Novel trypanosomatid species detected in Mongolian pikas (Ochotona pallasi) and their fleas in northwestern China.

BACKGROUND: In the family Trypanosomatidae, the genus Trypanosoma contains protozoan parasites that infect a diverse range of hosts, including humans, domestic animals, and wildlife. Wild rodents, as natural reservoir hosts of various pathogens, play an important role in the evolution and emergence of Trypanosomatidae. To date, no reports are available on the trypanosomatid infection of pikas (Lagomorpha: Ochotonidae). METHODS: In this study, Mongolian pikas and their fleas were sampled at the China-Mongolia border, northwestern China. The samples were analyzed with polymerase chain reaction (PCR) and sequencing for the presence of Trypanosomatidae on the basis of both the 18S ribosomal RNA (18S rRNA) gene and the glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) gene. The morphology of trypomastigotes was also observed in peripheral blood smears by microscopy. RESULTS: Molecular and phylogenetic analyses revealed a new genotype of the Trypanosoma lewisi clade that was found both in pika blood and flea samples. This genotype, which probably represents a new species, was provisionally designated as "Trypanosoma sp. pika". In addition, a novel genotype belonging to the genus Blechomonas of Trypanosomatidae was detected in fleas. On the basis of its molecular and phylogenetic properties, this genotype was named Blechomonas luni-like, because it was shown to be the closest related to B. luni compared with other flea-associated trypanosomatids. CONCLUSIONS: To the best of our knowledge, this is the first study to report any trypanosomatid species in Mongolian pikas and their fleas. Further studies are needed to investigate the epidemiology of these protozoan parasites, as well as to evaluate their pathogenicity for humans or domestic animals.

Dipylidium caninum in the twenty-first century: epidemiological studies and reported cases in companion animals and humans.

BACKGROUND: Dipilidiosis is a parasitic disease caused by the tapeworm Dipylidium caninum. Fleas and, less frequently, lice act as an intermediate host, and their ingestion is required for infection to occur. While the disease mainly affects domestic and wild carnivores, it is also considered a zoonotic disease, with most human cases reported in children. Dipylidium caninum is considered to be the most common tapeworm infesting companion animals, but dipilidosis in humans is rare. The aims of this review were to improve current understanding of the epidemiology of this parasitosis and its management by the medical and veterinary community. METHODS: A comprehensive review of the published literature during the last 21 years (2000-2021) on the epidemiology, clinical features, diagnosis, treatment and prevention measures of D. caninum infection and dipilidiosis in companion animals and humans was conducted. RESULTS: Using predefined eligibility criteria for a search of the published literature, we retrieved and screened 280 publications. Of these, 161 (141 epidemiological studies, 20 case reports [16 human cases]) were considered for inclusion in this review. This parasitosis is present worldwide; however, despite being the most frequent cestode infection in animals, it is often underdiagnosed using common coprological techniques. Its diagnosis in humans has also proved challenging, being frequently confused with pinworm infection, leading to inappropriate treatment and to the persistence of the disease over time. Prevention measures include control of ectoparasites in animals and the environment, as well as regular deworming of animals, most commonly with praziquantel. CONCLUSIONS: The diagnosis of dipilidiosis remains challenging in both animals and humans, primarily due to the low sensitivity of the diagnostic methods currently available and a lack of knowledge of the morphological characteristics of the parasite. Although treatment with the appropriate anti-cestode compounds is well tolerated and results in resolution of the infection, indiscriminate use of these compounds may predispose to an increase in resistance. Given the worldwide distribution of this parasite, it is essential to act on several fronts, with a focus on health education for children and animal owners and the control of intermediate hosts, both in animals and in the surrounding environment.

Detection of Trypanosoma lewisi DNA from Rattus norvegicus and Rattus rattus in Hanoi, Vietnam.

Trypanosoma lewisi is a worldwide nonpathogenic parasite that is exclusively found in rats. In general, T. lewisi infection in humans is an opportunistic infection from rats to humans through fleas. However, recently, infection with T. lewisi in humans, including a fatal case, has been reported. Notably, rats living close to a human settlement showed a higher prevalence of infection with T. lewisi than those living in other places. It is possible that the urbanization is associated with the prevalence of T. lewisi in rats and enhances the risk of T. lewisi transmission to humans through fleas. In this study, a total of 88 rats were captured from hospitals, markets, and a cargo station, of which 81 were identified as Rattus norvegicus and 7 as Rattus rattus in Hanoi, the urbanizing city of Vietnam. Of these, 55 rats (62.5%) harbored T. lewisi, of which 52 were R. norvegicus and 3 were R. rattus.

A community approach of pathogens and their arthropod vectors (ticks and fleas) in dogs of African Sub-Sahara.

BACKGROUND: Arthropod-borne pathogens and their vectors are present throughout Africa. They have been well-studied in livestock of sub-Saharan Africa, but poorly in companion animals. Given the socio-economic importance of companion animals, the African Small Companion Animal Network (AFSCAN), as part of the WSAVA Foundation, initiated a standardized multi-country surveillance study. METHODS: Macro-geographic variation in ectoparasite (ticks and fleas) and pathogen communities in dogs was assessed through molecular screening of approximately 100 infested dogs in each of six countries (Ghana, Kenya, Nigeria, Tanzania, Uganda and Namibia), both in rural and urban settings. The most important intrinsic and extrinsic risk factors within the subpopulation of infested dogs were evaluated. RESULTS: Despite the large macro-geographic variation in the dogs screened, there was no consistent difference between East and West Africa in terms of the diversity and numbers of ticks. The highest and lowest numbers of ticks were found in Nigeria and Namibia, respectively. Most often, there was a higher diversity of ticks in rural habitats than in urban habitats, although the highest diversity was observed in an urban Uganda setting. With the exception of Namibia, more fleas were collected in rural areas. We identified tick species (including Haemaphysalis spinulosa) as well as zoonotic pathogens (Coxiella burnetti, Trypanosoma spp.) that are not classically associated with companion animals. Rhipicephalus sanguineus was the most abundant tick, with a preference for urban areas. Exophilic ticks, such as Haemaphysalis spp., were more often found in rural areas. Several multi-host ticks occurred in urban areas. For R. sanguineus, housing conditions and additional pets were relevant factors in terms of infestation, while for a rural tick species (Haemaphysalis elliptica), free-roaming dogs were more often infested. Tick occurrence was associated to the use of endoparasiticide, but not to the use of ectoparasiticide. The most prevalent tick-borne pathogen was Hepatozoon canis followed by Ehrlichia canis. High levels of co-parasitism were observed in all countries and habitats. CONCLUSIONS: As dogs share a common environment with people, they have the potential to extend the network of pathogen transmission to humans. Our study will help epidemiologists to provide recommendations for surveillance and prevention of pathogens in dogs and humans.

Patterns of flea infestation in rodents and insectivores from intensified agro-ecosystems, Northwest Spain.

BACKGROUND: Fleas frequently infest small mammals and play important vectoring roles in the epidemiology of (re)emerging zoonotic disease. Rodent outbreaks in intensified agro-ecosystems of North-West Spain have been recently linked to periodic zoonotic diseases spillover to local human populations. Obtaining qualitative and quantitative information about the composition and structure of the whole flea and small mammal host coexisting communities is paramount to understand disease transmission cycles and to elucidate the disease-vectoring role of flea species. The aims of this research were to: (i) characterise and quantify the flea community parasiting a small mammal guild in intensive farmlands in North-West Spain; (ii) determine and evaluate patterns of co-infection and the variables that may influence parasitological parameters. METHODS: We conducted a large-scale survey stratified by season and habitat of fleas parasitizing the small mammal host guild. We report on the prevalence, mean intensity, and mean abundance of flea species parasitizing Microtus arvalis, Apodemus sylvaticus, Mus spretus and Crocidura russula. We also report on aggregation patterns (variance-to-mean ratio and discrepancy index) and co-infection of hosts by different flea species (Fager index) and used generalized linear mixed models to study flea parameter variation according to season, habitat and host sex. RESULTS: Three flea species dominated the system: Ctenophthalmus apertus gilcolladoi, Leptopsylla taschenbergi and Nosopsyllus fasciatus. Results showed a high aggregation pattern of fleas in all hosts. All host species in the guild shared C. a. gilcolladoi and N. fasciatus, but L. taschenbergi mainly parasitized mice (M. spretus and A. sylvaticus). We found significant male-biased infestation patterns in mice, seasonal variations in flea abundances for all rodent hosts (M. arvalis, M. spretus and A. sylvaticus), and relatively lower infestation values for voles inhabiting alfalfas. Simultaneous co-infections occurred in a third of all hosts, and N. fasciatus was the most common flea co-infecting small mammal hosts. CONCLUSIONS: The generalist N. fasciatus and C. a. gilcolladoi dominated the flea community, and a high percentage of co-infections with both species occurred within the small mammal guild. Nosopsyllus fasciatus may show higher competence of inter-specific transmission, and future research should unravel its role in the circulation of rodent-borne zoonoses.

Molecular detection of pathogens in ticks and fleas collected from companion dogs and cats in East and Southeast Asia.

BACKGROUND: Ticks and fleas are considered amongst the most important arthropod vectors of medical and veterinary concern due to their ability to transmit pathogens to a range of animal species including dogs, cats and humans. By sharing a common environment with humans, companion animal-associated parasitic arthropods may potentially transmit zoonotic vector-borne pathogens (VBPs). This study aimed to molecularly detect pathogens from ticks and fleas from companion dogs and cats in East and Southeast Asia. METHODS: A total of 392 ticks and 248 fleas were collected from 401 infested animals (i.e. 271 dogs and 130 cats) from China, Taiwan, Indonesia, Malaysia, Singapore, Thailand, the Philippines and Vietnam, and molecularly screened for the presence of pathogens. Ticks were tested for Rickettsia spp., Anaplasma spp., Ehrlichia spp., Babesia spp. and Hepatozoon spp. while fleas were screened for the presence of Rickettsia spp. and Bartonella spp. RESULT: Of the 392 ticks tested, 37 (9.4%) scored positive for at least one pathogen with Hepatozoon canis being the most prevalent (5.4%), followed by Ehrlichia canis (1.8%), Babesia vogeli (1%), Anaplasma platys (0.8%) and Rickettsia spp. (1%) [including Rickettsia sp. (0.5%), Rickettsia asembonensis (0.3%) and Rickettsia felis (0.3%)]. Out of 248 fleas tested, 106 (42.7%) were harboring at least one pathogen with R. felis being the most common (19.4%), followed by Bartonella spp. (16.5%), Rickettsia asembonensis (10.9%) and "Candidatus Rickettsia senegalensis" (0.4%). Furthermore, 35 Rhipicephalus sanguineus ticks were subjected to phylogenetic analysis, of which 34 ticks belonged to the tropical and only one belonged to the temperate lineage (Rh. sanguineus (sensu stricto)). CONCLUSION: Our data reveals the circulation of different VBPs in ticks and fleas of dogs and cats from Asia, including zoonotic agents, which may represent a potential risk to animal and human health.

Host distribution and pathogen infection of fleas (Siphonaptera) recovered from small mammals in Pennsylvania.

The number of recognized flea-borne pathogens has increased over the past decade. However, the true number of infections related to all flea-borne pathogens remains unknown. To better understand the enzootic cycle of flea-borne pathogens, fleas were sampled from small mammals trapped in central Pennsylvania. A total of 541 small mammals were trapped, with white-footed mice (Peromyscus leucopus) and southern red-backed voles (Myodes gapperi) accounting for over 94% of the captures. Only P. leucopus were positive for examined blood-borne pathogens, with 47 (18.1%) and ten (4.8%) positive for Anaplasma phagocytophilum and Babesia microti, respectively. In addition, 61 fleas were collected from small mammals and tested for pathogens. Orchopeas leucopus was the most common flea and Bartonella vinsonii subspecies arupensis, B. microti, and a Rickettsia felis-like bacterium were detected in various flea samples. To the best of our knowledge, this is the first report of B. microti DNA detected from a flea and the first report of a R. felis-like bacterium from rodent fleas in eastern North America. This study provides evidence of emerging pathogens found in fleas, but further investigation is required to resolve the ecology of flea-borne disease transmission cycles.

Zoonotic Trypanosomes in Rats and Fleas of Venezuelan Slums.

Rattus spp. are reservoirs of many human zoonoses, but their role in domestic transmission cycles of human trypanosomiasis is underestimated. In this study, we report trypanosome-infected Rattus norvegicus and Rattus rattus in human dwellings in slums neighboring Maracay, a large city near Caracas, the capital of Venezuela. Blood samples of R. norvegicus and R. rattus examined by PCR and FFLB (fluorescent fragment length barcoding) revealed a prevalence of 6.3% / 31.1% for Trypanosoma lewisi (agent of rat- and flea-borne human emergent zoonosis), and 10.5% / 24.6% for Trypanosoma cruzi (agent of Chagas disease). Detection in flea guts of T. lewisi (76%) and, unexpectedly, T. cruzi (21.3%) highlighted the role of fleas as carriers and vectors of these trypanosomes. A high prevalence of rats infected with T. lewisi and T. cruzi and respective flea and triatomine vectors poses a serious risk of human trypanosomiasis in Venezuelan slums. Anthropogenic activities responsible for growing rat and triatomine populations within human dwellings drastically increased human exposure to trypanosomes. This scenario has allowed for the reemergence of Chagas disease as an urban zoonosis in Venezuela and can propitiate the emergence of atypical T. lewisi infection in humans.

Effectiveness of deltamethrin-impregnated dog collars on the incidence of canine infection by Leishmania infantum: A large scale intervention study in an endemic area in Brazil.

To reduce morbidity and mortality caused by visceral leishmaniasis (VL), the Brazilian Visceral Leishmaniasis Control and Surveillance Program promotes the diagnosis and treatment of cases, vector control, euthanasia of seropositive dogs, and health education. Nevertheless, the effectiveness of these measures is questionable as they lead to little reduction in the transmission of the disease. Thus, the effectiveness of strategies such as insecticide-impregnated collars, spot-on insecticides, and immunization of dogs should be assessed. Herein, we evaluated the effectiveness of deltamethrin-impregnated collars on reducing the incidence of Leishmania infantum infection in dogs living in an endemic area of VL. An intervention study was conducted and a total 5,850 dogs were analyzed in baseline. Of these 3,742 seronegative dogs were divided into two groups: collared and uncollared (control). Dogs were followed for 12 months and three interventions were performed. The Cox regression model was used to evaluate the effectiveness of the collar. All analyzes were performed by Intention-to-treat and per-protocol. By intention-to-treat, the incidence rates of L. infantum infection were 7.5 and 7.9 in the collar group, and 6.5 and 13.2 per 1,000 dogs-months in the control group after 6 and 12 months, respectively. In the per-protocol analysis, the incidence rates in the control group were similar to those observed in the intention-to-treat analysis. In the collar group, the incidence rate was 5.1/1,000 dogs-months after 6 and 12 months. The effectiveness by intention-to-treat after adjustment by the multivariate Cox model was 48%. In the analysis per-protocol, the effectiveness increased to 63%. Although collar use was effective when it was evaluated by intention-to-treat, higher effectiveness was found in the per-protocol analysis after one year of follow-up. The data emphasize the importance of the uninterrupted use of deltamethrin-impregnated collars to increase protection against canine VL.

A randomized, blinded, controlled, multi-centered field study assessing the treatment of gastrointestinal nematode infections in cats with fluralaner plus moxidectin spot-on solution (Bravecto® Plus).

BACKGROUND: A spot-on formulation containing fluralaner (280 mg/ml) plus moxidectin (14 mg/ml) (Bravecto® Plus) was developed for the treatment of nematode infections as well as providing 12 weeks of protection against insect and acarine parasites in cats. The effectiveness and safety of this product against feline gastrointestinal nematodes was assessed in naturally-infested, client-owned cats under field conditions in Albania, Bulgaria, Germany and Hungary. METHODS: To be eligible for enrollment in this investigator-blinded study cats had to be at least 10 weeks-old, weigh at least 1.2 kg, be clinically healthy, and have a faecal sample testing positive for nematodes no more than eight days prior to treatment. Cats were stratified into blocks of three in order of presentation at each center and randomly allocated in a 2:1 ratio to be treated topically on Day 0 with fluralaner plus moxidectin (minimum dose rates 40 mg/kg and 2 mg/kg, respectively) or emodepside plus praziquantel (minimum dose rates 3 mg/kg and 12 mg/kg, respectively) (Profender®). Faecal samples were collected from cats prior to treatment and 14 ± 4 days later. RESULTS: There were 182 cats randomized to the fluralaner plus moxidectin group, and 91 to the emodepside plus praziquantel group. Prior to treatment the most commonly identified nematode egg was Toxocara cati, found in 79.1 and 82.4% of cats in the fluralaner plus moxidectin and emodepside plus praziquantel groups, respectively. Eggs of Toxascaris leonina were found in 8.2 and 6.6% of cats; of hookworms in 30.8 and 24.2%; and of Capillaria spp. in 7.1 and 4.3%, respectively. After treatment, faecal samples from 98.3% of fluralaner plus moxidectin treated and 96.6% of emodepside plus praziquantel-treated cats were free of nematode ova. Geometric mean faecal egg count reductions for T. cati, the only eggs found in post-treatment faecal samples, were 99.97% and 99.93%, respectively. Treatment with fluralaner plus moxidectin was non-inferior to emodepside plus praziquantel. Both products were safe and well tolerated by cats treated under field conditions. CONCLUSIONS: This field study confirms that, in addition to 12-week extended duration flea and tick control, fluralaner plus moxidectin provides broad spectrum treatment of nematodes in cats.

RNA Viruses in Blechomonas (Trypanosomatidae) and Evolution of Leishmaniavirus.

In this work, we analyzed viral prevalence in trypanosomatid parasites (Blechomonas spp.) infecting Siphonaptera and discovered nine species of viruses from three different groups (leishbunyaviruses, narnaviruses, and leishmaniaviruses). Most of the flagellate isolates bore two or three viral types (mixed infections). Although no new viral groups were documented in Blechomonas spp., our findings are important for the comprehension of viral evolution. The discovery of bunyaviruses in blechomonads was anticipated, since these viruses have envelopes facilitating their interspecific transmission and have already been found in various trypanosomatids and metatranscriptomes with trypanosomatid signatures. In this work, we also provided evidence that even representatives of the family Narnaviridae are capable of host switching and evidently have accomplished switches multiple times in the course of their evolution. The most unexpected finding was the presence of leishmaniaviruses, a group previously solely confined to the human pathogens Leishmania spp. From phylogenetic inferences and analyses of the life cycles of Leishmania and Blechomonas, we concluded that a common ancestor of leishmaniaviruses most likely infected Leishmania first and was acquired by Blechomonas by horizontal transfer. Our findings demonstrate that evolution of leishmaniaviruses is more complex than previously thought and includes occasional host switching.IMPORTANCE Flagellates belonging to the genus Leishmania are important human parasites. Some strains of different Leishmania species harbor viruses (leishmaniaviruses), which facilitate metastatic spread of the parasites, thus aggravating the disease. Up until now, these viruses were known to be hosted only by Leishmania Here, we analyzed viral distribution in Blechomonas, a related group of flagellates parasitizing fleas, and revealed that they also bear leishmaniaviruses. Our findings shed light on the entangled evolution of these viruses. In addition, we documented that Blechomonas can be also infected by leishbunyaviruses and narnaviruses, viral groups known from other insects' flagellates.

Fleas and Ticks of Red Foxes as Vectors of Canine Bacterial and Parasitic Pathogens, in Slovakia, Central Europe.

The present survey aimed to investigate flea and tick fauna parasitizing Slovak red fox populations with special emphasis on canine pathogens they transmit. A total of 407 fleas and 105 ticks were collected from 90 red foxes from two geographically distant regions. Seven flea species (Chaetopsylla globiceps, Pulex irritans, Archaeopsylla erinacei, Chaetopsylla rothschildi, Chaetopsylla trichosa, Ctenocephalides canis, and Ctenopthalmus assimilis) and three species of hard ticks (Ixodes ricinus, Ixodes hexagonus, Haemaphysalis concinna) were recorded on sampled animals. Consequently, the DNA of five different pathogen taxa was confirmed in collected arthropod vectors: Bartonella spp. (in P. irritans, Ch. globiceps, and Ct. assimilis), Rickettsia spp. (in A. erinacei, I. ricinus, I. hexagonus, and H. concinna), Anaplasma phagocytophilum (in I. ricinus), Theileria sp. (in Ch. globiceps and H. concinna), and Hepatozoon canis (in I. ricinus and I. hexagonus). Mycoplasma spp., Dipylidium caninum, and Acanthocheilonema reconditum were not found in fleas or ticks in this study.

Analysis of Dipylidium caninum tapeworms from dogs and cats, or their respective fleas - Part 1. Molecular characterization of Dipylidium caninum: genetic analysis supporting two distinct species adapted to dogs and cats.

A 28S rDNA PCR detection assay was previously developed to identify Dipylidium caninum DNA inside single fleas collected from both cats and dogs. Sequence analysis of the 28S rDNA fragment indicated two genetically distinct variations of the target region. The two genotypes, so-called "D. caninum canine genotype" and "D. caninum feline genotype", based on host origin, are further investigated and described in this paper. Restriction fragment length polymorphism (RFLP) analysis and hydrolysis probe-based genotyping assays were developed and validated for genotyping D. caninum DNA. The complete mitochondrial (mt) genome of the "feline genotype" was sequenced and compared to the D. caninum mt genome available in GenBank. The molecular characterization of D. caninum isolates collected from infected fleas, and also proglottids collected from dogs and cats, confirmed the existence of two distinct genotypes. These genotypes are related to host origin (dogs or cats), irrespective of their geographical origin, and they present a biological adaptation to their respective host, as confirmed by the comparison of biological development and host preference in another study. The genetic differences (Part 1, present paper) and biological observations (Part 2, in this journal) enabled us to suggest the existence of two distinct species within D. caninum, which will have to be clarified.

Analysis of Dipylidium caninum tapeworms from dogs and cats, or their respective fleas - Part 2. Distinct canine and feline host association with two different Dipylidium caninum genotypes.

Initial investigations suggested the existence of two distinct genotypes of Dipylidium caninum from infected cat fleas (Ctenocephalides felis). One genotype was found almost always (> 95%) in fleas collected from, and proglottids shed by, domestic dogs. The other was found almost always (> 95%) in fleas collected from, and proglottids shed by, domestic cats. Molecular investigations (Part 1, in this journal) confirmed the presence of two distinct genotypes. Due to the apparent host association observed, these were referred to as the "D. caninum canine genotype" and the "D. caninum feline genotype". The current article reports on an in vivo experimental infection study assessing the host-parasite interaction for each genotype. Mixed infections with the two genotypes in both dogs and cats were conducted. The specific genotyping of proglottids allowed us to assess the specific prepatent periods, prolificity, and longevity of each genotype in dogs versus cats. The possible hybridisation was also studied through molecular evaluation of the proglottids expelled by infected dogs and cats. Results demonstrate a clear distinct host interaction. The canine D. caninum genotype occurred at a higher frequency in dogs, with a shorter prepatent period and a longer lifespan; and the feline genotype occurred at a higher frequency in cats, with a shorter prepatent period and a longer lifespan. The absence of any hybrids in the mixed infections of both dogs and cats confirm the hypothesis of two distinct genotypes, suggesting the possibility of two distinct species within Dipylidium caninum.

Animal and human tungiasis-related knowledge and treatment practices among animal keeping households in Bugiri District, South-Eastern Uganda.

BACKGROUND: Zoonotic tungiasis caused by Tunga penetrans remains a serious public and animal health problem among endemic villages in Uganda and many sub Saharan African countries. Studies on human and animal tungiasis-related knowledge and treatment practices in endemic communities have never been undertaken, a limitation to development of sustainable control measures. METHODS: A cross sectional study using semi-structured questionnaires (Supplementary file S1) was conducted among 236 animal rearing households in 10 endemic villages in Bugiri District, South-Eastern Uganda. Focus group discussions and observation checklists were used to validate and clarify the findings. RESULTS: Most respondents knew the aetiology (89.4%), clinical signs (98%) and the ecology of T. penetrans as well as the major risk factors of human tungiasis (65.2%). In contrast, very few respondents were aware of animal tungiasis. Only 4.8% of those with infected animals on the compound knew that some of their animals were infected and 13.6% of the respondents had ever seen tungiasis-affected animals. Pigs (13.1%, n=31) and dogs (0.85%, n=2) were the only T. penetrans animal hosts known to animal owners. Affected humans were treated by extraction of embedded sand fleas using non-sterile sharp instruments in all households that reported occurrence of human tungiasis at least once (n=227). Also, affected animals were mainly treated by mechanical removal of embedded sand fleas in households that have ever experienced animal tungiasis (four out of 12; 33.3%). In a few instances, plant and animal pesticides (n=3) and other chemicals such as grease, paraffin and wood preservative (n=3) were also used to treat animal tungiasis. CONCLUSION: The study revealed a high level of knowledge on human tungiasis but inadequate knowledge on the zoonotic nature of tungiasis. Commonly applied methods for treatment of human and animal tungiasis are a health hazard by themselves. Concerted i.e. One Health-based efforts aiming at promoting appropriate treatment of tungiasis, adequate living conditions and increased awareness on tungiasis in the communities are indicated in order to eliminate tungiasis-associated disease.

Diversity of piroplasmids among wild and domestic mammals and ectoparasites in Pantanal wetland, Brazil.

Piroplasmoses are one of the most prevalent arthropod-borne diseases of animals. The present work aimed to investigate the occurrence of piroplasmid in wild mammals, domestic dogs and ectoparasites in southern Pantanal region, central-western Brazil. For that purpose, blood or tissue samples from 31 Nasua nasua, 78 Cerdocyon thous, 7 Leopardus pardalis, 42 dogs, 110 wild rodents, and 30 marsupials, and 1582 ticks were submitted to PCR assays for piroplasmid targeting 18SrRNA and hps70 genes. Seven dogs, one C. thous, five L. pardalis, three N. nasua, six wild rodents, eight Amblyomma parvum, two Amblyomma sculptum and one Amblyomma ovale were positive for piroplasmid-PCR assays. Genotypes closely related to Babesia vogeli were detected in six dogs and five wild rodents. While genotypes closely related to Babesia caballi were detected in one C. thous, one dog, one A. ovale and one A. sculptum, genotypes closely related to Babesia bigemina and Babesia bovis were detected in four A. parvum ticks. Four sequences obtained from A. parvum, three coatis and one wild rodent were closely related to Theileria equi. Cytauxzoon spp. was detected in four ocelots. The present study revealed that wild and domestic animals in Brazilian southern Pantanal are exposed to different piroplasmid species.

Diagnosis and genetic analysis of the worldwide distributed Rattus-borne Trypanosoma (Herpetosoma) lewisi and its allied species in blood and fleas of rodents.

Trypanosoma (Herpetosoma) lewisi is a cosmopolitan parasite of rodents strongly linked to the human dispersal of Rattus spp. from Asia to the rest of the world. This species is highly phylogenetically related to trypanosomes from other rodents (T. lewisi-like), and sporadically infects other mammals. T. lewisi may opportunistically infect humans, and has been considered an emergent rat-borne zoonosis associated to poverty. We developed the THeCATL-PCR based on Cathepsin L (CATL) sequences to specifically detect T. (Herpetosoma) spp., and assess their genetic diversity. This method exhibited high sensitivity using blood samples, and is the first molecular method employed to search for T. lewisi in its flea vectors. THeCATL-PCR surveys using simple DNA preparation from blood preserved in ethanol or filter paper detected T. lewisi in Rattus spp. from human dwellings in South America (Brazil and Venezuela), East Africa (Mozambique), and Southeast Asia (Thailand, Cambodia and Lao PDR). In addition, native rodents captured in anthropogenic and nearby human settlements in natural habitats harbored T. (Herpetosoma) spp. PCR-amplified CATL gene fragments (253bp) distinguish T. lewisi and T. lewisi-like from other trypanosomes, and allow for assessment of genetic diversity and relationships among T. (Herpetosoma) spp. Our molecular surveys corroborated worldwide high prevalence of T. lewisi, incriminating Mastomys natalensis as an important carrier of this species in Africa, and supported its spillover from invader Rattus spp. to native rodents in Brazil and Mozambique. THeCATL-PCR provided new insights on the accurate diagnosis and genetic repertoire of T. (Herpetosoma) spp. in rodent and non-rodent hosts, revealing a novel species of this subgenus in an African gerbil. Phylogenetic analysis based on CATL sequences from T. (Herpetosoma) spp. and other trypanosomes (amplified using pan-trypanosome primers) uncovered rodents harboring, beyond mammal trypanosomes of different subgenera, some species that clustered in the lizard-snake clade of trypanosomes.

Exposure to vector-borne pathogens in privately owned dogs living in different socioeconomic settings in Brazil.

This survey was conducted in four Brazilian cities, from three federative units (Pernambuco, Minas Gerais, and Federal District), representing different socioeconomic settings, to determine the presence of antibodies to or antigens and DNA of selected pathogens in privately owned dogs. From January to April 2015, blood and serum samples were collected and assayed using different tests. In particular, antibodies to Anaplasma spp., Borrelia burgdorferi sensu lato, and Ehrlichia spp., and antigens of Dirofilaria immitis were detected using a rapid enzyme-linked immunosorbent assay, whereas antibodies to Babesia spp. were detected by an immunofluorescence antibody assay. Moreover, the presence of Leishmania DNA in blood samples was assessed by real-time polymerase chain reaction. Overall, 208 (69.3%) out of 300 dogs were positive for at least one tested pathogen (intended here as antibodies, antigen or DNA, as abovementioned), with 139 (66.8%) of them being positive to two or more pathogens. No dog presented antibodies to B. burgdorferi s.l., and D. immitis antigens were detected exclusively in dogs from Pernambuco. Among positive dogs, the most common clinical signs were lymphadenomegaly (45.2%), onychogryphosis (41.3%), dermatitis (34.1%), pale mucous membranes (19.7%), weight loss (14.9%), fever (12.5%), alopecia (11.1%), and lethargy (4.8%). Tick and flea infestations were recorded in 41.7% and 29.3% of the dogs, respectively, with 49 (16.3%) dogs being co-infested by both ticks and fleas. Most of the tick- and flea-infested dogs presented high level of infestation (>10 ticks and >20 fleas). The level of tick infestation varied significantly among federative units, being highest in Minas Gerais (68.0%), followed by Pernambuco (36.0%) and Federal District (21.0%). On the other hand, the level of flea infestation was higher in Pernambuco (50.0%), followed by Minas Gerais (29.0%) and Federal District (9.0%). The number of dog owners reporting the use of ectoparasiticides (on dogs and/or in the environment) varied significantly, ranging from 6.0% in Pernambuco to 60.0% in Federal District. This study reveals disparate results in terms of dog exposure to fleas and ticks in the studied communities, which may be related to factors such as the owners' capability/willingness to afford the use of ectoparasiticides on their dogs and the dog's level of restriction. Further research is needed to establish the relationship between dog owners' socioeconomic situation and the level of exposure to ectoparasites and their transmitted pathogens.

Molecular detection of Hepatozoon spp. in domestic dogs and wild mammals in southern Pantanal, Brazil with implications in the transmission route.

Hepatozoon parasites comprise intracellular apicomplexan parasites transmitted to vertebrate animals by ingestion of arthropods definitive hosts. The present work aimed to investigate the occurrence of Hepatozoon spp. in wild animals, domestic dogs and their respective ectoparasites, in southern Pantanal region, central-western Brazil, by molecular techniques. Between August 2013 and March 2015, 31 coatis (Nasua nasua), 78 crab-eating foxes (Cerdocyon thous), seven ocelots (Leopardus pardalis), 42 dogs (Canis lupus familiaris), 110 wild rodents (77 Thichomys fosteri, 25 Oecomys mamorae, and 8 Clyomys laticeps), 30 marsupials (14 Thylamys macrurus, 11 Gracilinanus agilis, 4 Monodelphis domestica and 1 Didelphis albiventris), and 1582 ticks and 80 fleas collected from the sampled animals were investigated. DNA samples were submitted to PCR assays for Hepatozoon spp. targeting 18S rRNA gene. Purified amplicons were directly sequenced and submitted to phylogenetic analysis. A high prevalence of Hepatozoon among carnivores (C. thous [91.02%], dogs [45.23%], N. nasua [41.9%] and L. pardalis [71.4%]) was found. However, ticks and fleas were negative to Hepatozoon PCR assays. By phylogenetic analysis based on 18S rRNA sequences, Hepatozoon sequences amplified from crab-eating foxes, dogs, coatis and ocelots clustered with sequences of H. canis, H. americanum and H. felis. The closely related positioning of Hepatozoon sequences amplified from wild rodents and T. macrurus marsupial to Hepatozoon from reptiles and amphibians suggest a possible transmission of those Hepatozoon species between hosts by ectoparasites or by predation. Hepatozoon haplotypes found circulating in wild rodents seem to present a higher degree of polymorphism when compared to those found in other groups of animals. Although rodents seem not to participate as source of Hepatozoon infection to wild carnivores and domestic dogs, they may play an important role in the transmission of Hepatozoon to reptiles and amphibians in Pantanal biome.

Field Evaluation of Two Different Treatment Approaches and Their Ability to Control Fleas and Prevent Canine Leishmaniosis in a Highly Endemic Area.

This study investigated the efficacy of two collars for the treatment and prevention of flea infestations. Additionally the effect of these collars on the incidence of Leishmania infantum infection as compared with a group of vaccinated dogs was evaluated. A total of 224 young dogs from private animal shelters were enrolled in April/May into four groups: G1, 55 dogs treated with 10% imidacloprid + 4.5% flumethrin collar (Seresto, Bayer Animal Health); G2, 60 dogs treated with 4% deltamethrin collar (Scalibor protector band, MSD Animal Health); G3, 54 dogs vaccinated with CaniLeish (Virbac Animal Health); and G4, 55 dogs left non-treated as controls. Dogs were followed up at days 120 (September), 210 (December), and 360 (April-May). At those time points, clinical assessments, ectoparasite counts and blood, bone marrow and skin samples, to detect the presence of L. infantum, were performed. The efficacy of Seresto in protecting dogs from flea infestation was 100% (P < 0.01) on day 120 and 210, while animals treated with Scalibor showed a prevalence of the infestation ranging from 23.3% to 33.3% on day 120 and 210, respectively. At the end of the study, the incidence of L. infantum infection in collared dogs-based on animals being positive in any of the tests-was 5.5% in Seresto-treated dogs and 20% in Scalibor-treated dogs, resulting in overall efficacy of prevention of 88.3% for Seresto and 61.8% for Scalibor. No statistical difference was detected in L. infantum positive dogs for bone marrow PCR and/or cytology at day 360 between the CaniLeish (15.4%) and non-treated control dogs (10.0%). Both collars proved to be effective (P < 0.01) in preventing L. infantum infection throughout one transmission season, whereas no significant difference was recorded in the frequency of active infections between dogs vaccinated with CaniLeish and control dogs, emphasizing the importance of using repellent/insecticide actives as a priority measure for protection against canine leishmaniosis.