Pathological Characterization and Risk Factors of Splenic Nodular Lesions in Dogs (Canis lupus familiaris).

In dogs, the spleen is a secondary lymphoid organ that can be affected by both neoplastic and non-neoplastic nodules. In general, few studies relate histopathological diagnosis to tumor size and the number of nodules in spleen biopsies. Some of these studies are inconclusive regarding the difference between neoplastic and non-neoplastic lesions and have small sample sizes or do not consider all splenic lesions. This study aimed to characterize splenic masses and determine risk factors for spleen tumors in dogs. A total of 507 histological reports corresponding to the diagnosis of splenic lesions in dogs from a private laboratory of animal pathology in the Metropolitan Region, Chile, were used. Data were analyzed by descriptive statistics and multiple logistic regression. The most frequent neoplastic and non-neoplastic diagnoses were hemangiosarcoma and hyperplasia, respectively. Most of the cases occurred in male (265 cases, 52.3%), senior (421 cases, 83%), and purebred individuals (342 cases, 67.5%). The most affected breeds were the Cocker Spaniel, German Shepherd, and Labrador Retriever. The most frequent lesion was a single nodule. The variables that exhibited a greater risk for the presentation of splenic neoplasia were male sex (odds ratio (OR) = 16.21; 95% confidence interval (CI) 1.741-150.879; p = 0.014), the presence of two or more splenic nodules (OR = 3.94; 95% CI 2.168-7.177; p < 0.001), an increase in nodule size greater than 2 cm (OR for quartiles 2, 3 and 4 of 2.2; 95% CI 1.036-4.941; p = 0.041, 2.9; 95% CI 1.331-6.576; p = 0.008, and 3.6; 95% CI 1.562-8.499; p = 0.003, respectively), and increasing age (OR = 1.23; 95% CI 1.048-1.436; p = 0.011). On the other hand, males exhibited a lower risk as age increases (OR = 0.76; 95% CI 0.615-0.928; p = 0.008). In conclusion, this study identified that males, multinodular presentation, nodule size, and age are risk factors for the occurrence of splenic neoplasia in dogs, knowledge that will contribute to the diagnostic management of dogs with spleen lesions.

Spatial clusters, temporal behavior, and risk factors analysis of rabies in livestock in Ecuador.

Rabies, a globally distributed and highly lethal zoonotic neglected tropical disease, has a significant impact in South America. In Ecuador, animal rabies cases are primarily linked to livestock, and hematophagous bats play a crucial role in disease transmission. This study aims to identify temporal trends, spatial patterns, and risk factors for animal rabies in Ecuador between 2014 and 2019. Epidemiological survey reports from the official Animal Rabies Surveillance Program of the Phyto and Zoosanitary Regulation and Control Agency of Ecuador (AGROCALIDAD) were used. The Animal Rabies Surveillance Program from AGROCALIDAD consists of an official passive surveillance program that receives reports from farmers or individuals (both trained or untrained) who have observed animals with neurological clinical signs and lesions compatible with bat bites, or who have seen or captured bats on their farms or houses. Once this report is made, AGROCALIDAD personnel is sent for field inspection, having to confirm the suspicion of rabies based on farm conditions and compatibility of signs. AGROCALIDAD personnel collect samples from all suspicious animals, which are further processed and analyzed using the Direct Fluorescent Antibody (DFA) test for rabies confirmatory diagnosis. In this case, study data comprised 846 bovine farms (with intra-farm sample sizes ranging from 1 to 16 samples) located in different ecoregions of Ecuador; out of these, 397 (46.93%) farms tested positive for animal rabies, revealing six statistically significant spatial clusters. Among these clusters, three high-risk areas were identified in the southeast of Ecuador. Seasonality was confirmed by the Ljung-Box test for both the number of cases (p < 0.001) and the positivity rate (p < 0.001). The Pacific Coastal lowlands and Sierra regions showed a lower risk of positivity compared to Amazonia (OR = 0.529; 95% CI = 0.318 - 0.883; p = 0.015 and OR = 0.633; 95% CI = 0.410 - 0.977; p = 0.039, respectively). The breeding of non-bovine animal species demonstrated a lower risk of positivity to animal rabies when compared to bovine (OR = 0.145; 95% CI = 0.062 - 0.339; p < 0.001). Similarly, older animals exhibited a lower risk (OR = 0.974; 95% CI = 0.967 - 0.981; p < 0.001). Rainfall during the rainy season was also found to decrease the risk of positivity to animal rabies (OR = 0.996; 95% CI = 0.995 - 0.998; p < 0.001). This study underscores the significance of strengthening the national surveillance program for the prevention and control of animal rabies in Ecuador and other countries facing similar epidemiological, social, and geographical circumstances.

High Exposure to Livestock Pathogens in Southern Pudu (Pudu puda) from Chile.

A significant gap in exposure data for most livestock and zoonotic pathogens is common for several Latin America deer species. This study examined the seroprevalence against 13 pathogens in 164 wild and captive southern pudu from Chile between 2011 and 2023. Livestock and zoonotic pathogen antibodies were detected in 22 of 109 wild pudus (20.18%; 95% CI: 13.34-29.18) and 17 of 55 captive pudus (30.91%; 95% CI: 19.52-44.96), including five Leptospira interrogans serovars (15.38% and 10.71%), Toxoplasma gondii (8.57% and 37.50%), Chlamydia abortus (3.03% and 12.82%), Neospora caninum (0.00% and 9.52%), and Pestivirus (8.00% and 6.67%). Risk factors were detected for Leptospira spp., showing that fawn pudu have statistically significantly higher risk of positivity than adults. In the case of T. gondii, pudu living in "free-range" have a lower risk of being positive for this parasite. In under-human-care pudu, a Pestivirus outbreak is the most strongly suspected as the cause of abortions in a zoo in the past. This study presents the first evidence of Chlamydia abortus in wildlife in South America and exposure to T. gondii, L. interrogans, and N. caninum in wild ungulate species in Chile. High seroprevalence of livestock pathogens such as Pestivirus and Leptospira Hardjo in wild animals suggests a livestock transmission in Chilean template forest.

Specific and non-specific effects of Mycobacterium bovis BCG vaccination in dairy calves.

Bovine tuberculosis (bTB) is a chronic disease mainly caused by Mycobacterium bovis, a zoonotic pathogen with economic significance as it leads to reduced milk and meat production, and high costs for control measures. The Bacillus Calmette-Guérin (BCG) vaccine, primarily used to prevent tuberculosis in humans, has also been studied for controlling bTB. While showing effectiveness in preventing M. bovis infection and disease in cattle, the BCG vaccine can induce non-specific effects on the immune system, enhancing responses to infections caused by unrelated pathogens, and also having non-specific effects on lactation. The aim of this study is to describe both the specific and non-specific effects of BCG vaccination in calves from a commercial dairy herd in central Chile. Diagnosis of M. bovis infection was performed through the IFNγ release assay (IGRA) using ESAT6/CFP-10 and Rv3615c antigens. The records of milk production, somatic cell count (SCC), clinical mastitis (CM) and retained placenta (RP) during the first lactation were compared between vaccinated and non-vaccinated animals. The breed (Holstein Friesian [HF] v/s HF × Swedish Red crossbred [HFSR]) and the season (warm v/s cold) were also analyzed as categorical explanatory variables. Results of IGRA showed significant differences between vaccinated and control groups, indicating a vaccine efficacy of 58.5% at 18 months post vaccination in HFSR crossbred animals. Although milk production did not vary, SCC and CM showed differences between groups, associated to the breed and the season, respectively. When analyzing CM and RP as a whole entity of disease, BCG showed protection in all but the cold season variables. Overall, the BCG vaccine induced protective specific and non-specific effects on health parameters, which may be influenced by the breed of animals and the season. These results provide new features of BCG protection, supporting initiatives for its implementation as a complementary tool in bTB control.

Epidemiological Characterization of Isolates of Salmonella enterica and Shiga Toxin-Producing Escherichia coli from Backyard Production System Animals in the Valparaíso and Metropolitana Regions.

Backyard production systems (BPS) are distributed worldwide, rearing animals recognized as reservoirs of Salmonella enterica and Shiga toxin-producing Escherichia coli (STEC), both zoonotic pathogens. The aim of this study was to characterize isolates of both pathogens obtained from animals raised in BPS from two central Chile regions. The presence of pathogens was determined by bacterial culture and confirmatory PCR for each sampled BPS, calculating positivity rates. Multivariate logistic regression was used to determine risk factors. Additionally, phenotypic antimicrobial resistance was determined. A positivity rate of 2.88% for S. enterica and 14.39% for STEC was determined for the complete study region (Valparaíso and Metropolitana regions). Risk factor analysis suggests that the presence of ruminants (OR = 1.03; 95% CI = 1.002-1.075) increases the risk of STEC-positive BPS, and the presence of ruminants (OR = 1.05; 95% CI = 1.002-1.075) and the animal handlers being exclusively women (OR = 3.54; 95% CI = 1.029-12.193) increase the risk for S. enterica/STEC positivity. Eighty percent of S. enterica isolates were multidrug resistant, and all STEC were resistant to Cephalexin. This study evidences the circulation of multidrug-resistant zoonotic bacterial strains in animals kept in BPS and the presence of factors that modify the risk of BPS positivity for both pathogens.

Dental Disease in Rabbits (Oryctolagus cuniculus) and Its Risk Factors-A Private Practice Study in the Metropolitan Region of Chile.

Rabbits (Oryctolagus cuniculus) have elodont dentition, a characteristic that predisposes them to the development of Acquired Dental Disease (ADD), which is a multifactorial disease. The aim of this study was to assess the risk factors for ADD in domestic pet rabbits. To do this, a retrospective analysis of 1420 rabbits treated at a private practice during 2018-2021 was performed. For this, several variables were retrieved from clinical records, in addition to signology at the time of diagnosis. ADD was found on 25.4% of rabbits, mostly on their cheek teeth. In addition, age (OR = 1.029; 95% CI = 1.023-1.035; p < 0.001) and sex (male) (OR = 1.591; 95% CI = 1.226-2.064; p < 0.001) were found to be significant risk factors for ADD. In contrast, a free lifestyle (OR = 0.565; 95% CI = 0.362-0.882; p = 0.012) and consuming hay in the diet (OR = 0.323; 95% CI = 0.220-0.473; p < 0.001) were protective factors. In conclusion, ADD has a high prevalence and is usually underdiagnosed, highlighting the need for an exhaustive evaluation of patients during the clinical examination. This study improves our knowledge of ADD and its prevention.

Molecular and phylogenetic analysis of herpesviruses in endangered free-ranging cervids of Chile: ovine gammaherpesvirus-2-A novel threat to wild and domestic animal health in Chilean Patagonia.

Introduction: Herpesvirus infections have been highlighted as emerging diseases affecting wildlife health and the conservation of several taxa. Malignant catarrhal fever (MCF) and infectious keratoconjunctivitis (IKC) are two viruses that infect wild ruminants. Nevertheless, epidemiological data on herpesviruses in South American wild ruminants are limited. An outbreak of caprine gammaherpesvirus-2 (CpHV-2) that recently was suspected as the cause of MCF in southern pudus (Pudu puda) prompted the need to conduct molecular screenings in Chilean cervids to understand the epidemiology of herpesviruses. The aim of this study was to determine the occurrence and genetic diversity of herpesviruses in free-ranging cervids from Chile. Methods: Herpesvirus infection was assessed in antemortem blood samples (n = 86) from pudus (n = 81) and huemuls (Hippocamelus bisulcus) (n = 5), as well as postmortem samples of spleen (n = 24) and lung (n = 3) from pudus, using a nested pan-herpesvirus PCR assay. Results: Combining all suitable sample types, DNA of pudu gammaherpesvirus-1 was detected in five pudues and five huemuls, with an overall prevalence of 9.90% (n = 10/101; 95% CI = 5.11-17.87%). One pudu tested positive for ovine gammaherpesvirus-2 (n = 1/96; 1.04%; 95% CI = 0.05-6.49%), and one pudu tested positive for a Macavirus sequence with 98.63 similarity to ovine gammaherpesvirus-2 (n = 96; 1.04%; 95% CI = 0.05-6.49%). Discussion: To the best of our knowledge, this is the first report of a herpesvirus in huemul and of ovine gammaherpesvirus-2 in Chile. Our results also confirm the active circulation of herpesvirus in free-ranging cervids in Chilean Patagonia, and as such, MCF should be considered as a possible cause of disease in free-ranging Chilean pudus and livestock species. Further research is necessary to develop a plan of systematic monitoring (serological and pathological screening) of herpesviruses in Chilean wild and domestic ruminants to understand their diversity and impact on animal health and conservation.

Vaccination of Calves with the Mycobacterium bovis BCG Strain Induces Protection against Bovine Tuberculosis in Dairy Herds under a Natural Transmission Setting.

Bovine tuberculosis (bTB) is a zoonotic disease caused mainly by Mycobacterium bovis, which is associated with major economic losses for milk and meat producers. The objective of this trial was to assess the efficacy of the BCG Russia strain in a cohort study performed under field conditions, with the vaccination of calves in seven dairy farms from a high prevalence area in central Chile. The trial was performed with 501 animals, subcutaneously vaccinated with 2-8 × 105 colony-forming units of BCG, whilst 441 matched control animals received a saline placebo. Peripheral blood was collected at 6, 12 and 18 months post-vaccination, and infection status was determined using the IFNγ release assay in conjunction with the DIVA (Detecting Infected amongst Vaccinated Animals) antigens ESAT-6, CFP-10 and Rv3615c. The BCG vaccine showed a low but significant level of protection of 22.4% (95% CI 4.0 to 36.4) at the end of the trial. However, diverse levels of protection and a variable duration of immunity were observed between trial herds. This diverse outcome could be influenced by the general health condition of calves and their exposition to non-tuberculous mycobacteria. These results suggest that BCG vaccination of dairy calves in a natural transmission setting confers variable protection to animals against bTB in a high prevalence area.

Vaccination of Holstein heifers with Mycobacterium bovis BCG strain induces protection against bovine tuberculosis and higher milk production yields in a natural transmission setting.

Bovine tuberculosis (TB) is a chronic disease caused mainly by Mycobacterium bovis, a zoonotic pathogen that has a worldwide distribution causing serious economic losses for milk and meat producers. In Chile, the disease in dairy cattle has a heterogeneous distribution, where the Metropolitan Region concentrates the highest animal prevalence and the main challenge for the national control and eradication programme. In this epidemiological context, vaccination with the M. bovis Bacillus Calmette-Guerin (BCG) vaccine might be a useful strategy for disease prevention and control. The objective of this study was to assess the efficacy and impacts on productivity and fertility of vaccination with the BCG Russia strain in 11 month-old heifers from a dairy farm, under a natural transmission condition. Sixty-two animals were vaccinated via the subcutaneous route with the equivalent of one human dose of BCG, and 60 control animals received saline. Subsequently, blood sampling was performed at 3, 6, 9, 12, 15 and 18 months post-inoculation, and infection status was determined using the IFNγ release assay (IGRA) with the DIVA (differentiate infected from vaccinated animals) antigens ESAT-6, CFP-10 and Rv3615c. Efficacy was calculated as the percentage of reduction in the incidence of infection attributable to vaccination, which showed a statistically significant level of overall protection of 66.5%. No adverse effects on fertility and production were recorded. In contrast, we observed beneficial effects of vaccination on several milk production parameters, with the milk yield in the first 100 days after calving in the BCG group significantly higher compared to unvaccinated heifers (p < .05). These results suggest that BCG vaccination of heifers in a natural transmission setting might result in both sanitary and productive benefits, supporting its implementation as a new strategy for TB prevention in a high prevalence area of Chile.

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.

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".

Serological prevalence and risk factors of Toxoplasma gondii in Zoo Mammals in Chile.

Toxoplasma gondii is a zoonotic cosmopolitan protozoan that causes a high mortality rate among zoo mammals such as New World primates, meerkats, marsupials and Pallas' cat. It has been recently reported in Chile, mainly among wild populations, but also as the cause of death of a kangaroo and a mara. However, there has not been a T. gondii report at a Zoo population level in Chile in the last 35 years. The aim of the study was to estimate the seroprevalence and risk factors associated with T. gondii infection in mammals housed in a zoo located in the Metropolitan Region of Chile between 2011 and 2018. In this study, we analyzed 350 samples, from 324 animals, belonging to 57 species of carnivores, non-human primates, macropodids, ungulates and rodents to detect the presence of anti-T. gondii antibodies. Additionally, 20 animals were longitudinally sampled to evaluate intra-zoo infection. Using a commercial indirect Enzyme-Linked Immuno Sorbent Assay (ELISA) test, we detected T. gondii antibodies in 72 (22.2 %) samples. The overall seroprevalence estimates were 48.4 % in felines, 22.9 % in non-feline carnivores, 21.1 % in ungulates and 15.0 % in non-human primates. There were no positive samples from rodents or marsupials. Of animals sampled longitudinally, only a culpeo fox (Lycalopex cualpaeus) became seropositive along the study indicating exposition inside the facility. T. gondii seroprevalence differed significantly in taxonomic groups (p = 0.003), felines are statistically different from non-feline carnivores (NFC) (p = 0.040), ungulate (p = 0.027) and non-human primates (NHP) (p = 0.009). Annual prevalence comparison was performed showing no statistical difference (p = 0.941). A multivariable logistic regression was performed to ascertain the effect of taxonomic groups, proximity to water sources, diet, sex and type of housing on seropositivity. Only taxonomic group was statistically significant, indicating that NFC (OR = 0.35; 95 % CI = 0.15 - 0.83; p = 0.017), ungulates (OR = 0.30; 95 % CI = 0.13 - 0.69; p = 0.005), and NHP (OR = 0.25; 95 % CI = 0.09 - 0.72; p = 0.010) have lower risk of positivity to T. gondii compared to felines. Additionally, a black-faced spider monkey (Ateles chamek) and a siamang (Symphalangus syndactylus) were seropositive, being the first description of T. gondii infection in these species worldwide. As seen in previous studies, the widespread presence and exposure of T. gondii in zoo mammals was confirmed, and there may be contact with the agent and transmission within the zoo, which was confirmed by one animal became seropositive over the time. This fact could be a health problem for animals susceptible to fatal toxoplasmosis.

Urban public squares as potential hotspots of dog-human contact: A spatial analysis of zoonotic parasites detection in Gran Santiago, Chile.

Zoonotic parasites are a major public health problem globally, representing a hazard to human health. The infections occur through contact with different parasite forms. Public squares are a common meeting place for people and dogs, becoming a risk area for transmission to other dogs and humans. A spatial analysis of zoonotic parasite distribution in urban public squares in Gran Santiago, Chile was performed to elucidate its importance, exploring spatial aggregation on positivity rates at commune level. A total of 170 stool samples were analysed; 54 (31.7%) were positive for gastrointestinal-zoonotic parasites. Positivity was detected in 27 (79.4%) squares. Diversity of parasite species was variable across sites. Toxocara canis was the most frequently detected (27 samples, 21/34 squares), followed by Toxascaris leonina (13 samples, 10/34 squares), cestode eggs (10 samples, 9/34 squares), Giardia sp. (6 samples, 5/34 squares), Ancylostomatidae-like eggs (4 samples, 4/34 squares), Cryptosporidium spp. (2 samples, 2/34 squares) and Entamoeba spp. (1 sample, 1/34 square). Spatial autocorrelation (SA) was observed at commune level. The local indicators of the spatial association test showed statistical significance for cestode eggs (p < 0.001) with a negative SA (Moran's I Index = -0.2929, CI-95% = -0.3869--0.1989) and Ancylostomatidae (p = 0.046) with a positive SA (Moran's I Index = 0.1404, CI-95% = 0.0499-0.2309). Toxocara canis is the most prevalent geo-helminth in public places of Santiago, Chile. Spatial autocorrelation was detected for cestode eggs and Ancylostomatidae. Diagnosis and detection of parasites is key for the establishment of control/eradication of environmental dissemination following One Health guidelines.

Cryptosporidium spp. and Giardia spp. in Free-Ranging Introduced Monk Parakeets from Santiago, Chile.

Monk Parakeets (Myiopsitta monachus) are medium-sized parrots that due to international pet trade currently exist as invasive species in 19 countries globally. Such is the case of Chile, where Monk Parakeets have thrived in the city of Santiago. Cryptosporidium spp. and Giardia spp. are worldwide distributed gastrointestinal parasites whose potential hosts include birds and humans. The present study sought to determine the presence of these pathogens in Monk Parakeets from Santiago. During the austral summers of 2017 and 2018, 207 Monk Parakeet nestlings were captured, and fecal samples were studied via microscopical analyses. Environmental data related to the trees in which the nestlings were captured were analyzed to establish the existence of infection clusters. Associations between spatial clusters, environmental variables, and the presence or absence of these pathogens were explored. In total, 33 samples were positive to the presence of one or both protozoa. Of the 33, Cryptosporidium spp. oocysts were detected in 10 nestlings (30%) while Giardia spp. cysts were detected in 25 (76%). Two nestlings presented poly-parasitism (6%). Statistical analyses established pruned trees as a potential protective factor against infection with these parasites. The present study corresponds to the second report of Cryptosporidium spp. in Monk Parakeets in Chile and the first worldwide report of Giardia spp. in these birds, emphasizing Monk Parakeet's potential role as a reservoir and pathogen disseminator, especially in urban environments.

Phenotypic and Genotypic Antimicrobial Resistance in Non-O157 Shiga Toxin-Producing Escherichia coli Isolated From Cattle and Swine in Chile.

Non-O157 Shiga toxin-producing Escherichia coli (STEC) is a zoonotic pathogen that causes bloody diarrhea and hemolytic-uremic syndrome in humans, and a major cause of foodborne disease. Despite antibiotic treatment of STEC infections in humans is not recommended, the presence of antimicrobial-resistant bacteria in animals and food constitutes a risk to public health, as the pool of genes from which pathogenic bacteria can acquire antibiotic resistance has increased. Additionally, in Chile there is no information on the antimicrobial resistance of this pathogen in livestock. Thus, the aim of this study was to characterize the phenotypic and genotypic antimicrobial resistance of STEC strains isolated from cattle and swine in the Metropolitan region, Chile, to contribute relevant data to antimicrobial resistance surveillance programs at national and international level. We assessed the minimal inhibitory concentration of 18 antimicrobials, and the distribution of 12 antimicrobial resistance genes and class 1 and 2 integrons in 54 STEC strains. All strains were phenotypically resistant to at least one antimicrobial drug, with a 100% of resistance to cefalexin, followed by colistin (81.5%), chloramphenicol (14.8%), ampicillin and enrofloxacin (5.6% each), doxycycline (3.7%), and cefovecin (1.9%). Most detected antibiotic resistance genes were dfrA1 and tetA (100%), followed by tetB (94.4%), bla TEM-1 (90.7%), aac(6)-Ib (88.9%), bla AmpC (81.5%), cat1 (61.1%), and aac(3)-IIa (11.1%). Integrons were detected only in strains of swine origin. Therefore, this study provides further evidence that non-O157 STEC strains present in livestock in the Metropolitan region of Chile exhibit phenotypic and genotypic resistance against antimicrobials that are critical for human and veterinary medicine, representing a major threat for public health. Additionally, these strains could have a competitive advantage in the presence of antimicrobial selective pressure, leading to an increase in food contamination. This study highlights the need for coordinated local and global actions regarding the use of antimicrobials in animal food production.

Antimicrobial Usage Factors and Resistance Profiles of Shiga Toxin-Producing Escherichia coli in Backyard Production Systems From Central Chile.

Shiga toxin-producing Escherichia coli (STEC) is a zoonotic pathogen and important cause of foodborne disease worldwide. Many animal species in backyard production systems (BPS) harbor STEC, systems characterized by low biosecurity and technification. No information is reported on STEC circulation, antimicrobial resistance (AMR) and potential drivers of antimicrobial usage in Chilean BPS, increasing the risk of maintenance and transmission of zoonotic pathogens and AMR generation. Thus, the aim of this study was to characterize phenotypic and genotypic AMR and to study the epidemiology of STEC isolated in BPS from Metropolitana region, Chile. A total of 85 BPS were sampled. Minimal inhibitory concentration and whole genome sequencing was assessed in 10 STEC strain isolated from BPS. All strains were cephalexin-resistant (100%, n = 10), and five strains were resistant to chloramphenicol (50%). The most frequent serotype was O113:H21 (40%), followed by O76:H19 (40%), O91:H14 (10%), and O130:H11 (10%). The stx1 type was detected in all isolated strains, while stx2 was only detected in two strains. The Stx subtype most frequently detected was stx1c (80%), followed by stx1a (20%), stx2b (10%), and stx2d (10%). All strains harbored chromosomal bla AmpC. Principal component analysis shows that BPS size, number of cattle, pet and horse, and elevation act as driver of antimicrobial usage. Logistic multivariable regression shows that recognition of diseases in animals (p = 0.038; OR = 9.382; 95% CI: 1.138-77.345), neighboring poultry and/or swine BPS (p = 0.006; OR = 10.564; 95% CI: 1.996-55.894), visit of Veterinary Officials (p = 0.010; OR = 76.178; 95% CI: 2.860-2029.315) and close contact between animal species in the BPS (p = 0.021; OR = 9.030; 95% CI: 1.385-58.888) increase significantly the risk of antimicrobial use in BPS. This is the first evidence of STEC strains circulating in BPS in Chile, exhibiting phenotypic AMR, representing a threat for animal and public health. Additionally, we identified factors acting as drivers for antimicrobial usage in BPS, highlighting the importance of integration of these populations into surveillance and education programs to tackle the potential development of antimicrobial resistance and therefore the risk for ecosystemic health.

Can Responsible Ownership Practices Influence Hunting Behavior of Owned Cats?: Results from a Survey of Cat Owners in Chile.

The domestic cat (Felis catus) has become a worldwide threat to wildlife. The potential impact of owned cats on wildlife in Chile has not been documented at a large scale. The purpose of this study was to investigate the number and type of prey that owned cats bring back in Chile and its relation with responsible ownership practices. An online survey was distributed to 5216 households that included questions about the type of pet, responsible ownership practices, and in the case of cats, the type of prey they brought home. Descriptive statistics as well as univariate and multivariate logistic regression analysis were applied. The results showed that 94.3% of respondents had a pet, and from these, 49.9% had at least one cat. A total of 84.1% of owners reported that their cats had brought back prey. Birds were the most common type of prey, followed by mammals and insects. Not being registered with a microchip, not having a litter box, living in a house with access to a garden, not having a hiding place for the cats, and having free access to the outdoors significantly increased the odds of cats bringing back prey. Body condition score or providing ad libitum food to cats did not have an effect on bringing prey.

Virulence Genes, Shiga Toxin Subtypes, Serogroups, and Clonal Relationship of Shiga Toxin-Producing Escherichia Coli Strains Isolated from Livestock and Companion Animals.

Shiga toxin-producing Escherichia coli (STEC) is a zoonotic pathogen that causes severe illness in humans and is an important cause of foodborne disease. In Chile, there is limited information on the virulence characteristics of this pathogen in livestock, and none in companion animals. The aim of this study was to characterize STEC strains isolated from cattle, swine, dogs, and cats, in Chile, in terms of the presence of Shiga toxin types and subtypes, virulence genes, serogroups, and clonality. One-thousand two-hundred samples were collected, isolating 54 strains (4.5%), where stx1a (68.5%) and ehxA (74.1%) were the most frequently detected virulence genes. Only one strain belonging to the most clinically relevant serogroups was identified. Pulsed field gel electrophoresis analysis showed high clonal diversity among strains isolated from cattle, while those from swine showed the same pattern. This study provides further evidence regarding cattle and swine in Chile as a potential source of a wide variety of STEC strains that could potentially cause severe illness in humans, and that companion animals do not seem to represent a relevant reservoir. It also argues that preventive and control strategies should not be focused on detecting serogroups, but instead, on detecting their determinants of virulence.

Food Preferences in Cats: Effect of Dietary Composition and Intrinsic Variables on Diet Selection.

A ten-year database of food preference tests (n = 1021; period 2007-2017) was used to explore the feeding behavior of domestic cats. Principal component (PC) analysis and linear regression between food nutrients and preferences (for the most preferred diet of each test; Diet A) were performed. Intake and preference for Diet A were analyzed by intrinsic cats' variables and climate season. The PC1 (calcium (Ca), phosphorus (P), and ash), PC2 (lipids and ether extract) and PC4 (crude fiber; CF) had borderline significance (p < 0.06; ß = -1.42, ß = -1.56, and ß = 2.68, respectively). Ash and CF contents presented negative correlations with food preference (rho = -0.269, p = 0.031; rho = -0.338, p = 0.006, respectively), and Ca had borderline significance and negative correlation with food preference (rho = -0.241, p = 0.054). Body weight and sex influenced the intake of Diet A, being lower for females (ß = 11.758; p = 0.014) and heaviest cats (ß = -5.490; p < 0.001). However, only body weight affected food preferences, where the heaviest cats had greater preferences for Diet A. Hot season decreased food intake (ß = -2,117; p = 0.032), mostly in females (rho = -3.537; p = 0.002). Males had greater preferences for Diet A during hot seasons (ß = 10.216; p = 0.023) and females presented similar preferences throughout the year (p = 0.950). Mineral contents, body weight and sex affected food intake and preferences of cats under the influence of climate season, probably explained by adaptive changes in food detection.

Food Preferences in Dogs: Effect of Dietary Composition and Intrinsic Variables on Diet Selection.

A ten-year food preference database (2007-2017) was used to relate food selection in dogs to the nutritional components of diets by doing a principal component analysis (PCA) and a linear regression between components obtained and dogs' preferences. Intake and preference of preferred diets were analyzed by dogs' sex, breed, age, body weight, and the season of the year (hot or cold). The fourth component after PCA presented a relation with food preferences (OR = -2.699, p = 0.026), showing negative correlations with crude fiber (rho = -0.196; P = 0.038) and dry matter (rho = -0.184; p = 0.049). Weight (OR = -1.35; p < 0.001), breed, both Boxer (OR = 10.62; p = 0.003) and Labrador Retriever (OR = 26.30; p < 0.001), and season (hot season) (OR = -5.27; p < 0.001) all influenced animals' intake. Boxers presented a lower food preference compared to the other breeds (OR = -44.3; p < 0.001), while animals' weight influenced preferences only in Boxers (OR = 2.02; p < 0.001). Finally, age and sex did not affect dogs' preference or intake of preferred diets. Thus dry matter and fiber content have a negative impact on dogs' food choices. Dogs' weight, breed, and season affected food intake, but only breed affected dogs' preferences, which is probably explained by adaptive changes in the detection, metabolization, and learning of nutritive food cues.