High-resolution prediction models for Rhipicephalus microplus and Amblyomma cajennense s.l. ticks affecting cattle and their spatial distribution in continental Ecuador using bioclimatic factors.

In Ecuador, the main tick species affecting cattle are Rhipicephalus microplus and Amblyomma cajennense sensu lato. Understanding their spatial distribution is crucial. To assess their distribution, data from 2895 farms visited between 2012 and 2017 were utilized. Ticks were collected during animal inspections, with each farm's location georeferenced. Bioclimatic variables and vapor pressure deficit data were obtained from Climatologies at High resolution for the Earth´s Land Surface Areas (CHELSA) dataset. They were overlaid to develop predictive maps for each species using Random Forest (RF) models. The cross-validation results for RF prediction models showed high accuracy for both R. microplus and A. cajennense s.l. presence with values of accuracy = 0.97 and 0.98, sensitivity = 0.96 and 0.99, and specificity = 0.96 and 0.93, respectively. A carefully selected subset of bioclimatic variables was used to describe the presence of each tick species. Higher levels of precipitation had positive effect on the presence of R. microplus but a negative effect on A. cajennense s.l. In contrast, isothermality (BIO3) was more important for the presence of A. cajennense s.l. compared to R. microplus. As a result, R. microplus had a broader distribution across the country, while A. cajennense s.l. was mainly found in coastal areas with evident seasonality. The coexistence of both species in some regions could be attributed to transitional zones, whereas high altitudes limited tick presence. This information can aid in developing appropriate tick management plans, particularly considering A. cajennense s.l.'s broad host range species and R. microplus's specificity for cattle. Moreover, the predictive models can identify areas at risk of associated challenging hemoparasite, requiring special attention and mitigation measures.

First report of multiple Rickettsia sp., Anaplasma sp., and Ehrlichia sp. in the San Miguel Department of El Salvador from zoonotic tick vectors.

Neglected bacterial zoonoses are a group of Neglected Tropical Diseases (NTDs) that are commonly underdiagnosed and underreported due to their undifferentiated febrile illness symptomology. Spotted fever group rickettsioses (SFGR), a subset of tick-borne bacterial zoonoses, belong in this group. There is a dichotomy in the reporting and recognition of these pathogens in Central America: countries with reduced human development scores-like El Salvador-have little to no research or surveillance dedicated to these pathogens and the diseases they cause. This was the third-ever tick survey in El Salvador, highlighting the knowledge gap in this country. A total of 253 ticks were collected from 11 animals at two farm sites and one veterinary office. Standard and quantitative PCR were used to detect presence of SFGR, Ehrlichia, and Anaplasma sp. pathogens in ticks. Ehrlichia sp. were detected in 2.4% of all collected ticks and Anaplasma sp. were detected in 5.5% of all ticks. Rickettsia rickettsii was amplified in 18.2% of ticks, and amplicons similar to R. parkeri, and R. felis were found in 0.8% and 0.4%, of collected ticks, respectively. This is the first report of these pathogenic bacterial species in El Salvador. This study emphasizes the need for further surveillance and research including incorporating additional human seroprevalence and testing to understand the public health burden in this country.

Identification of Omega-class glutathione transferases in helminths of the Taeniidae family.

Glutathione transferase enzymes (GSTs) are believed to be a major detoxification system in helminth parasites and have been associated with immunomodulation of the host response. Echinococcus granulosus sensu lato (s.l.) is a cestode parasite known to express at least five different GSTs, but no Omega-class enzymes have been reported in this parasite or in any other cestode. Herein we report the identification of a new member of the GST superfamily in E. granulosus s.l., which is phylogenetically related to the Omega-class: EgrGSTO. Through mass spectrometry, we showed that the 237 amino acids protein EgrGSTO is expressed by the parasite. Moreover, we identified homologues of EgrGSTO in other eight members of the Taeniidae family, including E. canadensis, E. multilocularis, E. oligarthrus, Hydatigera taeniaeformis, Taenia asiatica, T. multiceps, T. saginata and T. solium. A manual sequence inspection and rational modification yielded eight Taeniidae's GSTO sequences, each one encoding for a 237 aa polypeptide showing 80.2% overall identity. To the best of our knowledge, this is the first description of genes encoding for Omega-class GSTs in worms belonging to the Taeniidae family -that at least in E. granulosus s.l. is expressed as a protein- suggesting the gene encodes for a functional protein.

Molecular detection of Rickettsia felis in fleas and ticks collected from dogs and cats of Puebla, Mexico.

Fleas and ticks represent the two main groups of ectoparasites that infest companion animals. In particular, the flea Ctenocephalides felis felis and several members of the Rhipicephalus sanguineus complex are the main vectors of a wide range of pathogens on the American continent. They are competent vectors for several members of the genus Rickettsia, which encompass at least 15 pathogenic obligate intracellular bacteria that colonize the endothelial cells of vertebrates. In Mexico, 10 species of Rickettsia belonging to three groups have been detected in six species of ectoparasites from dogs in 9 of the 32 states of the country. However, in some larger regions of the country, active epidemiological surveillance has not been carried out. For this reason, the aim of this study was to identify the presence of members of the genus Rickettsia in fleas and ticks of dogs and cats in the state of Puebla, Mexico. A cross-sectional study was carried out to collect ectoparasites of dogs and cats during August to November 2019. Samples were fixed in 70% ethanol and examined to identify the presence of Rickettsia DNA by the amplification and sequencing of specific fragments of the gltA and ompB genes using conventional PCR. The recovered sequences were compared with those deposited in GenBank, and phylogenetic analyses were carried out to identify the position of the pathogens detected with respect to the valid species previously reported worldwide. Additionally, ecological parameters of the ectoparasite infestations were also calculated. We recovered 196 ectoparasites belonging to two species, 33 C. felis felis and 163 R. sanguineus s.l. (Rhipicephalus linnaei), parasitizing 46 hosts (42 dogs and 4 cats) in 11 localities of the state of Puebla. We detected the presence of Rickettsia felis in three pools of C. felis felis, and five from R. sanguineus s.l. Our work provides the first record of R. felis in hard ticks of Mexico and Central America, with new collection localities for this pathogen in central Mexico.

Genomic and epidemiologic surveillance of SARS-CoV-2 in Southern Brazil and identification of a new Omicron-L452R sublineage.

Recently, SARS-CoV-2 Omicron variant (B.1.1.529) was first identified in Botswana in November 2021. In a short period of time, this highly mutated variant replaced the previous dominant Delta variant, causing an exponential increase in the number of COVID-19 cases, resulting in a new wave of pandemic. This current research article aims to analyze and summarize information about the genetic characteristics, amino acid mutations and epidemiological data providing scientific findings to enrich the SARS-CoV-2 knowledge. More importantly, we describe here, for the first time, the identification of a new Omicron variant of concern: Omicron-L452R in Brazil.

Immunodiagnosis of cystic echinococcosis in livestock: Development and validation dataset of an ELISA test using a recombinant B8/2 subunit of Echinococcus granulosus sensu lato.

The accuracy of screening tests for detecting cystic echinococcosis (CE) in livestock depends on characteristics of the host-parasite interaction and the extent of serological cross-reactivity with other taeniid species. The AgB8 kDa protein is considered to be the most specific native or recombinant antigen for immunodiagnosis of ovine CE. A particular DNA fragment coding for rAgB8/2 was identified, that provides evidence of specific reaction in the serodiagnosis of metacestode infection. We developed and validated an IgG Enzyme Linked Immunosorbent Assay (ELISA) test using a recombinant antigen B sub-unit EgAgB8/2 (rAgB8/2) of Echinoccocus granulosus sensu lato (s.l.) to estimate CE prevalence in sheep. A 273 bp DNA fragment coding for rAgB8/2 was expressed as a fusion protein (∼30 kDa) and purified by affinity chromatography. Evaluation of the analytical and diagnostic performance of the ELISA followed the World Organisation for Animal Health (OIE) manual, including implementation of serum panels from: uninfected lambs (n = 79); experimentally infected (with 2,000 E. granulosus s.l. eggs each) sheep with subsequent evidence of E. granulosus cysts by necropsy (n = 36), and animals carrying other metacestode/trematode infections (n = 20). The latter were used to assess the cross-reactivity of rAgB8/2, with these animals being naturally infected with Taenia hydatigena, Thysanosoma actinioides and/or Fasciola hepatica. EgAgB8/2 showed cross-reaction with only one serum sample from a sheep infected with Ta. hydatigena out of the 20 animals tested. Furthermore, the kinetics of the humoral response over time in five 6-month old sheep, each experimentally infected with 2,000 E. granulosus s.l. eggs, was evaluated up to 49 weeks (approximately one year) post infection (n = 5). The earliest detectable IgG response against rAgB8/2 was observed in sera from two and four sheep, 7 and 14 days after experimental infection, respectively. The highest immune response across all five animals was found 16 to 24 weeks post infection.

Expanding the family of Mu-class glutathione transferases in the cestode parasite Echinococcus granulosus sensu lato.

Glutathione transferases (GSTs) perform catalytic and non-catalytic activities, mostly involved in stress-response and cell detoxification. Helminth parasites express several GSTs of multiple classes that are involved in the neutralization of potentially harmful oxidants, and in the inactivation or removal of xenobiotics. Additionally, GSTs participate in immunomodulatory processes that facilitate the parasite establishment and survival within its host. In Echinococcus granulosus sensu lato (s.l.) -the cestode parasite responsible for cystic echinococcosis- only one Mu-class GST has been reported. In the present work, by using bioinformatic and proteomic approaches we searched for novel Mu-class GSTs potentially involved in the parasite oxidative-stress metabolism. In the genome of E. granulosus s.l., 6 GST-related sequences were found to constitute a strongly conserved phylogenetical clade with Mu-class members. Among them, 5 displayed conserved gene structure (exon/intron), as well as specific residues and motifs characteristic of Mu-class enzymes. By proteomic analysis, 3 Mu-GSTs were identified to be expressed in the protoscolex parasite stage, 2 of them being firstly described as Mu-class GSTs here. The existence of more than one productive Mu-GST gene expands the parasite xenobiotic phase II metabolism, which might have beneficial roles on E. granulosus s.l. ability to successfully infect its host.

Larval habitats, species composition and distribution of malaria vectors in regions with autochthonous and imported malaria in Roraima state, Brazil.

BACKGROUND: Malaria control requires local action. Assessing the vector diversity and abundance provides information on the local malariogenic potential or risk of transmission. This study aimed to determine the Anopheles species composition, habitats, seasonal occurrence, and distribution in areas with autochthonous and imported malaria cases in Roraima State. METHODS: A longitudinal study was conducted from January 2017 to October 2018, sampling larvae and adult mosquitoes in three municipalities of Roraima State: Boa Vista, Pacaraima and São João da Baliza. These areas have different risks of malaria importation. Four to six mosquito larval habitats were selected for larval sampling at each municipality, along with two additional sites for adult mosquito collection. All larval habitats were surveyed every two months using a standardized larval sampling methodology and MosqTent for adult mosquitoes. RESULTS: A total of 544 Anopheles larvae and 1488 adult mosquitoes were collected from the three municipalities studied. Although the species abundance differed between municipalities, the larvae of Anopheles albitarsis s.l., Anopheles nuneztovari s.l. and Anopheles triannulatus s.l. were collected from all larval habitats studied while Anopheles darlingi were collected only from Boa Vista and São João da Baliza. Adults of 11 species of the genus Anopheles were collected, and the predominant species in Boa Vista was An. albitarsis (88.2%) followed by An. darlingi (6.9%), while in São João da Baliza, An. darlingi (85.6%) was the most predominant species followed by An. albitarsis s.l. (9.2%). In contrast, the most abundant species in Pacaraima was Anopheles braziliensis (62%), followed by Anopheles peryassui (18%). Overall, the majority of anophelines exhibited greater extradomicile than peridomicile-biting preference. Anopheles darlingi was the only species found indoors. Variability in biting times was observed among species and municipalities. CONCLUSION: This study revealed the composition of anopheline species and habitats in Boa Vista, Pacaraima and São João da Baliza. The species sampled differed in their behaviour with only An. darlingi being found indoors. Anopheles darlingi appeared to be the most important vector in São João da Baliza, an area of autochthonous malaria, and An. albitarsis s.l. and An. braziliensis in areas of low transmission, although there were increasing reports of imported malaria. Understanding the diversity of vector species and their ecology is essential for designing effective vector control strategies for these municipalities.

Virome analysis of three Ixodidae ticks species from Colombia: A potential strategy for discovering and surveying tick-borne viruses.

Ticks are a group of obligate blood-sucking ectoparasites that play a critical role in transmitting several important zoonotic pathogens that can infect animals and humans. Viruses are part of the tick microbiome and are involved in the transmission of important diseases. Furthermore, the little information on these as etiological agents of zoonoses suggests the need to study these microorganisms. For this reason, in this study, we sought to characterize the virome in Rhipicephalus microplus, Dermacentor nitens, and Rhipicephalus sanguineus s.l., which were collected from different domestic animals in Antioquia, Colombia. RNA sequencing was used for virome characterization in these three tick species, using RNA-dependent polymerase as a marker gene. Forty-eight sequences corresponding to 14 different viruses were identified, some of which were previously identified in the tick's virome. Overall, these data indicate that ticks from domestic animals in cattle farms harbor a wide viral diversity at the local scale. Thus, the metatranscriptomic approach provides important baseline information for monitoring the tick virome and to develop future studies on their biology, host-virus interactions, host range, worldwide distribution, and finally, their potential role as emerging vector-borne agents.

Expression and distribution of glutathione transferases in protoscoleces of Echinococcus granulosus sensu lato.

Glutathione transferases (GSTs) belong to a diverse superfamily of multifunctional proteins involved in metabolic detoxification. In helminth parasite, GSTs are particularly relevant since they are also involved in host immunomodulation. Echinococcus granulosus sensu lato (s.l.) is a cestode parasite known to express at least three phylogenetically distant cytosolic GSTs: EgGST1 and EgGST2 previously grouped within Mu and Sigma classes, respectively; and EgGST3 related to both Omega and Sigma classes. To better characterize E. granulosus s.l. GSTs, herein their expression and distribution were assessed in the pre-adult protoscolex (PSC) parasite stage. Potential transcriptional regulatory mechanisms of the corresponding EgGST genes were also explored. Firstly, the transcription of the three EgGSTs was significantly induced during the early stages of the murine model of infection, suggesting a potential role during parasite establishment. EgGST1 was detected in the parenchyma of PSCs and its expression increased after H2O2 exposure, supporting its role in detoxification. EgGST2 was mainly detected on the PSCs tegument, strategically localized for potential immunoregulation functions due to its Sigma-class characteristics. In addition, its expression increased after anthelmintic treatment, suggesting a role in chemotherapy resistance. Finally, the Omega-related EgGST3 was localized throughout the entire PSC body, including suckers and tegument, and since its expression also increased after H2O2 treatment, a potential role in oxidative stress response could also be ascribed. On the other hand, known cis-acting regulatory motifs were detected in EgGST genes, suggesting similar transcription processes to other eukaryotes. The results herein reported provide additional data regarding the roles of EgGSTs in E. granulosus s.l. biology, contributing to a better understanding of its host-parasite interaction.

Prevalence of human cystic echinococcosis in the towns of Ñorquinco and Ramos Mexia in Rio Negro Province, Argentina, and direct risk factors for infection.

BACKGROUND: Cystic echinococcosis (CE) is a parasitic zoonosis caused by infection with the larval stage of Echinococcus granulosus (s.l.). This study investigated the prevalence and potential risk factors associated with human CE in the towns and rural areas of Ñorquinco and Ramos Mexia, Rio Negro province, Argentina. METHODS: To detect abdominal CE cysts, we screened 892 volunteers by ultrasound and investigated potential risk factors for CE using a standardized questionnaire. Prevalence ratio (PR) with 95% confidence intervals (CI) was used to measure the association between CE and the factors investigated, applying bivariate and multivariate analyses. RESULTS: Abdominal CE was detected in 42/892 screened volunteers (4.7%, 95% CI 3.2-6.1), only two of whom were under 15 years of age. Thirteen (30.9%) CE cases had 25 cysts in active stages (CE1, CE2, CE3a, according to the WHO Informal Working Group on Echinococcosis [WHO-IWGE] classification). The most relevant risk factors identified in the bivariate analysis included: living in rural areas (P = 0.003), age > 40 years (P = 0.000), always drinking water from natural sources (P = 0.007), residing in rural areas during the first 5 years of life (P = 0.000) and having lived more than 20 years at the current address (P = 0.013). In the multivariate final model, the statistically significant risk factors were: frequently touching dogs (P = 0.012), residing in rural areas during the first 5 years of life (P = 0.004), smoking (P = 0.000), age > 60 years (P = 0.002) and living in rural areas (P = 0.017). CONCLUSIONS: Our results point toward infection with CE being acquired since childhood and with constant exposure throughout life, especially in rural areas with a general environmental contamination.

DNA barcoding of Lutzomyia longipalpis species complex (Diptera: Psychodidae), suggests the existence of 8 candidate species.

The sand fly Lutzomyia (L.) longipalpis has been implicated as the primary vector of Leishmania infantum, the causative agent of visceral leishmaniasis VL. In addition, it has been associated with atypical cutaneous leishmaniasis transmission in the Neotropic and Central America, respectively. The existence of a L. longipalpis complex species has been suggested with important implications for leishmaniasis epidemiology; however, the delimitation of species conforming it remains a topic of controversy. The DNA Barcoding Initiative based on cox1 sequence variation was used to identify the MOTUs in L. longipalpis including previously described L. pseudolongipalpis. The genetic variation was analyzed based on tree and distance methods. Fifty-five haplotypes were obtained from 103 sequences which were assigned to MOTUs, with a clear separation and a high correspondence of individuals to the groups. Maximum likelihood and Bayesian phylogenetic analysis showed eight MOTUs (100% bootstrap) with high genetic divergence (12.6%). Data obtained in the present study suggest that L. longipalpis complex consists of at least 8 lineages that may represent species. It would be desirable perform additional morphological and molecular analysis of L. longipalpis from Colosó (Caribbean ecoregion) considering that specimens from that area were grouped with L. pseudolongipalpis one of the complex species previously described from Venezuela, which has not been registered in Colombia.

Molecular screening for tick-borne bacteria and hematozoa in Ixodes cf. boliviensis and Ixodes tapirus (Ixodida: Ixodidae) from western highlands of Panama.

The first molecular screening for Rickettsia, Anaplasma, Ehrlichia, Borrelia, Babesia and Hepatozoon was carried out in questing Ixodes cf. boliviensis and Ixodes tapirus from Talamanca Mountains, Panama, using specific primers, sequencing and phylogeny. Phylogenetic analyses for the microorganisms in Ixodes cf. boliviensis confirmed the presence of Rickettsia sp. strain IbR/CRC endosymbiont (26/27 ticks), three genotypes of the Borrelia burgdorferi (sensu lato) complex (4/27 ticks), Babesia odocoilei (1/27 ticks), and Hepatozoon sp. (2/27 ticks), tentatively designated Hepatozoon sp. strain Chiriquensis. Phylogenetic analyses for the microorganisms in I. tapirus revealed an undescribed Rickettsia sp., tentatively designated Rickettsia sp. strain Itapirus LQ (6/6 ticks), and Anaplasma phagocytophilum (2/6 ticks). To the best of our knowledge, this is the first report of B. burgdorferi (s.l.) complex, A. phagocytophilum, B. odocoilei, and Hepatozoon sp. in Ixodes ticks from Central America, and also the first detection of Rickettsia spp. in Ixodes species in Panama. In light of the importance of these findings, further studies are needed focusing on the role of I. tapirus and I. cf. boliviensis as vectors, and the vertebrates acting as reservoirs.

Respuesta a atrayentes químicos y actividad horaria de Anopheles Meigen spp. (Diptera: Culicidae) en un área malárica del estado Bolívar, Venezuela / Response to chemical attractants and host seeking behavior of Anopheles Meigen spp. (Diptera: Culicidae) in a malaria endemic area of Bolívar State, Venezuela

Se evaluó la eficiencia de la trampa Mosquito Magnet Liberty PlusTM (MMLP) cebada con los atrayentes químicos 1-octen-3-ol (octenol) y Lurex 3TM (L-ácido láctico) junto con dióxido de carbono (CO2) para recolectar anofelinos en la localidad Ye'kwana de Boca de Nichare, municipio Sucre, estado Bolívar, Venezuela. La trampa fue operada entre las 18:00 y 06:00 horas, durante 10 noches por mes, durante cuatro meses (Julio ­ Octubre, 2015), alternándose cada noche el atrayente. A fin de caracterizar la actividad nocturna de los anofelinos, cada cuatro horas se cambiaba la jaulita donde eran atrapados los mosquitos. Se capturaron cinco especies de Anopheles y cuatro de Culicinae, siendo las más abundantes, Anopheles (Nyssorhynchus) darlingi Root y Anopheles (Nyssorhynchus) oswaldoi (Peryassú) sensu lato. El ANOVA factorial no paramétrico de Transformación de Rangos Alineados para la variable atrayente y sus interacciones con especie e intervalo horario no fueron estadísticamente significativas (p > 0.05). Estas especies muestran comportamientos diferentes: An. darlingi tiene el pico de actividad nocturna entre las 22:00 y 02:00 horas, mientras que en An. oswaldoi s.l. se observa un incremento de la actividad nocturna progresivamente que se mantiene durante el intervalo 02:00-06:00 horas. Los resultados permiten concluir que la trampa MMLP cebada con octenol o Lurex es un método alternativo de recolección de mosquitos para la vigilancia entomológica en áreas maláricas remotas con población indígena. La actividad horaria de los vectores An. darlingi y An. oswaldoi s.l. sugiere que el uso de mosquiteros tratados con insecticidas puede resultar un método efectivo para el control de la transmisión de malaria en esta zona del país(AU)

The efficiency of the Mosquito Magnet Liberty PlusTM (MMLP) trap was evaluated baited with the chemical attractants 1-octen-3-ol (octenol) and Lurex3M (L-lactic acid) together with carbon dioxide (CO2) to collect anophelines in the Amerindian village Boca de Nichare, Sucre municipality, Bolívar State, Venezuela. The trap was operated between 18:00 and 06: hours, 10 nights per month for four months (July-October 2015), the attractants were alternated nightly. To determine the host seeking behavior of anophelines, the trap cage was changed every 4 hours. Five species of Anopheles and four of Culicinae were collected; Anopheles (Nyssorhynchus) darlingi Root and Anopheles oswaldoi (Peryassú) sensu lato were the most abundant species collected. The Align Rank Transform (ART) test for nonparametric ANOVA for the variable attractant and its interactions with species and hour interval were not significant (p > 0.05). An. darlingi and An. oswaldoi s.l. showed contrasting host seeking activity: An. darlingi showed a peak between 22:00 and 02:00 hours, while in An. oswaldoi s.l. there was a steady increase in activity up to the interval 02:00-06:00 hours. The MMLP trap baited either with octenol or Lurex is an alternative method for entomological surveillance in remote Amerindian malaria endemic areas. The biting activity of the vectors An. darlingi and An. oswaldoi s.l. suggests that the use of long-lasting insecticide treated nets could be an effective method to control the transmission of malaria in this area(AU)

Contribution of Anthropogenic Factors and Climate Variables to Human Cystic Echinococcosis Mortality in Chile (2001-2011).

Here we assess the effect of weather and anthropogenic environmental variables, particularly urbanization, on cystic echinococcosis mortality in Chile from 2001 to 2011 using a nonparametric regression model, multivariate adaptive regression splines, and Poisson nonlinear regression models. This study integrated data from various sources on weather and anthropogenic variables. The canine population had the greatest influence on human cystic echinococcosis mortality during the period analyzed. Urbanization among anthropogenic variables and temperature and precipitation among the weather-related variables were the main factors related to cystic echinococcosis deaths. Deaths decreased with urbanization level. Temperature showed a nonlinear impact on mortality, with an optimum value ∼11°C. Public policies aimed at improving safe management of companion animal populations are crucial in controlling the spread of this disease. Effective animal management strategies would have wide-ranging public health benefits, advance the welfare of companion animals and livestock, and decrease the number of human cystic echinococcosis cases.

Absence of knockdown mutations in pyrethroid and DDT resistant populations of the main malaria vectors in Colombia.

BACKGROUND: Knockdown resistance (kdr) is a well-characterized target-site insecticide resistance mechanism that is associated with DDT and pyrethroid resistance. Even though insecticide resistance to pyrethroids and DDT have been reported in Anopheles albimanus, Anopheles benarrochi sensu lato (s.l.), Anopheles darlingi, Anopheles nuneztovari s.l., and Anopheles pseudopunctipennis s.l. malaria vectors in Latin America, there is a knowledge gap on the role that kdr resistance mechanisms play in this resistance. The aim of this study was to establish the role that kdr mechanisms play in pyrethroid and DDT resistance in the main malaria vectors in Colombia, in addition to previously reported metabolic resistance mechanisms, such as mixed function oxidases (MFO) and nonspecific esterases (NSE) enzyme families. METHODS: Surviving (n = 62) and dead (n = 67) An. nuneztovari s.l., An. darlingi and An. albimanus mosquitoes exposed to diagnostic concentrations of DDT and pyrethroid insecticides were used to amplify and sequence a ~ 225 bp fragment of the voltage-gated sodium channels (VGSC) gene. This fragment spanning codons 1010, 1013 and 1014 at the S6 segment of domain II to identify point mutations, which have been associated with insecticide resistance in different species of Anopheles malaria vectors. RESULTS: No kdr mutations were detected in the coding sequence of this fragment in 129 samples, 62 surviving mosquitoes and 67 dead mosquitoes, of An. darlingi, An. nuneztovari s.l. and An. albimanus. CONCLUSION: Mutations in the VGSC gene, most frequently reported in other species of the genus Anopheles resistant to pyrethroid and DDT, are not associated with the low-intensity resistance detected to these insecticides in some populations of the main malaria vectors in Colombia. These results suggest that metabolic resistance mechanisms previously reported in these populations might be responsible for the resistance observed.

First detection and genetic identification of Hepatozoon canis in Rhipicephalus sanguineus sensu lato ticks collected from dogs of Taiwan.

We determined the prevalence of infection and genetic identity of Hepatozoon spp. harbored by Rhipicephalus sanguineus sensu lato ticks in Taiwan. A total of 1082 ticks were collected from dogs and DNA extraction was performed from individual tick specimens. Hepatozoon infection was detected by performing a nested-PCR assay based on the 18S small subunit ribosomal RNA (ssrRNA) gene. The genetic identity of detected Hepatozoon was identified by gene sequencing and phylogenetic analysis. Hepatozoon infection was detected in nymphs, males and females of R. sanguineus s. l. ticks with an infection rate of 20.8%, 22.8% and 29.4%, respectively. Sequence and phylogenetic analysis revealed that these Hepatozoon spp. of Taiwan were genetically affiliated to the same clade within the genospecies of H. canis and can be discriminated from other genospecies of H. americanum and H. felis. Intraspecies analysis based on the genetic distance (GD) values indicates a lower level (GD < 0.005) genetic divergence within the same genospecies of H. canis detected in Taiwan, Brazil and Spain. Interspecies analysis also reveals a higher heterogeneity of Taiwan strains distinguished from other genospecies of H. felis (GD > 0.040) and H. americanum (GD > 0.056). This study provides the first molecular evidence of H. canis detected and identified in various stages of R. sanguineus s. l. ticks in Taiwan. Detection of H. canis in unfed male ticks may imply the possible mechanism of transstadial survival in R. sanguineus s. l. ticks. Further investigations on Hepatozoon spp. harbored by various vector ticks in Taiwan may illustrate the epidemiological significance of this parasite.

Detection of Rickettsia spp. in Rhipicephalus sanguineus (sensu lato) collected from free-roaming dogs in Coahuila state, northern Mexico.

BACKGROUND: The aim of this study was to detect and molecularly identify Rickettsia spp. in Rhipicephalus sanguineus (sensu lato) collected from free-roaming dogs in 30 communities from five municipalities in the south of Coahuila State, northern Mexico, where Rocky Mountain spotted fever is endemic. METHODS: In total, 60 dogs from each municipality were examined for engorged ticks. DNA was isolated from tick pools and conventional PCR assays targeting the 23S-5S ribosomal RNA intergenic spacer and outer membrane protein (ompA) gene of Rickettsia spp. were performed. RESULTS: All ticks (n = 1238) were morphologically identified as R. sanguineus (s.l.). Six pools (each with six engorged females) from four municipalities were positive to Rickettsia spp. DNA sequencing and phylogenetic analyses confirmed the presence of R. rickettsii and R. rhipicephali in R. sanguineus (s.l.) in these ticks. CONCLUSIONS: This study confirms the presence of R. rickettsii and R. rhipicephali in R. sanguineus (s.l.) from stray dogs in the south of Coahuila. This suggests that stray dogs may play a role in the inter-municipal dissemination of infected ticks in this region. Further research is required to assess whether ticks from stray dogs could serve as good indicators for the molecular xenomonitoring of R. rickettsii in this region. Considering that R. sanguineus (s.l.) is a proven vector of R. rickettsii in Mexico, increased awareness regarding permanent tick control in dogs is warranted.

Especies de garrapatas (Acari: Ixodidae) parásitas de perros (Canis familiaris) en zonas urbanas del Gran Mendoza, Argentina

El conocimiento de las especies de ectoparásitos presentes en los animales de compañía tiene implicancias tanto en medicina veterinaria como en salud humana, ya que varios agentes tienen la potencialidad de transmitirse del animal al humano y viceversa. Se revisaron 466 garrapatas remitidas al Centro de Investigación en Parasitología Regional (CIPaR) durante los años 2010 y 2013, recolectadas en 217 perros de zonas urbanas de la provincia de Mendoza. El 83,5% (n=385) fueron identificadas como Rhipicephalus sanguineus sensu lato (84 machos y 199 hembras - relación 1:2,37 -, 65 ninfas y 37 larvas), mientras que el 16,5% (n=76) fue identificado como Amblyomma tigrinum (10 machos y 61 hembras - relación 1:6,1 -, y cinco ejemplares adultos en mal estado, imposibilitando su sexado). Adicionalmente, cinco ejemplares adultos sólo se identificaron como pertenecientes al género Amblyomma, ya que por encontrarse ingurgitados no se pudo realizar la determinación de especie. Éste registro es un importante aporte al conocimiento de las especies de garrapatas presentes en perros (Canis familiaris) en la provincia de Mendoza.

Knowledge of the ectoparasitic agents of pets has implications both in veterinary medicine and public health since several agents have the potential to spread from animal to human and vice versa. Ticks sent to the Centro de Investigación en Parasitología Regional (CIPaR) during 2010 to 2013 were analysed. 466 adult ticks were collected from 217 dog in urban areas from Mendoza province, Argentina. Of these, 83.5% (n=385) specimens were identified as Rhipicephalus sanguineus s.l. (84 male and 199 female - ratio 1:2.37 -, 65 nymphs and 37 larvae), while 16.5% (n = 76) were identified as Amblyomma tigrinum (10 males and 61 females - ratio 1:6.1 -, and five adults in bad condition, which precluded their sexing). In addition, five adult specimens, identified as Amblyomma, were not identified to the species level since they were fully engorged. This report is an important contribution to the knowledge of the species of ticks in dogs (Canis familiaris) in the province of Mendoza.

Evaluation of the vector competence of six ixodid tick species for Rangelia vitalii (Apicomplexa, Piroplasmorida), the agent of canine rangeliosis.

Rangelia vitalii is the etiologic agent of canine rangeliosis, a severe piroplasmosis that affects domestic dogs in Brazil, Uruguay and Argentina. While R. vitalii is one of the most pathogenic tick-borne pathogens for dogs in the world, its tick vector has remained unknown. The present study evaluated the vector competence of Rhipicephalus sanguineus sensu lato (both tropical and temperate species), Amblyomma aureolatum, Amblyomma ovale, Amblyomma tigrinum, and Amblyomma sculptum for R. vitalii. These six tick species were selected for the study because they comprise the main tick species infesting dogs within the distribution area of canine rangeliosis in South America. Acquisition feeding of the above six tick species was performed on domestic dogs showing clinical signs of canine rangeliosis, after being experimentally infected through intravenous inoculation or infestation with R. vitalii-infected ticks. Thereafter, engorged ticks were evaluated for transstadial and transovarial passages of R. vitalii through molecular analysis after molting or oviposition and egg hatching. The resultant ticks were evaluated for their competence to transmit R. vitalii to susceptible dogs. Among the six tick species, only A. aureolatum was able to acquire and perpetuate R. vitalii by transstadial and transovarial passages, as demonstrated by >5% infection rates of ticks after hatching or molting. When exposed to transmission feeding, only A. aureolatum ticks were competent to transmit R. vitalii to dogs, which became severely ill, and the results confirmed by molecular methods and blood smear examination to have acquired rangeliosis. Results of the present study, coupled with epidemiological data, indicate that A. aureolatum is a natural vector of R. vitalii. Our results also indicate that R. vitalii is the first Piroplasmorida agent to be transovarially transmitted in Amblyomma ticks.