1. Investigación

Visceral leishmaniosis is the one of the most important protozoal zoonoses in Europe, and it is caused by Leishmania infantum, an intracellular protozoan parasite. The disease is endemic in dogs in the Mediterranean area. The main goal of this work is to correlate the levels of several cytokines linked to immune response against L. infantum infection in two canine breeds. Thirty-one Boxer and twenty-eight Ibizan Hound dogs living in the Valencian Community (East coast of Spain) were analyzed for the presence of anti-Leishmania antibodies in serum by IFAT test. Cytokines IFN-γ, TNF-α, IL-2, IL-6, IL-8, and IL-18 were determined by ELISA commercial tests. The levels of IFN-γ, IL-2, and IL-18 in our study, cytokines linked to a cellular immune response, were higher (p < 0.05) in the Ibizan Hound breed; IL-6 levels were higher, although not significant, and only levels of IL-8 were higher in Boxer than in Ibizan Hound. No expression of TNF-α was found. These results corroborate that Ibizan Hound can develop a protective response against canine leishmaniosis, while Boxer is a susceptible breed. The study of immunological aspects in the different canine breeds may represent a useful tool in the prediction of the disease.

Background: The Ibizan Hound is a canine breed native to the Mediterranean region, where leishmaniosis is an endemic zoonosis. Several studies indicate low prevalence of this disease in these dogs but the underlying molecular mechanism remains unknown. Methods: In this study, qualitative immunological and genomic profiles of this breed have been analyzed. Results: Our analysis shows relevant differences between the cytokine serum profile of Ibizan Hound dogs and previously published data from other canine strains. Additionally, several genetic risk variants related to the immune response, regulation of the immune system, and genes encoding cytokines and their receptors have been studied. The most relevant genes that presented such fixed polymorphisms were IFNG and IL6R. Other variants with frequencies ≥ 0.7 were found in the genes ARHGAP18, DAPK1, GNAI2, MITF, IL12RB1, LTBP1, SCL28A3, SCL35D2, PTPN22, CIITA, THEMIS, and CD180. Epigenetic regulatory genes such as HEY2 and L3MBTL3 showed also intronic polymorphisms. Conclusions: Our analysis and results indicate that the regulation of immune responses is different in Ibizan Hounds compared to other breeds. Future studies are needed to elucidate whether these differences are related to the low prevalence of L. infantum infection in the Ibizan Hound.

Leishmaniosis, one of themost important zoonoses in Europe, is caused by Leishmania infantum, an intracellular protozoan parasite. This disease is endemic in theMediterranean area, where the main reservoir is the dog. Several studies indicate a possible susceptibility to L. infantum infection with clinical signs in some canine breeds. One of them is the boxer breed, which shows a high prevalence of disease. In this study, immunological and genomic characterization of serum samples from boxer dogs living in theMediterranean area were evaluated to analyze the immune response and the possible genetic explanation for this susceptibility. Serum levels of cytokines IFN- , IL-2, IL-6, IL-8, and IL-18 were determined by ELISA commercial tests, while the genotyping study was performed using the CanineHD DNA Analysis BeadChip. The results show relevant differences in the serum levels of cytokines compared to published data on other canine breeds, as well as sequence changes that could explain the high susceptibility of the boxer breed to the disease. Concretely, polymorphic variants in the CIITA, HSF2BP, LTBP1,MITF, NOXA1, PKIB, RAB38, RASEF, TLE1, and TLR4 genes were found, which could explain the susceptibility of this breed to L. infantum infection.

Canine leishmaniosis is caused by infection with parasite Leishmania infantum, which are transmitted by sandflies Phlebotomus. Canine leishmaniosis is an endemic disease in the Mediterranean region. The immune response could vary between hosts and determines the severity of the disease and clinical features. The aim of this study was to analyze the serum levels of cytokines TNF- , IFN- , IL-2, IL-6, and IL-8, which are related to the activation of Th1 or Th2 immune responses in dogs living in the L. infantum endemic region. Moreover, we intend to relate and correlate these levels with different factors, such as sex, age, diet, lifestyle, and breed. Epidemiological data and serum were recovered for seventy-eight dogs, and serum levels of cytokines described previously were analyzed by using the ELISA method. The results showed differences in serum levels of IFN- , IL-2, and IL-8 between breeds. The lifestyle also affected serum levels of IL-2. The main conclusion of this study is that Ibizan hounds and crossbred dogs have a serological profile of cytokines that seems to indicate certain protections against infection by L. infantum compared to boxer and purebred breeds.

Leishmaniosis is an important zoonotic protozoan disease primarily spread to the Mediterranean region by Leishmania infantum, the predominant protozoan species, which accounts for the majority of cases. Development of disease depends on the immune response of the definitive host and, predictably, their genetic background. Recent studies have revealed breed-typical haplotypes that are susceptible to the spread of the protozoan parasite. The objective of this study was to analyze the prevalence of leishmaniosis on a Mediterranean island and determine the relationship between disease prevalence and breed. In addition, information on seropositive animals was recorded to characterize animals affected by the disease. To study the prevalence, a total of 3141 dogs were analyzed. Of these, the 149 infected animals were examined for age, sex, antibody titer, and disease stage. We observed a prevalence of 4.74%, which varied between breeds (p < 0.05). The Doberman Pinscher and Boxer breeds had the highest prevalence of leishmaniosis. Significant differences were observed between breeds with common ancestors, emphasizing the important genetic component. Finally, regarding the characterization of seropositive animals, the distribution is similar to other studies. We discovered a relationship (p < 0.05) between the number of antibody titers and the clinical disease stage, which was also present in Leishmania infantum, suggesting that the development of the disease depends on the humoral or Th2 immune response with ineffective antibodies.