Effect of Vaccination against Leptospira on Shelter Asymptomatic Dogs Following a Long-Term Study.

(1) Background: Vaccination of dogs against leptospirosis is of paramount importance, as they ideally must provide not only long-term protection, but also against the renal carrier state of leptospires. This study assessed the post-vaccine humoral response against Leptospira in naturally exposed dogs and effects on renal carrier status. (2) Methods: A total of 118 dogs were studied for 365 days, separated into Group A (vaccinated, n = 94) and Group B (non-vaccinated, n = 24). Group A was subdivided into three groups: A1 with 32 dogs immunized with the vaccine #1; A2 by 32 dogs with #2; and A3 30 dogs with #3. Serology (MAT and IgG-ELISA) and urinary PCR were conducted. (3) Results: Seroreactivity increased at D15 post-vaccination and, regardless of vaccine brand, remained high up to D180, with antibody switch to IgG after D30. A total of 46.8% of animals from Group A were PCR-positive at least once, in contrast to 75% in Group B, regardless of vaccine brand (p < 0.05; OR: 0.3). (4) Conclusions: All commercial vaccines succeeded at eliciting a long-term IgG-based response and were partially effective at protecting against kidney infection.

Molecular epidemiology of Leptospira noguchii reveals important insights into a One Health context.

Leptospirosis presents a complex and dynamic epidemiology. Bovine leptospirosis has been described as a major infectious disease impairing reproductive efficiency. Although infections by Leptospira interrogans, L. santarosai and L. borgpetersenii are frequently reported in cattle, the presence of L. noguchii in these animals should not be neglected. In this study, we describe serological (MAT) and molecular characterization (rrs and secY gene sequencing, multilocus sequence typing [MLST] and pulsed-field gel electrophoresis [PFGE]) of eight L. noguchii strains obtained from slaughtered cows. Intraspecific genetic diversity was evaluated, and haplotype networks were constructed based on hosts and geographical localizations. Strains were characterized as belonging to serogroups Australis, Autumnalis and Panama, and molecular characterization showed a high heterogeneity of these strains. Ten different STs were found (including nine new STs and 39 novel alleles) as well as nine different pulsotypes. Two clonal complexes were found. Phylogenetic trees based on secY locus and concatenated MLST loci showed two main clusters, with sequences from the present study included in the first. In general, there was no relationship between the geographical origin and the secY phylogenetic clusters, as well as between secY phylogenetic clusters and serogroups. Molecular diversity indexes confirmed a high variability (H > 0.8). This high intraspecific variation observed may be related to differences in virulence, pathogenicity and antigenicity or even adaptability of the strains. In addition, haplotype networks clearly demonstrated the circulation of genotypes between humans and animals, confirming the zoonotic potential. The present study provides relevant data for the study of leptospirosis in the One Health context, where human, animal and environmental health is closely connected.

Neotropical wild rodents Akodon and Oligoryzomys (Cricetidae: Sigmodontinae) as important carriers of pathogenic renal Leptospira in the Atlantic forest, in Brazil.

Leptospirosis is an important worldwide zoonosis, caused by a bacterium of the genus Leptospira. For a better understanding of the disease, it is relevant the application of the one health concept. The Atlantic Forest is considered a biodiversity hotspot, with a great endemism of species and despite its importance and proximity to urban areas, the potential role of its fauna as carriers of infectious agents is still poorly understood. Although it is well-known that rats and mice are key reservoirs of leptospires, particularly Rattus norvegicus, wild rodents have also been revealed as reservoirs of leptospiral strains. Thus, the purpose of this study was to investigate the role of the sigmodontine rodents from genera Akodon and Oligoryzomys as pathogenic Leptospira carriers in the Atlantic Forest. We studied 52 Akodon spp. and 15 Oligoryzomys spp. from three areas. Overall 30% were PCR-positive for pathogenic Leptospira, 27% (14/52) Akodon spp. and 40% (6/15) Oligoryzomys spp. DNA sequencing of LipL32 gene confirmed nine species as pathogenic Leptospira. This remarkable incidence of leptospiral carriage within wild genera emphasizes the role of these rodents as carriers of leptospires throughout in this environment.

Nematode parasites of commercially important fish from the southeast coast of Brazil: Morphological and genetic insight.

Studies of nematofauna of teleost fish from the Brazilian coast are relatively scarce and limited to identification based on morphology. The objective of the present study was to determine the diversity and prevalence of nematode parasites in teleost fish from the southeast Atlantic coast of Rio de Janeiro, through morphological, molecular, and ecological approaches. Parasites were collected from sixty specimens each of Genypterus brasiliensis, Micropogonias furnieri, and Mullus argentinae obtained in winters and summers of 2012­2014. Morphological and genetic characterization was conducted using light microscopy and the molecular targets 18S rDNA, ITS1, and mtDNA cox2. Nematodes identified in M. furnieri were Cucculanus genypteri (n = 1575, P = 98.3%) and Hysterothylacium deardorffoverstreetorum (s.l.) (n = 2, P = 3.3%); in G. brasiliensis were Dichelyne (Cucullanellus) sciaenidicola (n = 99, P = 33.3%), Cucculanus pulcherrimus (n = 45, P = 18.3%), Hysterothylacium deardorffoverstreetorum (s.l.) (n = 3, P = 5%), and Anisakis typica (n = 1, P = 1.7%); and, in M. argentinae, were H. deardorffoverstreetorum (s.l.) (n = 146, P = 48.3%), and Procamallanus (Spirocamallanus) halitrophus (n = 4, P = 6.7%). DNA sequence data of C. genypteri, C. pulcherrimus, D. (C.) sciaenidicola, and P. (S.) halitrophus were reported for the first time. New host records are M. argentinae for P. (S.) halitrophus, M. furnieri for A. typica, while H. deardorffoverstreetorum (s.l.) was found in all three fish species. Intestine showed significantly higher intensity than other sites, and no significant seasonal variation in parasitological indices was observed. Hysterothylacium specimens (n = 6) were found in fish muscle, potentially a public health concern.

Integrative taxonomy of anisakid nematodes in stranded cetaceans from Brazilian waters: an update on parasite's hosts and geographical records.

Anisakidae are represented mainly by species of Anisakis Dujardin, 1845, Pseudoterranova Krabbe, 1878, and Contracaecum Railliet and Henry, 1913. Integrative taxonomy, based on morphological and genetic approaches, can provide a precise identification of species, increasing the knowledge of their systematics, biology, and evolution. In this study, integrative taxonomy was applied in order to identify, at generic and/or specific taxonomic level, 245 anisakids from 63 cetaceans of 12 different species, which stranded in the Brazilian coast. Parasites from nine hosts were preserved by freezing and, from 54 hosts, were fixed in 70% ethanol (n = 43) or alcohol-formalin-acetic (n = 11) for a period varying from 6 to 19 years. The morphological analysis allowed the identification of 171 specimens at genus level: 120 as Anisakis sp. clade I, 47 as Anisakis sp. clade II, and four as Pseudoterranova sp. From those 15, specimens were identified at species level, seven as Anisakis typica, seven as Anisakis paggiae and one as Anisakis brevispiculata. Moreover, 74 specimens were identified as belonging to Anisakidae family. The genetic analysis based on cox2 gene allowed the identification/confirmation of Anisakis typica (n = 33), Anisakis paggiae (n = 1), and for the first time in the Brazilian coast, Anisakis brevispiculata (n = 4) and Anisakis ziphidarum (n = 1). Additionally, a probably new Anisakis species, genetically close to A. paggiae, is reported infecting a dwarf sperm whale. The results clearly indicated that the preservation of nematodes influenced significantly on the level of taxonomic definition, in both morphological and, mainly, genetic analyses. Pseudoterranova sp. was identified for the first time in the spinner dolphin, Stenella longirostris. The detection of nine A. paggiae specimens, in three Kogia breviceps animals, substantiates the austral occurrence of this species. The identification of A. brevispiculata in Kogia sima and K. breviceps confirms Kogiidae whales as their most common hosts. Anisakis typica was corroborated as a generalist species, the most widely distributed in the Brazilian coast, infecting a broad variety of Delphinidae and Kogiidae, with two new geographical records in Stenella frontalis and Lagenodelphis hosei. The study showed a new scenery of parasite diversity of marine anisakids, updating the Anisakis species distribution.

Integrative taxonomy of Anisakidae and Raphidascarididae (Nematoda) in Paralichthys patagonicus and Xystreurys rasile (Pisces: Teleostei) from Brazil.

Thirty-six Paralichthys patagonicus and 30 Xystreurys rasile were collected in the state of Rio de Janeiro, Brazil to investigate the presence of anisakid and raphidascaridid nematodes. Anisakis typica, Terranova sp., Contracaecum sp., Hysterothylacium deardorffoverstreetorum, and Raphidascaris sp. were identified using integrative taxonomy of morphological and genetic data. Morphological and morphometric analysis was conducted using bright field microscopy with scanning electron microscopy for topographic characterization of the cuticular surface. Phylogenetic analysis, using ITS and cox2 molecular targets, clearly demonstrated the species identification of A. typica and H. deardorffoverstreetorum and the high diversity of H. deardorffoverstreetorum. This is the first report of A. typica, H. deardorffoverstreetorum, and Raphidascaris sp. parasitizing P. patagonicus and X. rasile.

Morphological and genetic identification of Anisakis paggiae (Nematoda: Anisakidae) in dwarf sperm whale Kogia sima from Brazilian waters.

Anisakid nematodes have been identified in a wide variety of fish and marine mammal species. In Brazil, Anisakis physeteris, A. insignis, A. typica, A. nascetti, and those of the A. simplex complex have been reported infecting fishes and cetaceans. In this study, specimens collected from a dwarf sperm whale Kogia sima (Owen, 1866) stranded on the northeastern coast of Brazil were identified through morphological and genetic analyses as A. paggiae. Anisakids were examined through differential interference contrast light and scanning electron microscopy (SEM). Morphological and morphometric analysis revealed that these specimens belonged to Anisakis sp. clade II and more specifically to A. paggiae, exhibiting a violin-shaped ventriculus and 3 denticulate caudal plates, which are taxonomic characters considered unique to this species. Genetic analysis based on the mtDNA cox2 gene confirmed our identification of A. paggiae. Phylogenetic trees using both maximum likelihood and neighbor-joining methods revealed a strongly supported monophyletic clade (bootstrap support = 100%) with all available A. paggiae sequences. Integrative taxonomic analysis allowed the identification of A. paggiae for the first time in Brazilian waters, providing new data about their geographical distribution. Moreover, here we present the first SEM images of this species.