Assessing the Microbial Quality of Shrimp (Xiphonaeus kroyeri) and Mussels (Perna perna) Illegally Sold in the Vitória Region, Brazil, and Investigating the Antimicrobial Resistance of Escherichia coli Isolates.

The consumption of seafood is crucial for food security, but poor hygiene along the food production chain can result in low microbiological quality, posing significant risks for public health and seafood quality. Thus, this study aimed to assess the microbiological quality and antimicrobial sensitivity of E. coli from 69 samples of illegally marketed shrimp and mussels in the Vitória Region, Brazil. These foods exhibited poor microbiological quality due to high counts of mesophilic, psychrotrophic, and enterobacteria microorganisms. While this issue is widespread in this area, shrimp samples displayed higher microbial counts compared to mussels, and fresh mussels had elevated counts of enterobacteria compared to frozen ones. Among the 10 E. coli isolates, none carried the genes blaCTX-M-1, blaCTX-M-2, blaCTX-M-3, blaCTX-M-15, mcr-1, mcr-2, mcr-3, mcr-4, and tet, associated with antibiotic resistance. Phenotypical resistance to tetracycline and fosfomycin was not observed in any isolate, while only 20% demonstrated resistance to ciprofloxacin. Regarding ampicillin and amoxicillin with clavulanic acid, 60% of isolates were resistant, 10% showed intermediate susceptibility, and 30% were sensitive. One isolate was considered simultaneously resistant to ß-lactams and quinolones, and none were conserved as ESBL producers. These findings highlight the inherent risks to local public health that arise from consuming improperly prepared seafood in this area.

The importance of the intestinal microbiota in humans and dogs in the neonatal period.

The neonatal period represents a critical stage for the establishment and development of the gut microbiota, which profoundly influences the future health trajectory of individuals. This review examines the importance of intestinal microbiota in humans and dogs, aiming to elucidate the distinct characteristics and variations in the composition between these two species. In humans, the intestinal microbiota contributes to several crucial physiological processes, including digestion, nutrient absorption, immune system development, and modulation of host metabolism. Dysbiosis, an imbalance or disruption of the gut microbial community, has been linked to various disorders, such as inflammatory bowel disease, obesity, and even neurological conditions. Furthermore, recent research has unveiled the profound influence of the gut-brain axis, emphasizing the bidirectional communication between the gut microbiota and the central nervous system, impacting cognitive function and mental health. Similarly, alterations in the canine intestinal microbiota have been associated with gastrointestinal disorders, including chronic enteropathy, such as inflammatory bowel disease, food allergies, and ulcerative histiocytic colitis. However, our understanding of the intricacies and functional significance of the intestinal microbiota in dogs remains limited. Understanding the complex dynamics of the intestinal microbiota in both humans and dogs is crucial for devising effective strategies to promote health and manage disease. Moreover, exploring the similarities and differences in the gut microbial composition between these two species can facilitate translational research, potentially leading to innovative therapeutic interventions and strategies to enhance the well-being of both humans and dogs.

Molecular analysis of enteropathogenic Escherichia coli (EPEC) isolates from healthy food-producing animals and humans with diarrhoea.

Enteropathogenic Escherichia coli (EPEC) is a pathogen associated with acute diarrhoea in humans. To determine whether EPEC isolated from healthy food-producing animals possesses the same virulence gene repertoire as EPEC isolated from human with diarrhoea, we compared six typical EPEC (tEPEC) and 20 atypical EPEC (aEPEC) from humans with diarrhoea and 42 aEPEC from healthy animals (swine, sheep and buffaloes), using pulsed-field gel electrophoresis (PFGE), virulence markers, serotyping and subtyping of eae and tir genes. We found that human and animal isolates shared virulence genes, including nleB, nleE and nleF, which were associated with human diarrhoea. Serogroups and serotypes identified in isolates of food-producing animals such as O26:H11, O128:H2, O76:H7, O103, O108, O111 and O145, have previously been implicated in human disease. The subtypes eae and tir were also shared between human and animal isolates, being eae-γ1 and eae-ß1 the most prevalent in both groups, while the most common tir subtypes were α and ß. Despite PFGE analysis demonstrating that EPEC strains are heterogeneous and there was no prevalent clone identified, EPEC isolated from humans and food-producing animals shared some characteristics, such as virulence genes associated with human diarrhoea, indicating that food-producing animals could play a role as reservoirs for those genes.

Comparative genomics study of Staphylococcus aureus isolated from cattle and humans reveals virulence patterns exclusively associated with bovine clinical mastitis strains.

Staphylococcus aureus causes nosocomial and intramammary infections in humans and cattle, respectively. A large number of virulence factors are thought to play important roles in the pathogenesis of this bacterium. Currently, genome-wide and data-analysis studies are being used to better understand its epidemiology. In this study, we conducted a genome wide comparison and phylogenomic analyses of S. aureus to find specific virulence patterns associated with clinical and subclinical mastitis strains in cattle and compare them with those of human origin. The presence/absence of key virulence factors such as adhesin, biofilm, antimicrobial resistance, and toxin genes, as well as the phylogeny and sequence type of the isolates were evaluated. A total of 248 genomes (27 clinical mastitis, 43 subclinical mastitis, 21 milk, 53 skin-related abscesses, 49 skin infections, and 55 pus from cellulitis) isolated from 32 countries were evaluated. We found that the cflA, fnbA, ebpS, spa, sdrC, coa, emp, vWF, atl, sasH, sasA, and sasF adhesion genes, as well as the aur, hglA, hglB, and hglC toxin genes were highly associated in clinical mastitis strains. The strains had diverse genetic origins (72 protein A and 48 sequence types with ST97, ST8 and ST152 being frequent in isolates from clinical mastitis, abscess, and skin infection, respectively). Further, our phylogenomic analyses suggested that zoonotic and/or zooanthroponotic transmission may have occurred. These findings contribute to a better understanding of S. aureus epidemiology and the relationships between adhesion mechanisms, biofilm formation, antimicrobial resistance, and toxins and could aid in the development of improved vaccines and strain genotyping methods.

An anatomical training model for cerebrospinal liquid collection and myelography in embalmed dog cadavers / Modelo anatômico para treinamento de colheita de líquido cerebroespinhal e mielografia em cadáveres de cães quimicamente preparados

In research and academic activities, guidelines are essential and imperative especially on the use of animals. Alternative methods that do not bring academic or scientific harm should also be sought. This study aimed to develop a training model for the collection of cerebrospinal fluid (CSF) and myelography in the cervical and lumbar regions in cadavers of embalmed dogs, using an alcoholic solution and curing salts for fixation and conservation. The dogs were divided into 4 grups of 8 animal each and stored between 2ºC and 6ºC, for 30, 60, 90, or 120 days. Durotomy was performed to implant two urethral catheters (one in the cranial direction and another in the caudal direction to the spinal cord access site), in the subduraracnoid space. This space was fixed via manual infusion of saline solution with a 20-mL syringe to simulate the presence of the CSF and the positive pressure, while the puncture was made. Four cadavers of each group were randomly selected for the CSF puncture from the atlantooccipital joint and in the lumbar region between L5 and L6, respectively, and four were used for CSF puncture training, in which radiographic contrast (myelography) was injected in the same locations. This model was cost-effective, did not utilize toxic products, and can preserve cadavers for up to 120 days. In this novel anatomical model, a maximum of 15 students can be trained on CSF puncture, allowing cervical and lumbar myelography and at least 30 perforations per cadaver.

É essencial e imperioso ter critério quanto ao uso de animais em pesquisa e atividades de ensino e, consequentemente, buscar métodos alternativos que não causem prejuízo acadêmico ou científico. Para que não ocorra deterioração dos tecidos, a fixação e conservação de peças anatômicas e cadáveres devem ser realizadas. Objetivou-se, com este estudo, desenvolver um modelo anatômico para treinamento de colheita de líquido cerebroespinhal (LCE) e mielografia, nas regiões cervical e lombar. Os cães foram divididos em quatro grupos contendo oito animais cada e armazenados entre 2ºC e 6ºC, por 30, 60, 90 ou 120 dias. Foi realizada durotomia para implantação de duas sondas uretrais, no espaço subaracnóide. A infusão manual de solução fisiológica com seringa de 20 mL foi utilizada para simular a presença do LCE e a pressão positiva, enquanto era feita a punção. Quatro cadáveres de cada grupo foram selecionados para a punção de LCE na articulação atlantooccipital e na região lombar entre L5 e L6, e quatro foram utilizados para o treinamento da punção de LCE e injeção de contraste radiográfico (mielografia). A técnica anatômica empregada possibilitou o desenvolvimento de um modelo visando ao ensino e pesquisa da radiologia em cadáveres de cães quimicamente preparados, a custo baixo e sem utilização de produtos tóxicos, mantidos sob refrigeração por 120 dias. Com isso, um máximo de 15 alunos podem ser treinados em punção do LCR, permitindo mielografia cervical e lombar com 30 perfurações por cadáver.

First Report of Bartonella spp. in Marsupials from Brazil, with a Description of Bartonella harrusi sp. nov. and a New Proposal for the Taxonomic Reclassification of Species of the Genus Bartonella.

The genus Bartonella (Rhizobiales: Bartonellaceae) encompasses facultative intracellular Gram-negative alphaproteobacteria that parasitize mainly erythrocytes and endothelial cells, as well as macrophages, monocytes and dendritic cells. Although they can infect numerous mammal species and arthropod vectors worldwide, reports of Bartonella infections in marsupials are scarce. In fact, such agents have only been detected in marsupials and/or associated ectoparasites in Australia and the United States of America until the present moment. The present study aimed to isolate and characterize molecularly, morphologically and phenotypically Bartonella infecting free-living marsupials sampled in the Brazilian Pantanal, the largest wetland in South America. Two marsupials were captured in December 2018 and six marsupials in February 2019, totaling eight small mammals sampled: five (62.5%) Thylamys macrurus and three (37.5%) Monodelphis domestica. All blood samples were submitted to qPCR for Bartonella spp. based on the nuoG gene, a pre-enrichment liquid culture and a chocolate agar solid culture. Bartonella sp. was isolated from 3 T. macrurus and one M. domestica. One Bartonella isolate obtained from a T. macrurus blood sample (strain 117A) that showed to be closely related to the Bartonella vinsonii complex and Bartonella machadoae was selected for whole genome sequencing using a hybrid approach based on Illumina NovaSeq and Nanopore sequencing platforms. This strain showed a genome of 2.35 Mbp, with an average C + G content of 38.8%, coding for 2013 genes, and a 29 kb plasmid with an average C + G content of 34.5%. In addition, this strain exhibited an average nucleotide identity (ANI) of 85% with Bartonella species belonging to the B. vinsonii group and 91% with B. machadoae. Phylogenomic analysis based on 291 protein coding genes shared by the genomes of 53 Bartonella species positioned this strain closely to B. machadoae. This new isolated species was named Bartonella harrusi sp. nov., which was characterized as having small capnophilic, microaerophilic and aerobic rods with an absence of pili and flagella. In conclusion, the present work describes the biochemical, phenotypic and genomic characteristics of Bartonella harrusi, a new species isolated from the T. macrurus blood samples of the Brazilian Pantanal. Finally, a review of the taxonomic classification of members of the genus Bartonella is proposed, based on the ANI values accessed by whole genome sequencing analyses.

Macroscopic, Histological, and Microbiological Characterization of Contact Lesions at the Tibiotarsal Region of Broilers.

Brazil is considered as a great broiler feet exporter, especially for the Chinese trade. Contact lesions at the tibiotarsal region are responsible for economic losses and there is no model for its classification, thereby this study presents a fast and practical grade system to be used in the poultry industry and proposes these lesion characterizations into three different grades. For this, correlation was made between macroscopic, histological findings and microbiological quantification (Escherichia coli, Staphylococcus spp., Streptococcus spp. and sulphite-reducing clostridia) from contact lesions in the tibiotarsal region of 112 broiler carcasses, divided in four groups (n=28), accordingly to the lesion's intensity. There were no significant differences in microbiological quantification among the groups (p>0.05) except for the grade 3 group, as grade 1 and 2 lesions were in the early stages and histopathological changes such as ulceration were not observed. In grade 3 lesion group, it was observed bacterial cocci grume and ulceration at the articular region and significantly higher microbiological count (p<0.05) for E. coli and Staphylococcus spp. In conclusion, the visual standard proposed in this work, correlated and confirmed by the histopathologic, and microbiologic characterization, allows to precise and fast ascertainment of the contact lesion grade in the tibiotarsal regions of broiler carcasses. Moreover, it should be highlighted that grades 1 and 2 alterations are not caused by an inflammatory process caused by pathogenic agents and should not be considered a public health risk.

Bartonella machadoae sp. nov. isolated from wild rodents in the Pantanal wetland.

It has been estimated that 75% of emerging infectious diseases comprise zoonoses, whose majority have free-living animals as reservoirs and are mainly transmitted by arthropod vectors. Although rodents represent important Bartonella reservoirs, there are few studies on the genotypic characterization of Bartonella species commonly found in this taxon and from different Brazilian biomes. Therefore, the present study aimed to investigate the occurrence, isolate and molecularly, morphologically and phenotypically characterize a new Bartonella species infecting free-living rodents sampled in the Brazilian Pantanal, the largest wetland in South America. For this purpose, 129 free-living rodents (79 Thrichomys fosteri, 4 Clyomys laticeps, and Oecomys mamorae) were captured. While blood samples were collected from 57 T. fosteri, 4 C. laticeps and 32 O. mamorae; spleen samples were collected from 22 T. fosteri and 14 O. mamorae. Blood and spleen samples were submitted to a qPCR for Bartonella spp. targeting the nuoG gene, using DNA samples extracted directly from blood/spleen, after passage in pre-enrichment liquid culture, and from colonies obtained from solid culture on chocolate agar. Combining all techniques, occurrence of 24.8% for Bartonella sp. was found among the sampled rodents. One Bartonella isolate (strain 56A) obtained from a T. fosteri's blood sample was closely related to the Bartonella vinsonii complex and selected for Whole Genome Sequencing (WGS) hybrid approach using Illumina NovaSeq and Nanopore sequencing platforms. This strain exhibits a circular 2.7 Mbp genome with an average C+G content of 39% and encoding to 2239 genes. In the phylogenomics based on 291 shared protein-coding genes, this strain was positioned in a unique clade, closely related to Bartonella vinsonii subsp. vinsonii, B. vinsonii subsp. berkhoffii and B. visonii subsp. arupensis. An Average Nucleotide Identity of 85% was found between the obtained isolate and Bartonella species belonging to B. vinsonii complex. These findings supported the separation of this strain, now formally named as Bartonella machadoae sp. nov., from the Bartonella vinsonii complex. In addition, Bartonella machadoae sp. nov. was characterized by capnophilic, microaerophilic and aerobic small rods with absence of pili and flagella. Phylogenetic and distance analyses based on five concatenated molecular markers suggest that Bartonella machadoae may parasite rodents from different Brazilian biomes. In conclusion, we described biochemical, phenotypic and genomic characteristics of Bartonella machadoae nov. sp. isolated from blood samples of T. fosteri rodents from the Brazilian Pantanal.

Propolis in the control of bacterial bovine mastitis: a tool for the production of organic milk / Própolis no controle de mastite bovina bacteriana: uma ferramenta para a produção de leite orgânico

Bovine mastitis is one of the main causes of economic damage in dairy farms. Therefore, the control and prevention of microorganisms involved in this disease, mainly Escherichia coli, Staphylococcus aureus, and Streptococcus agalactiae, are essential. One of the most important steps for the prevention of the disease is the use of antiseptic products before and after the milking process to avoid bacteria from infecting the udder of the animal. Currently, the most used antiseptic product in dairy farms is iodine-based, and organic dairy farms, which follow several strict regulations, including the use of natural products whenever possible, are often forced to adopt non-natural antiseptic products, such as iodine-based ones, because of the lack of natural alternatives. Propolis, a natural substance produced by honeybees, has been extensively studied for its various properties, one of which is antimicrobial activity. Therefore, a new natural antiseptic product containing 1% propolis in 10% hydroalcoholic solution for the pre-dipping, and 10% glycerol solution added with 0.2% citronella oil for the post-dipping was analyzed for its capacity to reduce bacteria in vivo in order to prevent bovine mastitis, allowing its use on organic dairy farms. A total of 128 samples were analyzed in terms of bacterial growth for Enterobacteriaceae and Staphylococcus spp. using the spreadplate technique. The reduction in the bacterial concentration after the application of the products was compared between two antiseptic solutions, an iodine-based solution as the control and a propolis-based one as the natural alternative. The results obtained show a similar efficiency for both products in terms of total bacterial reduction, indicating considerable antimicrobial activity against bacteria most commonly associated with bovine mastitis. Molecular analysis was carried out for the identification of Streptococcus agalactiae; the PCR results were negative for the presence of S. agalactiae in all samples, indicating that the animals most likely did not have any form of the disease. The efficiency of the natural antiseptic was satisfactory, indicating an important find facilitating organic milk production worldwide, showcasing a natural antiseptic solution with efficient antimicrobial activity.(AU)

A mastite bovina é uma das principais causas de prejuízo econômico na indústria leiteira, portanto o controle e prevenção de microrganismos envolvidos nessa doença, principalmente Escherichia coli, Staphylococcus aureus e Streptococcus agalactiae é essencial. Uma das principais etapas na prevenção dessa doença é o uso de produtos antissépticos antes e depois do processo de ordenha, a fim de evitar contaminação bacteriana no úbere do animal. Atualmente, o produto antisséptico mais utilizado na indústria leiteira é a base de iodo, e fazendas produtoras de leite orgânico, que precisam seguir uma série de regulações estritas, incluindo o uso de produtos naturais sempre que possível, são frequentemente forçadas a adotar antissépticos não-naturais, como os a base de iodo por falta de alternativas naturais. Própolis, uma substância natural produzidas por abelhas, tem sido extensivamente estudada por suas várias propriedades, sendo uma delas antimicrobiana. Portanto, um novo produto antisséptico natural contendo própolis à 1% em solução hidroalcóolica 10% para o pré-dipping, e glicerinada 10% adicionada de óleo de citronela à 0,2% para o pós-dipping, foi avaliada quanto a sua capacidade de reduzir bactérias in vivo e prevenir a mastite bovina, além de poder ser utilizado na indústria leiteira orgânica. Um total de 128 amostras foram analisadas em termos de crescimento bacteriano para Enterobacteriaceae e Staphylococcus spp. utilizando a técnica de plaqueamento em superfície, a redução da concentração bacteriana após a aplicação dos produtos foi comparada entre duas soluções antissépticas, uma solução a base de iodo servindo como controle, e uma solução a base de própolis como a alternativa natural. Os resultados obtidos mostraram uma eficiência similar entre os produtos à base de iodo e própolis em termos de redução bacteriana total, indicando uma grande atividade antibacteriana contra as bactérias mais comumente associadas com a mastite bovina. Realizou-se análise molecular para a identificação de Streptococcus agalactiae, os resultados da PCR foram negativos para a presença de S. agalactiae em todas as amostras, indicando que os animais provavelmente não possuíam nenhuma forma da doença. A eficiência do antisséptico natural foi satisfatória, indicando um achado importante para auxiliar o crescimento da indústria de leite orgânico de forma mundial, mostrando uma solução antisséptica natural com atividade antimicrobiana eficiente.(AU)

Contamination by antimicrobial-resistant enterobacteria isolated from cell phones and hands in a veterinary hospital.

Hospital infections are of great relevance in human and animal health, and fomites are important in the spread of pathogens in hospital units. The aim of this study was to investigate the frequency of enterobacteria in the operating room of a veterinary hospital, the potential cross-contamination of samples, and to characterise the susceptibility profile of the isolates to antimicrobials. Sixty-five samples were collected from five different surgical procedures. These samples came from the hands and cell phones of the surgical team and pet owners, operating tables, and patients. Species detection was performed through polymerase chain reaction, genetic diversity by pulsed-field gel electrophoresis (PFGE), and susceptibility to antimicrobials through an antibiogram. Escherichia coli and Proteus mirabilis isolates were obtained from eight samples, from the hands of the anaesthesiologist, the pet owner, and the surgeon; the surgeon's, the nurse's and the anaesthesiologist's cell phones, and two surgical tables. Furthermore, PFGE showed high genetic diversity among the isolates, which showed multidrug resistance. The identification of multidrug-resistant E. coli and P. mirabilis on cell phones of the surgical team is a major concern and, although no direct correlation was found, the isolation of these bacteria inside the clean area of the operating room shows the possibility of nosocomial transmission from cell phones to susceptible patients.

Occurrence and Molecular Characteristics of Extended-Spectrum Beta-Lactamase-Producing Enterobacterales Recovered From Chicken, Chicken Meat, and Human Infections in Sao Paulo State, Brazil.

This study aimed to investigate the phylogenetic diversity and epidemiology of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae from chicken, chicken meat, and human clinical isolates in Sao Paolo, Brazil, and characterize their respective ESBL-encoding plasmids. Three hundred samples from chicken cloaca, chicken meat, and clinical isolates were phenotypically and genotypically assessed for ESBL resistance. Isolates were identified by MALDI TOF-MS and further characterized by MLST analysis and phylogenetic grouping. ESBL genes were characterized and their location was determined by I-Ceu-I-PFGE and Southern blot, conjugation, transformation, and PCR-based replicon typing experiments. Thirty-seven ESBL-producing isolates (28 E. coli and 9 K. pneumoniae) that were positive for the bla CTX-M-1 or bla CTX-M-2 gene groups were obtained. Two isolates were negative in the transformation assay, and the chromosomal location of the genes was deduced by Southern blot. The bla CTX-M genes identified were carried on plasmid replicon-types X1, HI2, N, FII-variants, I1 and R. The E. coli isolates belonged to nine sequence types, while the K. pneumoniae isolates belonged to four sequence types. The E. coli isolates belonged to phylotype classification groups A, B1, D, and F. This study demonstrated that isolates from cloacal swabs, chicken meat, and human feces had genetic diversity, with a high frequency of bla CTX-M-15 among chickens, chicken meat, and human feces. Thus, this reinforces the hypothesis that chickens, as well as their by-products, could be an important source of transmission for ESBL-producing pathogens to humans in South America.

Evaluation of the Resistance Profile of Bacteria Obtained From Infected Sites of Dogs in a Veterinary Teaching Hospital in Brazil: A Retrospective Study.

The aim of this study was to evaluate the prevalence and antimicrobial resistance profile of bacterial species isolated from infected sites of canines. All samples were collected from canine patients who received clinical or surgical care at the veterinary teaching hospital between March 2016 and November 2017. The samples were analyzed in a private pathology laboratory. A descriptive analysis of 295 antimicrobial susceptibility test reports was performed. Staphylococcus spp. (104/295 [35.25%]), Escherichia coli (100/295 [33.90%]), Proteus spp. (44/295 [14.92%]), Pseudomonas spp. (25/295 [8.47%]), and Klebsiella spp. (20/295 [6.78%]) were more frequently isolated, and a high incidence of multidrug resistance was observed (69,83% [206/295]). Methicillin-resistant Staphylococcus spp. accounted for 33% (33/100) of the Staphylococcus strains. Enterobacteriaceae cefotaxime resistance constituted 22.82 ± 4.49% and Enterobacteriaceae imipenem resistance constituted 5% (1/20) for Klebsiella spp., 5% (5/100) for E coli, and 6.82% (3/44) for Proteus spp. Pseudomonas spp. strains accounted for 8% (2/25) of imipenem resistance and 45.45% (10/22) of polymyxin B resistance. Our findings revealed a high rate of multidrug-resistant bacteria involvement in the infectious process of dogs. From the perspective of the One Health scenario, our results showed alarming data, given the high risk of resistant-strain dissemination between animals, owners, and healthcare professionals. There is an urgent need for strategies to control and prevent the evolution of new multidrug-resistant bacteria in veterinary hospitals. It is also crucial to understand and emphasize the role of veterinary professionals in this public health battle.

Experimental inoculation and nose-to-nose transmission of Clostridioides difficile between weaned piglets.

In piglets, Clostridioides (C.) difficile infection presents mostly subclinical manifestation. As this agent became important in veterinary medicine due to a hypothesis of zoonosis, the objective of this study was to evaluate the transmission of C. difficile by nose-to-nose contact in young piglets. Six 20-day-old piglets were separated into three groups (infected, sentinel and control), and distributed in different isolation cabinets which allowed nose-to-nose contact only between infected and sentinel groups. The challenged group received an inoculum 106 CFU/mL of C. difficile 096 by oropharyngeal route. Rectal swab samples were daily collected for microbiological and molecular analysis. Euthanasia of all piglets was performed 18 days after challenge to evaluate anatomical, histological and microbiological lesions of the organs of these animals. The challenged and sentinel groups showed clinical signs of infection and genes encoding TcdB were detected by conventional PCR in both groups, confirming the transmission of the pathogen from the challenged to the sentinel piglets. At necropsy, tonsil, liver, spleen, mesenteric lymph nodes, ileocolic lymph nodes, jejunum, ileum, proximal colon, distal colon and cecum were collected for microbiological analysis; lesions were observed varying in degree and intensity. This study demonstrated a novel route of transmission of C. difficile between young piglets, which was proven to occur by nose-to-nose contact.

Antibiotic resistance and virulence factors among Escherichia coli isolates from avian organic fertilizer / Resistência a antibióticos e fatores de virulência entre isolados de Escherichia coli provenientes de fertilizante orgânico aviário

Brazilian poultry industry generates large amounts of organic waste, such as chicken litter, which is often used in agriculture. Among the bacteria present in organic fertilizer are members of the Enterobacteriaceae family, such as Escherichia coli. The aim of this study was to analyze the profile of virulence factors and antimicrobial resistance of E. coli isolates from avian organic fertilizer. A total of 47 E. coli strains were tested through Polymerase chain reaction to detect virulence genes (hlyF, iss, ompT, iutA and iroN). Fourteen antimicrobials were used to test antimicrobial susceptibility in the strains. Genes characteristic of Avian Pathogenic E. coli (APEC) were reported among the strains, with the hlyF, iss and ompT genes being the most prevalent. The isolates showed high resistance (˃50%) to tetracycline, gentamicin, cefotaxime, nitrofurantoin, trimethoprim-sulfamethoxazole and ampicillin. Multidrug resistance was reported in a great number of strains (>70%). The results showed the presence of APEC cells with virulence genes and antimicrobial resistance after 15 days of the windrowing process in poultry houses, it means this process should be improved to eliminate these cells.(AU)

A indústria avícola brasileira gera grandes quantidades de resíduos orgânicos, como a cama de frango, utilizada frequentemente na agricultura. Entre as bactérias presentes neste fertilizante orgânico estão os membros da família Enterobacteriaceae, entre eles a Escherichia coli. O objetivo deste estudo foi analisar o perfil de fatores de virulência e resistência antimicrobiana de isolados de E. coli provenientes do fertilizante orgânico aviário. Um total de 47 cepas de E. coli foram testadas por meio da reação em cadeia da polimerase para detectar genes de virulência (hlyF, iss, ompT, iutA e iroN). Quatorze antimicrobianos foram utilizados para testar a susceptibilidade antimicrobiana nos isolados. Genes característicos de E. coli Patogênica Aviária (APEC) foram encontrados entre os isolados, sendo os genes hlyF, iss e ompT os mais prevalentes. Os isolados apresentaram alta resistência (˃50%) à tetraciclina, gentamicina, cefotaxima, nitrofurantoína, trimetoprima-sulfametoxazole e ampicilina. Múltipla resistência a drogas antimicrobianas foi encontrada em grande número de isolados (>70%). Os resultados obtidos mostraram a presença de células APEC portando genes de virulência e resistência a antimicrobianos após 15 dias de processo de empilhamento nas granjas, indicando que o processo necessita de um aperfeiçoamento para eliminar estas células.(AU)

Antibiotic resistance and virulence factors among Escherichia coli isolates from avian organic fertilizer / Resistência a antibióticos e fatores de virulência entre isolados de Escherichia coli provenientes de fertilizante orgânico aviário

ABSTRACT: Brazilian poultry industry generates large amounts of organic waste, such as chicken litter, which is often used in agriculture. Among the bacteria present in organic fertilizer are members of the Enterobacteriaceae family, such as Escherichia coli. The aim of this study was to analyze the profile of virulence factors and antimicrobial resistance of E. coli isolates from avian organic fertilizer. A total of 47 E. coli strains were tested through Polymerase chain reaction to detect virulence genes (hlyF, iss, ompT, iutA and iroN). Fourteen antimicrobials were used to test antimicrobial susceptibility in the strains. Genes characteristic of Avian Pathogenic E. coli (APEC) were reported among the strains, with the hlyF, iss and ompT genes being the most prevalent. The isolates showed high resistance (˃50%) to tetracycline, gentamicin, cefotaxime, nitrofurantoin, trimethoprim-sulfamethoxazole and ampicillin. Multidrug resistance was reported in a great number of strains (>70%). The results showed the presence of APEC cells with virulence genes and antimicrobial resistance after 15 days of the windrowing process in poultry houses, it means this process should be improved to eliminate these cells.

RESUMO: A indústria avícola brasileira gera grandes quantidades de resíduos orgânicos, como a cama de frango, utilizada frequentemente na agricultura. Entre as bactérias presentes neste fertilizante orgânico estão os membros da família Enterobacteriaceae, entre eles a Escherichia coli. O objetivo deste estudo foi analisar o perfil de fatores de virulência e resistência antimicrobiana de isolados de E. coli provenientes do fertilizante orgânico aviário. Um total de 47 cepas de E. coli foram testadas por meio da reação em cadeia da polimerase para detectar genes de virulência (hlyF, iss, ompT, iutA e iroN). Quatorze antimicrobianos foram utilizados para testar a susceptibilidade antimicrobiana nos isolados. Genes característicos de E. coli Patogênica Aviária (APEC) foram encontrados entre os isolados, sendo os genes hlyF, iss e ompT os mais prevalentes. Os isolados apresentaram alta resistência (˃50%) à tetraciclina, gentamicina, cefotaxima, nitrofurantoína, trimetoprima-sulfametoxazole e ampicilina. Múltipla resistência a drogas antimicrobianas foi encontrada em grande número de isolados (>70%). Os resultados obtidos mostraram a presença de células APEC portando genes de virulência e resistência a antimicrobianos após 15 dias de processo de empilhamento nas granjas, indicando que o processo necessita de um aperfeiçoamento para eliminar estas células.

Microbiological analysis in the fixation and preservation of dog cadavers with ethyl alcohol and sodium chloride solution / Análise microbiológica na fixação e conservação de cadáveres de cães com álcool etílico e solução de cloreto de sódio

There are several fixative or preservative solutions for use on cadavers, and formaldehyde is the most widely used. However, this chemical may present negative effects for professionals who manipulate it. Therefore, this study aimed to identify and quantify the main microorganisms related to the fixation and preservation of dog cadavers using an alternative and formaldehyde-free solution. After arterial injection (120 mL kg−1 95% 96° GL ethyl alcohol and 5% pure glycerin), cadavers were placed in 96° GL ethyl alcohol for 30 (group 1), 60 (group 2), 90 (group 3), and 120 days (group 4). After the fixation period, they remained under preservation in a 30% aqueous sodium chloride solution for 120 days. Bacterial quantification was performed by the pour plate method. The bacterial population was present in all groups during fixation, except for group 1, but never exceeded 9 × 101 CFU mL−1 in total aerobes and 7 × 101 CFU mL−1 in total anaerobes. The microbial population was present in all groups in at least two moments during preservation and never exceeded 7 × 101 CFU mL−1 in total aerobes and anaerobes. The presence of fungi was observed in 8 out of 34 analyses. Pseudomonas sp., Escherichia coli, and Bacillus sp. were identified in the analyzed samples. Microbiological counting was low, and no signs of contamination were observed in the vats at visual inspection.(AU)

Várias são as soluções fixadoras ou conservantes de cadáveres, e o formaldeído é o mais utilizado. Entretanto, esse agente pode apresentar efeitos negativos para os profissionais que o manipulam. Portanto, o objetivo deste trabalho foi identificar e quantificar os principais microrganismos relacionados a fixação e conservação de cadáveres de cães utilizando-se solução alternativa e livres de formaldeído. Após injeção arterial (120 mL kg−1, de 95% de álcool etílico 96° GL e 5% de glicerina pura), foram colocados em álcool etílico 96° GL por 30 dias (grupo 1), 60 dias (grupo 2), 90 dias (grupo 3) e 120 dias (grupo 4). Após o período de fixação, permaneceram sob conservação em solução aquosa de cloreto de sódio 30% por 120 dias. A quantificação bacteriana foi realizada por plaqueamento em profundidade (Pour Plate). Durante a fixação, houve presença de população bacteriana em todos os grupos, exceto no grupo 1, e nunca foi ultrapassado o valor de 9 × 101 UFC mL−1 (Unidade Formadora de Colônia) nos aeróbios totais e 7 × 101 UFC mL−1 nos anaeróbios totais. Durante a conservação, houve presença de população microbiana em todos os grupos em pelo menos dois momentos, e nunca foi ultrapassado o valor de 7 × 101 UFC mL−1 nos aeróbios totais e nos anaeróbios totais. A presença de fungos foi observada em 8 das 34 análises. Houve identificação de Pseudomonas sp., E. coli e Bacillus sp. nas amostras analisadas. A contagem microbiológica foi baixa e não foram observados, à inspeção visual, sinais de contaminação nas cubas.(AU)

Microbiological analysis in the fixation and preservation of dog cadavers with ethyl alcohol and sodium chloride solution / Análise microbiológica na fixação e conservação de cadáveres de cães com álcool etílico e solução de cloreto de sódio

There are several fixative or preservative solutions for use on cadavers, and formaldehyde is the most widely used. However, this chemical may present negative effects for professionals who manipulate it. Therefore, this study aimed to identify and quantify the main microorganisms related to the fixation and preservation of dog cadavers using an alternative and formaldehyde-free solution. After arterial injection (120 mL kg−1 95% 96° GL ethyl alcohol and 5% pure glycerin), cadavers were placed in 96° GL ethyl alcohol for 30 (group 1), 60 (group 2), 90 (group 3), and 120 days (group 4). After the fixation period, they remained under preservation in a 30% aqueous sodium chloride solution for 120 days. Bacterial quantification was performed by the pour plate method. The bacterial population was present in all groups during fixation, except for group 1, but never exceeded 9 × 101 CFU mL−1 in total aerobes and 7 × 101 CFU mL−1 in total anaerobes. The microbial population was present in all groups in at least two moments during preservation and never exceeded 7 × 101 CFU mL−1 in total aerobes and anaerobes. The presence of fungi was observed in 8 out of 34 analyses. Pseudomonas sp., Escherichia coli, and Bacillus sp. were identified in the analyzed samples. Microbiological counting was low, and no signs of contamination were observed in the vats at visual inspection.

Várias são as soluções fixadoras ou conservantes de cadáveres, e o formaldeído é o mais utilizado. Entretanto, esse agente pode apresentar efeitos negativos para os profissionais que o manipulam. Portanto, o objetivo deste trabalho foi identificar e quantificar os principais microrganismos relacionados a fixação e conservação de cadáveres de cães utilizando-se solução alternativa e livres de formaldeído. Após injeção arterial (120 mL kg−1, de 95% de álcool etílico 96° GL e 5% de glicerina pura), foram colocados em álcool etílico 96° GL por 30 dias (grupo 1), 60 dias (grupo 2), 90 dias (grupo 3) e 120 dias (grupo 4). Após o período de fixação, permaneceram sob conservação em solução aquosa de cloreto de sódio 30% por 120 dias. A quantificação bacteriana foi realizada por plaqueamento em profundidade (Pour Plate). Durante a fixação, houve presença de população bacteriana em todos os grupos, exceto no grupo 1, e nunca foi ultrapassado o valor de 9 × 101 UFC mL−1 (Unidade Formadora de Colônia) nos aeróbios totais e 7 × 101 UFC mL−1 nos anaeróbios totais. Durante a conservação, houve presença de população microbiana em todos os grupos em pelo menos dois momentos, e nunca foi ultrapassado o valor de 7 × 101 UFC mL−1 nos aeróbios totais e nos anaeróbios totais. A presença de fungos foi observada em 8 das 34 análises. Houve identificação de Pseudomonas sp., E. coli e Bacillus sp. nas amostras analisadas. A contagem microbiológica foi baixa e não foram observados, à inspeção visual, sinais de contaminação nas cubas.

Shigatoxigenic and atypical enteropathogenic Escherichia coli in fish for human consumption

Shigatoxigenic and enteropathogenic Escherichia coli with virulence and multidrug resistance profile were isolated from Nile tilapia. This study finding is of great importance to public health because they help understand this pathogen epidemiology in fish and demonstrate how these animals can transmit E. coli related diseases to humans.(AU)

Shigatoxigenic and atypical enteropathogenic Escherichia coli in fish for human consumption

ABSTRACT Shigatoxigenic and enteropathogenic Escherichia coli with virulence and multidrug resistance profile were isolated from Nile tilapia. This study finding is of great importance to public health because they help understand this pathogen epidemiology in fish and demonstrate how these animals can transmit E. coli related diseases to humans.