serotypes (O8:H19, O22:H8, O22:H16, O74:H42, O113:H21, O141:H21, O157:H7, O171:H2 and ONT:H21). The prevalent serotypes were O157:H7 (12.4%), O113:H21 (6.7%) and O8:H19 (5.7%). Virulence genes were identified by polymerase chain reaction (PCR). E-hlyA (77.1%) was the more prevalent virulence marker, followed by espP (64.8%), saa (39%), eae (24.8%) and astA (21.9%). All O157:H7 strains carried the γ (gamma) variant of the LEE genes and the stx2c gene, while the stx1/stx2 genotype prevailed among the eae-negative strains. None of the eae-positive STEC produced the localized adherence (LA) phenotype in HEp-2 or Caco-2 cells. However, intimate attachment (judged by the FAS test) was detected in some eae-positive strains, both in HEp-2 (23.1%) and in Caco-2 cells (11.5%). Most strains (87.5%) showed “peripheral association” (PA) adherence phenotype to undifferentiated Caco-2 cells. Twenty-five (92.6%) of 27 strains invaded Caco-2 cells. The highest average value of invasion (9.6%) was observed among the eae-negative bovine strains from serotypes described in human disease. Healthy dairy cattle is a reservoir of STEC carrying virulence genes and properties associated with human disease. Although reports of human disease associated with STEC are scarce in Brazil, the colonization of the animal reservoir by potentially pathogenic strains offers a significant risk to our population.