2. Universidad Cardenal Herrera-CEU

Antimicrobial resistance (AMR) and the increase in multi-resistant bacteria are among the most important threats to public health worldwide, according to the World Health Organisation (WHO). Moreover, this issue is underpinned by the One Health perspective, due to the ability of AMR to be transmitted between animals and humans living in the same environment. Therefore, since 2014 different surveillance and control programmes have been established to control AMR in commensal and zoonotic bacteria in production animals. However, public health authorities’ reports on AMR leave out companion animals, due to the lack of national programmes and data collection by countries. This missing information constitutes a serious public health concern due to the close contact between companion animals, humans and their surrounding environment. This absence of control and harmonisation between programmes in European countries leads to the ineffectiveness of antibiotics against common diseases. Thus, there is a pressing need to establish adequate surveillance and monitoring programmes for AMR in companion animals and further develop alternatives to antibiotic use in this sector, considering the impact this could have on the gut microbiota. In this context, the aim of this review is to evaluate the current control and epidemiological situations of AMR in companion animals in the European Union (EU), as well as the proposed alternatives to antibiotics.

Background: Giardia duodenalis is one of the most common enteric parasites in domestic animals including dogs. Young animals are more prone to the infection, with clinical manifestations ranging from asymptomatic to acute or chronic diarrhoea. Dogs are primarily infected by canine-specific (C-D) assemblages of G. duodenalis. However, zoonotic assemblages A and B have been increasingly documented in canine isolates, raising the question of whether and to which extent dogs can act as natural reservoirs of human giardiosis. Methods: In this cross-sectional epidemiological survey we assessed the molecular diversity of G. duodenalis in dogs in the province of Castellón, Eastern Spain. A total of 348 individual faecal samples from sheltered (n = 218), breeding (n = 24), hunting (n = 68), shepherd (n = 24), and pet (n = 14) dogs were collected between 2014 and 2016. Detection of G. duodenalis cysts in faecal material was carried out by direct fluorescence microscopy as a screening test, whereas a qPCR targeting the small subunit ribosomal RNA gene of the parasite was subsequently used as a confirmatory method. Results: Giardia duodenalis was detected in 36.5% (95% CI: 31.6–41.7%) of dogs. No significant differences in prevalence rates could be demonstrated among dogs according to their sex and geographical origin, but breeding (45.8%; 95% CI: 27.9–64.9%) and sheltered (40.4%; 95% CI: 34.1–47.0%) dogs harboured significantly higher proportions of G. duodenalis. Multi-locus sequence-based genotyping of the glutamate dehydrogenase and β-giardin genes of G. duodenalis allowed the characterization of 35 canine isolates that were unambiguously assigned to assemblages A (14.3%), B (22.9%), C (5.7%), and D (37.1%). A number of inter-assemblage mixed infections including A + B (11.4%), A + D (2.9%), and A + B + D (5.7%) were also identified. Conclusions: Data presented here are strongly indicative of high infection pressures in kennelled animals. Zoonotic sub-assemblages AII, BIII, and BIV were responsible for a considerable proportion of the G. duodenalis infections detected, but very few of the genotypes identified have been previously documented in Spanish human populations. Although possible, zoonotic transmission between dogs and humans seems an infrequent event in this Spanish region.

Salmonella spp. es descrita como la principal causa de los brotes declarados de toxiinfecciones alimentarias. Y la carne de cerdo es una de las principales fuentes de salmonelosis humana de origen alimentario. Debido a que el sector porcino es un sector importante de producción en España, se ve la necesidad de establecer programas de control para Salmonella con el fin de evitar importantes pérdidas económicas en el futuro. De forma previa a la publicación de la prevalencia de Salmonella en cerdos de cebo (EFSA, 2008) y al establecimiento de los objetivos comunitarios para el control de Salmonella en explotaciones porcinas, se han realizado estudios para estimar la situación de Salmonella en la producción porcina de España. En este sentido, la Concejalía de Agricultura, Pesca y Alimentación financió el presente proyecto de investigación para el estudio de las principales fuentes de contaminación de Salmonella en las explotaciones porcinas de la Comunidad Valenciana y el estudio de la epidemiología de Salmonella en condiciones de campo. Dichos estudios fueron planificados para anticipar el Programa Nacional de Control de Salmonella en la producción porcina, con el objetivo de aplicar medidas adecuadas en línea con los futuros objetivos comunitarios. Este estudio empezó en abril de 2007 y se terminó en noviembre de 2009. En este periodo de dos años, se muestrearon 89 lotes de cerdos pertenecientes a 51 explotaciones porcinas de engorde de la Comunidad Valenciana. Se analizaron un total de 3.255 muestras mediante la técnica microbiológica de referencia ISO 6579:2002 (Anexo D) y las muestras positivas fueron serotipadas mediante la técnica de Kauffman-White-Le-Minor. El 20,7% de las muestras, que fueron recogidas en diferentes momentos del ciclo productivo de los cerdos, resultaron positivas a Salmonella spp. y el mayor número de muestras positivas se observó en otoño. Se evaluó la limpieza y desinfección en los 89 lotes pertenecientes a explotaciones porcinas de engorde de la Comunidad Valenciana frente a Salmonella, permaneciendo positivas el 70,8%, tras la limpieza y desinfección, en alguna de las muestras recogidas. Las muestras más contaminadas fueron los restos de heces del lote anterior, la superficie del pasillo y la superficie de la tolva. En 47 de esos lotes se realizó una monitorización a lo largo de todo el ciclo productivo y se cumplimentaron unas encuestas epidemiológicas con el objetivo de describir las características más importantes de las explotaciones estudiadas, así como su posible relación con el estatus de contaminación de Salmonella en los lotes de los cerdos y de la explotación al final del periodo de engorde. Se determinaron las principales fuentes de contaminación: las heces de los cerdos, las superficies del corral y del pasillo, las manos y botas del granjero, el polvo y los vectores. El serotipo aislado con mayor frecuencia fue S. Typhimurium. Además, se analizaron los principales factores de riesgo para la contaminación en heces de Salmonella de los lotes de cerdos al final del periodo de engorde: el estatus de contaminación en heces de Salmonella del lote anterior, la superficie del corral antes de la limpieza y desinfección tras la salida del lote anterior y la superficie de la tolva a la entrada de los lechones en la explotación. Tras analizar la detección de Salmonella en heces a lo largo del periodo de engorde de los 47 lotes de cerdos se observó que, independientemente de si los lotes llegaban a la granja excretando la bacteria en heces o si se infectaban durante el periodo de engorde, presentaban un porcentaje similar de detección de Salmonella a partir del mes y medio de la entrada en la explotación y experimentaban un aumento justo a la salida (70,8% y 55,0%, respectivamente). El patrón de los serotipos más prevalentes en heces varió a lo largo del periodo de engorde. Por otro lado, se analizó el transporte a matadero en la detección de Salmonella en heces. Se observó que la presencia de camiones contaminados con la bacteria incrementó la detección de Salmonella en heces de un 60% a un 80%. El patrón de los serotipos aislados durante el periodo de engorde cambió después del transporte a matadero. Además, se analizaron las instalaciones de los mataderos estudiados. En el 100% de las visitas a las instalaciones al menos una muestra de corral o de pasillo resultó positiva a Salmonella. Por último, se determinó la capacidad de desarrollo de biofilm de las cepas aisladas en el estudio de la limpieza y desinfección. El 69,0% de las cepas mostró la capacidad de producir biofilm. Además, se evaluaron los desinfectantes utilizados y se observó que el desinfectante cuyo principio activo es el glutaraldehido resultó el menos eficaz frente a estas cepas productoras de biofilm. Salmonella spp. is described as the leading cause of reported outbreaks of food poisoning. And pork is one of the main sources of human foodborne salmonellosis. Because the pig sector is an important sector of production in Spain, is the need to establish monitoring programs for Salmonella in order to avoid major economic losses in the future. Previously to the publication of the prevalence of Salmonella in finishing pigs (EFSA, 2008) and the establishment of the Community objectives for the control of Salmonella in pork production, studies were performed to estimate the status of Salmonella in pig production of Spain. In this regard, the Department of Agriculture, Fisheries and Food funded this research project for the study of the main sources of contamination of Salmonella in pig farms of Valencia and the study of the epidemiology of Salmonella in field conditions. These studies were planned to anticipate the National Programme for Control of Salmonella in swine production, in order to apply appropriate sollutions acoording to future EU targets. This study began in April 2007 and was completed in November 2009. During this period of two years, 89 batches of pigs belonging to 51 fattening pig farms Valencia were sampled. A total of 3,255 samples were analyzed by microbiological reference technique ISO 6579:2002 (Annex D) and positive samples were serotyped by the technique of Kauffman-White-Le-Minor. 20.7% of the samples, which were collected at different times of the production cycle of pigs were positive for Salmonella spp. and the largest number of positive samples was observed in autumn. Cleaning and disinfection was evaluated in all 89 batches of a fattening pig farms in Valencia against Salmonella, 70.8% remained positive after cleaning and disinfection, in any of the samples. The most contaminated samples were the remains of feces previous batch, the gangway surface and the surface of the hopper. In 47 of these agent, a monitoring throughout the entire production cycle was performed and some epidemiological surveys were completed in order to describe the most important characteristics of the studied farms and their possible relation to the status of contamination of Salmonella in lots of pigs and exploitation at the end of the fattening period. The main sources of pollution were identified: the faeces of pigs, poultry and surfaces of the hall, hands and boots of the farmer, dust and vectors. The most frequently isolated serotype was S. Typhimurium. Moreover, the major risk factors for Salmonella contamination in feces of lots of pigs at the end of the fattening period were analyzed: the status of fecal contamination of Salmonella previous batch, pen area before cleaning and disinfection after the departure of the previous batch and the surface of the hopper to the inlet of piglets on the farm. After analyzing the detection of Salmonella in feces throughout the fattening period of 47 batches of pigs was observed that regardless of whether the lots came to the farm to excrete the bacteria in stool or if infected during the fattening period, had a similar percentage of Salmonella from the month and a half from the entrance on the farm and were experiencing right off (70.8% and 55.0%, respectively) increase. The pattern of the most prevalent serotypes in feces ranged along the fattening period. Furthermore, transportation to the slaughterhouse in the detection of Salmonella in feces was analyzed. It was observed that the presence of bacteria contaminated trucks increased faecal Salmonella 60% to 80%. The pattern of serotypes isolated during the fattening period changed after transport to slaughter. In addition, slaughter facilities studied were analyzed. In 100% of site visits at least one sample of poultry or hallway tested positive for Salmonella. Finally, the ability of biofilm development of strains isolated in the study of the cleaning and disinfection was determined. 69.0% of the strains were able to produce biofilm. Furthermore, the disinfectants used were evaluated and it was observed that the active substance disinfectant glutaraldehyde was less effective against biofilm these producing strains.