Carriage of Clostridium perfringens in domestic and farm animals across the central highlands of Colombia: implications for gut health and zoonotic transmission.

Clostridium perfringens inhabits the guts of humans and animal species. C. perfringens can proliferate and express an arsenal of toxins, promoting the development of multiple gut illnesses. Healthy animals carrying C. perfringens represents a risk of transmission to other animals or humans through close contact and an increased likelihood of acquisition of toxin plasmids. The aim of this study was to evaluate the frequency of C. perfringens carriage in domestic and farm animals in the central highlands of Colombia. C. perfringens was detected in six animal species using PCR targeting alpha toxin (cpa) and 16S ribosomal RNA (16S-rRNA) genes from 347 fecal samples collected in two Departments: 177 from farm animals of Boyacá and 170 from domestic animals of both Cundinamarca and Boyacá. The overall frequency of C. perfringens detection was 22.1% (n = 77/347), with the highest frequency observed in cats 34.2% (n = 41/120), followed by dogs 30.0% (n = 15/50). The lowest frequency was detected in ruminants: goats 11.1% (n = 3/27), sheep 8.0% (n = 4/50) and cattle 6.0% (n = 6/50). Domestic animals showed a higher frequency of C. perfringens carriage than farm animals. This difference could be associated with dietary patterns, as domestic animals have diets rich in proteins and carbohydrates, while ruminants have low-carbohydrate diets, resulting in high production of endopeptidase-type enzymes and differences in pH due to the anatomy of gastrointestinal tract, which can influence bacterial proliferation. These findings indicate a potential risk of transmission of C. perfringens among animals and from animals to humans through close contact.

Fungos filamentosos isolados da pelagem de coelhos e porquinhos-da-índia clinicamente saudáveis na cidade de Salvador / Bahia, Brasil / Filamentous fungi isolated from the coat of clinically healthy rabbits and guinea pigs in the city of Salvador, Bahia, Brazil

Fungos zoonóticos associados aos tecidos dos animais domésticos são potenciais riscos à saúde pública. O objetivo deste trabalho é identificar os principais fungos filamentosos associados ao pelo de coelhos (Oryctolagus cuniculus) e porquinhos-da-índia (Cavia porcellus) clinicamente saudáveis da cidade de Salvador, Bahia, Brasil. As amostras de pelo foram coletadas de 18 porquinhos-da-índia e 6 coelhos domiciliados, de juvenis à adultos e oriundos de diferentes residências. As coletas foram decorrentes da região dorsal, ventral e caudal a pina da orelha, por meio da técnica de arrancamento do pelo manualmente. O cultivo foi realizado em meio Dermatobac® de acordo com a rotina laboratorial. Os fungos isolados nos porquinhos-da-índia foram o Tricophyton mentagrophytes (66,6%), o Aspergillus spp. (44,4%), o Penicillium spp. (5,5%), o Mucor spp. (5,5%) e o Litcheimia (5,5%); e em coelhos Tricophyton mentagrophytes (50%), Curvularia spp. (50%), Penicillium spp. (11,1%) e Aspergillus spp. (11,1%). Todos os fungos observados possuem potencial zoonótico, o que deve ser considerado como critério de avaliação à saúde pública e para criação por contactantes imunocomprometidos.

Zoonotic fungi associated with tissues of domestic animals are potential risks to public health. The objective of this work was to identify the main filamentous fungi associated with the hair of clinically healthy rabbits (Oryctolagus cuniculus) and guinea pigs (Cavia porcellus) in the city of Salvador, Bahia, Brazil. The hair samples were collected from 18 guinea pigs and 6 domiciled rabbits, from juveniles to adults and from different homes. Samples were collected from the dorsal, ventral and caudal of the ear tip, using the technique of manual plucking. Cultivation was performed in Dermatobac® medium according to the laboratory routine. The fungi isolated from guinea pigs were Tricophyton mentagrophytes (66.6%), Aspergillus spp. (44.4%), Penicillium spp. (5.5%), Mucor spp. (5.5%) and Litcheimia (5.5%); and in rabbits Tricophyton mentagrophytes (50%), Curvularia spp. (50%), Penicillium spp. (11.1%) and Aspergillus spp. (11.1%). All fungi isolated have zoonotic potential, which should be considered as a criterion for assessing public health and for breeding by immunocompromised contacts.

Evaluation of three qPCR for the detection of pathogenic leptospires in domestic animals in Nicaragua / [Evaluación de tres PCR cuantitativas para la detección de leptospiras patógenas en animales domésticos en Nicaragua].

Introduction: Molecular biology diagnostic methods such as real-time PCR should be used in Nicaragua to improve the diagnosis of leptospirosis in humans and animals. Objective: To evaluate three qPCR methods for pathogenic Leptospira detection in domestic animals. Materials and methods: Real-time PCR primers were designed for the amplification of specific regions from the Lip 32 gene of Leptospira in SYBER Green (SYBER Green-A) and TaqMan, as well in SYBER Green-B as previously published. The sequences of 12 strains obtained from the database of the National Center for Biotechnology Information (NCBI) were aligned to select probes and primers. The analytical sensitivity was determined by calculating the detectable genomic equivalent while 18 pathogenic references strains and 28 negative controls were used to evaluate the sensitivity and specificity of each one of the three sets in 129 urine samples of domestic animals. Results: The detection limit of four genomic equivalents per reaction was obtained from SYBR Green-A. The specificities were 94.4% (95% CI: 81.1-100.0) for TaqMan, 77.8% (95% CI: 55.8-99.8) for SYBR Green-A, while for SYBR Green-B it was 61.1% (95% CI: 35.8-86.4). In the three tests, we obtained a specificity of 100% (95% CI: 98.2-100.0). In the field samples, 26.4% were positive with SYBR Green-A and 6.1% with SYBR Green-B. Conclusion: SYBR Green-A presented the lowest detection limit while the three techniques under evaluation showed high specificity while TaqMan was the most sensitive.

Introducción. En Nicaragua es necesario estandarizar pruebas moleculares como la PCR en tiempo real (quantitative Polymerase Chain Reaction, qPCR) que mejoren el diagnóstico de leptospirosis en humanos y animales. Objetivo. Evaluar tres qPCR para la detección de leptospiras patógenas en animales domésticos de Nicaragua. Materiales y métodos. Se diseñaron cebadores para la amplificación del gen LipL32 en SYBR Green (SYBR Green-A) y TaqMan, y en otros descritos previamente (SYBR Green-B). Las secuencias de 12 cepas obtenidas de la base de datos del National Center for Biotechnology Information (NCBI) se alinearon para la búsqueda de sondas y cebadores. La sensibilidad analítica se determinó calculando el equivalente genómico detectable, se utilizaron 18 cepas de referencia para la sensibilidad diagnóstica y 28 controles negativos para la especificidad. Los métodos se aplicaron en 129 muestras de orina de animales domésticos. Resultados. En SYBR Green-A se obtuvo un límite de detección de cuatro equivalentes genómicos; en TaqMan, la sensibilidad fue del 94,4 % (IC95% 81,1-100,0). Con SYBR Green-A, se obtuvo una sensibilidad del 77,8 % (IC95% 55,8-99,8), en tanto que con SYBR Green-B fue del 61,1 % (IC95% 35,8-86,4). En las tres pruebas se logró una especificidad del 100 % (IC95% 98,2-100,0). El 26,4 % de las muestras de animales domésticos fueron positivas con SYBR Green-A y el 6,2 % con SYBR Green-B. Conclusiones. El SYBR Green-A presentó un límite de detección bajo, en tanto que las tres técnicas evaluadas mostraron alta especificidad, en tanto que la TaqMan tuvo la mayor sensibilidad.

Flea-borne Rickettsia species in fleas, Caldas department, Colombia.

INTRODUCTION: Rickettsioses are zoonotic diseases caused by pathogenic bacteria of the genus Rickettsia and transmitted to man by means of arthropod vectors such as ticks, fleas, mites and lice. Historically, Caldas Department has reported a significant number of cases of murine typhus to the Colombian national health surveillance system, and consequent studies of flea-borne rickettsiosis identified the circulation of Rickettsia typhi and Rickettsia felis in multiple municipalities. Our aim was to genotype species of Rickettsia detected in fleas collected from domestic and wild mammals in Caldas. METHODOLOGY: Flea samples were taken by convenience sampling from dogs, cats and wild mammals (rodents and marsupials) in 26 municipalities. Specimens were classified by current taxonomic keys and pooled for DNA extraction and molecular screening for Rickettsia spp. by PCR amplification of gltA, htrA and sca5 genes. Positive samples were genotyped by enzyme digestion (htrA) and sequencing. RESULTS: A total of 1388 flea samples were collected. Rickettsia DNA was amplified in 818 (gltA), 883 (htrA) and 424 (sca5) flea pools. Alignment analysis with available Rickettsia DNA sequences showed greater similarity with R. asembonensis (gltA) and with R. felis (sca5 and htrA). Restriction pattern was compatible with R. felis. R. typhi was not identified. CONCLUSION: The present study confirms the presence and high prevalence of R. asembonensis and R. felis in fleas from domestic and wild animals in different municipalities from Caldas Department.

Antimicrobial resistance genes in bacteria from animal-based foods.

Antimicrobial resistance is a worldwide public health threat. Farm animals are important sources of bacteria containing antimicrobial resistance genes (ARGs). Although the use of antimicrobials in aquaculture and livestock has been reduced in several countries, these compounds are still routinely applied in animal production, and contribute to ARGs emergence and spread among bacteria. ARGs are transmitted to humans mainly through the consumption of products of animal origin (PAO). Bacteria can present intrinsic resistance, and once antimicrobials are administered, this resistance may be selected and multiply. The exchange of genetic material is another mechanism used by bacteria to acquire resistance. Some of the main ARGs found in bacteria present in PAO are the bla, mcr-1, cfr and tet genes, which are directly associated to antibiotic resistance in the human clinic.

Detection of Antimicrobial Resistance Genes in Escherichia coli Isolated from Black Howler Monkeys (Alouatta pigra) and Domestic Animals in Fragmented Rain-Forest Areas in Tabasco, Mexico.

The appearance and spread of antimicrobial resistance (AMR) in bacteria in natural environments and wildlife are related to agricultural and livestock activities and are a global health and conservation problem. We assessed the presence of AMR genes in Escherichia coli isolated from black howler monkeys (Alouatta pigra), sheep (Ovis aries), cattle (Bos taurus), and horses (Equus caballus) from a highly fragmented forest in southern Mexico. Fresh fecal samples were collected using swabs, seeded on eosin-methylene blue agar, and E. coli colonies identified by PCR; multiplex-PCR was performed on E. coli DNA for the detection of 10 AMR genes from four families (sulfonamides, tetracycline, ß-lactamase, and chloramphenicol). We detected E. coli in 94% (48/51) of fecal samples, of which 33% (16/48) tested positive for at least one AMR gene. We detected AMR genes in at least one individual from each sampled animal species, with the most prevalent genes being tet(B) 18% (9/48), sul2 14% (7/48), sul1, and blaTEM 12% (6/48). Sheep samples contained AMR genes from the four families of antibiotics detected in this study and 50% (5/10) tested positive for the presence of at least one gene. A total of 12% (2/16) of fecal samples from black howler monkeys tested positive for AMR genes. The presence of AMR genes in A. pigra and domestic animals has not been reported in the Balancán area of Tabasco, Mexico. Transmission of AMR bacteria from domestic animals to monkeys is rare; however, this is a potential health risk for wildlife and species conservation.

Factors Obscuring the Role of E. coli from Domestic Animals in the Global Antimicrobial Resistance Crisis: An Evidence-Based Review.

Recent studies have found limited associations between antimicrobial resistance (AMR) in domestic animals (and animal products), and AMR in human clinical settings. These studies have primarily used Escherichia coli, a critically important bacterial species associated with significant human morbidity and mortality. E. coli is found in domestic animals and the environment, and it can be easily transmitted between these compartments. Additionally, the World Health Organization has highlighted E. coli as a "highly relevant and representative indicator of the magnitude and the leading edge of the global antimicrobial resistance (AMR) problem". In this paper, we discuss the weaknesses of current research that aims to link E. coli from domestic animals to the current AMR crisis in humans. Fundamental gaps remain in our understanding the complexities of E. coli population genetics and the magnitude of phenomena such as horizontal gene transfer (HGT) or DNA rearrangements (transposition and recombination). The dynamic and intricate interplay between bacterial clones, plasmids, transposons, and genes likely blur the evidence of AMR transmission from E. coli in domestic animals to human microbiota and vice versa. We describe key factors that are frequently neglected when carrying out studies of AMR sources and transmission dynamics.

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.

Diverse Escherichia coli lineages from domestic animals carrying colistin resistance gene mcr-1 in an Ecuadorian household.

OBJECTIVE: The aim of this study was to detect potential animal reservoirs of Escherichia coli carrying the mcr-1 gene in an Ecuadorian household. METHODS: The mobile colistin-resistance gene, mcr-1, was first detected in Ecuador in a commensal E. coli isolate from a boy. A cross-sectional study was performed to detect the possible source of colistin-resistant E. coli in the boy's household. Faecal swabs and soil faecal samples were collected from companion animals. Samples were plated on selective media to isolate colistin-resistant E. coli and isolates were submitted to PCR detection of mcr-1, pulsed field gel electrophoresis (PFGE), and multi-locus sequences typing (MLST). Moreover, the genomes of all the isolates were sequenced. RESULTS: Three different colistin-resistant E. coli sequence types (ST3941, 1630 and 2170), corresponding to three PFGE patterns, were obtained from a chicken and two dogs; these isolates were different from the human isolate (ST609). By whole-genome sequencing, the mcr-1.1 gene was found on IncI2 plasmids with very high nucleotide identity. CONCLUSIONS: Our results indicate a polyclonal dissemination of mcr-1.1 in the environment surrounding the first MCR-producing E. coli strain reported in Ecuador. Our findings support the idea of lateral dissemination of mcr-1.1 gene between unrelated E. coli isolates.

Molecular characterization of multidrug-resistant Shiga toxin-producing Escherichia coli harboring antimicrobial resistance genes obtained from a farmhouse.

Shiga toxin-producing Escherichia coli (STEC) colonize the gastrointestinal tract of animals; however, STEC may also cause severe diarrheal diseases. Food-producing animals have been acting as reservoirs and disseminators of multidrug-resistant (MDR) bacteria and antimicrobial resistance genes (ARGs); however, there are few studies characterizing molecularly bacterial isolates from sheep. Therefore, this study aimed to characterize E. coli isolates obtained from feces of sheep in a Brazilian farmhouse. A total of 14 MDR E. coli isolates were obtained from 100 feces samples, six of which were classified as non-O157 STEC (stx1, stx2 and ehxA). MDR E. coli isolates presented different ARGs [blaCTX-M-Gp9, blaCMY, blaSHV, qnrS, oqxB, aac(6')-Ib, tet(A), tet(B), tet(C), sul1, sul2, and cmlA] and plasmids (IncI1, IncFrepB, IncFIB, IncFIA, IncHI1, IncK, and ColE-like). In addition, mutations in the quinolone-resistance determining region of GyrA (Ser83Leu; Asp87Asn) and ParC (Glu84Asp) were detected. PFGE showed a high genetic diversity (30.9 to 83.9%) and thirteen STs were detected (ST25, ST48, ST155, ST162, ST642, ST1247, ST1518, ST1725, ST2107, ST2522, ST3270, ST5036, and ST7100). Subtyping of the fimH gene showed seven fimH-type (25, 32, 38, 41, 54, 61, and 366). The results found in the present study showed high genetic diversity among MDR ARGs-producing E. coli obtained from a farmhouse. This study reports for the first time, the presence of MDR STEC and non-STEC belonging to ST25, ST162, ST642, ST1247, ST1518, ST1725, ST2107, ST3270, ST5036, and ST7100 in sheep, and contributes to the surveillance studies associated with One Health concept.

Assessing cross-species transmission of hemoplasmas at the wild-domestic felid interface in Chile using genetic and landscape variables analysis.

The co-occurrence of domestic cats (Felis silvestris catus) and wild felids in rural landscapes can facilitate pathogen transmission. However, in the relatively-isolated regions of southern South America there have been no comprehensive studies to assess disease transmission risks between domestic cats and forest-dwelling wild felids such as guigna (Leopardus guigna). We evaluated hemoplasma infection and the possibility of transmission between domestic cats and guignas by comparing spatial and phylogenetic patterns of pathogen prevalence. Blood/spleen samples were collected from 102 wild guignas and 262 co-occurring rural domestic cats across the entire distribution range of guigna in Chile. Hemoplasma infection was assessed by direct sequencing of the 16S RNA gene. Infection with hemoplasmas was common and geographically widespread across different bioclimatic areas for both species. The most common feline Mycoplasma species in guigna and domestic cats were Candidatus M. haemominutum (CMhm) (15.7% guigna; 10.3% domestic cat) and Mycoplasma haemofelis (Mhf) (9.8% guigna, 6.1% domestic cat). A previously undescribed Mycoplasma sp. sequence was found in two guignas and one cat. Continuous forest-landscapes were associated with higher hemoplasma-prevalence in guignas. Shared hemoplasma nucleotide sequence types between guigna and domestic cats were rare, suggesting that cross-species transmission between guignas and domestic cats may occur, but is probably uncommon. Ectoparasites, which have been linked with hemoplasma transmission, were not found on guignas and were infrequent on domestic cats. Our results suggest that transmission pathways vary among hemoplasma species and, contrary to our predictions, domestic cats did not appear to be the main driver of hemoplasma infection in guignas in these human-dominated landscapes.

Characterization of a bacteriophage with broad host range against strains of Pseudomonas aeruginosa isolated from domestic animals.

BACKGROUND: Pseudomonas aeruginosa is an opportunistic pathogen and one of the leading causes of nosocomial infections. Moreover, the species can cause severe infections in cystic fibrosis patients, in burnt victims and cause disease in domestic animals. The control of these infections is often difficult due to its vast repertoire of mechanisms for antibiotic resistance. Phage therapy investigation with P. aeruginosa bacteriophages has aimed mainly the control of human diseases. In the present work, we have isolated and characterized a new bacteriophage, named Pseudomonas phage BrSP1, and investigated its host range against 36 P. aeruginosa strains isolated from diseased animals and against P. aeruginosa ATCC strain 27853. RESULTS: We have isolated a Pseudomonas aeruginosa phage from sewage. We named this virus Pseudomonas phage BrSP1. Our electron microscopy analysis showed that phage BrSP1 had a long tail structure found in members of the order Caudovirales. "In vitro" biological assays demonstrated that phage BrSP1 was capable of maintaining the P. aeruginosa population at low levels for up to 12 h post-infection. However, bacterial growth resumed afterward and reached levels similar to non-treated samples at 24 h post-infection. Host range analysis showed that 51.4% of the bacterial strains investigated were susceptible to phage BrSP1 and efficiency of plating (EOP) investigation indicated that EOP values in the strains tested varied from 0.02 to 1.72. Analysis of the phage genome revealed that it was a double-stranded DNA virus with 66,189 bp, highly similar to the genomes of members of the genus Pbunavirus, a group of viruses also known as PB1-like viruses. CONCLUSION: The results of our "in vitro" bioassays and of our host range analysis suggested that Pseudomonas phage BrSP1 could be included in a phage cocktail to treat veterinary infections. Our EOP investigation confirmed that EOP values differ considerably among different bacterial strains. Comparisons of complete genome sequences indicated that phage BrSP1 is a novel species of the genus Pbunavirus. The complete genome of phage BrSP1 provides additional data that may help the broader understanding of pbunaviruses genome evolution.

Molecular detection and characterization of Anaplasma platys and Ehrlichia canis in dogs from northern Colombia.

Ehrlichia canis and Anaplasma platys are intracellular tick-transmitted bacteria that infect dogs; there is evidence for limited zoonotic potential as well. The prevalence of E. canis in Colombia has been evaluated in different regions; however little is known about the prevalence or distribution of A. platys. Neither pathogen has been studied in the Magdalena region, thus the purpose of our study was to assess the prevalence of these pathogens in dogs attending veterinary clinics from the cities of Santa Marta and Ciénaga, and to assess possible associated risk factors for infection. A. platys and E. canis infections in blood were evaluated by Taqman PCR assays. E. canis was detected in 26/170 (15.3%, 95% CI 10.4%-21.8%) and A. platys in 34/168 (20.2%, 95% CI 14.6%-27.3%) of all dogs tested. Eleven dogs (6.5%, 95% CI 3.4-11.7%) were coinfected with both pathogens. Sequencing results showed low diversity within E. canis and within A. platys strains, however a strain of E. canis detected in our study area is genetically distinct from strains reported in another city of Colombia. Our results suggest that for A. platys, Santa Marta dogs were at greater risk than Ciénaga dogs, and that purebred dogs were at slightly lower risk in both areas. The confirmation of these pathogens in northern Colombia should cause concern for the possible co-transmission of these agents to humans or animals in the region.

Diverse Commensal Escherichia coli Clones and Plasmids Disseminate Antimicrobial Resistance Genes in Domestic Animals and Children in a Semirural Community in Ecuador.

The increased prevalence of antimicrobial resistance (AMR) among Enterobacteriaceae has had major clinical and economic impacts on human medicine. Many of the multidrug-resistant (multiresistant) Enterobacteriaceae found in humans are community acquired, and some of them are possibly linked to food animals (i.e., livestock raised for meat and dairy products). In this study, we examined whether numerically dominant commensal Escherichia coli strains from humans (n = 63 isolates) and domestic animals (n = 174 isolates) in the same community and with matching phenotypic AMR patterns were clonally related or shared the same plasmids. We identified 25 multiresistant isolates (i.e., isolates resistant to more than one antimicrobial) that shared identical phenotypic resistance patterns. We then investigated the diversity of E. coli clones, AMR genes, and plasmids carrying the AMR genes using conjugation, replicon typing, and whole-genome sequencing. All of the multiresistant E. coli isolates (from children and domestic animals) analyzed had at least 90 or more whole-genome SNP differences between one another, suggesting that none of the strains was recently transferred. While the majority of isolates shared the same antimicrobial resistance genes and replicons, DNA sequencing indicated that these genes and replicons were found on different plasmid structures. We did not find evidence of the clonal spread of AMR in this community: instead, AMR genes were carried on diverse clones and plasmids. This presents a significant challenge for understanding the movement of AMR in a community.IMPORTANCE Even though Escherichia coli strains may share nearly identical phenotypic AMR profiles and AMR genes and overlap in space and time, the diversity of clones and plasmids challenges research that aims to identify sources of AMR. Horizontal gene transfer appears to play a more significant role than clonal expansion in the spread of AMR in this community.

Comparison of six methods of DNA extraction for the diagnosis of bovine brucellosis by real-time PCR.

Brucellosis is an infectious disease caused by bacteria of the genus Brucella, which affects domestic animals and is transmissible to humans. The objective of this study was to evaluate six methods of DNA extraction directly from bovine tissue to detect Brucella spp. The Cq values for all samples were above 30 and varied according to the extraction kit used, but four kits showed no statistical difference in sensitivity. This work demonstrates the importance of choosing the best extraction kit before validation of a molecular diagnostic technique.

Perfil de resistência antimicrobiana de Klebsiella pneumoniae isoladas de animais domésticos e silvestres / Antimicrobial resistance profile of Klebsiella pneumoniae isolated from domestic and wild animals

Klebsiella pneumoniae é um patógeno oportunista, responsável por diversos tipos de infecções nosocomiais, e é considerado um microrganismo multirresistente. Dados na literatura que forneçam informações a respeito da resistência desse microrganismo a antimicrobianos em amostras de animais são escassos. Dessa forma, o objetivo deste trabalho foi avaliar o perfil e o seu aumento das resistências a antimicrobianos dentro da medicina veterinária. Um total de 67 isolados de K. pneumoniae, provenientes de diferentes sítios de isolamento de animais domésticos (39/67) e silvestres (28/67), foi confirmado por sequenciamento do gene 16S rRNA. O maior percentual de isolamento de K. pneumoniae foi de amostras de urina, com 16% (11/67), fezes, com 15% (10/67), e pulmão, com 13,5% (09/67). No perfil de resistência, foram testadas 11 categorias de antibióticos, sendo a maior taxa de resistência ao metronidazol 97% (65/67), à ampicilina 94% (63/67), à amoxicilina 93% (62/67), às sulfonamidas 93% (62/67), à colistina 93% (62/67) e à nitrofurantoína 88% (59/67). Aqueles que apresentaram menor taxa de resistência foram: meropenem 3% (2/67), imipenem 6% (4/67) e amicacina 16% (11/67). Todos os isolados foram considerados bactérias multirresistentes (MRD), com o índice de resistência múltipla aos antibióticos (IRMA) variando de 0,15 a 0,85 e com 60 tipos de padrões de resistência. O resultado deste estudo reforça que os animais são reservatórios de K. pneumoniae multirresistentes.(AU)

Klebsiella pneumoniae is an opportunistic pathogen responsible for several types of nosocomial infections and is considered a multiresistant microorganism. Data in the literature that provide information regarding the resistance of this microorganism to antimicrobials in animal samples are scarce. Thus, the objective of this work was to evaluate the profile and its increase of antimicrobial resistance within Veterinary Medicine. A total of 67 K. pneumoniae isolates from different domestic (39/67) and wild (28/67) isolation sites were confirmed by sequencing the 16S rRNA gene. The highest percentage of K. pneumoniae isolation was from urine samples with 16% (11/67), faeces 15% (10/67) and lung 13.5% (09/67). In the resistance profile, 11 categories of antibiotics were tested, with the highest resistance to metronidazole being 97% (65/67), ampicillin 94% (63/67), amoxicillin 93% (62/67), sulfonamides 93% (62 / 67), 93% colistin (62/67), and 88% nitrofurantoin (59/67). The ones with the lowest resistance were: meropenem 3% (2/67), imipenem 6% (4/67) and amikacin 16% (11/67). All isolates were considered multiresistant bacteria (MDR), with the Multiple Resistance to Antibiotics Index (IRMA) ranging from 0.15 to 0.85 and with 60 types of resistance patterns. The result of this study reinforces that the animals are reservoirs of multiresistant K. pneumoniae.(AU)

Perfil de resistência antimicrobiana de Klebsiella pneumoniae isoladas de animais domésticos e silvestres / Antimicrobial resistance profile of Klebsiella pneumoniae isolated from domestic and wild animals

Klebsiella pneumoniae é um patógeno oportunista, responsável por diversos tipos de infecções nosocomiais, e é considerado um microrganismo multirresistente. Dados na literatura que forneçam informações a respeito da resistência desse microrganismo a antimicrobianos em amostras de animais são escassos. Dessa forma, o objetivo deste trabalho foi avaliar o perfil e o seu aumento das resistências a antimicrobianos dentro da medicina veterinária. Um total de 67 isolados de K. pneumoniae, provenientes de diferentes sítios de isolamento de animais domésticos (39/67) e silvestres (28/67), foi confirmado por sequenciamento do gene 16S rRNA. O maior percentual de isolamento de K. pneumoniae foi de amostras de urina, com 16% (11/67), fezes, com 15% (10/67), e pulmão, com 13,5% (09/67). No perfil de resistência, foram testadas 11 categorias de antibióticos, sendo a maior taxa de resistência ao metronidazol 97% (65/67), à ampicilina 94% (63/67), à amoxicilina 93% (62/67), às sulfonamidas 93% (62/67), à colistina 93% (62/67) e à nitrofurantoína 88% (59/67). Aqueles que apresentaram menor taxa de resistência foram: meropenem 3% (2/67), imipenem 6% (4/67) e amicacina 16% (11/67). Todos os isolados foram considerados bactérias multirresistentes (MRD), com o índice de resistência múltipla aos antibióticos (IRMA) variando de 0,15 a 0,85 e com 60 tipos de padrões de resistência. O resultado deste estudo reforça que os animais são reservatórios de K. pneumoniae multirresistentes.(AU)

Klebsiella pneumoniae is an opportunistic pathogen responsible for several types of nosocomial infections and is considered a multiresistant microorganism. Data in the literature that provide information regarding the resistance of this microorganism to antimicrobials in animal samples are scarce. Thus, the objective of this work was to evaluate the profile and its increase of antimicrobial resistance within Veterinary Medicine. A total of 67 K. pneumoniae isolates from different domestic (39/67) and wild (28/67) isolation sites were confirmed by sequencing the 16S rRNA gene. The highest percentage of K. pneumoniae isolation was from urine samples with 16% (11/67), faeces 15% (10/67) and lung 13.5% (09/67). In the resistance profile, 11 categories of antibiotics were tested, with the highest resistance to metronidazole being 97% (65/67), ampicillin 94% (63/67), amoxicillin 93% (62/67), sulfonamides 93% (62 / 67), 93% colistin (62/67), and 88% nitrofurantoin (59/67). The ones with the lowest resistance were: meropenem 3% (2/67), imipenem 6% (4/67) and amikacin 16% (11/67). All isolates were considered multiresistant bacteria (MDR), with the Multiple Resistance to Antibiotics Index (IRMA) ranging from 0.15 to 0.85 and with 60 types of resistance patterns. The result of this study reinforces that the animals are reservoirs of multiresistant K. pneumoniae.(AU)

Species richness and seasonal dynamics of ticks with notes on rickettsial infection in a Natural Park of the Cerrado biome in Brazil.

This study evaluated the species richness and seasonal dynamics of ticks and rickettsial agents infecting ticks in the largest natural Reserve of the Cerrado biome of Brazil, the Grande Sertão Veredas National Park. During 2012-2014, a total of 9531 host-seeking ticks were collected by dry ice traps and dragging, whereas 1563 ticks were collected from small mammals, and 1186 ticks from domestic animals. Overall, the following 12 tick species were identified: Amblyomma auricularium, Amblyomma dubitatum, Amblyomma naponense, Amblyomma ovale, Amblyomma parvum, Amblyomma sculptum, Amblyomma tigrinum, Amblyomma triste, Dermacentor nitens, Rhipicephalus microplus, Rhipicephalus sanguineus sensu lato, and Ornithodoros mimon. The three most abundant tick species, A. sculptum, A. parvum, and A. triste, are likely to develop one generation per year, with adults predominating between spring and autumn, and immature ticks during autumn-winter. Small mammals seem to be important hosts for immature stages of A. parvum, and A. triste, but not for A. sculptum. Molecular analyses revealed the presence of the human pathogen Rickettsia parkeri in 10% of the A. triste ticks, whereas two agents of unknown pathogenicity, Rickettsia bellii and 'Candidatus Rickettsia andeanae' were found in 7 and 5%, respectively, of the A. parvum ticks. A fourth rickettsial agent, Rickettsia amblyommatis, was found in a single A. sculptum tick. Several Vero cell-established isolates of R. parkeri and R. bellii were obtained from A. triste and A. parvum, respectively. Serological analyses of small mammals suggest that they have been infected by R. parkeri and R. bellii, possibly via natural infestations by A. triste and A. parvum, respectively. Because the Park has suffered low anthropic alterations, our results should provide baseline data that shall be used for future comparisons with other Cerrado areas with higher degree of anthropic changes.

Genomic features of a highly virulent, ceftiofur-resistant, CTX-M-8-producing Escherichia coli ST224 causing fatal infection in a domestic cat.

OBJECTIVES: Escherichia coli strains producing extended-spectrum ß-lactamases (ESBLs), especially CTX-M-type, have been largely described in companion animals; however, genomic data are lacking to clarify the clinical impact of ESBL-producing isolates in these hosts. The aim of this study was to present the genomic features of a highly virulent, ceftiofur-resistant, CTX-M-8-producing E. coli isolate from a case of pneumonia in a domestic cat with fatal outcome. METHODS: Genomic DNA was sequenced using an Illumina NextSeq 500 platform and was assembled using CLC Genomic Workbench. Genomic data were analysed using online bioinformatics tools. RESULTS: The genome size was evaluated at 5.1Mb, with 5334 protein-coding sequences. The strain was assigned to sequence type 224 (ST224) and presented genes conferring resistance to ß-lactams (blaCTX-M-8), sulphonamides (sul2), tetracycline (tetA) and trimethoprim (dfrA14) as well as chromosomal point mutations in ParC (S80I), GyrA (S83L) and GyrB (D87N). In addition, the presence of the virulence genes cba, gad, ipfA, iroN, iss, mchF and tsh was detected. CONCLUSION: This draft genome sequence might provide important data for a better understanding of genomic aspects regarding the dissemination of CTX-M-8-producing E. coli in the human-animal-environment interface.

Detection of Bartonella species, including Candidatus Bartonella ovis sp. nov, in ruminants from Mexico and lack of evidence of Bartonella DNA in saliva of common vampire bats (Desmodus rotundus) predating on them.

Bartonella spp. have been identified in many bat species worldwide, including the zoonotic species, Candidatus Bartonella mayotimonensis. The common vampire bat (Desmodus rotundus) preys preferentially on livestock in Latin America and is frequently infected with Bartonella spp. To determine the potential role of D. rotundus in transmitting Bartonella to livestock, common vampire bats and bat-bitten domestic ruminants from Mexico were tested for Bartonella infection by blood culture or conventional PCR. Furthermore, to explore the possibility of bite transmission during blood feeding, saliva swabs from 35 D. rotundus known to be either Bartonella bacteremic (N = 17) or blood culture negative (N = 18) were tested by PCR to detect the presence of Bartonella DNA. Twenty (17.1%) of 117 sheep and 16 (34.8%) of 46 cattle were Bartonella bacteremic by PCR testing. However, none of them were infected with Bartonella strains previously isolated from vampire bats and none of the 35 D. rotundus saliva swabs tested were PCR positive for Bartonella. All but two animals among those which were Bartonella culture and/or PCR positive, were infected with either B. bovis (cattle) or B. melophagi (sheep). Two sheep were infected by a possible new species, Candidatus Bartonella ovis, being phylogenetically closer to B. bovis than B. melophagi. This study does not support the role of D. rotundus as a reservoir of Bartonella species infecting livestock, which could be transmitted via bite and blood feeding and therefore suggest limited risk of zoonotic transmission of Bartonella from common vampire bats to humans.