RESUMO
1. Epidemiological surveillance of Salmonella spp. serves as a primary tool for maintaining the health of poultry flocks. Characterising circulating serotypes is crucial for implementing control and prevention measures. This study conducted phenotypic and molecular characterisation of S. enterica Pullorum, S. enterica Heidelberg, and S. enterica Corvalis isolated from broiler chickens during slaughtering.2. All strains were susceptible to gentamicin, neomycin and norfloxacin. However, resistance rates exceeded 50% for ciprofloxacin and tiamulin, irrespective of the serotype. Approximately 64% of strains were classified as multidrug-resistant, with S. enterica Heidelberg strains exhibiting significantly higher overall resistance. The isolates demonstrated the ability to adhere and produce biofilm at a minimum of three temperatures, with S. enterica Pullorum capable of biofilm production at all temperatures encountered during poultry rearing.3. Each strain possessed between two and seven different virulence-associated genes. Genetic similarity, as indicated by pulsed field gel electrophoresis, exceeded 90% for all three serotypes and strains were classified in the R5 ribotype by PCR, regardless of serotype. Sequencing revealed high similarity among all strains, with homology ranging from 99.61 to 100% and all were classified to a single cluster.4. The results suggested a clonal relationship among the strains, indicating the possible circulation of a unique clonal group of S. enterica Pullorum in the southern region of Brazil.
Assuntos
Antibacterianos , Galinhas , Doenças das Aves Domésticas , Salmonelose Animal , Salmonella enterica , Animais , Salmonelose Animal/microbiologia , Salmonelose Animal/epidemiologia , Galinhas/microbiologia , Doenças das Aves Domésticas/microbiologia , Doenças das Aves Domésticas/epidemiologia , Salmonella enterica/genética , Salmonella enterica/fisiologia , Salmonella enterica/efeitos dos fármacos , Salmonella enterica/isolamento & purificação , Antibacterianos/farmacologia , Farmacorresistência Bacteriana Múltipla , Biofilmes , Fenótipo , Virulência , Salmonella/genética , Salmonella/fisiologia , Salmonella/efeitos dos fármacos , Salmonella/isolamento & purificação , Testes de Sensibilidade Microbiana/veterinária , Eletroforese em Gel de Campo Pulsado/veterinária , SorogrupoRESUMO
In view of growing concerns, in a One Health context, regarding the transport and dissemination of pathogenic microorganisms among seabirds and other vertebrate animals, including humans, the aim of this study was to identify Salmonella spp. in stranded and non-stranded resident and migratory wild seabirds from the Brazilian coast. Antimicrobial susceptibility and molecular profiles, quinolone resistance genes and antigenic characterization of the isolates were also carried out. Fresh faeces and cloacal swabs were obtained totaling 122 seabirds sampled throughout different Brazilian coast regions. At the laboratory, sample culturing, Salmonella spp. isolation and biochemical identification were performed, followed by antigenic profile identification by serum agglutination, susceptibility profile characterization by the agar disc diffusion technique, detection of quinolone resistance genes (qnrA, qnrB, qnrS) using the multiplex polymerase chain reaction technique (multiplex PCR) and, finally, isolates profiles identification by pulsed field gel electrophoresis (PFGE). Salmonella enterica subsp. enterica was identified in 7% of the studied birds, comprising three different serovars: Panama (63 %), Typhimurium (25 %) and Newport (13 %). The most important findings reported herein are the first description of Salmonella panama in seabirds and the totality of isolates being resistant (or intermediate) to at least one tested antimicrobial, with emphasis on quinolone resistance. The molecular results suggest that the observed resistance cannot be explained by the presence of plasmid-mediated quinolone resistance genes. The PFGE suggests that the Panama and Newport profiles detected herein are not yet widespread in Brazil, unlike Typhimurium, which is already well distributed throughout the country. Considering this finding, we suggest that seabirds are an important link in the epidemiological chain of this serovar. The monitoring of these bacteria in seabirds, as well as of their susceptibility profiles to antimicrobials, must be continuous, strengthening the role of these animals as environmental health indicators and sentinels.
Assuntos
Aves/microbiologia , Farmacorresistência Bacteriana , Farmacorresistência Bacteriana Múltipla , Salmonella , Animais , Antibacterianos/farmacologia , Brasil/epidemiologia , Farmacorresistência Bacteriana/efeitos dos fármacos , Eletroforese em Gel de Campo Pulsado/veterinária , Lindera/microbiologia , Testes de Sensibilidade Microbiana/veterinária , Salmonella/classificação , Salmonella/efeitos dos fármacos , Salmonella enterica , Salmonella typhimuriumRESUMO
Brazil is the largest exporter of chicken meat and poultry farming is one of the most important productive segments, despite major losses due to the bacterium Escherichia coli, which is also a zoonotic microorganism. The objetive of this study was to isolate E. coli and to evaluate its transmissibility potential from the field to chicken meat using the Pulsed Field Gel Electrophoresis (PFGE) technique. Environmental samples (poultry litter, soil and water) were collected from broiler farms located in the South of Brazil where the majority of the Brazilian poultry production occurs. In addition, chicken meat (gizzard, heart, drumette and tulip) samples were collected from local supermarkets. As results, 47.36% of the samples were positives for E. coli. Furthermore, 10 pairs of clones of E. coli were found always in the same substrate (two water-water pairs; three soil-soil pairs and five meat-meat pairs) using PFGE. These findings suggest that certain strains of E. coli may have habitat preferences, making the transfer from one substrate type to another more difficult to occur. Moreover, since no clones were found between environmental samples and chicken meat, it is possible to imply a low risk of E. coli transmissibility throughout the chicken meat production chain.(AU)
Assuntos
Animais , Galinhas/genética , Galinhas/microbiologia , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Substratos para Tratamento Biológico/análise , Eletroforese em Gel de Campo Pulsado/veterinária , Saúde Pública VeterináriaRESUMO
Brazil is the largest exporter of chicken meat and poultry farming is one of the most important productive segments, despite major losses due to the bacterium Escherichia coli, which is also a zoonotic microorganism. The objetive of this study was to isolate E. coli and to evaluate its transmissibility potential from the field to chicken meat using the Pulsed Field Gel Electrophoresis (PFGE) technique. Environmental samples (poultry litter, soil and water) were collected from broiler farms located in the South of Brazil where the majority of the Brazilian poultry production occurs. In addition, chicken meat (gizzard, heart, drumette and tulip) samples were collected from local supermarkets. As results, 47.36% of the samples were positives for E. coli. Furthermore, 10 pairs of clones of E. coli were found always in the same substrate (two water-water pairs; three soil-soil pairs and five meat-meat pairs) using PFGE. These findings suggest that certain strains of E. coli may have habitat preferences, making the transfer from one substrate type to another more difficult to occur. Moreover, since no clones were found between environmental samples and chicken meat, it is possible to imply a low risk of E. coli transmissibility throughout the chicken meat production chain.
Assuntos
Animais , Eletroforese em Gel de Campo Pulsado/veterinária , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Galinhas/genética , Galinhas/microbiologia , Substratos para Tratamento Biológico/análise , Saúde Pública VeterináriaRESUMO
We characterized Shiga toxin-producing Escherichia coli (STEC) O157 (n = 20) and non-O157 (n = 68) isolated from carcasses (n = 54), the environment (n = 20), head meat (n = 3) and viscera washing and chilling water (n = 11) in provincial abattoirs before and after implementing improvement actions. The strains were tested for eae, saa, ehxA and fliCH7 genes. Variants stx1 and stx2 were also determined. Pulsed-field gel electrophoresis (PFGE) was carried out with restriction enzymes XbaI and BlnI. All twenty O157 STEC strains [H7; H21; HNM] carried genes rfbO157 and ehxA; 90.0 % were positive for eae and 15.0 % were negative for fliCH7 and positive for saa. Results of PFGE showed 17 XbaI patterns, of which 14 were unique and three formed clusters. From the 68 non-O157 STEC strains, 66.2 %, 55.9 % and 2.9 % were positive for ehxA, saa and eae genes, respectively. Fifty-three XbaI patterns were obtained (49 unique and four forming clusters). Cross-contamination between products and between the environment and products was confirmed in all abattoirs. While the proposed improvements reduced the risk of contamination, Good Hygiene Practices and Good Manufacturing Practices should be implemented in provincial abattoirs, stressing the importance of having a uniform national food safety standard.
Assuntos
Doenças dos Bovinos/epidemiologia , Infecções por Escherichia coli/veterinária , Escherichia coli Shiga Toxigênica/isolamento & purificação , Matadouros , Animais , Argentina/epidemiologia , Bovinos , Doenças dos Bovinos/microbiologia , Eletroforese em Gel de Campo Pulsado/veterinária , Infecções por Escherichia coli/epidemiologia , Infecções por Escherichia coli/microbiologia , Escherichia coli O157/isolamento & purificaçãoRESUMO
This study evaluated 250 animals from 25 different processing lots, processed in four slaughterhouses in São Paulo state, Brazil for the presence of Salmonella in the mesenteric lymph nodes (10â¯g sample of each animal) and characterized the antibiotics resistance profile, the Pulsed Field Gel Electrophoresis - PFGE and Multi Locus Sequence Typing - MLST profiles of selected strains. The pathogen was present in 36.4% (nâ¯=â¯91, CL 95% 30.4-43.4) of samples and 72% (nâ¯=â¯18, CL 95% 50.6-87.9%) of the analyzed lots. The main serovars were S. Typhimurium (nâ¯=â¯23), Salmonella enterica subsp. enterica 1.4,5,12:i:- (nâ¯=â¯17), followed by S. Infantis (nâ¯=â¯12) and S. Havana (nâ¯=â¯11). Twenty-eight strains (30%) were classified as other serovars. Sixty-eight percent of the strains were resistant to Streptomycin and tetracycline, followed by ampicillin and sulphonamides (62.6%), chloramphenicol (56.0%), trimethoprim-sulfamethoxazole (41.8%) and nalidixic acid (40.7%). The antibiotics with lower resistance rates were cephalothin and aztreonam (both with 3.3% resistant), and ceftriaxone and cefepime (both with 7.7%). Multidrug-resistant strains (MDR) accounted for 70.3% of the isolates. Eight strains were submitted to MLST: four S. Typhimurium and one S.1.4,5,12:i:-, all belonging to the ST 19, two Salmonella Infantis, belonging to the ST 32 and one S. Derby, belonging to ST 40. Twenty-one isolates with different antibiotics resistance profiles from the most prevalent serovars were selected for PFGE analysis. Serovar S. Typhimurium (nâ¯=â¯11) revealed 4 pulsotypes and 1 cluster and S. 1.4,5,12:i:- (nâ¯=â¯10) revealed 5 pulsotypes and 4 clusters. The high prevalence of the pathogen, with its high rates of antibiotics resistance and belonging to genetic groups that are often associated with disease in humans, shows that the production chain of pork is a potential source of infection in salmonellosis cases. Therefore, effective preventive measures for pathogen control are needed to reduce the risk of foodborne diseases.
Assuntos
Resistência Microbiana a Medicamentos , Linfonodos/microbiologia , Salmonelose Animal/epidemiologia , Salmonella/fisiologia , Doenças dos Suínos/epidemiologia , Animais , Brasil/epidemiologia , Eletroforese em Gel de Campo Pulsado/veterinária , Tipagem de Sequências Multilocus/veterinária , Prevalência , Salmonella/efeitos dos fármacos , Salmonelose Animal/microbiologia , Sus scrofa , Suínos , Doenças dos Suínos/microbiologiaRESUMO
Leptospirosis presents a complex and dynamic epidemiology. Bovine leptospirosis has been described as a major infectious disease impairing reproductive efficiency. Although infections by Leptospira interrogans, L. santarosai and L. borgpetersenii are frequently reported in cattle, the presence of L. noguchii in these animals should not be neglected. In this study, we describe serological (MAT) and molecular characterization (rrs and secY gene sequencing, multilocus sequence typing [MLST] and pulsed-field gel electrophoresis [PFGE]) of eight L. noguchii strains obtained from slaughtered cows. Intraspecific genetic diversity was evaluated, and haplotype networks were constructed based on hosts and geographical localizations. Strains were characterized as belonging to serogroups Australis, Autumnalis and Panama, and molecular characterization showed a high heterogeneity of these strains. Ten different STs were found (including nine new STs and 39 novel alleles) as well as nine different pulsotypes. Two clonal complexes were found. Phylogenetic trees based on secY locus and concatenated MLST loci showed two main clusters, with sequences from the present study included in the first. In general, there was no relationship between the geographical origin and the secY phylogenetic clusters, as well as between secY phylogenetic clusters and serogroups. Molecular diversity indexes confirmed a high variability (H > 0.8). This high intraspecific variation observed may be related to differences in virulence, pathogenicity and antigenicity or even adaptability of the strains. In addition, haplotype networks clearly demonstrated the circulation of genotypes between humans and animals, confirming the zoonotic potential. The present study provides relevant data for the study of leptospirosis in the One Health context, where human, animal and environmental health is closely connected.
Assuntos
Doenças dos Bovinos/epidemiologia , Leptospira/genética , Leptospirose/veterinária , Saúde Única , Animais , Técnicas de Tipagem Bacteriana/veterinária , Bovinos , Doenças dos Bovinos/microbiologia , Eletroforese em Gel de Campo Pulsado/veterinária , Feminino , Genótipo , Humanos , Leptospira/classificação , Leptospira/imunologia , Leptospira/patogenicidade , Leptospirose/epidemiologia , Leptospirose/microbiologia , Epidemiologia Molecular , Tipagem de Sequências Multilocus/veterinária , Panamá/epidemiologia , Filogenia , Sorogrupo , Virulência , ZoonosesRESUMO
Objective: The study was conducted to determine geneticrelatedness of Salmonella serotypes by pulsed field gel electrophoresis (PFGE). Design and Methodology: A total of 1503 caecal samples of freshly slaughtered poultry were randomly collected from 'pluck shops' across the country. The samples were screened for Salmonella by biochemical, serological and molecular methods. The Salmonella serotypes were analyzed for genetic relatedness for phenotypic antimicrobial resistance, resistance and virulence gene profiles by PFGE generated by digestion with XbaI. Results: Ten different serotypes were detected from all 91 Salmonella isolates. PFGE fingerprinting profiles showed that the Salmonella serotypes in general, were genetically diverse with the detection of a total of 20 PFGE groups. The antibiograms of the isolates were also clearly very variable, which suggest that genotypic antimicrobial resistance may not relate to the phenotypic antibiograms in dendrograms, except for qnrB gene. Results demonstrated a varied spectrum of antimicrobial resistance and PFGE patterns among Salmonella isolates and signify the importance of sustained surveillance of foodborne pathogens in retail poultry pluck shops. Conclusions: The findings provide evidence that poultry from pluck shops are colonized by pathogenic Salmonella harboring antimicrobial resistance genes. The study also reported for the first time in Trinidad, molecular characterization of Salmonella isolates from poultry regarding the relatedness of antibiograms, possession of resistance and virulence genes using their PFGE profiles. The epidemiological surveillance of these serotypes would be necessary to evaluate their possible impact on human health in the country and possibly in the Caribbean region.
Assuntos
Animais , Salmonelose Animal , Trinidad e Tobago , Eletroforese em Gel de Campo Pulsado/veterinária , Região do Caribe/etnologia , GenéticaRESUMO
Campylobacter is regarded as the most common bacterial cause of gastroenteritis throughout the world and most cases of human campylobacteriosis can be traced back to the consumption of poultry meat. In Brazil, few studies evaluated the genetic relatedness among Campylobacter isolates. The aim of this research was to evaluate the genetic diversity of Campylobacter spp. isolated from poultry meat products sold on the retail market in Southern Brazil. The presumptive identification of Campylobacter was performed using traditional microbiological analysis, followed by molecular confirmation by PCR. The genetic diversity of isolates was analyzed by pulsed-field gel electrophoresis (PFGE). Campylobacter spp. was isolated from 91.7% (33/36) of the samples, totaling 48 isolates. Campylobacter jejuni was the most prevalent species isolated (90.8%). PFGE data revealed 26 pulsotypes and 18 PFGE patterns composed of only 1 isolate. Campylobacter isolates exhibited high genetic diversity; however, some clones were recurrent in the poultry meat products sold on the retail market. As the south region of Brazil is an important producer and exporter of chicken meat, our results highlight the need to control this pathogen in the food chain in this area of the world to reduce the risks of exposing consumers to campylobacteriosis.
Assuntos
Campylobacter coli/genética , Campylobacter jejuni/genética , Microbiologia de Alimentos , Variação Genética , Produtos Avícolas/microbiologia , Animais , Brasil , Campylobacter coli/isolamento & purificação , Campylobacter jejuni/isolamento & purificação , Galinhas , Eletroforese em Gel de Campo Pulsado/veterinária , Reação em Cadeia da Polimerase/veterináriaRESUMO
Escherichia albertii is a recently discovered species with a limited number of well characterized strains. The aim of this study was to characterize four of the E. albertii strains, which were among 41 identified Escherichia strains isolated from the feces of living animals on James Ross Island, Antarctica, and Isla Magdalena, Patagonia. Sequencing of 16S rDNA, automated ribotyping, and rep-PCR were used to identify the four E. albertii isolates. Phylogenetic analyses based on multi-locus sequence typing showed these isolates to be genetically most similar to the members of E. albertii phylogroup G3. These isolates encoded several virulence factors including those, which are characteristic of E. albertii (cytolethal distending toxin and intimin) as well as bacteriocin determinants that typically have a very low prevalence in E. coli strains (D, E7). Moreover, E. albertii protein extracts caused cell cycle arrest in human cell line A375, probably because of cytolethal distending toxin activity.