2. Universidad Cardenal Herrera-CEU

Despite the enormous efforts made to achieve effective tools that fight against Staphylococcus aureus, the results have not been successful. This failure may be due to the absence of truly representative experimental models. To overcome this deficiency, the present work describes and immunologically characterizes the infection for 28 days, in an experimental low-dose (300 colony-forming units) intradermal model of infection in rabbits, which reproduces the characteristic staphylococcal abscess. Surprisingly, when mutant strains in the genes involved in virulence (JΔagr, JΔcoaΔvwb, JΔhla, and JΔpsmα) were inoculated, no strong effect on the severity of lesions was observed, unlike other models that use high doses of bacteria. The inoculation of a human rabbitized (FdltBr) strain demonstrated its capacity to generate a similar inflammatory response to a wild-type rabbit strain and, therefore, validated this model for conducting these experimental studies with human strains. To conclude, this model proved reproducible and may be an option of choice to check both wild-type and mutant strains of different origins.

Staphylococcus aureus is an opportunistic multi-host pathogen that threatens both human and animal health. Animals can act as a reservoir of S. aureus for humans, but very little is known about wild animals’ epidemiological role. Therefore, in this study, we performed a genomic characterization of S. aureus isolates from wildlife, hunters, and their auxiliary hunting animals of Eastern Spain. Of 20 different species, 242 wild animals were examined, of which 28.1% were S. aureus carriers. The common genet, the Iberian ibex, and the European hedgehog were the species with the highest S. aureus carriage. We identified 30 different sequence types (STs), including lineages associated with wild animals such as ST49 and ST581, multispecies lineages such as ST130, ST398, and ST425, and lineages commonly isolated from humans, including ST1 and ST5. The hunters and the single positive ferret shared ST5, ST398, or ST425 with wild animals. In wildlife isolates, the highest resistance levels were found for penicillin (32.8%). For virulence factors, 26.2% of them carried superantigens, while 14.8% harbored the immune evasion cluster (IEC), which indicates probable human origin. Our findings suggest that wild animals are a reservoir of clinically relevant genes and lineages that could have the potential to be transmitted to humans. These data support the notion that wildlife surveillance is necessary to better understand the epidemiology of S. aureus as a pathogen that circulates among humans, animals, and the environment.

Chickens intensively selected for accelerating growth rate and enhanced muscle mass has resulted in the occurrence of breast alterations with a negative consumers perception. Together with the already known breast abnormalities (White Striping, Wooden Breast and Spaghetti Meat) a new one has been described by the consumers as a “long and thin worm”. For this reason, the aim of this work was to macroscopically and microscopically describe this structure in broiler breast. To this end, 2 different experiments were performed. In the first one, 4 broiler breasts from a supermarket were analysed. Thereafter, a second experiment was carried out on an experimental farm simulating field management conditions in order to confirm the results of the first experiment. To this end, 120 chickens were reared in an experimental farm up to 42 d. Breast samples were collected and analysed macroscopically and microscopically to assess the abnormality on the broiler breast. In both experiments, the results of the analysis showed the finding of tubular structures located on the surface of pectoralis major muscle compatible with arteries. So far, currently, this occurrence has not been described in the literature and we propose calling this new broiler breast feature as breast-vessel.

Staphylococcal mastitis is a major health problem in humans and livestock that leads to economic loss running in millions. This process is currently one of the main reasons for culling adult rabbit does. Surprisingly, the two most prevalent S. aureus lineages isolated from non-differentiable natural clinical mastitis in rabbits (ST121 and ST96) generate different immune responses. This study aimed to genetically compare both types of strains to search for possible dissimilarities to explain differences in immune response, and to check whether they showed similar virulence in in vitro tests as in experimental intramammary in vivo infection. The main differences were observed in the enterotoxin gene cluster (egc) and the immune-evasion-cluster (IEC) genes. While isolate ST121 harboured all six egc cluster members (seg, sei, selm, seln, selo, selu), isolate ST96 lacked the egc cluster. Strain ST96 carried a phage integrase Sa3 (Sa3int), compatible with a phage integrated into the hlb gene (β-haemolysin-converting bacteriophages) with IEC type F, while isolate ST121 lacked IEC genes and the hlb gene was intact. Moreover, the in vitro tests confirmed a different virulence capacity between strains as ST121 showed greater cytotoxicity for erythrocytes, polymorphonuclear leukocytes and macrophages than strain ST96. Differences were also found 7 days after experimental intramammary infection with 100 colony-forming units. The animals inoculated with strain ST121 developed more severe gross and histological mastitis, higher counts of macrophages in tissue and of all the cell populations in peripheral blood, and a significantly larger total number of bacteria than those infected by strain ST96.

The appearance of methicillin-resistant strains of Staphylococcus aureus (MRSA) in several animal species (including rabbits) has set o alarms for their capacity to act as reservoirs for this bacterium. This is especially important in wild animals given its epidemiological implications. The objectives of this study were to identify and characterize S. aureus, specifically MRSA, strains in wild lagomorph high-density areas. Ten hares and 353 wild rabbits from 14 towns with a high rabbit density in the Valencian region (eastern Spanish coast) were sampled. Swabs from the nasal cavity, ears, perineum and lesions (when present) were taken for microbiological studies. The detection of di erent genes and antibiotic susceptibility studies were also carried out. Of all the animals, 41.3% were positive for S. aureus, of which 63.3% were MRSA. Ears were the anatomical location with more S. aureus and MRSA strains. The more frequently identified MLST type was ST1945 (97.1%, 136/140). The mecA gene was found only in one sample. The rest (n = 139) carried the mecC gene and were included in CC130, except one. Penicillin resistance was detected in 28 mec-negative isolates and, in one case, bacitracin resistance. mecA isolate presented resistance to enrofloxacin and tetracycline, and 10 mecC isolates also showed bacitracin resistance. No MRSA isolate was positive for genes chp, sea, tst and PVL. Two ST1945 isolates contained IEC type E (comprising genes scn and sak). mecA-isolate was positive for blaZ. Of the 28 MSSA strains showing resistance to penicillin, 22 carried the blaZ gene. These surprising results highlight the marked presence of MRSA strains in wild rabbits in high-density areas.

In recent decades, concern about rabbit welfare and sustainability has increased. The housing system is a very important factor for animal welfare. However, information about how different available housing types for female rabbits affect their health status is scarce, but this is an important factor for their welfare. Hence, the objective of this study was to evaluate the health status of female rabbits in five common housing systems: three different single-housing systems with distinct available surfaces and heights; a single-housing system with a platform; a collective system. Female rabbits in the collective and platform cages had greater cortisol concentrations in hair than those in the single-housing system with no platform. Haptoglobin concentrations and kit mortality rates during lactation were greater for the collective-cage female rabbits. The collective group had more culled females and more lesions than in the other groups. The main reasons for culling in all the groups were reproduction problems and presence of abscesses, and the collective group of females was the most affected. In conclusion, it appears that keeping females together in collective systems negatively affects their health status and welfare, while single-housing systems imply lower kit mortality rates during lactation and cortisol concentrations, and fewer lesions in female rabbits.

The main purpose of this study was to find several early factors affecting stayability in rabbit females. To reach this goal, 203 females were used from their first artificial insemination to their sixth parturition. Throughout that period, 48 traits were recorded, considered to be performance, metabolic and immunological indicators. These traits were initially recorded in females’ first reproductive cycle. Later, removed females due to death or culling and those that were non-removed were identified. A first analysis was used to explore whether it was possible to classify females between those reaching and those not reaching up to the mean lifespan of a rabbit female (the fifth reproductive) cycle using information from the first reproductive cycle. The analysis results showed that 97% of the non-removed females were classified correctly, whereas only 60% of the removed females were classified as animals to be removed. The reason for this difference lies in the model’s characteristics, which was designed using early traits and was able to classify only the cases in which females would be removed due to performance, metabolic or immunologic imbalances in their early lives. Our results suggest that the model defines the necessary conditions, but not the sufficient ones, for females to remain alive in the herd. The aim of a second analysis was to find out the main early differences between the non-removed and removed females. The live weights records taken in the first cycle indicated that the females removed in their first cycle were lighter, while those removed in their second cycle were heavier with longer stayability (−203 and þ202 g on average, respectively; P < 0.05). Non-removed females showed higher glucose and lower beta-hydroxybutyrate concentrations in the first cycle than the removed females (þ4.8 and −10.7%, respectively; P < 0.05). The average lymphocytes B counts in the first cycle were 22.7% higher in the non-removed females group (P < 0.05). The females removed in the first reproductive cycle presented a higher granulocytes/lymphocytes ratio in this cycle than those that at least reached the second cycle (4.81 v. 1.66; P < 0.001). Consequently, non-removed females at sixth parturition offered adequate body development and energy levels, less immunological stress and a more mature immune function in the first reproductive cycle. The females that deviated from this pattern were at higher risk of being removed from the herd.

Background: Primary corneal tumors are unusual in dogs although there has been a rise in the prevalence of canine corneal squamous cell carcinoma in the past decades due to different factors. Exposure to ultraviolet radiation, presence of chronic keratitis or history of superficial trauma are some of them. We report for the first time a highly infiltrative corneal neoplasia with both exophytic and deep stromal growth, which presented atypical histologic features of a squamous cell carcinoma. Case presentation: An adult male French bulldog was referred with an exophytic, pink to white gelatinous mass occupying approximately 70% of the central cornea on the right eye. Histological findings from the excisional biopsy were consistent with corneal carcinoma and transconjunctival enucleation was performed at the request of the owner. A final diagnosis of primary corneal squamous cell carcinoma was done based on the squamous differentiation observed in the neoplastic cells of the superficial layers. However, cells in deeper layers were less differentiated, showed pseudoacinar formations and did not expressed marker for stratified squamous epithelium (i.e., cytokeratin 5/6). The dramatic thickening of the cornea and the fact of observing neoplastic cells almost at the level of the Descemet’s membrane make this case very unusual as the squamous cell carcinoma in dogs tends to involve the superficial stroma or colonize the corneal surface as an exophytic lesion. Conclusions: Based on the histological findings, a high infiltrative and poorly differentiated corneal squamous cell carcinoma was diagnosed. In terms of clinical relevance, our results suggest that corneal lesions compatible with neoplasia need an early diagnosis in order to prevent the aggressive growth of the tumor and the enucleation of the eye.