Detection of pathogenic, heteropathogenic and hybrid Escherichia coli strains in psittacines from zoos and breeders in the state of Ceará, Brazil.

The current study aimed to detect virulence, hetero-pathogenicity, and hybridization genes in Escherichia coli strains, previously isolated from cloacal swabs in commercial breeding psittacines and zoological collections, via multiplex PCR. A total of 68 strains of E. coli, previously isolated from psittacines in zoos and commercial breeding facilities in Ceará, Brazil, were assessed for the presence of the following genes and/or probes: eae, bfpA (EPEC - Enteropathogenic E. coli), CVD432 (EAEC - Enteroaggregative E. coli); LT gene and ST gene (ETEC - Enterotoxigenic E. coli); ipaH (EIEC - Enteroinvasive E. coli); stx1 and stx2 (STEC - Shiga toxin-producing E. coli); iroN, ompT, hlyF, iss, and iutA (APEC - Avian pathogenic E. coli). Of the 68 E. coli strains analyzed, 61 (98.7 %) were positive for the following genes and/or probes: Stx1 (61/98.7 %), ST gene (54/79.4 %), CVD432 (49/72 %), bfpA (44/64.7 %), eae (42/61.8 %), Stx2 (41/60.3 %), ipaH (34/50 %), LT gene (33/48.5 %), iroN (21/30.9 %), hlyF (11/6.2 %), iss (06/8.8 %) and iutA (06/8.8 %). The following diarrheagenic pathotypes were identified: 66 (97 %) from STEC, 49 (72 %) from EAEC, 35 (52 %) from EIEC, 25 (37 %) from ETEC, and one (1.5 %) from EPEC. Regarding hetero-pathogenicity, 50 (74 %) heterogeneous strains were identified. Positivity for APEC was seen in four (6 %) strains, all characterized as pathogenic hybrids. This study describes significant associations of virulence factors in E. coli strains DEC/DEC and DEC/APEC, which were isolated from psittacines and may be potentially harmful to One Health.

Detection of Mycoplasma spp. in free-living seabirds.

This study aimed to investigate the presence of Mycoplasma spp. and identify the species of mycoplasma isolates obtained from seabirds found on Brazilian coastal beaches. Tracheal and cloacal swab samples were collected from 50 seabirds rescued by three conservation and marine animal rehabilitation centers located in Brazil. The tracheal and cloacal samples were subjected to mycoplasma culture and the isolates were identified through PCR. A "Mollicutes-specific" 16S rRNA PCR reaction was employed for triage. Four species-specific PCR reactions were used to detect Mycoplasma gallisepticum, Mycoplasma synoviae, Mycoplasma meleagridis, or M. gallinarum. The Mollicutes positive and species negative samples were submitted do 16S rRNA sequencing. Eighteen (36%) of 50 seabirds tested positive for mycoplasma by culture. In the PCR for the genus, 28 (56%) of 50 seabirds were positive for Mycoplasma spp., with 13 (26%) detected in the trachea, one (2%) in the cloaca, and 14 (28%) in both sites. In the species-specific PCR, M. gallisepticum was detected in 17.8%, and M. meleagridis in 17.8%. Both species were detected in 14.3%. Of the isolates not characterized at species level, we obtained ten sequences and they were divided into three clusters. The first cluster was closely related to M. meleagridis, the second to M. synoviae, and the third grouped M. tully, M. gallisepticum, and M. imitans. Four and five of nine species of seabirds studied had mycoplasma detected by culture or PCR, respectively. Mycoplasmas were found in the majority of the animals studied, with the highest prevalence proportionally found in Sula leucogaster, and the lowest in Fregata magnificens. The phylogenetic analysis identified Mycoplasma spp. adapted to aquatic birds.

Population Dynamics of Thermotolerant Campylobacter in Broilers Reared on Reused Litter.

A study using sentinel broiler chickens was performed to address Campylobacter persistence in litter that was reused for successive flocks. Cloacal swabs, litter, drag swabs, darkling beetles, feed, and drinking water were weekly sampled and analyzed by standard microbiological procedures. Thermotolerant Campylobacter isolated strains were confirmed by polymerase chain reaction and subtyped by pulsed-field gel electrophoresis analysis. Campylobacter was not detected in samples collected immediately after downtime between broiler flocks. However, Campylobacter-positive samples were first detected at 21 d. After Campylobacter was initially isolated from the cloacal swabs, reused litter, drag swabs, or darkling beetles, these samples remained Campylobacter positive in the following weeks until the end of the rearing period. Campylobacter-positive cloacal swabs obtained from sentinel broilers ranged from 97.3% to 100% at 42 d. All isolated strains were identified as Campylobacter jejuni. Among the subtypes identified, an indistinguishable C. jejuni strain was predominant in sentinel broilers and was also detected in the other environmental samples analyzed, suggesting a common and persistent contamination source within the flocks. Sentinel broilers may have contributed to amplify the Campylobacter level, maintaining flock and broiler house contamination until the end of the production cycle.

Changes in dominant Escherichia coli and antimicrobial resistance after 24 hr in fecal matter.

Intestinal bacteria carry antimicrobial resistance (AMR) genes in mobile genetic elements which have the potential to spread to bacteria in other animal hosts including humans. In fecal matter, Escherichia coli can continue to multiply for 48 hr after being excreted, and in certain environments, E. coli survive long periods of time. It is unclear the extent to which AMR in E. coli changes in the environment outside of its host. In this study, we analyzed changes in the population structure, plasmid content, and AMR patterns of 30 E. coli isolates isolated from 6 chickens (cloacal swabs), and 30 E. coli isolates from fecal samples (from the same 6 chickens) after 24 hr of incubation. Clonality of isolates was screened using the fumC gene sequence and confirmed in a subset of isolates (n = 14) by multi-locus sequence typing. Major shifts in the population structure (i.e., sequence types) and antibiotic resistance patterns were observed among the numerically dominant E. coli isolates after 24 hr. Four E. coli clones isolated from the cloaca swabs and the corresponding fecal samples (after 24 hr incubation) showed different antibiotic resistance patterns. Our study reveals that fecal matter in the environment is an intermediate habitat where rapid and striking changes occur in E. coli populations and antibiotic resistance patterns.

Scavenger birds exploiting rubbish dumps: Pathogens at the gates.

Different bacteria are present in rubbish dumps used as food resources by various bird species. Birds may be good indicators of the presence of zoonotic diseases in these sites since they can be infected with zoonotic pathogens by foraging on organic waste, and can also act as carriers. We studied if foraging in rubbish dumps increases the occurrence of Salmonella spp. and Chlamydia psittaci in American black vultures (Coragyps atratus, hereafter black vultures) from northwest Patagonia. We compared these pathogens isolated from or detected in cloacae and oropharynx swabs in two different groups of black vultures: individuals trapped in (a) the Patagonian wild steppe and (b) in a rubbish dump. We found that black vultures are colonized by Salmonella spp. (particularly Salmonella enterica serotypes Typhi, Paratyphi A, Salmonella enterica subsp. arizonae) and Chlamydia psittaci. Interestingly, there were differences in the prevalence of Salmonella spp., especially Salmonella Typhi, between individuals foraging in the rubbish dump and the steppe, but not in the prevalence of Chlamydia psittaci. The pathogens isolated from black vultures may impact their health status but could also have health impacts in other bird species and even humans. In fact, Salmonella Typhi can cause severe disease in humans leading to death. Our results are globally relevant given that bacterial infections from rubbish dumps may affect different species exploiting these sites around the world. There is a need to better control pathogens in rubbish dumps to avoid the risk of infecting wildlife, which could act as potential dispersers and reservoirs of these pathogens.

Extended-spectrum cephalosporin-resistant Escherichia coli isolated from chickens and chicken meat in Brazil is associated with rare and complex resistance plasmids and pandemic ST lineages.

Objectives: Brazil is the greatest exporter of chicken meat (CM) in the world. It is of utmost importance to monitor resistance to extended-spectrum cephalosporins (ESCs) in this sector because resistance to ESCs in Escherichia coli isolated from food-producing animals may contaminate humans through the food chain. Thus, the aim of this study was to characterize and compare ESC-resistant E. coli isolated from chickens and retail CM produced in south-eastern Brazil. Methods: Five CM samples and 117 chicken cloacal swabs (CCSs) were inoculated on MacConkey agar supplemented with cefotaxime. Presumptive E. coli colonies were identified and antimicrobial susceptibility was tested. Virulence and acquired blaESBL and blaAmpC genes were sought and genetic environments characterized. Isolates were typed by phylogenetic grouping, XbaI-PFGE and MLST. Results: All five CM samples and 36 CCSs (30.8%) were positive for the presence of ESC-resistant E. coli, leading to the selection of 58 resistant isolates. ESC resistance was mostly due to the presence of the chromosome-encoded blaCTX-M-2 gene, but plasmid-mediated blaCTX-M-2, blaCTX-M-8, blaCTX-M-15, blaCTX-M-55 and blaCMY-2 were also detected. Multireplicon plasmids were sporadically identified, such as IncHI2/P-blaCTX-M-2 and IncFII/N-blaCTX-M-55. Phylogroup D predominated, while PFGE and MLST revealed a high genetic diversity. Conclusions: Live Brazilian chickens and CM act as reservoirs of ESC-resistant E. coli and resistance genes are located on highly diverse genetic determinants. Potentially pathogenic strains, which may represent a threat to human health and a source of environmental contamination, were also identified. Active surveillance is therefore essential in Brazil's chicken production line.

Oral and Cloacal Helicobacter Detection in Wild and Captive Orinoco Crocodiles (Crocodylus intermedius) in Venezuela.

Helicobacter species can colonize digestive tract of animals and humans and have been associated with gastrointestinal diseases; however, this genus has not been studied in crocodiles. Our objective was to detect by PCR Helicobacter genus and Helicobacter pylori in oral and cloacal swabs from Orinoco crocodiles of two wild (Cojedes River System and Capanaparo River) and two captive breeding centers (CBCs; Masaguaral Ranch and UNELLEZ) populations. Bacterial DNA was found in 100% of oral samples (10 wild and 10 captives), and in the 95% of cloacal samples (10 wild and 9 captives). In wild populations, Helicobacter spp. was not detected, whereas in CBCs, Helicobacter was detected in 10% of the oral samples, and 66.7% of cloacal samples. H. pylori was detected in two Orinoco crocodiles. Two cloacal non-pylori Helicobacter amplicons were sequenced, showing low similarity (≤97%) to Helicobacter sequences reported. This is the first report of Helicobacter species, including H. pylori in Crocodylus intermedius from CBCs.

Wild birds and urban pigeons as reservoirs for diarrheagenic Escherichia coli with zoonotic potential.

In order to describe the role of wild birds and pigeons in the transmission of shiga toxigenic Escherichia coli (STEC) and enteropathogenic Escherichia coli (EPEC) to humans and other animals, samples were collected from cloacae and oropharynx of free-living wild birds and free-living pigeons. Two STEC (0.8%) and five EPEC strains (2.0%) were isolated from wild birds and four EPEC strains (2.0%) were recovered from pigeons. Serogroups, sequence types (STs) and virulence genes, such as saa, iha, lpfA O113, ehxA, espA, nleB and nleE, detected in this study had already been implicated in human and animal diseases. Multidrug resistance (MDR) was found in 25.0% of the pigeon strains and in 57.0% of the wild bird strains; the wild birds also yielded one isolate carrying extended-spectrum ß-lactamases (ESBLs) gene bla CTX-M-8. The high variability shown by PFGE demonstrates that there are no prevalent E. coli clones from these avian hosts. Wild birds and pigeons could act as carriers of multidrug-resistant STEC and EPEC and therefore may constitute a considerable hazard to human and animal health by transmission of these strains to the environment.

Isolamento e determinação de sensibilidade e resistência a antimicrobianos de cepas bacterianas presentes na cloaca de Trachemys scripta elegans (Wied, 1839) criadas em cativeiro em Petrolina, PE / Isolation and determination of antimicrobials sensitivity and resistance of bacterial strains present in the cloaca Trachemys scripta elegans (Wied, 1839) raised in captivity in Petrolina/PE, Brazil

Este estudo isolou e determinou o perfil de sensibilidade e de resistência a antimicrobianos de cepas bacterianas isoladas da cloaca de Trachemys scripta elegans (T. s. elegans) criadas em cativeiro. Após 120 dias de adaptação, amostras de swab cloacal obtidas de 20 animais adultos foram cultivadas e, após a identificação dos patógenos através de testes bioquímicos, submetidas ao teste de suscetibilidade a nove antimicrobianos. Enterobacter aerogenes (85%); Shigella spp. (10%) e Edwadsiella spp. (5%) foram isolados e identificados. Os isolados de E. aerogenes foram sensíveis à gentamicina (86%), enrofloxacina (79%), estreptomicina (50%), sulfazotrim (36%) e ampicilina (29%) e resistentes a penicilina (100%), eritromicina (93%), cefalexina (86%) ampicilina (71%) e sulfazotrim (64%). Isolados de Shigella spp. apresentaram sensibilidade à gentamicina (100%), enrofloxacina (50%), doxicilina (50%), estreptomicina (50%), ampicilina (50%), penicilina (50%) e sulfazotrim (50%) e resistência a doxicilina (50%), estreptomicina (50%), ampicilina (50%), penicilina (100%), cefalexina (50%) e sulfazotrim (50%), enquanto que os de Edwardsiella spp. foram sensíveis apenas à gentamicina (100%) e altamente resistentes (100%) aos demais antimicrobianos. Os resultados sugerem a participação de T. s. elegans na cadeia epidemiológica, como reservatório de patógenos importantes, como E. aerogenes, Shigella spp. e Edwardisiella spp., tornando importante a adoção de medidas preventivas pelo risco zoonótico que apresentam e corretas de tratamento e de controle em cativeiros e domicílios, assim como de estudos que enfoquem as características de sensibilidade e de resistência antimicrobiana dos isolados cloacais, pois a multirresistência a drogas pode ser transmitida aos humanos e comprometer o tratamento de indivíduos com doenças graves.(AU)

This study isolated and determined the profile of susceptibility and antimicrobials resistance of bacterial strains isolated from the cloaca Trachemys scripta elegans (T. s. elegans) raised in captivity. After 120 days of adaptation, cloacal swab samples obtained from 20 adults animals were grown and, after the pathogens identification through biochemical tests, submitted to the test of susceptibility to nine antimicrobials. Enterobacter aerogenes (85%); Shigella spp. (10%) and Edwadsiella spp. (5%) were isolated and identified. Isolates from E. aerogenes were sensitive to gentamicin (86%), enrofloxacin (79%), streptomycin (50%), sulfazotrim (36%) and ampicillin (29%) and resistant to penicillin (100%), erythromycin (93%), cephalexin (86%), ampicillin (71%) and sulfazotrim (64%). Isolates from Shigella spp. showed sensitivity to gentamicin (100%), enrofloxacin (50%), doxycycline (50%), streptomycin (50%), ampicillin (50%), penicillin (50%) and sulfazotrim (50%) and resistance to doxycycline (50 %), streptomycin (50%), ampicillin (50%), penicillin (100%), cephalexin (50%) and sulfazotrim (50%), while the Edwardsiella spp. were sensitive only to gentamicin (100%) and were highly resistant (100%) to other antibiotics. The results suggest the participation of T. s. elegans in the epidemiological chain, as reservoir of important pathogens, such as E. aerogenes, Shigella spp. and Edwardisiella spp., making it important to adopt preventive measures for zoonotic risk that present and correct treatment and control in captivity and households, as well as studies that address the sensitivity characteristics and antimicrobial resistance of isolates from cloaca, as it multidrug resistance to drugs can be transmitted to humans and compromise the treatment of patients with serious diseases.(AU)

Captive and free-living urban pigeons (Columba livia) from Brazil as carriers of multidrug-resistant pathogenic Escherichia coli.

Thirty Escherichia coli isolates from captive and free-living pigeons in Brazil were characterised. Virulence-associated genes identified in pigeons included those which occur relatively frequently in avian pathogenic E. coli (APEC) from commercial poultry worldwide. Eleven of 30 E. coli isolates from pigeons, belonging mainly to B1 and B2 phylogenetic groups, had high or intermediate pathogenicity for 1-day-old chicks. The frequency of multi-drug resistant (MDR) E. coli in captive pigeons was relatively high and included one isolate positive for the extended-spectrum ß-lactamase (ESBL) gene blaCTX-M-8. Pulsed field gel electrophoresis (PFGE) showed high heterogeneity among isolates. There is potential for pigeons to transmit antibiotic resistant pathogenic E. coli to other species through environmental contamination or direct contact.

Detection of Salmonella enterica Serovar Montevideo and Newport in Free-ranging Sea Turtles and Beach Sand in the Caribbean and Persistence in Sand and Seawater Microcosms.

Salmonellae are Gram-negative zoonotic bacteria that are frequently part of the normal reptilian gastrointestinal flora. The main objective of this project was to estimate the prevalence of non-typhoidal Salmonella enterica in the nesting and foraging populations of sea turtles on St. Kitts and in sand from known nesting beaches. Results suggest a higher prevalence of Salmonella in nesting leatherback sea turtles compared with foraging green and hawksbill sea turtles. Salmonella was cultured from 2/9 and identified by molecular diagnostic methods in 3/9 leatherback sea turtle samples. Salmonella DNA was detected in one hawksbill turtle, but viable isolates were not recovered from any hawksbill sea turtles. No Salmonella was detected in green sea turtles. In samples collected from nesting beaches, Salmonella was only recovered from a single dry sand sample. All recovered isolates were positive for the wzx gene, consistent with the O:7 serogroup. Further serotyping characterized serovars Montevideo and Newport present in cloacal and sand samples. Repetitive-element palindromic PCR (rep-PCR) fingerprint analysis and pulsed-field gel electrophoresis of the 2014 isolates from turtles and sand as well as archived Salmonella isolates recovered from leatherback sea turtles in 2012 and 2013, identified two distinct genotypes and four different pulsotypes, respectively. The genotyping and serotyping were directly correlated. To determine the persistence of representative strains of each serotype/genotype in these environments, laboratory-controlled microcosm studies were performed in water and sand (dry and wet) incubated at 25 or 35°C. Isolates persisted for at least 32 days in most microcosms, although there were significant decreases in culturable bacteria in several microcosms, with the greatest reduction in dry sand incubated at 35°C. This information provides a better understanding of the epizootiology of Salmonella in free-ranging marine reptiles and the potential public health risks associated with human interactions with these animals in the Caribbean.

Detection and Molecular Characterization of Escherichia coli Strains Producers of Extended-Spectrum and CMY-2 Type Beta-Lactamases, Isolated from Turtles in Mexico.

Multidrug-resistant bacteria are a growing problem in different environments and hosts, but scarce information exists about their prevalence in reptiles. The aim of this study was to analyze the resistance mechanisms, molecular typing, and plasmid content of cefotaxime-resistant (CTX(R)) Escherichia coli isolates recovered from cloacal samples of 71 turtles sheltered in a herpetarium in Mexico. CTX(R)-E. coli were recovered in 11 of 71 samples (15.5%), and one isolate/sample was characterized. Extended-spectrum ß-lactamase (ESBL)-producing E. coli isolates were detected in four samples (5.6%): two strains carried the blaCTX-M-2 gene (phylogroup D and ST2732) and two contained the blaCTX-M-15 gene (phylogroup B1 and lineages ST58 and ST156). The blaCMY-2 gene was detected by PCR in E. coli isolates of eight samples (9.8%) (one of them also carried blaCTX-M-2); these isolates were distributed into phylogroups A (n = 1), B1 (n = 6), and D (n = 1) and typed as ST155, ST156, ST2329, and ST2732. Plasmid-mediated quinolone resistance (PMQR) genes were detected in five isolates [aac(6')Ib-cr, qnrA, qnrB19, and oqxB]. From three to five replicon plasmids were detected among the strains, being IncFIB, IncI1, IncFrep, and IncK the most prevalent. ESBL or pAmpC genes were transferred by conjugation in four strains, and the blaCTX-M-15 and blaCMY-2 genes were localized in IncFIB or IncI1 plasmids by Southern blot hybridization assays. Class 1 and/or class 2 integrons were detected in eight strains with six different structures of gene cassette arrays. Nine pulsed-field gel electrophoresis patterns were found among the 11 studied strains. To our knowledge, this is the first detection of ESBL, CMY-2, PMQR, and mobile determinants of antimicrobial resistance in E. coli of turtle origin, highlighting the potential dissemination of multidrug-resistant bacteria from these animals to other environments and hosts, including humans.

Cloacal enterobacteria isolated from captive roadside hawks (Rupornis magnirostris, GMELIN, 1788) and their antimicrobial susceptibility profile / Enterobactérias isoladas da cloaca de Gaviões-carijós (Rupornis magnirostris, GMELIN, 1788) cativos e seu perfil de susceptibilidade a antimicrobianos

Knowledge of the enterobacteria present in the roadside hawk can bring about an understanding of infectious diseases that can affect this bird, as well as other animals and/or humans, while also adding information of great ecological importance. Thus, the aim of the study was to determine the enterobacteria present in the cloaca of captive roadside hawks and antimicrobial susceptibility profile. Initially, cloacal samples from nine specimens were collected with the aid of swabs. Samples were placed in petri dishes with MacConkey agar, Hektoen agar, EMB agar and Salmonella-Shigella (SS) agar and incubated for 24 h at 35C. After incubation, the microorganisms were submitted to biochemical testing to confirm the presence of enterobacteria. Thereafter, the susceptibility profile of bacteria to antimicrobial agents was evaluated by a disk diffusion test according to the Clinical and Laboratory Standards Institute (CLSI). Escherichia coli (77.8%), Klebsiella oxytoca (11.1%), Klebsiella pneumoniae (11.1%) and Salmonella spp. (55.6%) were isolated from the collected samples. Among the isolates, some bacteria showed resistance to up to three antimicrobial agents. This study has brought greater insight about the enterobacteria present in the roadside hawk (Rupornis magnirostris), exhibiting a significant percentage of enterobacteria important to public health; also, it showed the occurrence of strains with resistance profile to antimicrobial agents...

O conhecimento das enterobactérias presentes em Gavião-carijó pode trazer uma compreensão sobre as doenças infecciosas que podem acometer essa ave, como também outros animais e/ou humanos, além de trazer mais informações sobre essa espécie de grande importância ecológica. Desta forma, o objetivo do estudo foi determinar as enterobactérias presentes na cloaca de Gaviões-carijós cativos e seu perfil de susceptibilidade a antimicrobianos. Inicialmente, foram coletadas amostras cloacais de nove espécimes com o auxílio de swabs. As amostras foram plaqueadas em ágar MacConkey, ágar Hektoen, ágar EMB e ágar SS e incubadas por 24 h a 35C. Após incubação, as colônias foram submetidas às provas bioquímicas para confirmação da presença de enterobactérias. Posteriormente, o perfil de susceptibilidade das bactérias frente a agentes antimicrobianos foi avaliado através do teste da difusão em disco de acordo com o Clinical and Laboratory Standards Institute (CLSI). Escherichia coli (77,8%), Klebsiella oxytoca (11,1%), Klebsiella pneumoniae (11,1%) e Salmonella spp. (55,6%) foram isoladas das amostras coletadas. Dentre as cepas isoladas, algumas apresentaram resistência a até três antimicrobianos. O presente estudo trouxe um maior conhecimento sobre as enterobactérias presentes no Gavião-carijó (Rupornis magnirostris), mostrando um percentual significativo de enterobactérias de importância na saúde pública, evidenciando também a ocorrência de cepas com perfil de resistência a agentes antimicrobianos...

Identificação de bacterias presentes na cloaca das espécies Amazona Aestiva e Amazona Amazonica mantidos em cativeiro no criatório conservacionista do Centro Universitário CESMAC, Alagoas / Identification of cloacal bacteria in Amazona Aestiva and Amazona amazonica kept in captivity in CESMAC University, Alagoas

Os animais selvagens, tanto na natureza quanto no cativeiro, podem ser reservatórios de zoonoses. O estudo da microbiota bacteriana de aves silvestres clinicamente saudáveis é um passo importante para a compreensão da epidemiologia das doenças bacterianas. Objetivou-se traçar o perfil de bactérias cloacais presentes em Psitacídeos das espécies Amazona aestiva e Amazona amazonica, mantidos em cativeiro no Criatório Conservacionista do Centro Universitário Cesmac, Marechal Deodoro, Alagoas. Foram colhidas 20 amostras de mucosa clocais de psitacídeos (10 da espécie Amazona aestiva e 10 da espécie Amazona amazonica) com o auxílio de suabes. As amostras foram semeadas em placas com Ágar Sangue (5%) e Ágar MacConkey, incubadas em estufa bacteriológica em aerobiose a 37°C por 24 horas, e em seguida avaliadas as características morfológicas das colônias. Para a avaliação microscópica as amostras foram coradas em Gram e, em seguida, observadas em microscópio. Todas as côlonias que cresceram, foram submetidas a testes bioquímicos. Na análise microbiológica foram identificadas 07 espécies de bactérias. Nas 10 amostras colhidas de suabes cloacais na espécie Amazona amazonica foram isoladas 07 espécies de bactérias, sendo 2 Klebsiella oxytoca, 1 Shigella sonnei, 5 Enterobacter aerogenes, 2 Enterobacter sakazakiie, 2 Yersinia pseudotuberculosis. Nas 10 amostras colhidas de cloaca na espécie Amazona aestiva, foram identificadas 04 espécies bacterianas distintas, sendo 2 Enterobacter aerogenes, 1 Salmonella typhi, 1 Klebsiella pneumoniae e 1 Shigella sonnei. A espécie bacteriana de maior frequência neste estudo foi Enterobacter aerogenes, encontrada nas duas espécies de psitacídeos estudadas. Como medida de controle, devam ser coletados suabes cloacais periodicamente, assim como aplicar cuidados com higiene, limpeza correta e frequente do recinto ou gaiola, bem como disponibilizar alimentação adequada para cada espécie. Os Psitacídeos em cativeiro têm o trato gastrointestinal colonizado por diferentes tipos de Enterobacteriaceae, sendo que algumas dessas espécies bacterianas possuem fundamental importância para a saúde única. Um risco para estes animais, que são submetidos ao estresse do cativeiro, que em condições adversas estas bactérias podem produzir infecção secundária, quase sempre fatal. Além disso, as duas espécies de psitacídeos estudadas são comumente criadas como pets em residências e podem apresentar bactérias com alto potencial zoonótico.

Cloacal enterobacteria isolated from captive roadside hawks (Rupornis magnirostris, GMELIN, 1788) and their antimicrobial susceptibility profile / Enterobactérias isoladas da cloaca de Gaviões-carijós (Rupornis magnirostris, GMELIN, 1788) cativos e seu perfil de susceptibilidade a antimicrobianos

Knowledge of the enterobacteria present in the roadside hawk can bring about an understanding of infectious diseases that can affect this bird, as well as other animals and/or humans, while also adding information of great ecological importance. Thus, the aim of the study was to determine the enterobacteria present in the cloaca of captive roadside hawks and antimicrobial susceptibility profile. Initially, cloacal samples from nine specimens were collected with the aid of swabs. Samples were placed in petri dishes with MacConkey agar, Hektoen agar, EMB agar and Salmonella-Shigella (SS) agar and incubated for 24 h at 35C. After incubation, the microorganisms were submitted to biochemical testing to confirm the presence of enterobacteria. Thereafter, the susceptibility profile of bacteria to antimicrobial agents was evaluated by a disk diffusion test according to the Clinical and Laboratory Standards Institute (CLSI). Escherichia coli (77.8%), Klebsiella oxytoca (11.1%), Klebsiella pneumoniae (11.1%) and Salmonella spp. (55.6%) were isolated from the collected samples. Among the isolates, some bacteria showed resistance to up to three antimicrobial agents. This study has brought greater insight about the enterobacteria present in the roadside hawk (Rupornis magnirostris), exhibiting a significant percentage of enterobacteria important to public health; also, it showed the occurrence of strains with resistance profile to antimicrobial agents(AU)

O conhecimento das enterobactérias presentes em Gavião-carijó pode trazer uma compreensão sobre as doenças infecciosas que podem acometer essa ave, como também outros animais e/ou humanos, além de trazer mais informações sobre essa espécie de grande importância ecológica. Desta forma, o objetivo do estudo foi determinar as enterobactérias presentes na cloaca de Gaviões-carijós cativos e seu perfil de susceptibilidade a antimicrobianos. Inicialmente, foram coletadas amostras cloacais de nove espécimes com o auxílio de swabs. As amostras foram plaqueadas em ágar MacConkey, ágar Hektoen, ágar EMB e ágar SS e incubadas por 24 h a 35C. Após incubação, as colônias foram submetidas às provas bioquímicas para confirmação da presença de enterobactérias. Posteriormente, o perfil de susceptibilidade das bactérias frente a agentes antimicrobianos foi avaliado através do teste da difusão em disco de acordo com o Clinical and Laboratory Standards Institute (CLSI). Escherichia coli (77,8%), Klebsiella oxytoca (11,1%), Klebsiella pneumoniae (11,1%) e Salmonella spp. (55,6%) foram isoladas das amostras coletadas. Dentre as cepas isoladas, algumas apresentaram resistência a até três antimicrobianos. O presente estudo trouxe um maior conhecimento sobre as enterobactérias presentes no Gavião-carijó (Rupornis magnirostris), mostrando um percentual significativo de enterobactérias de importância na saúde pública, evidenciando também a ocorrência de cepas com perfil de resistência a agentes antimicrobianos(AU)

Evidence of Fluconazole-Resistant Candida Species in Tortoises and Sea Turtles.

The aim of this study was to evaluate the antifungal susceptibility of Candida spp. recovered from tortoises (Chelonoidis spp.) and sea turtles (Chelonia mydas, Caretta caretta, Lepidochelys olivacea, Eretmochelys imbricata). For this purpose, material from the oral cavity and cloaca of 77 animals (60 tortoises and 17 sea turtles) was collected. The collected specimens were seeded on 2% Sabouraud dextrose agar with chloramphenicol, and the identification was carried out by morphological and biochemical methods. Sixty-six isolates were recovered from tortoises, out of which 27 were C. tropicalis, 27 C. famata, 7 C. albicans, 4 C. guilliermondii and 1 C. intermedia, whereas 12 strains were obtained from sea turtles, which were identified as Candida parapsilosis (n = 4), Candida guilliermondii (n = 4), Candida tropicalis (n = 2), Candida albicans (n = 1) and Candida intermedia (n = 1). The minimum inhibitory concentrations for amphotericin B, itraconazole and fluconazole ranged from 0.03125 to 0.5, 0.03125 to >16 and 0.125 to >64, respectively. Overall, 19 azole-resistant strains (14 C. tropicalis and 5 C. albicans) were found. Thus, this study shows that Testudines carry azole-resistant Candida spp.

Detection of bacteria and fungi and assessment of the molecular aspects and resistance of Escherichia coli isolated from confiscated passerines intended for reintroduction programs.

Many native bird species are currently considered rare in Brazil because they have been indiscriminately collected by animal traffickers and commercialized, leading to dwindling numbers in their natural habitats. Confiscated animals are at times destined for reintroduction programs that must ensure these animals do not pose a risk to native populations. Healthy or sick wild passerines may carry a great diversity of microorganisms. Therefore, knowledge of the sanitary status of confiscated animals destined for reintroduction is critical to assess whether these animals act as microorganism carriers and to investigate the epidemiology of transmissible diseases, a crucial aspect for animal and human health preservation. This study examined the occurrence of aerobic and facultative anaerobic bacteria and fungi in cloacal swabs collected from wild confiscated passerines intended for reintroduction programs. In vitro susceptibility tests of the most frequent isolates as well as studies of the molecular aspects of Escherichia coli isolates were also performed. There was microorganism growth in 62.5% of 253 samples. The microorganisms that were most frequently isolated were Staphylococcus spp. (15.0%), Micrococcus spp. (11.5%), E. coli (10.7%) and Klebsiella spp. (10.7%). Fifteen bacteria genera and seven fungi genera were isolated. Multidrug-resistance to antimicrobials was observed in Staphylococcus spp., Micrococcus spp., E. coli and Klebsiella spp. isolates. The high occurrence of Enterobacteria observed is possibly related to the sanitary conditions in which confiscated animals are usually kept. One E. coli sample (out of 27 isolates) was positive for the S-fimbrial adhesion encoding gene (sfa). Considering the low occurrence of genes that encode virulence factors, confiscated passerines may represent a low risk for the potential transmission of EPEC, APEC, UPEC and NMEC isolates to other animals or humans. The potential risk of intra- or inter-specific transmission of multidrug-resistant isolates and the introduction of these microorganisms into the environment must be considered, although there are still therapeutic alternatives for treatment of these animals among the antimicrobials which were tested. The stress and poor hygiene conditions imposed on animals during trafficking may have caused their contamination by multidrug-resistant agents transmitted by humans or by the precarious environment to which they were subjected. Risks related to the dissemination of Salmonella spp., Cryptococcus spp. and Candida spp. are low when reintroduction programs are considered.

Identification and antimicrobial resistance of members from the Enterobacteriaceae family isolated from canaries (Serinus canaria) / Identificação e resistência antimicrobiana de membros da família Enterobacteriaceae isolados de canários (Serinus canaria)

The Enterobacteriaceae family contains potentially zoonotic bacteria, and their presence in canaries is often reported, though the current status of these in bird flocks is unknown. Therefore, this study aimed to identify the most common genera of enterobacteria from canaries (Serinus canaria) and their antimicrobial resistance profiles. From February to June of 2013, a total of 387 cloacal swab samples from eight domiciliary breeding locations of Fortaleza city, Brazil, were collected and 58 necropsies were performed in canaries, which belonged to the Laboratory of Ornithological Studies. The samples were submitted to microbiological procedure using buffered peptone water and MacConkey agar. Colonies were selected according to their morphological characteristics on selective agar and submitted for biochemical identification and antimicrobial susceptibility. A total of 61 isolates were obtained, of which 42 were from cloacal swabs and 19 from necropsies. The most isolated bacteria was Escherichia coli with twenty five strains, followed by fourteen Klebsiellaspp., twelve Enterobacterspp., seven Pantoea agglomerans, two Serratiaspp. and one Proteus mirabilis. The antimicrobial to which the strains presented most resistance was sulfonamides with 55.7%, followed by ampicillin with 54.1% and tetracycline with 39.3%. The total of multidrug-resistant bacteria (MDR) was 34 (55.7%). In conclusion, canaries harbor members of the Enterobacteriaceae family and common strains present a high antimicrobial resistance rate, with a high frequency of MDR bacteria.

A família Enterobacteriaceae possui bactérias com potencial zoonótico e a presença destas bactérias em canários é relatada na literatura, porém a realidade dos plantéis de criadores de canários é desconhecida. Portanto, este trabalho teve como objetivo isolar enterobactérias de canários belga (Serinus canarius) com o intuito de conhecer os gêneros mais comuns nestas aves e suas respectivas resistências a antimicrobianos. De fevereiro a junho de 2013 foram coletadas 387 amostras de swabs cloacais de canários de oito propriedades da cidade de Fortaleza, Brasil e de 58 necropsias de aves do acervo próprio do Laboratório de Estudos Ornitológicos. As amostras foram submetidas a isolamento microbiológico utilizando-se água peptonada e ágar MacConkey. As colônias foram selecionadas de acordo com suas características morfológicas nas placas, submetidas à tipificação bioquímica para identificação e ao teste de sensibilidade a antimicrobianos. Foram isoladas 61 cepas, sendo 42 de suabes cloacais e 19 de necropsias. A bactéria mais isolada foi Escherichia coli com vinte e cinco cepas, seguida por catorze Klebsiella spp., doze Enterobacter spp., sete Pantoea agglomerans, duas Serratiaspp. e uma cepa de Proteus mirabilis. As cepas apresentaram maior resistência a sulfonamidas com 55,7%, seguidas por ampicilina com 54,1% e tetraciclina com 39,3%. Além disso, o total de cepas resistentes a múltiplas drogas (RMD) foi 34 (55,7%). Portanto, conclui-se que os canários albergam enterobactérias e que as cepas apresentam alto índice de resistência a antimicrobianos, com alta frequência de cepas RMD.

Identification and antimicrobial resistance of members from the Enterobacteriaceae family isolated from canaries (Serinus canaria) / Identificação e resistência antimicrobiana de membros da família Enterobacteriaceae isolados de canários (Serinus canaria)

The Enterobacteriaceae family contains potentially zoonotic bacteria, and their presence in canaries is often reported, though the current status of these in bird flocks is unknown. Therefore, this study aimed to identify the most common genera of enterobacteria from canaries (Serinus canaria) and their antimicrobial resistance profiles. From February to June of 2013, a total of 387 cloacal swab samples from eight domiciliary breeding locations of Fortaleza city, Brazil, were collected and 58 necropsies were performed in canaries, which belonged to the Laboratory of Ornithological Studies. The samples were submitted to microbiological procedure using buffered peptone water and MacConkey agar. Colonies were selected according to their morphological characteristics on selective agar and submitted for biochemical identification and antimicrobial susceptibility. A total of 61 isolates were obtained, of which 42 were from cloacal swabs and 19 from necropsies. The most isolated bacteria was Escherichia coli with twenty five strains, followed by fourteen Klebsiellaspp., twelve Enterobacterspp., seven Pantoea agglomerans, two Serratiaspp. and one Proteus mirabilis. The antimicrobial to which the strains presented most resistance was sulfonamides with 55.7%, followed by ampicillin with 54.1% and tetracycline with 39.3%. The total of multidrug-resistant bacteria (MDR) was 34 (55.7%). In conclusion, canaries harbor members of the Enterobacteriaceae family and common strains present a high antimicrobial resistance rate, with a high frequency of MDR bacteria.(AU)

A família Enterobacteriaceae possui bactérias com potencial zoonótico e a presença destas bactérias em canários é relatada na literatura, porém a realidade dos plantéis de criadores de canários é desconhecida. Portanto, este trabalho teve como objetivo isolar enterobactérias de canários belga (Serinus canarius) com o intuito de conhecer os gêneros mais comuns nestas aves e suas respectivas resistências a antimicrobianos. De fevereiro a junho de 2013 foram coletadas 387 amostras de swabs cloacais de canários de oito propriedades da cidade de Fortaleza, Brasil e de 58 necropsias de aves do acervo próprio do Laboratório de Estudos Ornitológicos. As amostras foram submetidas a isolamento microbiológico utilizando-se água peptonada e ágar MacConkey. As colônias foram selecionadas de acordo com suas características morfológicas nas placas, submetidas à tipificação bioquímica para identificação e ao teste de sensibilidade a antimicrobianos. Foram isoladas 61 cepas, sendo 42 de suabes cloacais e 19 de necropsias. A bactéria mais isolada foi Escherichia coli com vinte e cinco cepas, seguida por catorze Klebsiella spp., doze Enterobacter spp., sete Pantoea agglomerans, duas Serratiaspp. e uma cepa de Proteus mirabilis. As cepas apresentaram maior resistência a sulfonamidas com 55,7%, seguidas por ampicilina com 54,1% e tetraciclina com 39,3%. Além disso, o total de cepas resistentes a múltiplas drogas (RMD) foi 34 (55,7%). Portanto, conclui-se que os canários albergam enterobactérias e que as cepas apresentam alto índice de resistência a antimicrobianos, com alta frequência de cepas RMD.(AU)