Phenotypic and genotypic characterizations of bacteria isolated from the respiratory microbiota of healthy turkeys with potential for probiotic composition.

Desirable characteristics of Staphylococcus sp., Streptococcus sp., Bacillus sp., Klebsiella sp., Escherichia coli, and Pseudomonas pseudoalcaligenes isolated from the trachea of healthy turkeys were evaluated as probiotic candidates in the search for new alternatives to solve antimicrobial resistance issues in poultry. In current study phenotypic and genotypic capacity to produce bacteriocin-like substances, efficacy to inhibit the growth of avian pathogens, susceptibility to antimicrobials of bacteria isolated from the respiratory microbiota of healthy turkeys, and the presence of virulence-associated genes (VAGs) predictors of Avian Pathogenic Escherichia coli (APEC) were evaluated. Nine E. coli and one Klebsiella sp. strains produced bacteriocin-like substances, and all harbored the cvaA gene. Some strains also showed antagonistic activity against APEC. Multidrug-resistant profile was found in 54% of the strains. Six strains of bacteriocin-like substances producing E. coli also harbored 3-5 VAGs. The study showed that two bacterial genuses (Klebsiella sp. and E. coli) present desirable probiotic characteristics. Our results identified strains with potential for poultry's respiratory probiotic.

Characterization of avian pathogenic Escherichia coli isolates based on biofilm formation, ESBL production, virulence-associated genes, and phylogenetic groups.

Escherichia coli is a part of both animal and human commensal microbiota. Avian pathogenic E. coli (APEC) is responsible for colibacillosis in poultry, an economically important disease. However, the close similarities among APEC isolates make it difficult to differentiate between pathogenic and commensal bacteria. The aim of this study was to determine phenotypic and molecular characteristics of APEC isolates and to compare them with their in vivo pathogenicity indices. A total of 198 APEC isolates were evaluated for their biofilm-producing ability and extended-spectrum ß-lactamase (ESBL) production phenotypes. In addition, 36 virulence-associated genes were detected, and the isolates were classified into seven phylogenetic groups using polymerase chain reaction. The sources of the isolates were not associated with biofilms, ESBL, genes, or phylogroups. Biofilm and ESBL production were not associated with pathogenicity. Group B2 had the highest pathogenicity index. Groups B2 and E were positively associated with high-pathogenicity isolates and negatively associated with low-pathogenicity isolates. In contrast, groups A and C were positively associated with apathogenic isolates, and group B1 was positively associated with low-pathogenicity isolates. Some virulence-associated genes showed positive or negative associations with specific phylogenetic groups. None of the individual techniques produced results that correlated with the in vivo pathogenicity index. However, the combination of two techniques, namely, detection of virulence-associated genes and the phylogenetic groups, could help the classification of the isolates as pathogenic or commensal.

Respiratory microbiota of healthy broilers can act as reservoirs for multidrug-resistant Escherichia coli.

This study aimed at evaluate the presence and to study characteristics of Escherichia coli in the respiratory system microbiota of healthy broilers. Trachea, air sacs, and lungs of 20 broilers were analyzed at 21 days of age, reared in experimental conditions, without receiving antimicrobials. E. coli strains were isolated and identified using conventional bacteriology through morphological and biochemical characterization. The production of bacteriocin-like substances, the presence of virulence-associated genes (VAGs) of APEC (Avian Pathogenic Escherichia coli) predictors, and the antimicrobial susceptibility were evaluated. E. coli was found in 85 % of the animals (17/20), in the trachea, air sacs or lungs; and it was not found in 15 % of the animals (3/20). A total of 34 isolates were recovered, 13 from the air sacs, 13 from the lungs, and 8 from the trachea, which showed no production of bacteriocin-like substances nor virulence genes associated with APEC. Most isolates, 59 % (20/34), showed resistance to at least one of the tested antimicrobials, and six multiresistant strains were identified. The results demonstrated that strains of E. coli were commensal of the respiratory microbiota, and that they did not present pathogenicity to the host, since there were no clinical signs of disease, macroscopic lesions in the organs of the evaluated broilers, production of bacteriocin-like substances, nor virulence-associated genes considered as predictors of APEC in bacteria. These strains of E. coli were mostly susceptible to antimicrobials. However, the occurrence of multidrug-resistant strains suggests that these animals can act as reservoirs of resistant to antimicrobials E. coli.

Occurrence of genes associated with virulence in Escherichia coli isolates from chicken carcasses at different stages of processing at a slaughterhouse.

Escherichia coli is a bacterium frequently found in chicken carcasses, causing carcass condemnation with losses to the industry and when present in food, it carries a risk to public health as there is evidence that some strains pathogenic to birds (APEC - Avian Pathogenic E. coli) have zoonotic potential. Carcass contamination can occur at the slaughterhouse, but the influence of the different stages of processing in the selection of potential extraintestinal pathogenic E. coli strains is unknown. This study aimed to analyze the influence of the processing steps in the slaughterhouse on the detection of E. coli isolates carrying APEC predictor's virulence-associated genes (VAGs), and to relate their presence with post-mortem condemnation. A sample consisted of four pooled carcasses collected at seven different stages of slaughter (before scalding, after scalding, after plucking, before evisceration/after shower wash, after evisceration, after pre-coolers, and after packing) from 15 batches of broilers. The total samples obtained was 105 pools with four carcasses each, totaling 420 carcasses analyzed. Enterobacteriaceae were counted from each pool and E. coli were subsequently selected, which were submitted to pentaplex PCR to identify the five VAG APEC predictor's: iroN, ompT, hlyF, iss, and iutA. The Enterobacteriaceae count demonstrated a reduction of 4.25 log CFU per gram of carcass from the first to the last stage analyzed, with scalding and pre-cooling by immersion being the procedures that contributed most to this reduction. The presence of VAGs and potential APEC (presence of two or more of these gene predictors) was observed at all points evaluated in the slaughterhouse, which suggested that bacteria carrying these genes could reach the consumer.

Characterization of multidrug-resistant avian pathogenic Escherichia coli: an outbreak in canaries.

The canary (Serinus canaria) is appreciated for its beautiful song, colors, and docile temperament and drives a lucrative business. However, diseases caused by avian pathogenic Escherichia coli (APEC) compromise the health of canaries, and the inadequate antimicrobial treatment can lead to the emergence of resistant strains. This study aimed to characterize 21 isolates of E. coli obtained from canaries infected with colibacillosis during an outbreak in northern Paraná State, Brazil. APEC and diarrheagenic E. coli (DEC) virulence genes were screened for by polymerase chain reaction (PCR). All isolates were positive for the hlyF, iss, and ompT genes, which are characteristic of APEC. The iroN gene was found in 95.2% of isolates, and none had the iutA gene. The ipaH gene, characteristic of enteroinvasive E. coli (EIEC), was found in 71.4% of isolates, all belonging to the phylogenetic group B1. High genetic similarity (>95%) was found using enterobacterial repetitive intergenic consensus PCR (ERIC-PCR). The isolates belonged to serotypes O117:H4 (71.4%) and O1:H20 (23.8%). This is the first report of a clonal colibacillosis outbreak in canaries caused by APEC. All isolates were resistant to ampicillin, nalidixic acid, ciprofloxacin, enrofloxacin, norfloxacin, and tetracycline. The high rate of multidrug resistance in our study shows the importance of avoiding the inadequate antibiotic treatment. We suggest that further studies should be conducted to contribute to the understanding of colibacillosis in canaries since the health of animals is linked to human and environmental health, as defined by the concept of One Health.

Antimicrobial susceptibility and detection of virulence-associated genes of Escherichia coli and Salmonella spp. isolated from domestic pigeons (Columba livia) in Brazil.

Overpopulation of domestic pigeons is considered to be one of the major problems of urban centers, as these birds are responsible for the dissemination of relevant pathogens to animal and human health. The aim of this study was to detect potentially pathogenic Escherichia coli and Salmonella spp. in domestic pigeons captured in areas near silos used for grain and feed storage, analyzing the antimicrobial sensitivity and the presence of virulence-associated genes. We evaluated 41 pigeons. From each bird, cecal contents and a pool of viscera (heart, spleen, and liver) were collected. Fifty strains of E. coli and three strains of S. Typhimurium were isolated. The antimicrobial susceptibility assay showed that 2% of the isolates of E. coli were resistant to chloramphenicol and the combination of sulfamethoxazole + trimethoprim and 4% to tetracycline, doxycycline, and sulfonamide. The three S. Typhimurium strains were sensitive to all antimicrobials tested. The pathogenicity profile demonstrated that no E. coli isolates showed a STEC compatible profile. Regarding the APEC pathotype, all genes were observed in 8% of E. coli, 6% had only the iss gene and 4% presented ompT, hlyF, and iutA genes. invA, hilA, avrA, and lpfA genes were detected in 100% of Salmonella isolates. The sitC and pefA genes were only present in one strain and the remaining genes were detected in two. In conclusion, it was found that pigeons living in the vicinity of silos are carriers of important pathogens, and control measures should be taken to minimize animal and human health risks.

Detection of ESBL/AmpC-Producing and Fosfomycin-Resistant Escherichia coli From Different Sources in Poultry Production in Southern Brazil.

This study discussed the use of antimicrobials in the commercial chicken production system and the possible factors influencing the presence of Extended-spectrum ß-lactamase (ESBL)/AmpC producers strains in the broiler production chain. The aim of this study was to perform longitudinal monitoring of ESBL-producing and fosfomycin-resistant Escherichia coli from poultry farms in southern Brazil (Paraná and Rio Grande do Sul states) and determine the possible critical points that may be reservoirs for these strains. Samples of poultry litter, cloacal swabs, poultry feed, water, and beetles (Alphitobius sp.) were collected during three distinct samplings. Phenotypic and genotypic tests were performed for characterization of antimicrobial resistant strains. A total of 117 strains were isolated and 78 (66%) were positive for ESBL production. The poultry litter presented ESBL positive strains in all three sampled periods, whereas the cloacal swab presented positive strains only from the second period. The poultry litter represents a significant risk factor mainly at the beginning poultry production (odds ratio 6.43, 95% confidence interval 1-41.21, p < 0.05). All beetles presented ESBL positive strains. The predominant gene was bla CTX-M group 2, which occurred in approximately 55% of the ESBL-producing E. coli. The cit gene was found in approximately 13% of the ESBL-producing E. coli as AmpC type determinants. A total of 19 out of 26 fosfomycin-resistant strains showed the fosA3 gene, all of which produced ESBL. The correlation between fosA3 and bla CTX-M group 1 (bla CTX-M55 ) genes was significant among ESBL-producing E. coli isolated from Paraná (OR 3.66, 95% CI 1.9-9.68) and these genetic determinants can be transmitted by conjugation to broiler chicken microbiota strains. Our data revealed that poultry litter and beetles were critical points during poultry production and the presence of fosfomycin-resistant strains indicate the possibility of risks associated with the use of this antimicrobial during production. Furthermore, the genetic determinants encoding CTX-M and fosA3 enzymes can be transferred to E. coli strains from broiler chicken microbiota, thereby creating a risk to public health.

An Online Survey of Occupational Hazards in Brazilian Aquaculture.

Information on occupational health and safety practices in Brazilian aquaculture is limited. This paper reports preliminary results from an online survey based on research questions to identify occupational hazards, risk assessment practices, and prevention measures adopted in Brazilian aquaculture. Data were collected through an online questionnaire, comprising 25 questions, on a voluntary and anonymous basis. Aquaculture stakeholders were invited to participate in the study through email and social media channels. The demographic data demonstrated that the majority of respondents were men (72%) and having a higher education (95%). Most respondents employed administrative controls and personal protective equipment (PPE) rather than substitution and other risk elimination measures to reduce exposure. The most commonly adopted measures were PPE use (87%), adequate handling of chemicals (86%), and imparting knowledge of risks (90%). However, only 12% of participants reported the presence of safety protocols at their workplace, and 17% had some form of immunization of workers. In this study, it was possible to identify a lack of hazard signage in the workplace and lack of occupational health and safety training. The results further indicate that risk management in aquaculture continues to be a challenge in low-income countries. Aquaculture farmers should be encouraged and supported in adopting measures and appropriate technologies to eliminate risks in Brazilian aquaculture.

Distribution of ExPEC Virulence Factors, bla CTX-M, fosA3, and mcr-1 in Escherichia coli Isolated From Commercialized Chicken Carcasses.

Pathogenic Escherichia coli found in humans and poultry carcasses harbor similar virulence and resistance genes. The present study aimed to analyze the distribution of extraintestinal pathogenic E. coli (ExPEC) virulence factors (VF), bla CTX-M groups, fosA3, and mcr-1 genes in E. coli isolated from commercialized chicken carcasses in southern Brazil and to evaluate their pathogenic risk. A total of 409 E. coli strains were isolated and characterized for genes encoding virulence factors described in ExPEC. Results of antimicrobial susceptibility testing confirmed that the strains were resistant to ß-lactams, fosfomycin, colistin, and others resistance groups. The highest prevalence of VFs was observed in isolates belonging to the CTX-M groups, especially the CTX-M-2 group, when compared to those in other susceptible strains or strains with different mechanisms of resistance. Furthermore, ESBL strains were found to be 1.40 times more likely to contain three to five ExPEC virulence genes than non-ESBL strains. Our findings revealed the successful conjugation between ESBL-producing E. coli isolated from chicken carcass and the E. coli recipient strain J53, which suggested that genetic determinants encoding CTX-M enzymes may have originated from animals and could be transmitted to humans via food chain. In summary, chicken meat is a potential reservoir of MDR E. coli strains harboring resistance and virulence genes that could pose serious risks to human public health.

Occupational Health and Safety in Aquaculture: Insights on Brazilian Public Policies.

Aquaculture has many occupational hazards, including those that are physical, chemical, biological, ergonomic, and mechanical. The risks in aquaculture are inherent, as this activity requires particular practices. The objective of the present study was to show the risks associated with the aquaculture sector and present a critical overview on the Brazilian public policies concerning aquaculture occupational health. Methods include online research involved web searches and electronic databases including Pubmed, Google Scholar, Scielo and government databases. We conducted a careful revision of Brazilian labor laws related to occupational health and safety, rural workers, and aquaculture. The results and conclusion support the idea that aquaculture requires specific and well-established industry programs and policies, especially in developing countries. Aquaculture still lacks scientific research, strategies, laws, and public policies to boost the sector with regard to occupational health and safety. The establishment of a safe workplace in aquaculture in developing countries remains a challenge for all involved in employer-employee relationships.

Comparison of Antibiotic Resistance and Virulence Factors among Escherichia coli Isolated from Conventional and Free-Range Poultry.

Microbiological contamination in commercial poultry production has caused concerns for human health because of both the presence of pathogenic microorganisms and the increase in antimicrobial resistance in bacterial strains that can cause treatment failure of human infections. The aim of our study was to analyze the profile of antimicrobial resistance and virulence factors of E. coli isolates from chicken carcasses obtained from different farming systems (conventional and free-range poultry). A total of 156 E. coli strains were isolated and characterized for genes encoding virulence factors described in extraintestinal pathogenic E. coli (ExPEC). Antimicrobial susceptibility testing was performed for 15 antimicrobials, and strains were confirmed as extended spectrum of ß-lactamases- (ESBLs-) producing E. coli by phenotypic and genotypic tests. The results indicated that strains from free-range poultry have fewer virulence factors than strains from conventional poultry. Strains from conventionally raised chickens had a higher frequency of antimicrobial resistance for all antibiotics tested and also exhibited genes encoding ESBL and AmpC, unlike free-range poultry isolates, which did not. Group 2 CTX-M and CIT were the most prevalent ESBL and AmpC genes, respectively. The farming systems of poultries can be related with the frequency of virulence factors and resistance to antimicrobials in bacteria.

Prevalence of ColV Plasmid-Linked Genes and In Vivo Pathogenicity of Avian Strains of Escherichia coli.

Avian pathogenic Escherichia coli (APEC) causes extraintestinal infections in birds, leading to an increase in the cost of poultry production. The ColV plasmid-linked genes iroN, ompT, hlyF, iss, and iutA have previously been suggested to be predictors of the virulence of APEC. In this research, we analyzed the frequencies of these genes in a Brazilian collection of E. coli isolated from birds with colibacillosis (APEC) and from apparently healthy birds (avian fecal [A(fecal)]), as well as from the litter of poultry houses of apparently healthy flocks (avian litter [A(litter)]). All the isolates that harbored ompT also harbored hlyF, so they were considered as one trait for statistical analysis. The relationship between in vivo virulence in 1-day-old chicks, expressed as a pathogenicity score, and the number of genes in each isolate showed that isolates with less than two of the four genes were rarely pathogenic, while most pathogenic isolates contained two or more genes. Nevertheless, about half of the nonpathogenic isolates also harbored two or more genes, in agreement with previous observations that commensal E. coli isolates from the birds' microbiota can serve as a reservoir of virulence genes. Thus, the pentaplex polymerase chain reaction can be used to indicate that a strain carrying none or only one gene would be nonpathogenic, but it cannot be used to indicate that a strain with two to four genes would be an APEC. Isolates allocated to phylogenetic group B2, which is frequently associated with extraintestinal infections, had the highest pathogenicity scores, while isolates allocated to group B1 had the lowest.

Evaluation of the Antibiotic Resistance and Virulence of Escherichia coli Strains Isolated from Chicken Carcasses in 2007 and 2013 from Paraná, Brazil.

The frequent use of antimicrobials in commercial poultry production has raised concerns regarding the potential impact of antimicrobials on human health due to selection for resistant bacteria. Several studies have reported similarities between extraintestinal pathogenic Escherichia coli (ExPEC) strains isolated from birds and humans, indicating that these contaminant bacteria in poultry may be linked to human disease. The aim of our study was to analyze the frequency of antimicrobial resistance and virulence factors among E. coli strains isolated from commercial chicken carcasses in Paraná, Brazil, in 2007 and 2013. A total of 84 E. coli strains were isolated from chicken carcasses in 2007, and 121 E. coli strains were isolated in 2013. Polymerase chain reaction was used to detect virulence genes (hlyF, iss, ompT, iron, and iutA) and to determine phylogenetic classification. Antimicrobial susceptibility testing was performed using 15 antimicrobials. The strains were also confirmed as extended-spectrum ß-lactamase (ESBL)-producing E. coli with phenotypic and genotypic tests. The results indicated that our strains harbored virulence genes characteristic of ExPEC, with the iutA gene being the most prevalent. The phylogenetic groups D and B1 were the most prevalent among the strains isolated in 2007 and 2013, respectively. There was an increase in the frequency of resistance to a majority of antimicrobials tested. An important finding in this study was the large number of ESBL-producing E. coli strains isolated from chicken carcasses in 2013, primarily for the group 2 cefotaximase (CTX-M) enzyme. ESBL production confers broad-spectrum resistance and is a health risk because ESBL genes are transferable from food-producing animals to humans via poultry meat. These findings suggest that our strains harbored virulence and resistance genes, which are often associated with plasmids that can facilitate their transmission between bacteria derived from different hosts, suggesting zoonotic risks.

Molecular characterization and clonal relationships among Escherichia coli strains isolated from broiler chickens with colisepticemia.

This study characterized 52 Escherichia coli isolates from distinct diseased organs of 29 broiler chickens with clinical symptoms of colibacillosis in the Southern Brazilian state of Rio Grande do Sul. Thirty-eight isolates were highly virulent and 14 were virtually avirulent in 1-day-old chicks, yet all isolates harbored virulence factors characteristic of avian pathogenic E. coli (APEC), including those related to adhesion, iron acquisition, and serum resistance. E. coli reference collection phylogenetic typing showed that isolates belonged mostly to group D (39%), followed by group A (29%), group B1 (17%), and group B2 (15%). Phylogenetic analyses using the Amplified Ribosomal DNA Restriction Analysis and pulse-field gel electrophoresis methods were used to discriminate among isolates displaying the same serotype, revealing that five birds were infected with two distinct APEC strains. Among the 52 avian isolates, 2 were members of the pandemic E. coli O25:H4-B2-ST131 clone.

Detecção de Salmonella spp e Listeria monocytogenes através de técnica de PCR / Detection of Salmonella spp and Listeria monocytogenes through PCR / Detección de Salmonella spp y Listeria monocytogenes a través de la técnica de PCR

A maioria dos casos de doenças transmitidas por alimentos é causada por bactérias como Salmonella spp, Listeria monocytogenes, Campylobacter spp e Escherichia coli, entre outros. A contaminação decorre do consumo de alimentos contaminados preparados sob condições impróprias de higiene, manipulação e conservação. Para a detecção dessas bactérias em alimentos, o método convencional de isolamento é o mais utilizado, porém a técnica é demorada e laboriosa. A Reação em Cadeia da Polimerase (PCR) é um método rápido e sensível na detecção de agentes patogênicos e tem sido bastante estudada e gradativamente empregada nas indústrias de alimentos. O objetivo deste trabalho foi avaliar o limite de detecção da técnica de PCR, para Salmonella spp e L. monocytogenes. O limite para detecção das bactérias foram 2 e 5 UFC para Salmonella spp e L. monocytogenes, respectivamente, demonstrando uma boa sensibilidade da técnica de PCR...

Most cases of foodborne illnesses are caused by bacteria such as Salmonella spp, Listeria monocytogenes, Campylobacter spp, Escherichia coli, among others. Contamination stems from the consumption of contaminated food prepared under improper hygiene, handling and storage conditions. For the detection of bacteria in food, the conventional isolation method is the most commonly used, but the technique is labor-intensive and time consuming. The Polymerase Chain Reaction (PCR) is a rapid and sensitive method for the detection of pathogen agents, and has been extensively studied and gradually used in food industries. The objective of this study is to evaluate the sensitivity of the PCR method for the detection of Salmonella spp and L. monocytogenes. The threshold for the detection of bacteria was 2 and 5 CFU for Salmonella spp and L. monocytogenes, respectively, presenting a high sensitivity to PCR...

La mayoría de los casos de enfermedades transmitidas por alimentos es causada por bacterias como la Salmonella spp, Listeria monocytogenes, Campylobacter spp y Escherichia coli, entre otros. La contaminación se produce por el consumo de alimentos contaminados y preparados bajo condiciones inadecuadas de higiene, manipulación y conservación. Para la detección de esas bacterias en alimentos, el método convencional de aislamiento es el más utilizado, pero la técnica es laboriosa y consume mucho tiempo. La Reacción en Cadena de la Polimerasa (PCR) es un método rápido y sensible para la detección de patógenos y ha sido ampliamente estudiado y gradualmente empleada en las industrias de alimentos. El objetivo de este estudio fue evaluar el límite de detección de la técnica de PCR, para L. monocytogenes y Salmonella spp. El límite para detección de las bacterias fueron 2 y 5 UFC para Salmonella spp y L. monocytogenes, respectivamente, demostrando buena sensibilidad a la técnica de PCR...

Genotypes and pathogenicity of cellulitis isolates reveal traits that modulate APEC virulence.

We characterized 144 Escherichia coli isolates from severe cellulitis lesions in broiler chickens from South Brazil. Analysis of susceptibility to 15 antimicrobials revealed frequencies of resistance of less than 30% for most antimicrobials except tetracycline (70%) and sulphonamides (60%). The genotyping of 34 virulence-associated genes revealed that all the isolates harbored virulence factors related to adhesion, iron acquisition and serum resistance, which are characteristic of the avian pathogenic E. coli (APEC) pathotype. ColV plasmid-associated genes (cvi/cva, iroN, iss, iucD, sitD, traT, tsh) were especially frequent among the isolates (from 66.6% to 89.6%). According to the Clermont method of ECOR phylogenetic typing, isolates belonged to group D (47.2%), to group A (27.8%), to group B2 (17.4%) and to group B1 (7.6%); the group B2 isolates contained the highest number of virulence-associated genes. Clonal relationship analysis using the ARDRA method revealed a similarity level of 57% or higher among isolates, but no endemic clone. The virulence of the isolates was confirmed in vivo in one-day-old chicks. Most isolates (72.9%) killed all infected chicks within 7 days, and 65 isolates (38.1%) killed most of them within 24 hours. In order to analyze differences in virulence among the APEC isolates, we created a pathogenicity score by combining the times of death with the clinical symptoms noted. By looking for significant associations between the presence of virulence-associated genes and the pathogenicity score, we found that the presence of genes for invasins ibeA and gimB and for group II capsule KpsMTII increased virulence, while the presence of pic decreased virulence. The fact that ibeA, gimB and KpsMTII are characteristic of neonatal meningitis E. coli (NMEC) suggests that genes of NMEC in APEC increase virulence of strains.

Characterization of extraintestinal Escherichia coli isolated from a peacock (Pavo cristatus) with colisepticemia.

Extraintestinal infections by avian pathogenic strains of Escherichia coli (APEC) are commonly reported in poultry, but there is little information on infections by APEC in other bird species. Here we report on the characterization of extraintestinal E. coli isolated from a domesticated peacock, from the south of Brazil, that died of colisepticemia. Necropsy examination revealed congested liver, hypertrophied kidneys, peritonitis, severe typhlitis suggestive of coligranuloma, pneumonia, and airsacculitis--typical signs of colisepticemia. The isolates from lungs, kidney, heart, intestine, liver, and bone marrow all harbored the same virulence-associated factors (iucD, colV, iss, mat, fimC, ompA, traT crl, csgA vgrG, and hcp), yielded the same band pattern in amplified ribosomal DNA restriction analysis, and were allocated to the Escherichia coli Reference Collection group B1. The isolates were resistant to bacitracin, trimethoprim, and tetracycline, but displayed slight differences in their resistance to other antimicrobials. The isolates also differed in their virulence in 1-day-old chickens, but none displayed high virulence in vivo. We conclude that the peacock died of colisepticemia after it was infected with an extraintestinal E. coli strain of low virulence that nevertheless harbored virulence factors generally associated with APEC. This study represents the first characterization of an APEC isolated from a nonpoultry bird species.

Virulence factors and clonal relationships among Escherichia coli strains isolated from broiler chickens with cellulitis.

In this study, we compared Escherichia coli isolates from chickens with avian cellulitis with those from feces of healthy chickens. Cellulitis-derived strains presented phenotypic and genotypic characteristics of greater virulence than did the fecal isolates. Phylogenetic analysis by repetitive extragenic palindromic-PCR showed that, in agreement with their virulence characteristics, the cellulitis isolates form two clonal groups distinct from the fecal isolates.

Virulence-associated genes in Escherichia coli isolates from poultry with colibacillosis.

Avian pathogenic Escherichia coli, the causative agent of colibacillosis, harbors several putative virulence genes. In this study we examined by polymerase chain reaction (PCR) the presence of 16 of those genes in 200 colibacillosis isolates from our region. The seven virulence genes iutA, iss, cvaC, tsh, papC, papG and felA were detected significantly more often amongst colibacillosis isolates than in fecal isolates from healthy birds, thereby confirming their worldwide occurrence and possible pathogenic role in colibacillosis. However, several of those genes were not detected in many colibacillosis isolates, and none of them were detected in 27.5% of those isolates, which suggests that variants of those genes and yet undetected virulence factors should be searched for.