A preliminary report on critical antimicrobial resistance in Escherichia coli, Enterococcus faecalis, and Enterococcus faecium strains isolated from healthy dogs in Chile during 2021-2022.

Antimicrobial Resistance (AMR) represents one of the main current threats to global public health; where production animals, companion animals, humans, and the environment play a significant role in its dissemination. However, little attention has been given to companion animals as reservoirs and disseminators of relevant antimicrobial resistant bacteria, especially in South American countries such as Chile. For this reason, this research aimed to estimate the prevalence of AMR to different critical antibiotics at a screening level in commensal bacteria such as E. coli and Enterococcus spp., isolated from healthy pet dogs in the Metropolitan Region of Chile, studying their geographical distribution and evaluating associations of phenotypic resistance to different antibiotics. Thus, in E. coli we detected AMR to all critical drugs assessed, including 34.1% to amoxicillin, 20.1% to colistin, 15.7% to enrofloxacin, and 9.2% to cefotaxime. On the other hand, AMR prevalence in E. faecalis was 8.1% for ampicillin and 3.4% for vancomycin; while for E. faecium the AMR prevalence was 19.1% for ampicillin and 10.2% for vancomycin. Additionally, significant differences in prevalence of the different possible AMR were detected according to their geographical distribution, suggesting the existence of various risk factors and stressing the need to establish mitigation measures specific to the differences identified.

Phenotypic and genotypic antimicrobial resistance in Escherichia coli strains isolated from household dogs in Chile.

Introduction: Antimicrobial resistance (AMR) is a major threat to animal and public health worldwide; consequently, several AMR surveillances programs have been implemented internationally in both human and veterinary medicine, including indicator bacteria such as Escherichia coli. However, companion animals are not typically included in these surveillance programs. Nevertheless, there have been reports of increasing levels of antimicrobial resistance in E. coli strains isolated from dogs worldwide. In Chile, there is limited information available on AMR in E. coli isolated from companion animals, which prevents the establishment of objective prevention and control measures. Methods: For this reason, the aim of this study was to characterize the phenotypic and genotypic AMR of E. coli strains isolated from healthy household dogs in Chile. For this purpose, a multi-stage sampling was carried out in the Metropolitan Region of Chile, obtaining samples from 600 healthy dogs. These samples were processed using traditional bacteriology and molecular techniques to isolate E. coli strains. We assessed the minimal inhibitory concentration of 17 antimicrobials and conducted a search of six antimicrobial resistance genes, as well as class 1 and 2 integrons, in the isolated strains. Results: Two-hundred and twenty-four strains of E. coli were recovered, and 96.9% (n = 217) showed resistance to at least one drug and only 3.1% (n = 7) were susceptible to all analyzed antimicrobials. Most strains were resistant to cefalexin (91.5%, n = 205, 1st-generation cephalosporin), followed by ampicillin (68.3%, n = 153) and cefpodoxime (31.3%, n = 70, 3rd-generation cephalosporin). Moreover, 24.1% (n = 54) tested positive for extended-spectrum-ß-lactamases and 34.4% (n = 77) were multidrug resistant. As for the AMR genes, the most detected was qnrB (28.1%, n = 63), followed by blaCTX-M (22.3%, n = 50), and blaTEM-1 (19.6%, n = 44). Additionally, 16.1% (n = 36) harbored class 1 integrons. Our study shows that E. coli strains isolated from healthy household dogs exhibit resistance to several relevant drugs and also antimicrobial resistance genes considered critical for human health. These results can be used as a starting point for the prevention and control of antimicrobial resistance from companion animals. This background should be considered when formulating future resistance surveillance programs or control plans in which companion animals must be included.

Host-pathogen interaction involving cytoskeleton changes as well as non-coding regulation as primary mechanisms for SRS resistance in Atlantic salmon.

The salmonid rickettsial syndrome (SRS) is a systemic bacterial infection caused by Piscirickettsia salmonis that generates significant economic losses in Atlantic salmon (Salmo salar) aquaculture. Despite this disease's relevance, the mechanisms involved in resistance against P. salmonis infection are not entirely understood. Thus, we aimed at studying the pathways explaining SRS resistance using different approaches. First, we determined the heritability using pedigree data from a challenge test. Secondly, a genome-wide association analysis was performed following a complete transcriptomic profile of fish from genetically susceptible and resistant families within the challenge infection with P. salmonis. We found differentially expressed transcripts related to immune response, pathogen recognition, and several new pathways related to extracellular matrix remodelling and intracellular invasion. The resistant background showed a constrained inflammatory response, mediated by the Arp2/3 complex actin cytoskeleton remodelling polymerization pathway, probably leading to bacterial clearance. A series of biomarkers of SRS resistance, such as the beta-enolase (ENO-ß), Tubulin G1 (TUBG1), Plasmin (PLG) and ARP2/3 Complex Subunit 4 (ARPC4) genes showed consistent overexpression in resistant individuals, showing promise as biomarkers for SRS resistance. All these results together with the differential expression of several long non-coding RNAs show the complexity of the host-pathogen interaction of S. salar and P. salmonis. These results provide valuable information on new models describing host-pathogen interaction and its role in SRS resistance.

Developing Methods for Maintaining Genetic Diversity in Novel Aquaculture Species: The Case of Seriola lalandi.

Developing sound breeding programs for aquaculture species may be challenging when matings cannot be controlled due to communal spawning. We developed a genotyping-by-sequencing marker panel of 300 SNPs for parentage testing and sex determination by using data from an in-house reference genome as well as a 90 K SNP genotyping array based on different populations of yellowtail kingfish (Seriola lalandi). The minimum and maximum distance between adjacent marker pairs were 0.7 Mb and 13 Mb, respectively, with an average marker spacing of 2 Mb. Weak evidence of the linkage disequilibrium between adjacent marker pairs was found. The results showed high panel performance for parental assignment, with probability exclusion values equaling 1. The rate of false positives when using cross-population data was null. A skewed distribution of genetic contributions by dominant females was observed, thus increasing the risk of higher rates of inbreeding in subsequent captive generations when no parentage data are used. All these results are discussed in the context of breeding program design, using this marker panel to increase the sustainability of this aquaculture resource.

Virulence and antimicrobial resistance factors in Salmonella enterica serotypes isolated from pigs and chickens in central Chile.

Salmonella enterica is a food-borne pathogen with a wide host-range that during decades has been of public health concern in developed and developing countries. In Chile, the poultry and pig industries represent the biggest contribution of meat consumption in the population, and sanitary regulations have been imposed for Salmonella control. The aim of this work was to determine and characterize Salmonella strains isolated from pigs and chicken raised on commercials farms in Chile. For this, isolates belonging to pigs (n = 46) and poultry (n = 57) were genotyped by two multiplex PCR reactions and virulotyped by the PCR detection of virulence-associated genes. In addition, isolates were serotyped and analyzed by the Kirby Bauer assay to determine their antimicrobial resistance phenotypes. From these analyses 52 genotypes, six serotypes and several multidrug resistance phenotypes and different combinations of virulence-associated genes were detected. These results suggest that S. enterica in pigs and poultry in central Chile should be monitored due to potential consequences in public and animal health.

Characterization and antimicrobial susceptibility of coagulase-positive Staphylococcus isolated in a veterinary teaching hospital in Chile / Caracterización y sensibilidad antimicrobiana de Staphylococcus coagulasa-positivos aislados en un hospital clínico veterinario en Chile

Abstract There is limited information about the prevalence and antimicrobial susceptibility of coagulase-positive Staphylococcus (CoPS) strains in veterinary settings in Chile. The aim of this observational study was to identify and characterize CoPS strains from dogs, owners, veterinary professionals and surfaces in a veterinary teaching hospital at Universidad de Chile to determine the presence of methicillin-resistant strains and evaluate the genetic relationship among the strains. Veterinarians (n = 24), surfaces (n = 10), and healthy dogs (n = 40) and their respective owners (n = 40) were sampled for CoPS. Isolates were identified by PCRand antimicrobial susceptibility was assessed by the disk diffusion method and MIC. The presence of the mecA gene was evaluated by PCR, and the genetic relationship among the strains was established by PFGE. A total of 45 CoPS strains were obtained, eight from veterinary professionals, three from hospital surfaces, eight from owners and 26 from dogs. Nine of the strains were resistant to methicillin (20%), and all of them carried the mecA gene. A high percentage of the strains was resistant to clindamycin (33.3%). Additionally, the isolated CoPS showed high genetic diversity. This study suggests that veterinarians are in high risk of harboring methicillin-resistant CoPS (25% versus 2.5% from owners) and our results provide evidence that clindamycin could not be an empiric alternative for CoPS in the analyzed hospital. This is the first report of methicillin-resistant CoPS in veterinary settings in Chile, considering humans, pets and surfaces.

Resumen Existe información limitada sobre prevalencia y sensibilidad antimicrobiana de cepas de Staphylococcus coagulasa-positivas (CoPS) en entornos veterinarios en Chile. El objetivo de este estudio observacional fue identificar y caracterizar cepas CoPS de perros, duenos, veterinarios y superficies de un hospital veterinario de la Universidad de Chile, determinar la presencia de cepas meticilino-resistentes y evaluar la relación genética entre las cepas. Se colectaron muestras de veterinarios (n = 24), de superficies hospitalarias (n = 10) y de perros sanos (n =40) y sus respectivos duenos (n = 40). Los aislamientos se identificaron mediante PCR y la sensibilidad antimicrobiana se evaluó por difusión en discos y CIM. También se empleó PCR para detectar la presencia del gen mecA; la relación genética entre las cepas se estableció mediante electroforesis de campos pulsantes (PFGE). Se obtuvo un total de 45 cepas de CoPS, 8 de veterinarios, 3 de superficies hospitalarias, 8 de duenos y 26 de perros. Nueve cepas fueron meticilino-resistentes (20%), todas portadoras del gen mecA. Un porcentaje importante de cepas fue resistente a clindamicina (33,3%). Además, las cepas aisladas mostraron una alta diversidad genética. Este estudio sugiere que los veterinarios tienen alto riesgo de portar CoPS resistentes a meticilina (25% versus 2,5% propietarios). Asimismo, nuestros resultados proporcionan evidencia de que la clindamicina podría no ser una alternativa empírica para CoPS en el hospital analizado. Este es el primer estudio de CoPS meticilino-resistentes en entornos veterinarios en Chile que considera humanos, mascotas y superficies.

Advances in Integrated Antimicrobial Resistance Surveillance and Control Strategies in Asia-Pacific Economic Cooperation Economies: Assessment of a Multiyear Building Capacity Project.

Antimicrobial resistance (AMR) is a growing global health concern for both animal and public health, and collaborative strategies are needed to combat the threat. The level of awareness and funding for policies focused on reducing AMR varies between countries. The aim of this study was to compare the integrated surveillance systems for AMR in high and low-middle economies of the Asia-Pacific Economic Cooperation and determine whether there was any improvement from 2015 to 2018. We conducted a survey with a group of 21 countries at different development levels. Associations between the economic development level and the questions of AMR awareness and funding were established using Fisher's exact test. Improvements were identified where countries established public policies for integrated surveillance of AMR. High economies showed greater advancement in several topics related to AMR than low-middle economies. The survey revealed that there is a better understanding surrounding the implications of the emergence of AMR in human medicine than in veterinary medicine, agriculture, and food production. Our results show that countries enhanced overall AMR surveillance over the 4-year-period; however, more research is needed concerning these advances, especially in low-middle economies and the food production sector.

Characterization and antimicrobial susceptibility of coagulase-positive Staphylococcus isolated in a veterinary teaching hospital in Chile.

There is limited information about the prevalence and antimicrobial susceptibility of coagulase-positive Staphylococcus (CoPS) strains in veterinary settings in Chile. The aim of this observational study was to identify and characterize CoPS strains from dogs, owners, veterinary professionals and surfaces in a veterinary teaching hospital at Universidad de Chile to determine the presence of methicillin-resistant strains and evaluate the genetic relationship among the strains. Veterinarians (n=24), surfaces (n=10), and healthy dogs (n=40) and their respective owners (n=40) were sampled for CoPS. Isolates were identified by PCR and antimicrobial susceptibility was assessed by the disk diffusion method and MIC. The presence of the mecA gene was evaluated by PCR, and the genetic relationship among the strains was established by PFGE. A total of 45 CoPS strains were obtained, eight from veterinary professionals, three from hospital surfaces, eight from owners and 26 from dogs. Nine of the strains were resistant to methicillin (20%), and all of them carried the mecA gene. A high percentage of the strains was resistant to clindamycin (33.3%). Additionally, the isolated CoPS showed high genetic diversity. This study suggests that veterinarians are in high risk of harboring methicillin-resistant CoPS (25% versus 2.5% from owners) and our results provide evidence that clindamycin could not be an empiric alternative for CoPS in the analyzed hospital. This is the first report of methicillin-resistant CoPS in veterinary settings in Chile, considering humans, pets and surfaces.

Estimates of Effective Population Size in Commercial and Hatchery Strains of Coho Salmon (Oncorhynchus kisutch (Walbaum, 1792)).

Understanding the genetic status of aquaculture strains is essential for developing management guidelines aimed at sustaining the rates of genetic gain for economically important traits, as well as securing populations that will be robust to climate change. Coho salmon was the first salmonid introduced to Chile for commercial purposes and now comprises an essential component of the country's aquaculture industry. Several events, such as admixture, genetic bottlenecks, and rapid domestication, appear to be determinants in shaping the genome of commercial strains representing this species. To determine the impact of such events on the genetic diversity of these strains, we sought to estimate the effective population size (Ne) of several of these strains using genome-wide approaches. We compared these estimates to commercial strains from North America and Japan, as well as a hatchery strain used for supportive breeding of wild populations. The estimates of Ne were based on a method robust to assumptions about changes in population history, and ranged from low (Ne = 34) to relatively high (Ne = 80) in the Chilean strains. These estimates were higher than those obtained from the commercial North American strain but lower than those observed in the hatchery population and the Japanese strain (with Ne over 150). Our results suggest that some populations require measures to control the rates of inbreeding, possibly by using genomic information and incorporating new genetic material to ensure the long-term sustainability of these populations.

SEROSURVEY FOR SELECTED PARASITIC AND BACTERIAL PATHOGENS IN DARWIN'S FOX (LYCALOPEX FULVIPES): NOT ONLY DOG DISEASES ARE A THREAT.

The Darwin's fox (Lycalopex fulvipes) is one of the most endangered carnivores worldwide, with the risk of disease spillover from domestic dogs being a major conservation threat. However, lack of epidemiologic information about generalist, non-dog-transmission-dependent protozoal and bacterial pathogens may be a barrier for disease prevention and management. To determine the exposure of some of these agents in Darwin's fox populations, 54 serum samples were collected from 47 Darwin's foxes in Southern Chile during 2013-18 and assessed for the presence of antibodies against Brucella abortus, Brucella canis, Coxiella burnetii, pathogenic Leptospira (serovars Grippotyphosa, Pomona, Canicola, Hardjo, and Copehageni), Toxoplasma gondii, and Neospora caninum. The highest seroprevalence was detected for T. gondii (78%), followed by pathogenic Leptospira (14%). All the studied Leptospira serovars were confirmed in at least one animal. Two foxes seroconverted to Leptospira and one to T. gondii during the study period. No seroconversions were observed for the other pathogens. No risk factors, either intrinsic (sex, age) or extrinsic (season, year, and degree of landscape anthropization), were associated with the probability of being exposed to T. gondii. Our results indicate that T. gondii exposure is widespread in the Darwin's fox population, including in areas with minimal anthropization, and that T. gondii and pathogenic Leptospira might be neglected threats to the species. Further studies identifying the causes of morbidity and mortality in Darwin's fox are needed to determine if these or other pathogens are having individual or population-wide effects in this species.

Risk Factors for Positivity to Shiga Toxin-Producing Escherichia coli and Salmonella enterica in Backyard Production Systems Animals from Metropolitana Region, Chile: A Threat to Public Health?

In the Metropolitana region of Chile there are 3836 backyard production systems (BPS), characterized as small-scale systems. They act as a source of zoonotic pathogens, such as Salmonella enterica and Shiga toxin-producing Escherichia coli (STEC), whose prevalence in BPS has not been fully described. The objective of this study was to determine the positivity for both agents in BPS and to establish the risk factors related to their presence. In each BPS, an epidemiological survey was undertaken, and stool samples were collected to detect these pathogens via bacteriological culture and conventional PCR techniques. Subsequently, multivariable logistic regression models were applied to establish the risk factors associated with their presence. BPS positivity rates of 11.76% for STEC and 4.7% for S. enterica were observed. The systems showed poor welfare standards and a lack of biosecurity measures. The risk factor analysis concluded that the Gini-Simpson index (p = 0.030; OR = 1.717) and the presence of neighboring intensive poultry or swine production systems (p = 0.019; OR = 20.645) act as factors that increased the risk of positivity with respect to STEC. In the case of S. enterica, exchanging embryonated eggs (p = 0.021; OR = 39) and the presence of debeaked chickens (p = 0.001; OR = 156) were determined as factors that increased the risk of positivity for this agent. For positivity with respect to both pathogens, the Gini-Simpson index (p = 0.030; OR = 1.544) and being INDAP/PRODESAL users (p = 0.023; OR = 15.026) were determined as factors that increased the risk, whereas the type of confinement (p = 0.002; OR = 0.019) decreased it. Epidemiological surveillance of these neglected populations is lacking, highlighting the fact that STEC and S. enterica maintenance on BPS represents a potential threat to public health.

Survey of Zoonotic Bacterial Pathogens in Native Foxes in Central Chile: First Record of Brucella canis Exposure.

Brucella abortus, B. canis, and pathogenic Leptospira are zoonotic pathogens that infect humans, as well as domestic and wild animals. In wild canids, they may affect their fertility and reproduction, threatening their conservation. Wild canids play a crucial role in the environment as meso- and top-predators and environmental sentinels for zoonotic pathogens. In Chile, three species of wild canids are present, and due to changes in land use and environmental dynamics, it is of utmost relevance to determine the role of these species in the epidemiology of brucellosis and leptospirosis. This study aimed to detect the exposure to B. abortus, B. canis, and pathogenic Leptospira by serologic, bacteriologic, and molecular techniques in native foxes from rehabilitation and exhibition centers in Central Chile. Forty-six blood samples were obtained from Lycalopex culpaeus and L. griseus, detecting 10.9% of seropositivity to B. canis and 7.7% to L. Javanica. No seropositivity was seen for B. abortus. Exposure was not registered by culture and qPCR in any of the sampled animals. Our findings are the first register of exposure to any Brucella species in wild canids in Chile and highlight the need to establish surveillance programs of these emerging pathogens.

Genomic Epidemiology of Shiga Toxin-Producing Escherichia coli Isolated from the Livestock-Food-Human Interface in South America.

Shiga toxin-producing Escherichia coli (STEC) are zoonotic pathogens responsible for causing food-borne diseases in humans. While South America has the highest incidence of human STEC infections, information about the genomic characteristics of the circulating strains is scarce. The aim of this study was to analyze genomic data of STEC strains isolated in South America from cattle, beef, and humans; predicting the antibiotic resistome, serotypes, sequence types (STs), clonal complexes (CCs) and phylogenomic backgrounds. A total of 130 whole genome sequences of STEC strains were analyzed, where 39.2% were isolated from cattle, 36.9% from beef, and 23.8% from humans. The ST11 was the most predicted (20.8%) and included O-:H7 (10.8%) and O157:H7 (10%) serotypes. The successful expansion of non-O157 clones such as ST16/CC29-O111:H8 and ST21/CC29-O26:H11 is highlighted, suggesting multilateral trade and travel. Virulome analyses showed that the predominant stx subtype was stx2a (54.6%); most strains carried ehaA (96.2%), iha (91.5%) and lpfA (77.7%) genes. We present genomic data that can be used to support the surveillance of STEC strains circulating at the livestock-food-human interface in South America, in order to control the spread of critical clones "from farm to table".

Detection of Antimicrobial Residues in Poultry Litter: Monitoring a Risk through a Selective and Sensitive HPLC-MS/MS Method.

Tetracyclines, sulphonamides, and quinolones are families of antimicrobials (AMs) widely used in the poultry industry and can excrete up to 90% of AMs administrated, which accumulate in poultry litter. Worryingly, poultry litter is widely used as an agriculture fertilizer, contributing to the spread AMs residues in the environment. The aim of this research was to develop a method that could simultaneously identify and quantify three AMs families in poultry litter by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Samples of AMs free poultry litter were used to validate the method according to 657/2002/EC and VICH GL49. Results indicate that limit of detection (LOD) ranged from 8.95 to 20.86 µg kg-1, while limits of quantitation (LOQ) values were between 26.85 and 62.58 µg kg-1 of tetracycline, 4-epi-tetracycline, oxytetracycline, 4-epi-oxytetracycline, enrofloxacin, ciprofloxacin, flumequine, sulfachloropyridazine, and sulfadiazine. Recoveries obtained ranged from 93 to 108%. The analysis of field samples obtained from seven commercial poultry flocks confirmed the adequacy of the method since it detected means concentrations ranging from 20 to 10,364 µg kg-1. This provides us an accurate and reliable tool to monitor AMs residues in poultry litter and control its use as agricultural fertilizer.

Evaluation of Antibiotic Dissemination into the Environment and Untreated Animals, by Analysis of Oxytetracycline in Poultry Droppings and Litter.

Oxytetracycline (OTC) is widely used in broiler chickens. During and after treatment a fraction of OTC is excreted in its original form and as its epimer, 4-epi-OTC in droppings. To address the transfer of OTC into the environment, we evaluated the dissemination of OTC and 4-epi-OTC from treated birds to the environment and sentinels, through the simultaneous analysis of broiler droppings and litter. Male broiler chickens were bred in controlled conditions. One group was treated by orogastric tube with 80 mg kg-1 of OTC and two groups received no treatment (sentinels). OTC+4-epi-OTC were analyzed and detected by a HPLC-MS/MS post the end of treatment. The highest concentrations of OTC+4-epi-OTC were detected in the droppings of treated birds 14-days following the end of treatment (2244.66 µg kg-1), and one day following the end of treatment in the litter (22,741.68 µg kg-1). Traces of OTC+4-epi-OTC were detected in the sentinels' droppings and litter (<12.2 µg kg-1). OTC+4-epi-OTC can be transferred from treated birds to the environment and to other untreated birds. The presence and persistence of OTC+4-epi-OTC in litter could contribute to the selection of resistant bacteria in the environment, increasing the potential hazard to public and animal health.

Genomic features and antimicrobial resistance patterns of Shiga toxin-producing Escherichia coli strains isolated from food in Chile.

Shiga toxin-producing Escherichia coli (STEC) is a zoonotic pathogen that causes severe illness in humans, often associated with foodborne outbreaks. Antimicrobial resistance among foodborne E. coli has increased over the last decades becoming a public health issue. In this study, the presence and features of STEC were investigated in samples of meat, seafood, vegetables and ready-to-eat street-vended food collected in Chile, using a genomic and microbiological approach. Phenotypic and genotypic antimicrobial resistance profiles were determined, and serotype, phylogroup, sequence type (ST) and phylogenomics were predicted using bioinformatic tools. Three thousand three hundred samples collected in 2019 were screened, of which 18 were positive for STEC strains (0.5%), with stx2a (61.1%) being the predominant stx subtype. The presence of the virulence genes lpfA (100%), iha and ehaA (94.4%), and ehxA, hlyA and saa (83.3%) was confirmed among the STEC strains; the Locus of adhesion and autoaggregation (LAA) was predicted in 14 (77.8%) strains. Strains displayed resistance to colistin (100%), and intermediate resistance to enrofloxacin (11.1%) and chloramphenicol (5.6%). In this regard, mutations in the two-component regulatory system genes pmrA (S29G), pmrB (D283G) and phoP (I44L), and the presence of the qnrB19 gene were confirmed. STEC strains belonged to ST11231 (38.9%), ST297 and ST58 (16.7% each), and ST1635, ST11232, ST446, ST442 and ST54 (5.6% each), and the most frequently detected serotypes were O113:H21 (44.4%), O130:H11 and O116:H21 (16.7% each), and O174:H21 (11.1%). Strains belonging to the international ST58 showed genomic relatedness with worldwide strains from human and non-human sources. Our study reports for the first time the genomic profile of STEC strains isolated from food in Chile, highlighting the presence of international clones and sequence types commonly associated with human infections in different geographical regions, as well as the convergence of virulence and resistance in STEC lineages circulating in this country.

Antimicrobial Use in Companion Animals: Assessing Veterinarians' Prescription Patterns through the First National Survey in Chile.

Although the relationship between the use of antimicrobials and the development of resistant bacteria is well established, information about the use of antimicrobials in companion animals has been poorly described, which represents a risk to public health. The aim of this study was to describe the antimicrobial prescription patterns of Chilean companion animal veterinarians. A nationally distributed survey targeted at companion animal veterinarians was designed. The survey included questions about the veterinarian's demographics, bacterial diseases treated, prescribed antimicrobials, and the use of laboratory diagnostic tools. Three hundred twenty-three responses were collected, most of the respondents were female (59.4%). The most frequently reported bacterial diseases were pyoderma (17.2%), followed by otitis and abscesses (7.4%). The antimicrobials most used corresponded with critically or highly important drugs for veterinary and human use, including ß-lactams (65.3%), quinolones (36.2%) and tetracyclines (23.2%). Only 15% of the veterinarians reported the use of laboratory diagnostic tests, although 67% declared they were aware of the official antimicrobial classification schemes. Our results describe for the first time the usage of antimicrobials by veterinarian practitioners in Chile to treat different diseases in companion animals. The data presented here provide a baseline that could help to promote the implementation of clinical guidelines and regulations in order to improve current treatments.

First genome sequence of Chilean Brucella canis SCL strain provides insights on the epidemiology and virulence factors, explaining differences between geographical origins

BACKGROUND: Brucella canis is the etiological agent of canine brucellosis, a worldwide neglected zoonosis that constitutes one of the major infectious causes of infertility and reproductive failure in dogs. Although genomic information available for this pathogen has increased in recent years, here we report the first genome sequencing of a B. canis strain in Chile, and the differences in virulence genes with other B. canis strains. RESULTS: Genome assembly produced a total length of 3,289,216 bp, N50 of 95,163 and GC% of 57.27, organized in 54 contigs in chromosome I, and 21 contigs in chromosome II. The genome annotation identified a total of 1981 CDS, 3 rRNA and 36 tRNA in chromosome I, and 1113 CDS and 10 tRNA in chromosome II. There is little variation between the different strains and the SCL isolate. Phylogenetic analysis showed that the Chilean SCL strain is closely related to B. canis and B. suis strains. Small differences were found when compared to the Serbian isolate, but all strains shared the same recent common ancestor. Finally, changes in the sequence of some virulence factors showed that the SCL strain is similar to other South American B. canis strains. CONCLUSIONS: This work sequenced and characterized the complete genome of B. canis strain SCL, evidencing the complete presence of all the genes of the virB operon, and minor changes in outer membrane proteins and in the urease operon. Our data suggest that B. canis was introduced from North America and then spread throughout the South American continent.

A household case evidences shorter shedding of SARS-CoV-2 in naturally infected cats compared to their human owners.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been detected in domestic and wild cats. However, little is known about natural viral infections of domestic cats, although their importance for modelling disease spread, informing strategies for managing positive human-animal relationships and disease prevention. Here, we describe the SARS-CoV-2 infection in a household of two human adults and sibling cats (one male and two females) using real-time RT-PCR, an ELISA test, viral sequencing, and virus isolation. On May 5th, 2020, the cat-owners tested positive for SARS-CoV-2. Two days later, the male cat showed mild respiratory symptoms and tested positive. Four days after the male cat, the two female cats became positive, asymptomatically. Also, one human and one cat showed antibodies against SARS-CoV-2. All cats excreted detectable SARS-CoV-2 RNA for a shorter duration than humans and viral sequences analysis confirmed human-to-cat transmission. We could not determine if cat-to-cat transmission also occurred.

Prevalence and Genomic Characterization of Brucella canis Strains Isolated from Kennels, Household, and Stray Dogs in Chile.

Canine brucellosis caused by Brucella canis is a zoonotic disease that causes reproductive alterations in dogs, such as infertility, abortion, and epididymitis. This pathogen is especially prevalent in South America, and due to the lack of official control programs and the growing trend of adopting dogs it constitutes a public health risk that must be addressed. The aim of this study was to determine the prevalence of B. canis infection in kennel, shelter, and household dogs and to characterize the genomic properties of circulating strains, including ure and virB operons and omp25/31 genes. Samples from 771 dogs were obtained, and the infection was detected by blood culture and/or serology in 7.0% of the animals. The complete ure and virB operons and the omp25/31 genes were detected. Interestingly, we found different single-nucleotide polymorphisms (SNPs) in some of the analyzed genes, which could mean a change in the fitness or virulence of these strains. This study provides further evidence about dogs as a source of B. canis strains that can infect people. This also highlights the need to implement official control programs, including the mandatory testing of dogs, especially stray dogs, before adoption.