Wild mammals involved in the transmission of Trypanosoma cruzi and food sources of Triatoma sherlocki in an endemic region of northeastern Brazil.

The present study, carried out in the municipality of Gentio do Ouro, Bahia, Brazil aimed to evaluate which wild mammals may be involved in the transmission of T. cruzi and which are the blood sources for triatomines collected in the study area. PCR analysis of 31 wild mammals captured revealed T. cruzi infection in 6.4% (2/31): one specimen of the opossum Didelphis albiventris (1/3) and one of the rodent Kerodon rupestris (1/5); despite being more frequent in the area, no specimen of the rodent Thrichomys sp. (0/23) was infected. A total of 169 triatomines were captured. The conclusive detection of food sources was possible only for Triatoma sherlocki Papa et al., 2002 (n = 56), with evidence for: K. rupestris (35.7%), Gallus (17.9%), D. albiventris (14.3%), Homo sapiens (14.3%), Tropidurus hispidus (7.1%), Leopardus geoffroyi (5.3%), Conepatus semistriatus (1.8%), Thrichomys inermis (1.8%) and Rattus norvegicus (1.8%). Triatomines of the species T. sherlocki showed food eclecticism, including feeding on humans, with some of them being captured at dwellings. These facts make this triatomine a potential link for the transmission of T. cruzi between wild and anthropic environments, highlighting a latent risk of the reemergence of Chagas disease outbreaks.

Identification and molecular characterization of Wolbachia strains and natural infection for Leishmania sp. in neotropical Phlebotominae (Diptera: Psychodidae) species, leishmaniasis vectors.

Recently, Wolbachia infection has been described in leishmaniasis vector sandflies. This endosymbiont bacterium is present in 60% of insects, and has been suggested as a mechanism of biological control of vector insects, because it causes a series of changes in the invertebrate host. In addition, recent studies have shown that this bacterium can prevent the development of parasites in vector insects. In this context, the present study aims to molecularly characterize the circulating strain of this bacterium in sandflies in the State of Bahia, Brazil, as well as the natural infection rate of Leishmania sp., and to evaluate the coinfection between Wolbachia and Leishmania. Seven hundred and forty-five (745) specimens of sandflies were collected in nine municipalities of Bahia, belonging to two species, Lutzomyia longipalpis (Lutz and Neiva, 1912) and Nyssomyia whitmani (Antunes and Coutinho, 1939). The results confirm infection by the protozoan Leishmania infantum and Wolbachia in both species collected. The identified strain of Wolbachia in sandflies was wStv MI, known to lead to a phenotype of cytoplasmic incompatibility in vector insects.

Ticks and Associated Pathogens From Rescued Wild Animals in Rainforest Fragments of Northeastern Brazil.

The Ixodidae family comprises ticks that are hematophagous ectoparasites and are considered vectors of several hemoparasites from the Anaplasmataceae family and the genus Hepatozoon, Babesia, and Rickettsia. These ectoparasites parasitize domestic and wild animals belonging to several vertebrate groups. Ticks are highly adapted to different biomes and thus possess a wide geographical distribution. In the Brazilian state of Bahia, localized in the Northeast region, there are large rainforest fragments. Studies have rarely been carried out on ticks, and their hemoparasites, that parasitize wild animals in this region. Thus, this study aimed to identify the tick species parasitizing wild animals rescued in rainforest fragments of Bahia and investigate the presence of hemoparasites in tick tissues. During a 2-year period, 238 ticks were collected from 41 wild mammalians, reptiles, and amphibians. These ectoparasites were taxonomically classified according to their morphological characteristics. The ticks identified belonged to five different species from the Ixodidae family: Amblyomma varium, Amblyomma rotundatum, Amblyomma nodosum, Ixodes loricatus, and Rhipicephalus sanguineus. For the first time, an A. rotundatum parasitizing the Mesoclemmys tuberculata turtle was described. PCR assays using DNA extracted from salivary glands or midgut of the ticks were performed to detect specific DNA fragments of hemoparasites from the genus Rickettsia, Ehrlichia, Babesia, Hepatozoon, and from the Anaplasmataceae family. The results showed positive detection of the Rickettsia genus (7.9%), Anaplasmataceae family (15.8%), and Hepatozoon genus (15.8%). Specific DNA from the Ehrlichia and Babesia genera were not detected in these samples. Specific DNA from members of the Anaplasmataceae family was detected in A. varium for the first time. The present work showed that amphibians, reptiles, and mammals from Bahia's Atlantic Forest areparasitized by different tick species, and that these ectoparasites present pathogens in their tissues that impact both humans and animals due to their zoonotic potential.

Molecular detection of benzimidazole resistance levels associated with F167Y and F200Y polymorphisms in Haemonchus contortus of goats from Mozambique.

Benzimidazole (BZ) resistance of Haemonchus contortus has been associated with single nucleotide polymorphisms (SNPs) in codons 200 (F200Y) and 167 (F167Y) and, to a lesser extent, in codon E198A, of the ß-tubulin isotype 1 gene. The present study was undertaken to survey the status of BZ resistance in naturally infected goats in smallholder farms in southern Mozambique by real-time PCR (qPCR) using TaqMan® assays. H. contortus-infective larvae (L3; n = 432) from 12 populations were individually genotyped for F200Y and F167Y SNPs to detect BZ resistance. For the F200Y SNP, the results revealed an overall mean percentages of 18.8% homozygous resistant (RR), 47.8% homozygous susceptible (SS) and 33.4% heterozygous (RS) H. contortus. For the F167Y SNP, the overall mean percentages were 1.6% RR, 94.9% SS and 3.5% RS. The percentage of resistant alleles (%R) for the F200Y and F167Y SNPs was 35.7 and 3.4%, respectively. Genotype combinations of the two mutations indicate resistant percentages ranging from 0.0 to 52.9%. From the four herds with high RR individuals, three farms dewormed the animals monthly, while the fourth farm dewormed the animals every 3 months. In farms where animals were dewormed every 6 months, low percentages of RR individuals were found, whereas no RR individuals were discovered in herds where animals were dewormed annually. These results suggest that the F200Y SNP is more significant in BZ resistance development of the surveyed population compared with the F167Y SNP.

Genotypic profile of benzimidazole resistance associated with SNP F167Y and F200Y beta-tubulin gene in Brazilian populations of Haemonchus contortus of goats.

Benzimidazoles are the most common anthelminthic used for control of gastrointestinal nematodes of goats in the Brazilian semi-arid region. Resistance to these compounds in the nematode Haemonchus contortus has been associated with single nucleotide polymorphisms (SNP) in codons 167 (F167Y) and 200 (F200Y) on the ß-tubulin isotype 1 gene. To determine the resistance profile to benzimidazoles of populations of H. contortus of goats of Brazilian semi-arid region, larvae of 29 populations of these nematodes were individually genotyped by real time PCR using a Taqman assay. The percentage of larvae homozygous (RR) for SNP F200Y was relatively low (18.9%), particularly when compared to SNP F167Y (32.7%), indicating that the latter has more relevance in this region. However, the associations between these two SNP demonstrate percentages of resistance ranging from 34.7% to 100% between populations, being the highest percentages for homozygous individuals resistant for the mutation 167 and susceptible to mutation 200 (RR-F167Y/F200Y-SS: 26.7%), followed by combination of heterozygous for both mutations (F167Y-SR/F200Y-SR: 22.8%). These results indicate high levels of resistance in populations of H. contortus of goats in the Brazilian semi-arid region, and thus ineffective antiparasitic control with the use of benzimidazoles in the region.

Galectins and collectinis expression are increased in Haemonchus contortus-infected corriedale sheep.

Galectins and collectins are proteins classified in the lectin family that have the ability to recognize molecular patterns associated with pathogens. Studies on cattle have demonstrated high expression of these proteins during infection with gastrointestinal nematodes. The aim of this study was to investigate whether the level of Haemonchus contortus infection would alter the expression of galectins (Gal11 and Gal14) and collectins (SPA and CGN) in sheep. Twelve Corriedale sheep exposed to natural infection with nematodes were divided into two groups: group 1 (G1, n = 7) and group 2 (G2, n = 5), with low and high parasite burdens, respectively, based on fecal egg counts and abomasal parasite counts. The fecal egg counts and abomasal parasite counts were significantly different (p < 0.05) between the groups. Galectin and collectin gene expression was observed in all sheep abomasal samples. However, animals with lower infection levels showed lower expression of the genes Gal14, SPA and CGN (p < 0.05). Expression of lectins was associated with the abomasal H. contortus burden, thus suggesting that these proteins may have a role in controlling of this infection.

Galectins and collectinis expression are increased in Haemonchus contortus-infected corriedale sheep / Aumento da expressão gênica de colectinas e galectinas em ovinos corriedale infectados por Haemonchus contortus

Galectins and collectins are proteins classified in the lectin family that have the ability to recognize molecular patterns associated with pathogens. Studies on cattle have demonstrated high expression of these proteins during infection with gastrointestinal nematodes. The aim of this study was to investigate whether the level of Haemonchus contortus infection would alter the expression of galectins (Gal11 and Gal14) and collectins (SPA and CGN) in sheep. Twelve Corriedale sheep exposed to natural infection with nematodes were divided into two groups: group 1 (G1, n = 7) and group 2 (G2, n = 5), with low and high parasite burdens, respectively, based on fecal egg counts and abomasal parasite counts. The fecal egg counts and abomasal parasite counts were significantly different (p < 0.05) between the groups. Galectin and collectin gene expression was observed in all sheep abomasal samples. However, animals with lower infection levels showed lower expression of the genes Gal14, SPA and CGN (p < 0.05). Expression of lectins was associated with the abomasal H. contortus burden, thus suggesting that these proteins may have a role in controlling of this infection.

Colectinas e galectinas são proteínas da família das lectinas que possuem a capacidade de reconhecer padrões moleculares associados aos patógenos. Estudos em bovinos têm demonstrado a alta expressão dessas proteínas durante a infecção por nematoides gastrintestinais. O objetivo deste estudo foi investigar se o nível de infecção de Haemonchus contortus altera a expressão de colectinas (SPA e CGN) e galectinas (Gal11 e Gal14) de ovinos. Doze ovinos da raça Corriedale expostos a infecção natural com nematoides foram separados em dois grupos: grupo 1 (G1, n=7) com menor grau de parasitismo; e grupo 2 (G2, n=5) com maior grau, a partir da contagem do número de parasitos recuperados do abomaso e OPG. A contagem de OPG e de parasitos recuperados do abomaso dos grupos G1 e G2 apresentaram diferença estatística (p<0,05). A expressão dos genes de colectinas e galectina foi observada em todas as amostras de abomaso dos ovinos, porém animais com menor grau de infecção apresentaram menor expressão dos genes de Gal14, SPA e CGN (p<0,05). A expressão de lectinas foi associada ao número de H. contortus encontrados no abomaso de ovinos, indicando um possível papel dessas proteínas no controle da infecção.

Comparison of two commercial vaccines against visceral leishmaniasis in dogs from endemic areas: IgG, and subclasses, parasitism, and parasite transmission by xenodiagnosis.

BACKGROUND: The incidence of zoonotic canine visceral leishmaniasis (CVL) would decrease if dogs were effectively vaccinated; however, additional data on the efficacy of canine vaccines are required for their approved preventative use. PURPOSE: To prospectively evaluate vaccination outcomes using two products commercially available in Brazil, with respect to adverse reactions (reactogenicity), humoral response, disease signs, parasitism, and parasite infectiousness in naturally exposed pet dogs in an endemic area of visceral leishmaniasis (VL). METHODS: From 2010 to 2012, healthy dogs were vaccinated with Leishmune(®) (50 animals) or Leish-Tec(®) (50 animals). Each dog was examined to identify clinical signs during peri- and post-vaccination procedures every 2 months for 11 months to identify the presence of parasites or parasite DNA in splenic samples using culturing or PCR, respectively. Levels of anti-Leishmania IgG, IgG1, and IgG2 were quantified in sera by ELISA and infectiousness was assessed by xenodiagnosis. RESULTS: Adverse effects occurred in 2.2% (1/45) and 13.0% (6/46) of the animals in the Leishmune(®) and Leish-Tec(®) groups, respectively. IgG levels peaked on the 21st day following the first dose of Leishmune(®) and on the 21st day after the second dose of Leish-Tec(®). The final seropositivity rate for IgG was 32.5% (13/40) and 30.9% (13/42) in the Leishmune(®) and Leish-Tec(®) groups, respectively. The Leishmune(®) group presented higher levels of IgG1 and IgG2 compared to the Leish-Tec(®) group (p<0.001), and ELISA reactivity in both vaccinated groups was significantly lower (p<0.001) than in infected positive control dogs. Parasitism was observed in 12.2% (5/41) of the Leishmune(®) group, and 7.9% (3/38) of the Leish-Tec(®) group, with xenodiagnostic transmission rates of Leishmania to Lutzomyia longipalpis of 5.1% (2/39), and 5.4% (2/37), respectively. CONCLUSIONS: No significant differences were observed in dogs vaccinated with Leishmune(®) or Leish-Tec(®), with respect to LVC clinical aspects, parasitism, IgG seropositivity, or dog infectiousness. The Leishmune(®)-vaccinated animals presented higher levels of IgG, IgG1, and IgG2. The animals vaccinated with Leish-Tec(®) exhibited adverse reactions with greater frequency and severity.

Prevalence of ehrlichial infection among dogs and ticks in Northeastern Brazil.

This study investigated the epidemiology of canine ehrlichiosis in Northeastern Brazil, focusing the identification of the Ehrlichia species and vectors involved. Samples were collected from 472 domestic dogs residing in the health districts of Cajazeiras and Itapuã of Salvador city. The average prevalence of antibodies reactive to E. canis by immunofluorescent antibody test (IFAT) (titer ≥ 1:80) was 35.6% (168/472). Blood samples from the E. canis-seropositive animals were tested by nested PCR in order to identify the Ehrlichia species responsible for the infection. Among the seropositives, 58 (34.5%) were found to be PCR-positive for E. canis. Ticks were found in 32 dogs. Nested-PCR analysis showed that 21.9% (7/32) of the Rhipicephalus sanguineus were infected by E. canis. In both dogs and Rhipicephalus sanguineus, nested-PCR for E. ewingii and E. chaffeensis was negative, with no amplification of DNA fragment.

Prevalence of ehrlichial infection among dogs and ticks in Northeastern Brazil / Prevalência da infecção por Ehrlichia em cães e carrapatos no Nordeste do Brasil

This study investigated the epidemiology of canine ehrlichiosis in Northeastern Brazil, focusing the identification of the Ehrlichia species and vectors involved. Samples were collected from 472 domestic dogs residing in the health districts of Cajazeiras and Itapuã of Salvador city. The average prevalence of antibodies reactive to E. canis by immunofluorescent antibody test (IFAT) (titer > 1:80) was 35.6 percent (168/472). Blood samples from the E. canis-seropositive animals were tested by nested PCR in order to identify the Ehrlichia species responsible for the infection. Among the seropositives, 58 (34.5 percent) were found to be PCR-positive for E. canis. Ticks were found in 32 dogs. Nested-PCR analysis showed that 21.9 percent (7/32) of the Rhipicephalus sanguineus were infected by E. canis. In both dogs and Rhipicephalus sanguineus, nested-PCR for E. ewingii and E. chaffeensis was negative, with no amplification of DNA fragment.

Este estudo objetivou pesquisar a epidemiologia da erliquiose canina no Nordeste do Brasil, com especial atenção na identificação da espécie de Ehrlichia envolvida nas infecções caninas e vetoriais detectadas. Para isso foram coletadas amostras de 472 cães domiciliados nos distritos sanitários de Cajazeiras e Itapuã. A prevalência de anticorpos anti-E. canis, pela imunofluorescência indireta (título > 1:80), em cães foi de 35,6 por cento (168/472). Os animais soropositivos foram analisados por uma nested-PCR para identificação da espécie de Ehrlichia responsável pela infecção. Dentre os positivos, 58 (34,5 por cento) cães foram PCR-positivos para E. canis. Foram coletados e classificados os carrapatos em 32 cães. A nested-PCR de Rhipicephalus sanguineus resultou em 21,9 por cento (7/32) de infecção por E. canis. A nested-PCR de amostras de sangue de cães e Rhipicephalus sanguineus para E. chaffeensis e E. ewingii foi negativa, não havendo amplificação de fragmento de DNA.