An assessment of the owned canine and feline demographics in Chile: registration, sterilization, and unsupervised roaming indicators.

The global rise in companion animal populations, particularly dogs and cats, is driven by emotional and social benefits for owners, and their population management is becoming critically important to avoid a plethora of adverse effects on themselves, humans, and wildlife. We estimated the size and density of the owned canine and feline population in Chile and evaluated the status of microchipping, registration, sterilization rates, and the proportion of owned animals that roam unsupervised. A cross-sectional household survey in 36 districts was conducted and standard inferential statistics was employed to analyze differences between cats and dogs, sexes within each species, and between rural and urban areas. Additionally, two negative binomial models with mixed effects were developed to predict the number of dogs and cats per households. Two methods were used to compare population size estimates at the country level, multiplying: (1) the estimated mean number of companion animals per household by the estimated number of households at the country level, and (2) the estimated human:dog and human:cat ratios by the total human population. The study involved 6333 respondents, of which 76% (74% urban; 83% rural) owned companion animals (dogs and/or cats). Individuals in rural multi-person households increase the probability of owning dogs and/or cats. Additionally, women exhibit a greater inclination towards cat and dog ownership compared to men, while those over 30 years old demonstrate lower rates of companion animal ownership in contrast to the 18-30 age group for both species. The overall human:dog and human:cat ratios estimated were 2.7:1, and 6.2:1, respectively. The estimated total number of owned dogs and cats in Chile ranged from 9.6 to 10.7 million, depending on the methodological approach, while national median density of companion animals was 12 dogs per km2 (ranging from 0.02 to 7232) and 5 cats per km2 (ranging from 0.01 to 3242). This nationwide study showed one of the highest percentages of households with companion animals in Latin America and relatively low registration and sterilization rates, highlighting the need to strength long-term public policies to control populations of companion animals and promote responsibility in pet ownership.

Postharvest shelf life extension of minimally processed kale at ambient and refrigerated storage by use of modified atmosphere.

Kale is becoming an important vegetable worldwide, mainly due to its nutritional properties. Kale leaves can be marketed whole, although minimal processing is also in demand. In this article, it was analyzed the effect of packaging in a modified atmosphere of fresh-cut kale leaves stored at 20 °C and 4 °C. Kale leaves were cut into 4 × 4 cm strips and stored in low-density polyethylene bags. Samples processed in the same way but stored in PVC were used as controls. Leaves kept in a modified atmosphere showed a delay in color change with Hue values from about 130 to 120 under PMA against 130 to 100 in control group (CTR) leaves. Chlorophyll degradation was also delayed in both storage temperatures. Samples stored under PMA showed about two times the levels of total chlorophylls with respect to CTR samples at the end of the storage. No changes in total sugar content were detected during storage and no differences were detected between control and modified atmospheres stored samples. Samples maintained in a modified atmosphere showed a lower decrement in soluble proteins and a lower rate of RUBISCO degradation at both temperatures. The relation of RUBISCO content PMA/CTR ranged from 1 to about 3 toward the end of storage No changes in phenols content were found when comparing control and treated samples. However, flavonoid and the antioxidant contents increased in samples stored in modified atmospheres with respect to their controls. We demonstrated that storage in modified atmospheres could be an adequate and simple methodology to extend postharvest life of this minimally processed product at both ambient and refrigerated storage.

Loss-of-function mutations in WRKY22 and WRKY25 impair stomatal-mediated immunity and PTI and ETI responses against Pseudomonas syringae pv. tomato.

Plants defend themselves against pathogens using a two-layered immune system. The first response, pattern-triggered immunity (PTI), is activated upon recognition of microbe-associated molecular patterns (MAMPs). Virulent bacteria such as Pseudomonas syringae pv. tomato (Pst), deliver effector proteins into the plant cell to promote susceptibility. However, some plants possess resistance (R) proteins that recognize specific effectors leading to the activation of the second response, effector-triggered immunity (ETI). Resistant tomatoes such as Río Grande-PtoR recognize two Pst effectors (AvrPto and AvrPtoB) through the host Pto/Prf complex and activate ETI. We previously showed that the transcription factors (TF) WRKY22 and WRKY25 are positive regulators of plant immunity against bacterial and potentially non-bacterial pathogens in Nicotiana benthamiana. Here, the CRISPR-Cas9 technique was used to develop three knockout tomato lines for either one or both TFs. The single and double mutants were all compromised in Pto/Prf-mediated ETI and had a weaker PTI response. The stomata apertures in all of the mutant lines did not respond to darkness or challenge with Pst DC3000. The WRKY22 and WRKY25 proteins both localize in the nucleus, but we found no evidence of a physical interaction between them. The WRKY22 TF was found to be involved in the transcriptional regulation of WRKY25, supporting the idea that they are not functionally redundant. Together, our results indicate that both WRKY TFs play a role in modulating stomata and are positive regulators of plant immunity in tomato.

Preventive healthcare among dogs and cats in Chile is positively associated with emotional owner-companion animal bond and socioeconomic factors.

Global companion animal population has been increasing as well as the number of dogs and cats being considered as a family member. However, it is unclear whether this close relationship could be associated with higher preventive healthcare in companion animals. Using data from 7,048 questionnaires of dogs and 3,271 of cats from the First National Study on Responsible Companion Animal Ownership, we estimated the proportion of preventive healthcare in companion animals of Chile. We also conducted a general linear mixed-effect regression model to identify socioeconomic factors and indicators of the emotional owners-companion animal bond that could influence owners' practices related to vaccination, parasite control, and veterinary visits. Based on the owner's answers, Chile has a satisfactory overall rates of parasite control (71%) and annual veterinary visits (65%) but a low vaccination coverage of both dogs (39%) and cats (25%). 'Purebred', 'live in urban areas', 'acquired by monetary compensation', and 'dog species' were associated with a higher probability of preventive healthcare in companion animals. Conversely, this probability was lower among senior animals compared to adults, males, and those owned by the Silent Generation or Baby Boomers (i.e., owners born before 1964). 'Sleeping inside', 'acquired for an emotional reason' (e.g., companionship), and 'considered a family member' were positively associated with at least one of assessed preventive measures. Our findings suggest that emotional owner-companion animal bonds could positively impact the frequency and quality of preventive healthcare in dogs and cats. However, owners who totally disagreed that a companion animal is a "family member" were also associated with a higher likelihood of vaccination uptake and veterinary visits for their animals. This highlights that owner's compliance with veterinary preventive healthcare is multifactorial. Chile has a high prevalence of infectious diseases circulating among dogs and cats and increasingly close contacts between owners and companion animals due to emotional bonds. Thus, our study calls for One Health approaches to reduce the risks of cross-species disease transmission. Specifically, increasing vaccination coverage of companion animals in Chile is the most urgent preventive measure needed, especially among cats, males, and older animals. Expand preventive healthcare among dogs and cats will promote public and animal health, including local wildlife that is susceptible to infectious diseases circulating in companion animals.

Environmental and anthropogenic factors associated with the likelihood of detecting Salmonella in agricultural watersheds.

Surface water is one of the primary sources of irrigation water for produce production; therefore, its contamination by foodborne pathogens, such as Salmonella, may substantially impact public health. In this study, we determined the presence of Salmonella in surface water and characterized the relationship between Salmonella detection and environmental and anthropogenic factors. From April 2019 to February 2020, 120 samples from 30 sites were collected monthly in four watersheds located in two different central Chile agricultural regions (N = 1080). Water samples from rivers, canals, streams, and ponds linked to each watershed were obtained. Surface water (10 L) was filtrated in situ, and samples were analyzed for the presence of Salmonella. Salmonella was detected every month in all watersheds, with a mean detection percentage of 28% (0%-90%) across sampling sites, regardless of the season. Overall, similar detection percentages were observed for both regions: 29.1% for Metropolitan and 27.0% for Maule. Salmonella was most often detected in summer (39.8% of all summer samples tested positive) and least often in winter (14.4% of winter samples). Random forest analysis showed that season, water source, and month, followed by latitude and river, were the most influential factors associated with Salmonella detection. The influences of water pH and temperature (categorized as environmental factors) and factors associated with human activity (categorized as anthropogenic factors) registered at the sampling site were weakly or not associated with Salmonella detection. In conclusion, Salmonella was detected in surface water potentially used for irrigation, and its presence was linked to season and water source factors. Interventions are necessary to prevent contamination of produce, such as water treatment before irrigation.

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.

Genome-wide analysis uncovers tomato leaf lncRNAs transcriptionally active upon Pseudomonas syringae pv. tomato challenge.

Plants rely on (in)direct detection of bacterial pathogens through plasma membrane-localized and intracellular receptor proteins. Surface pattern-recognition receptors (PRRs) participate in the detection of microbe-associated molecular patterns (MAMPs) and are required for the activation of pattern-triggered immunity (PTI). Pathogenic bacteria, such as Pseudomonas syringae pv. tomato (Pst) deploys ~ 30 effector proteins into the plant cell that contribute to pathogenicity. Resistant plants are capable of detecting the presence or activity of effectors and mount another response termed effector-triggered immunity (ETI). In order to investigate the involvement of tomato's long non-coding RNAs (lncRNAs) in the immune response against Pst, we used RNA-seq data to predict and characterize those that are transcriptionally active in leaves challenged with a large set of treatments. Our prediction strategy was validated by sequence comparison with tomato lncRNAs described in previous works and by an alternative approach (RT-qPCR). Early PTI (30 min), late PTI (6 h) and ETI (6 h) differentially expressed (DE) lncRNAs were identified and used to perform a co-expression analysis including neighboring (± 100 kb) DE protein-coding genes. Some of the described networks could represent key regulatory mechanisms of photosynthesis, PRR abundance at the cell surface and mitigation of oxidative stress, associated to tomato-Pst pathosystem.

WRKY22 and WRKY25 transcription factors are positive regulators of defense responses in Nicotiana benthamiana.

KEY MESSAGE: NbWRKY22 and NbWRKY25 are required for full activation of bacteria-associated pattern- and effector-triggered immunity as well as for the response to other non-bacterial defense elicitors. Plants defend themselves against pathogens using a two-layered immune system. Pattern-triggered immunity (PTI) can be activated upon recognition of epitopes from flagellin including flg22. Pseudomonas syringae pv. tomato (Pst) delivers effector proteins into the plant cell to promote host susceptibility. However, some plants express resistance (R) proteins that recognize specific effectors leading to the activation of effector-triggered immunity (ETI). Resistant tomato lines such as Rio Grande-PtoR (RG-PtoR) recognize two Pst effectors, AvrPto and AvrPtoB, and activate ETI through the Pto/Prf protein complex. Using RNA-seq, we identified two tomato WRKY transcription factor genes, SlWRKY22 and SlWRKY25, whose expression is increased during Pst-induced ETI. Silencing of the WRKY25/22 orthologous genes in Nicotiana benthamiana led to a delay in programmed cell death normally associated with AvrPto recognition or several non-bacterial effector/R protein pairs. An increase in disease symptoms was observed in silenced plants infiltrated with Pseudomonas syringae pv. tabaci expressing AvrPto or HopQ1-1. Expression of both tomato WRKY genes is also induced upon treatment with flg22 and callose deposition and cell death suppression assays in WRKY25/22-silenced N. benthamiana plants supported their involvement in PTI. Our results reveal an important role for two WRKYs as positive regulators of plant immunity against bacterial and potentially non-bacterial pathogens.

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.

Transcriptome-based identification and validation of reference genes for plant-bacteria interaction studies using Nicotiana benthamiana.

RT-qPCR is a widely used technique for the analysis of gene expression. Accurate estimation of transcript abundance relies strongly on a normalization that requires the use of reference genes that are stably expressed in the conditions analyzed. Initially, they were adopted from those used in Northern blot experiments, but an increasing number of publications highlight the need to find and validate alternative reference genes for the particular system under study. The development of high-throughput sequencing techniques has facilitated the identification of such stably expressed genes. Nicotiana benthamiana has been extensively used as a model in the plant research field. In spite of this, there is scarce information regarding suitable RT-qPCR reference genes for this species. Employing RNA-seq data previously generated from tomato plants, combined with newly generated data from N. benthamiana leaves infiltrated with Pseudomonas fluorescens, we identified and tested a set of 9 candidate reference genes. Using three different algorithms, we found that NbUbe35, NbNQO and NbErpA exhibit less variable gene expression in our pathosystem than previously used genes. Furthermore, the combined use of the first two is sufficient for robust gene expression analysis. We encourage employing these novel reference genes in future RT-qPCR experiments involving N. benthamiana and Pseudomonas spp.

Abdomen abierto y contenido: sistema "no comercial - hand made" en pacientes traumatizados. Alternativa terapéutica en situaciones con bajos recursos. ¿Es una opción segura y eficiente? / Open abdomen and content in "non commercial hand made system" in trauma patients Therapeutic alternative in situations with low resources. Is it a safe and efficient option?

Introducción: En la década del '80 se iniciaron los conceptos de laparotomía y relaparotomías. La hipertensión intraabdominal, la cirugía del control del daño, la necesidad de reexploración por packing hemostáticos o las colecciones abdominales son las indicaciones para considerar un abdomen abierto y contenido. A lo largo de los años distintos sistemas de contención del abdomen fueron comercializados, reduciendo la mortalidad a 7 %. Objetivos: El problema es la disponibilidad del recurso dado su alto costo en instituciones con bajo presupuesto. El objetivo es analizar la efectividad y seguridad del método propuesto para aplicar en medios donde el recurso no está disponible en su forma comercial. Material y Método: Realizado entre el 01 de enero 2015 y Julio de 2016, 10 pacientes fueron tratados con abdomen abierto y contenido "no comercial ­hand made". Consiste en la aplicación de un film de polivinilo, multifenestrado, ubicado en la cavidad abdominal por debajo del peritoneo. Sobre ésta, una cubierta de poliuretano y gasas que envuelve una sonda tipo K9. Un film adhesivo cubre toda la incisión, aplicándose una presión negativa de 100 a 150 cm de agua. En un solo caso se utilizó el Wittmann Patch. Resultados: En 7 pacientes se hallaron lesiones combinadas. Lesión hepática: 7; 2 presentaban hipertensión intraabdominal con dificultad para el afrontamiento abdominal. Packing pancreático duodenal: 1. Contaminación de la cavidad: 1. La cantidad de exploraciones fue de 9 y de 3 lavados en promedio. Conclusiones: El sistema "no comercial ­hand made" es un método seguro, de bajo costo y con baja de tasa de complicaciones

Introduction: In the decade of the'80 the concept of laparotomy and relaparotomies began. Intra-abdominal hypertension, damage control surgery, need for re ­examination by hemostatic packing or abdominal collections are indications to consider perfoming the open and contained abdomen. Throughout the years different systems of containment of the abdomen were commercialized. reducing mortality to 7%. Objectives: To analyze the effectiveness and safety of the method proposed to be applied where the resource is not available in its commercial way Material and Methods: In the period between January 1, 2015 and July 2016, 10 patients were treated with open abdomen and "non-commercial -hand made" contents. Consists in the application of a multilayer polyvinyl film of 80 microns, located in the abdominal cavity below the peritoneum, surpassing it inwards by 5 cm. A layer of polyurethane and gauze is placed over this layer, which envelops a polyvinyl probe type K9, until it reaches the edge of the wound. An adhesive film covers the whole incision and passes it by 15 cm, while applying a negative pressure of 100 to 150 cm of water. In only one case the technique described by Dietmar Witmann, the Wittmann Patch was used. Results: Combined lesions were found in 7 patients. Liver injury: 7; 2 also presented HIA with difficulty for abdominal coping. Packing Pancreatic duodenal 1. Cavity contamination: 1. The number of scans was 9 and 3 washes on average. Conclusions: The "non -commercial -hand made" system is a safe, low -cost and low complication rate.