1. Investigación

Antimicrobial resistance (AMR) poses a major threat to health worldwide. Poultry products are one of the main threats, due to the transmission of antimicrobial resistance genes throughout the food chain. Escherichia coli is the main cause of mortality in the poultry industry, mainly mitigated with antibiotics, but due to the high genetic strain variability, recurrent outbreaks of multidrug resistant E. coli take place. The major challenge to tackling AMR is understanding the burden of resistance. For this reason, one of the main strategies is monitoring AMR by phenotypic characterisation. Our study aimed to monitor the resistance of E. coli strains isolated from the poultry sector over a period of three years (2019–2021) to provide information on the resistance magnitude and trends. Promising results have been found concerning the low frequency of resistance to cephalosporins, polymyxin, and fluoroquinolones. However, levels of resistance found to antimicrobials such as erythromycin (100%), tylosin (98%), or penicillin (97%) suggest the need to continue working on the limitation of use of antimicrobials in poultry to achieve the demise of MDR.

Newcastle disease is a devasting disease in poultry production worldwide, thus it is important to implement control measures to avoid entrance of the disease and its spread in the field. In this context, the aim of this study was to design and implement a seroprevalence map based on business intelligence for avian paramyxovirus type 1 (APMV-1) in broilers in the Valencia Community (eastern Spain). This tool consists in software mapping based on data collection, data analysis and data representation. In order to obtain the serological data, 12 495 sera from 131 broiler farms over 5 years were analysed (2008–2012). The data were represented on a map of the Valencian Community including geographical information of flock locations to facilitate disease monitoring. No clinical signs of APMV-1 were reported in the studied flocks. The data from this study showed no evidence contact with APMV-1 in broiler flocks and the novel software mapping tool as a valuable method for easily monitoring the serological response to avian paramyxovirus type 1 (APMV-1) including geographical information.

Infectious Bronchitis Virus (IBV) is one of the most important viral diseases which causes important economic losses in poultry industry. This study aimed to assess the seroprevalence, prevalence, and variants of IBV in broilers, layers, and broiler breeders farms of Gallus gallus species in Eastern Spain. Thus, 29, 16, and 14 flocks of broilers, layers and broiler breeders, respectively were analyzed. To assess seroprevalence, sera samples were analyzed by ELISA. Tracheal swabs and tissue samples were tested by PCR to know the prevalence and detect specific variants. An IBV seroprevalence of 100% was detected in the 3 productive orientations. According to PCR results, a prevalence of 38% in broilers, 44% in layers and 43% in broiler breeders was obtained. The variant-specific RT-PCR analysis showed that 4/91, Massachusetts, QX, Italy- 02 and D274 strains were present in commercial flocks in eastern Spain, being 4/91 the most prevalent in all the productive orientations. In layers 100% of QX prevalence, 14% of Italy 02 and 14% of D274 was detected. Regarding broilers, a prevalence of 18% of Massachusetts strain was also detected. In contrast, in broiler breeders farms only 4/91 strain was found. In conclusion, our findings showed the presence of IBV in eastern Spain and the changing situation of the IBV variants prevalence, being different according to the productive orientation. The continuous emergence of new variants emphasizes the importance of continuous IBV monitoring in order to optimize vaccination strategies.

Salmonella spp. is one of the most important zoonotic pathogens with economic impact in public health worldwide. The relevance of Salmonella increases with the appearance of resistant strains. The aim of this study was to determine the level of antimicrobial resistance in 332 Salmonella isolates selected from 3 different poultry productive orientations in Eastern Spain during 3 yr (2015−2017). Antimicrobial susceptibly was evaluated by broth microdilution method using 14 antibiotics. Epidemiological cut-off values (ECOFF) were used to evaluate the microbiological resistance to antibiotics. The rates of Salmonella resistance at least to one antibiotic were 96, 98, and 56% in broilers, turkeys, and layers, respectively. Regarding multidrug resistance, all productive orientations seems to present a decreasing trend along the study, being the mean rates 80% in turkeys followed by broilers (40%) and layers (6%). Throughout the study, the highest percentage of resistance was found to sulfamethoxazole in all productive orientations. Strains from broilers showed the highest resistance rates to sulfamethoxazole (73%), gentamicin (57%), ciprofloxacin (50%), nalidixic acid (29%), and tetracycline (24%). Relative to turkeys the highest resistance rates were to sulfamethoxazole (76%), ciprofloxacin (69%), tetracycline (75%), nalidixic acid (63%), and ampicillin (63%). Layers presented the most elevated resistance rates to sulfamethoxazole (39%) and tetracycline (13%). Regarding serovars the most MDR common serovars to the 3 productive orientations were S. Kentucky and S. Hadar. In the other hand, high MDR rates were found in other serovars like S. Infantis and S. Typhimurium in broilers and turkeys. Results shown in the present study suggest that the reduction in the use of antibiotics begins to be reflected in the reduction of the number of MDRs, especially in layers, with no MDR Salmonella strains in the last period. However, the level of resistances found in this study suggests the necessity of continuing working on the limitation of the use of antimicrobials in poultry to achieve (as in layers) the control of MDRs.

Campylobacteriosis was the most frequently reported foodborne infection in humans in the European Union in the last years. Campylobacter spp. in broiler flocks from Spain was monitored at farm level during 12-month period (2020−2021). Feces samples were analyzed according to ISO (International Standard Organization) 10272-2:2018. From all samples collected, 54% were Campylobacter spp. positive. Regarding the age, Campylobacter spp. was isolated in 36% of the flocks during thinning and 64% in flocks at slaughter age. In addition, Campylobacter spp. counts increased with the age of the animals. On the other hand, the presence of Campylobacter showed statistical differences between the months of the year (P-value <0.05) in flocks at thinning age that exceeded the ≥1,000 CFU/g limit. The highest rates were found from June to December coinciding with the seasons of summer and autumn. In conclusion, our study shows the situation of Campylobacter spp. in broiler flocks in Spain considering age and season effects. This way, it was found higher rates and counts in broilers close to slaughter age and peaking during the summer to autumn period.

Mycoplasma species are worldwide recognized poultry pathogens, with Mycoplasma synoviae being the second most important species from the clinical point of view that causes considerable economic losses in the poultry industry. The aim of this study was to assess the seroprevalence, prevalence, and phylogenetic variants of M. synoviae present in layers and broiler breeders’ farms of Gallus gallus species located in eastern Spain. Thus, 19 and 23 flocks of layers and broiler breeders, respectively, were analyzed at 3 different ages. To assess seroprevalence, sera samples were analyzed by ELISA. Tracheal swabs were tested by PCR to assess the prevalence. A M. synoviae seroprevalence of 95 and 74% was detected in layers and broiler breeders, respectively. Regarding age-wise analysis, the positive rates obtained seemed to be higher as the age of sampling increased. As per PCR results, a prevalence of 95% in layers and 35% in broiler breeders was obtained. The genetic analysis showed that the strains present in broilers breeders were vaccine strains (MS H strain). In contrast, 6 different field strains were detected in layer hens. In conclusion, this study carried out in eastern Spain showed a higher seroprevalence and prevalence of M. synoviae field strains in layer flocks regarding broiler breeders, highlighting the usefulness of monitoring flocks to control this poultry pathogen. Moreover, our findings suggest M. synoviae vaccination in broiler breeders could be an effective prevention strategy.

Background: Salmonella enterica serovar Infantis is a zoonotic pathogen isolated in broilers causing great economic losses in the European poultry sector. It is demonstrated that an investment in management measures at farm level could directly affect the control of food chain microorganisms. The aim of this study was to investigate the development of S. Infantis antimicrobial resistance (AMR) patterns during the growing period, according to flock density and ventilationmanagement, without antibiotic administration. Methods: The experiment was performed in two identical poultry houses, evaluating commercial and optimal farm conditions. At 24 h of rearing, 20% of the animals were orally infected with a S. Infantis strain susceptible to all the antibiotics tested. To study Salmonella shedding, faeces samples from each experimental groupwere takenweekly and analysed as per ISO/TS 6579- 2:2017. Antibiotic susceptibilitywas assessed according toDecision 2013/653. Results: Salmonella shedding showed that the lowest counts were observed in the first week post-infection and highest at slaughter day for both groups. Moreover, 100% of the isolates were multi-resistant. Conclusion: The acquisition of AMR by S. Infantis starts at the onset of the production cycle and is maintained until the end, demonstrating the importance of transmission of AMR in zoonotic bacteria at farmlevel.

Poultry is one of the main agricultural sub-sectors worldwide. However, public concern regarding animal welfare and antimicrobial resistance has risen in recent years. Due to the influence of management practices on microbiota, it might be considered to evaluate poultry welfare and health. Therefore, the objective of this research was to analyse the influence on microbiota balance of broilers under commercial and optimal farm conditions, using 16S rRNA sequencing analysis. The research was performed in two identical poultry houses (commercial vs. optimal). Results showed a higher level of microbiota complexity in the group reared under optimal farm conditions at the end of rearing. Regarding microbiota composition, Firmicutes was the dominant phylum during the entire growing period. However, the second most prevalent phylum was Proteobacteria at the arrival day, and Bacteroidetes from the mid-period onward in both groups. Moreover, the most predominant genera identified were Oscillospira, Ruminococcus, Bacteroides, and Coprococcus. In conclusion, it is necessary to optimize farm management as much as possible. Using gut microbiota diversity and composition as biomarkers of animal health could be an important tool for infectious disease control, with the aim of reducing the administration of antibiotics at field level.

New measures applied to reduce antimicrobial resistances (AMR) at field level in broiler production are focused on improving animals’ welfare and resilience. However, it is necessary to have better knowledge of AMR epidemiology. Thus, the aim of this study was to evaluate AMR and multidrug resistance (MDR) dynamics during the rearing of broilers under commercial (33 kg/m2 density and max. 20 ppm ammonia) and improved (17 kg/m2 density and max. 10 ppm ammonia) farm conditions. Day-old chicks were housed in two poultry houses (commercial vs. improved), and no antimicrobial agents were administered at any point. Animals were sampled at arrival day, mid-period and at slaughter day. High AMR rates were observed throughout rearing. No statistical differences were observed between groups. Moreover, both groups presented high MDR at slaughter day. These results could be explained by vertical or horizontal resistance acquisition. In conclusion, AMR and MDR are present throughout rearing. Moreover, although a lower level of MDR was observed at mid-period in animals reared under less intensive conditions, no differences were found at the end. In order to reduce the presence of AMR bacteria in poultry, further studies are needed to better understand AMR acquisition and prevalence in differing broiler growing conditions.