2. Universidad Cardenal Herrera-CEU

Listeria monocytogenes is a major human and animal foodborne patho-gen. However, data from environmental reservoirs remain scarce. Here, we usedwhole-genome sequencing to characterize Listeria species isolates recovered over 1year from wild animals in their natural habitats in Spain. Three different Listeria spp.(L. monocytogenes [n = 19], Listeria ivanovii subsp. londoniensis [n = 4], and Listeriainnocua [n = 3]) were detected in 23 animal tonsils (9 deer, 14 wild boars) and 2feeding troughs. No Listeria species was detected in feces. L. monocytogenes wasdetected in tonsils of 44.4% (8 out of 18) of deer and 40.7% (11 out of 27) of wildboars. L. monocytogenes isolates belonged to 3 different core genome multilocussequence typing (cgMLST) types (CTs) of 3 distinct sublineages (SL1, SL387, andSL155) from lineages I and II. While cgMLST type L1-SL1-ST1-CT5279 (IVb; clonalcomplex 1 [CC1]) occurred only in one animal, types L1-SL387-ST388-CT5239 (IVb;CC388) and L2-SL155-ST155-CT1170 (IIa; CC155) were retrieved from multiple ani-mals. In addition, L1-SL387-ST388-CT5239 (IVb; CC388) isolates were collected 1 yearapart, revealing their long-term occurrence within the animal population and/orenvironmental reservoir. The presence of identical L. monocytogenes strains in deerand wild boars suggests contamination from a common food or environmentalsource, although interhost transmission cannot be excluded. Pathogenicity islandsLIPI-1, LIPI-3, and LIPI-4 were present in 100%, 5%, and 79% of the L. monocytogenesisolates, respectively, and all L. monocytogenes lineage II isolates (n = 3) carried SSI-1stress islands. This study highlights the need for monitoring L. monocytogenes envi-ronmental contamination and the importance of tonsils as a possible L. monocyto-genes intrahost reservoir.

Background: Listeria monocytogenes is a foodborne bacterial pathogen that causes listeriosis, an infectious disease in animals and people, with pigs acting as asymptomatic reservoirs. In August 2019 an outbreak associated with the consumption of pork meat caused 222 human cases of listeriosis in Spain. Determining the diversity as well as the virulence potential of strains from pigs is important to public health. Methods: The behaviour of 23 L monocytogenes strains recovered from pig tonsils, meat and skin was compared by studying (1) internalin A, internalin B, listeriolysin O, actin assembly-inducing protein and PrfA expression levels, and (2) their invasion and intracellular growth in eukaryotic cells. Results: Marked differences were found in the expression of the selected virulence factors and the invasion and intracellular replication phenotypes of L monocytogenes strains. Strains obtained from meat samples and belonging to serotype 1/2a did not have internalin A anchored to the peptidoglycan. Some strains expressed higher levels of the studied virulence factors and invaded and replicated intracellularly more efficiently than an epidemic L monocytogenes reference strain (F2365). Conclusion: This study demonstrates the presence of virulent L monocytogenes strains with virulent potential in pigs, with valuable implications in veterinary medicine and food safety.

Leishmania is a genus of parasitic protozoa that causes a disease called leishmaniasis. Leishmaniasis is transmitted to humans through the bites of infected female sandflies. There are several different species of Leishmania that can cause various forms of the disease, and the symptoms can range from mild to severe, depending on species of Leishmania involved and the immune response of the host. Leishmania parasites have a variety of reservoirs, including humans, domestic animals, horses, rodents, wild animals, birds, and reptiles. Leishmaniasis is endemic of 90 countries, mainly in South American, East and West Africa, Mediterranean region, Indian subcontinent, and Central Asia. In recent years, cases have been detected in other countries, and it is already an infection present throughout the world. The increase in temperatures due to climate change makes it possible for sandflies to appear in countries with traditionally colder regions, and the easy movement of people and animals today, facilitate the appearance of Leishmania species in new countries. These data mean that leishmaniasis will probably become an emerging zoonosis and a public health problem in the coming years, which we must consider controlling it from a One Health point of view. This review summarizes the prevalence of Leishmania spp. around the world and the current knowledge regarding the animals that could be reservoirs of the parasite.

Leishmaniasis is a zoonotic parasitic disease, and the main reservoir of the parasite is the dog, although recent years have seen an increase in other mammalian species. In the Mediterranean region, where it is an endemic disease, it is caused by the species Leishmania infantum. The Ibizan hound, an autochthonous breed of this region, appears to have a genetic resistance to parasitic infection, whereas other canine breeds, such as the Boxer, are susceptible to infection. These differences are related to the differentiated activation of the immune response, with the Ibizan hound activating the Th1 immune response, whereas the Boxer breed triggers the Th2 immune response. Cytokine levels and genomic haplotypes of several genes involved in the immune response were analysed in twenty-eight Ibizan hound (resistant canine breed model) and twenty-four Boxer (susceptible canine breed) without clinical signs in the Mediterranean region. Cytokine levels were analysed by ELISA commercial kits and haplotypes were studied using CanineHD DNA Analysis BeadChip including 165,480 mapped positions. The results show 126 haplotypes associated with differential immune response in dogs. Specifically, haplotypes in IL12RB1, IL6R, CIITA, THEMIS, NOXA1, HEY2, RAB38, SLC35D2, SLC28A3, RASEF and DAPK1 genes are associated with serum levels of IFN-γ, IL-2, IL-8, and IL-18. These results suggest that the resistance or susceptibility to Leishmania infantum infection could be a consequence of haplotypes in several genes related to immune response. Future studies are needed to elucidate the relationship of these haplotypes with immune response and gene expression regulation.