Leishmaniases in a tropical area of Brazil: a spatio-temporal analysis.

Leishmaniases are a group of tropical and neglected diseases caused by the protozoa of the genus Leishmania which are transmitted by insect sandflies. Despite of the major efforts undertaken at global level for the control of these diseases, a recent spreading to non-endemic areas been observed. In this study we analyzed the spatio-temporal distribution of Visceral Leishmaniasis (VL) and Tegumentary Leishmaniasis (TL) cases notified in a tropical region of Brazil. A retrospective evaluation was carried out using secondary data (e.g., demographics, origin, age and area of occurrence) of human patients obtained from 2009 to 2019. Choropleth maps with levels of color intensity represented the spatial distribution of VL and TL cases. Overall, 116 (10.5 ± 8.5) cases of leishmaniases were registered in the study area during the whole period. Of those, 61 (5.5 ± 5.5) and 55 (5 ± 4.3) were diagnosed as VL and TL, respectively. VL cases were recorded in 61.9% (13/21) and TL cases in 42.8% (9/21) of the municipalities. Most of the cases were predominantly notified in men from rural areas, and specifically for TL with an age range of 30-59 years old. These findings will be useful to drive strategies of control (e.g., vector control, dog surveillance and notification of suspected cases) against these neglect diseases, preventing the spreading to non-endemic areas.

Effective immuno-therapeutic treatment of Canine Leishmaniasis.

BACKGROUND: Canine Leishmaniasis (CanL) caused by the L. infantum species is one of the biggest threats to the health of the South American canine population. Chemotherapeutics currently used for the treatment of CanL fail to induce a total parasite clearance while inducing numerous side effects. As CanL is an immunomodulated disease, the use of immuno-treatments should strengthen the deficient immune response of infected dogs. In this study, we evaluated a nasally administered immunotherapy in dogs naturally infected with L. infantum (stage 2), with both visceral and cutaneous manifestations. Noteworthy, some of them were also infected by other parasites (E. canis, D. immitis, A. platys), what worsen their chance of survival. METHODOLOGY/PRINCIPAL FINDINGS: The treatment was based on 2 intranasal (IN.) administrations of a killed L. infantum parasite loaded into maltodextrin nanoparticles, which treatment was compared with the classical oral administration of Miltefosine (2 mg/kg) for 28 days, as well as a combination of these 2 treatments. The results showed that two IN administrations significantly reduced the serology, and were at least as efficient as the chemotherapy to reduce the skin and bone marrow parasite burden, as well as clinical scores, and that unlike Miltefosine treatments, this nasally administered nanoparticle vaccine was without side effects. CONCLUSIONS: These results confirm the feasibility of a simple therapeutic immuno-treatment against L. infantum infected dogs, which is a promising tool for future developments.

Molecular detection of vector-borne pathogens in cats tested for FIV and FeLV.

The aim of this study was to detect molecularly vector borne pathogens (VBPs) in domiciled cats tested for Feline immunodeficiency virus (FIV) and Feline leukemia virus (FeLV). Blood samples (n = 119) were analyzed microscopically and molecularly through PCR and sequenced for the detection of the following pathogens: piroplasmids., Bartonella henselae, Cytauxzoon felis, Ehrlichia canis, Leishmania spp., hemotropic Mycoplasma spp., Trypanosoma spp., and Ricketssia spp. Animals were also serological assessed for detection of antibodies against FIV and FeLV. Out of all animals, 20.16% (24/119) tested positive for at least one VBPs at molecular examination. Conversely, no animal resulted positive at microscopic analysis. The most prevalent pathogen was hemotropic Mycoplasma haemofelis (8.40%; 10/119), followed by Candidatus Mycoplasma haemominutum (5.88%; 7/119), E. canis (5.04%; 6/119), C. felis (0.84%; 1/119) and B. henselae (0.84%; 1/119). One animal (0.84%; 1/119) was co-infected with. E. canis and B. henselae. A total of 5.88% (7/119) and 1.68% (2/119) tested positive for FIV and FeLV, respectively. Data of this study demonstrate that owned cats can be at risk of hemotropic Mycoplasma spp., E. canis, C. felis and B. henselae. Therefore, preventive measures against vectors of these pathogens should be implemented in order to reduce the risk of exposition and consequently infection. Additionally, aggressive behaviors among cats should be avoided, especially because hemotropic Mycoplasma spp. may be transmitted through the bite of animals.

Vector-borne pathogens in dogs from areas where leishmaniosis is endemic.

Many vector-borne pathogens (VBPs), including Ehrlichia canis and Dirofilaria immitis, may infect simultaneously dogs in areas where Leishmania infantum is endemic, especially in the tropics, where highly abundant arthropod vectors thrive. The aim of this study was to compare the frequency of simultaneous VBPs infection in Leishmania-positive and Leishmania-negative dogs. Animals enrolled in this study were divided in two groups (G1 and G2), G1 being comprised of L. infantum-infected dogs (n = 58) and the G2 of L. infantum-negative dogs (n = 58). Blood samples were screened using a qualitative ELISA test (SNAP® 4Dx® Plus, IDEXX Laboratory, Westbrook, Maine, USA) for detection of antibodies against Anaplasma spp., Borrelia burgdorferi sensu lato, Ehrlichia spp. and antigens of Dirofilaria immitis. Overall, 89.7% (52/58) of dogs from G1 were positive for at least one VBP, whereas 50.0% (29/58) of dogs from G2 dogs were positive as well. The highest positivity was to E. canis (67.2%; 78/116), followed by D. immitis (12.9%; 15/116), and A. platys (6.0%; 7/116). None of the animals scored positive for B. burgdorferi s.l.. There was a statistically significant difference for the simultaneous positivity to E. canis plus D. immitis between groups. Furthermore, 43.1% (25/58) of dogs from G1 were infested by ectoparasites (ticks, fleas, or both), compared to 20.6% (12/58) of dogs from G2. In conclusion, Leishmania-infected dogs were more co-infected with other VBPs than Leishmania-negative animals. Therefore, it is pivotal to increase the awareness of veterinarian and dog owners about the importance of testing Leishmania-infected dogs for other VBPs, as this may directly affect treatment decisions and management.

Occurrence of Dirofilaria immitis in Stray Dogs from Nepal.

BACKGROUND: Nematodes of the genus Dirofilaria are widespread vector-borne helminths (VBH) of veterinary relevance. Dirofilariosis caused by D. immitis is frequently diagnosed in dogs often showing a severe clinical condition known as heartworm disease. Assessing the distribution pattern of canine dirofilariosis is pivotal to undertake appropriate control measures and define the risk of infection in animals and humans. This study provides original data on the occurrence of D. immitis in naive stray dogs from Nepal. METHODS: An epidemiological study was performed on stray dogs from Siddharthanagar, Lumbini region, Nepal. Alive animals were blood sampled and examined for microfilariae using direct microscopy, buffy coat centrifugation and modified Knott's method. In addition, hematobiochemical parameters, including packed cell volume (PCV), alanine aminotransferase (SGPT), aspartate aminotransferase (SGOT), alkaline phosphate (ALP), creatinine, blood urea nitrogen (BUN), urea and total protein were analysed, along with a clinical scoring (body temperature and respiratory rate). During the observation period, data on autopsies performed on stray dogs showing canine heartworm disease were also included. RESULTS: The overall prevalence of D. immitis in both alive and necropsied dogs was 22.49% (38/169). Out of 150 alive dogs, 29 (19.3%) were positive for D. immitis microfilariae. The prevalence of the infection varied according to the technique used, ranging from 16.0 to 19.3% based on direct blood smear microscopy and modified Knott's technique, respectively. The infection rate was significantly higher (p < 0.05) in dogs aging more than 3 years. A positive correlation was observed between SGPT and SGOT and the presence of microfilariae (p < 0.05). Among the 19 dead stray dogs, 9 animals showed adult D. immitis nematodes. CONCLUSION: Results of this survey account for the presence of D. immitis in stray dogs from Nepal, thus supporting the existence of a local transmission cycle of diroflariosis in the region. Further investigations are warranted to accurately define the prevalence of the infection in the canine population and to undertake appropriate control measures to reduce the burden on the infection in dogs and, accordingly, the risk of transmission to humans.

Zoonotic parasites in wild animals such as carnivores and primates that are traded illegally in Brazil.

Brazil accounts for around 20% of all animal species, but these are constantly threatened by illegal anthropic activities. Unfortunately, animal dealers are totally unaware of the sanitary risks among wild animals, or that occurrences of parasites in these animals are bioindicators for their current sanitary status within the ecosystem in which they live. This status is an important parameter with regard to assessing the spreading of pathogens. Therefore, the aim of this study was to perform a survey of zoonotic parasites in carnivores and non-human primates that are illegally traded in Brazil. Between June 2016 and July 2017, 43 wild animals (20 carnivores and 23 non-human primates) were presented at the Wild Animal Screening Center of Sergipe (CETAS/SE). Fecal and blood samples were obtained and analyzed to detect occurrences of pathogens of medical and veterinary importance, such as Cryptosporidium spp., Giardia spp., Dirofilaria immitis, Leishmania infantum, Leishmania braziliensis, Toxoplasma gondii, Trypanosoma cruzi and gastrointestinal helminths. Out of all the animals analyzed, 55.8% (24/43) were found to be positive for at least one parasite species, i.e. 41.7% and 58.3% of the carnivores and non-human primates, respectively. However, all the animals were negative for D. immitis, L. braziliensis and T. cruzi. These findings demonstrate that illegally traded wild animals may represent a risk to public health because of absence of sanitary control during their transportation. Therefore, preventive measures might be employed to avoid infection of these animals and people in close contact with them.

O Brasil abriga cerca de 20% de todas as espécies animais existentes no mundo que são continuamente ameaçadas pelas ações antrópicas. Infelizmente, os comerciantes de animais não são suficientemente esclarecidos em relação à ameaça sanitária que representam a vida silvestre, assim como sobre a ocorrência de parasitos que nestes animais funcionam como bio-indicadores do status sanitário do ecossistema, visto serem importantes parâmetros para avaliar a dispersão de patógenos. Portanto, objetivaram-se neste estudo a pesquisa de parasitos zoonóticos em carnívoros e primatas ilegalmente comercializados no Brasil. Entre Junho de 2016 e Julho de 2017, 43 animais silvestres (20 carnívoros e 23 primatas) foram recebidos no Centro de Triagem de Animais Silvestres de Sergipe (CETAS/SE). Amostras fecais e sanguíneas foram obtidas e analisadas para detectar a presença de patógenos de importância médico-veterinária como espécies de Cryptosporidium e Giardia, Dirofilaria immitis, Leishmania infantum, Leishmania braziliensis, Toxoplasma gondii, Trypanosoma cruzi e helmintos gastrintestinais. De todas as amostras analisadas, 55,8% (24/43) foram positivas a pelo menos um agente etiológico testado, sendo 41,7% e 58,3% carnívoros e primatas, respectivamente. Todas as amostras foram negativas a D. immitis, L. braziliensis e T. cruzi. Estes achados demonstram que o comércio ilegal de animais silvestres pode representar risco a saúde publica e a saúde destes animais devido à ausência de medidas sanitárias durante o deslocamento. Por fim, medidas preventivas devem ser propostas para evitar a infecção destes animais e consequentemente das pessoas que os manipulam.

Occurrence of zoonotic gastrointestinal parasites of rodents and the risk of human infection in different biomes of Brazil.

Rodents are synanthropic mammals adapted to several ecosystems, where they can contribute to the transmission of zoonotic pathogens, including gastrointestinal parasites. The aim of this study was to study the occurrence of gastrointestinal parasites from rodents and discuss the risk of transmission to humans. Fecal samples (n = 110) from different rodent species, namely, Cerradomys subflavus (n = 4), Mus musculus (n = 14), Rattus norvegicus (n = 80), Rattus rattus (n = 8) and Thrichomys apereoides (n = 4), were analyzed using the FLOTAC technique. Of the samples examined, 73.6% (81/110) tested positive for at least one gastrointestinal parasite. The most commonly identified parasites were Aspiculuris sp., Hymenolepis nana, Moniliformis sp., Syphacia sp., Strongyloides spp., Taenia spp., and Trichuris spp. eggs, Angiostrongylus cantonensis larvae and Entamoeba spp. cysts. The findings of this study demonstrate that rodents living in different Brazilian biomes are parasitized by a wide range of parasites, including some of public health concern. Therefore, the proximity of rodents to human settlements may represent a tangible risk of infection for people living in these areas.

Roedores são mamíferos sinantrópicos adaptados a vários ecossistemas, onde podem contribuir para a transmissão de patógenos zoonóticos, incluindo parasitas gastrointestinais. O objetivo deste estudo foi descrever a ocorrência de parasitos gastrointestinais de roedores e discutir o risco de transmissão ao ser humano. Amostras fecais (n = 110) de diferentes espécies de roedores, a saber, Cerradomys subflavus (n = 4), Mus musculus (n = 14), Rattus norvegicus (n = 80), Rattus rattus (n = 8) e Thrichomys apereoides (n = 4), foram analisados pela técnica FLOTAC. Das amostras examinadas, 73,6% (81/110) apresentaram resultado positivo para pelo menos um parasito gastrointestinal. Os parasitos mais comumente identificados foram ovos de Aspiculuris sp., Hymenolepis nana, Moniliformis sp., Syphacia sp., Strongyloides spp., Taenia spp., Trichuris spp., larvas de Angiostrongylus cantonensis e cistos de Entamoeba spp. Os resultados deste estudo demonstraram que os roedores que vivem em diferentes biomas brasileiros são parasitados por uma ampla variedade de parasitos, incluindo alguns problemas de saúde pública. Portanto, a proximidade de roedores com assentamentos humanos pode representar um risco de infecção nessas áreas.

Detection of Rickettsia felis in ectoparasites collected from domestic animals.

Ticks and fleas are arthropods widely distributed around the world involved in the transmission of various vector-borne diseases (VBDs), including Brazilian Spotted Fever (BSF), Baggio-Yoshinari Syndrome and the plague, with outstanding consequences for the public health. The aim of this study was to investigate the presence of Rickettsia spp., Borrelia spp. and Yersinia pestis in arthropods collected from dogs, cats and horses living in the state of Pernambuco, Northeastern Brazil. From January 2017 to April 2019, ectoparasites were collected, morphologically identified and molecularly analysed through PCR and sequencing. In total 401 specimens were collected from 86 animals, being 68% (n = 273) and 32% (n = 128) from rural and urban areas, respectively. The most commonly detected species were the ticks Dermacentor nitens, Amblyomma sculptum, Rhipicephalus sanguineus sensu lato (s.l.), Rhipicephalus microplus, and Amblyomma ovale, and the fleas Ctenocephalides felis and Ctenocephalides canis. DNA of Rickettsia felis was detected in D. nitens collected from horses, and C. felis, and R. sanguineus s.l. collected from dogs. All samples scored negative for Borrelia spp. and Y. pestis DNA. This study provides valuable data on ectoparasite fauna from domestic animals and identifies the circulation of a zoonotic pathogen (i.e., R. felis) in the population of the arthropods assessed. Therefore, preventive measures should be adopted in order to reduce the risk of occurrence of neglected VBD caused by this pathogen in animal and human hosts.

Spatial distribution of triatomine bugs in a Chagas disease endemic region in Brazil.

INTRODUCTION: Trypanosoma cruzi is the etiological agent of Chagas disease (CD), a zoonotic infection transmitted by triatomine bug vectors to human beings. Although the story of this parasitic infection was born in Brazil and here this has made major step forward information, the same cannot be said about the actual distribution of the triatomine vector in several areas of this country. The aim of this study was to assess the occurrence of triatomine species in an endemic region for CD in Northeastern Brazil. METHODS: A retrospective study was performed using data obtained from 2008 to 2017. All information was provided by the V Gerência Regional de Saúde of the state of Pernambuco. The spatial distribution of triatomine species was analyzed by drawing a map using the Quantum geographic information system. RESULTS: A total of 4,694 triatomine specimens (469.4 ± 221.2 per year) were collected during the period 2008-2017, with 94.5% (4,434/4,694) at the intradomicile and 5.5% (260/4,694) at peridomicile environment. Of all arthropods collected, 92.5% (4,340/4,694) and 7.5% (354/4,694) were adults and nymphs, respectively. The species most frequently detected were Panstrongylus lutzi (30.36%), Triatoma brasiliensis (26.12%), Triatoma pseudomaculata (22.43%), and Panstrongylus megistus (20.54%). CONCLUSIONS: These data contribute to a better understanding of the distribution of T. cruzi infection in the Northeastern region of Brazil. Preventive measures based on vector control should be implemented in the study area in order to reduce the burden this neglected tropical disease.

An Additional Asset for the FLOTAC Technique: Detection of Gastrointestinal Parasites in Vegetables.

INTRODUCTION: Gastrointestinal parasites are considered amongst the most important threats to veterinary and human health worldwide. The transmission of these parasitic diseases usually involves the accidental ingestion of oocysts, cysts, eggs, and larvae, whose detection is generally achieved by classical coproparasitological methods, based on sedimentation and/or flotation techniques. These procedures might be inefficient or display a low sensitivity when performed on food matrices. This study aimed to assess the value of the FLOTAC technique for the detection of different parasitic stages of gastrointestinal parasites of domestic animals in fresh lettuce. MATERIAL AND METHODS: Samples (n = 100) were purchased from public markets located in the metropolitan region of Recife (Brazil). RESULTS: A total of 79% scored positive for eggs and/or cysts of gastrointestinal parasites. Protozoa and nematodes were detected in 32% (32/100) and 64% (64/100) of samples, respectively, with cysts of the genus Entamoeba (32%) and eggs of nematodes of the order Strongylida (30%) being the most frequently diagnosed. CONCLUSIONS: The findings herein reported demonstrate that the FLOTAC technique can be successfully applied for recovering food-borne parasites of medical and veterinary concern in food matrices.

LONGRANGE® (eprinomectin 5% w/v extended-release injection) efficacy against Hypoderma lineatum in an endemic area in southern Italy.

BACKGROUND: Despite intensive control measures including governmental campaigns using highly-efficacious systemic insecticides, there is evidence for persisting or recurring bovine Hypoderma species populations in parts of Europe, the USA and Canada. The present study evaluated the efficacy of LONGRANGE® (eprinomectin 5% w/v extended-release injection) against the infestation of cattle with Hypoderma lineatum, which is considered to be the predominant bovine warble fly in southern Europe and in North America. METHODS: Thirty-six local breed cattle sourced in an endemic area in southern Italy and confirmed positive for Hypoderma exposure by ELISA were randomly assigned to three groups of 12 animals each. Cattle of one group served as control and received saline injectable solution, whereas those in the two other groups received LONGRANGE® by subcutaneous injection. LONGRANGE® was administered once, either when Hypoderma larvae were expected to be first-instars (L1) or after warbles development, with Hypoderma larvae moulting to the second-(L2) and third-(L3) instars. Cattle were checked at intervals for warbles and Hypoderma larvae were collected, examined for their viability and morphologically identified. The detection of Hypoderma on cattle was terminated when warbles were no longer emerging. RESULTS: All intact larvae collected were identified as H. lineatum. No live larvae were collected from any animal treated with LONGRANGE® while live specimens were sampled from nine of the 12 control cattle (1 to 9 larvae per animal) (P = 0.0001 at α = 0.05). LONGRANGE® treatment was well accepted and no adverse events related to treatment or other health problems were observed. CONCLUSIONS: This study confirmed the continued 'preventive' (efficacy against migrating L1) and 'therapeutic' (efficacy against L2 and L3 in warbles) efficacy of LONGRANGE® against H. lineatum infestation of cattle under contemporary field conditions.

Spatial distribution of triatomine bugs in a Chagas disease endemic region in Brazil

Abstract INTRODUCTION: Trypanosoma cruzi is the etiological agent of Chagas disease (CD), a zoonotic infection transmitted by triatomine bug vectors to human beings. Although the story of this parasitic infection was born in Brazil and here this has made major step forward information, the same cannot be said about the actual distribution of the triatomine vector in several areas of this country. The aim of this study was to assess the occurrence of triatomine species in an endemic region for CD in Northeastern Brazil. METHODS: A retrospective study was performed using data obtained from 2008 to 2017. All information was provided by the V Gerência Regional de Saúde of the state of Pernambuco. The spatial distribution of triatomine species was analyzed by drawing a map using the Quantum geographic information system. RESULTS: A total of 4,694 triatomine specimens (469.4 ± 221.2 per year) were collected during the period 2008-2017, with 94.5% (4,434/4,694) at the intradomicile and 5.5% (260/4,694) at peridomicile environment. Of all arthropods collected, 92.5% (4,340/4,694) and 7.5% (354/4,694) were adults and nymphs, respectively. The species most frequently detected were Panstrongylus lutzi (30.36%), Triatoma brasiliensis (26.12%), Triatoma pseudomaculata (22.43%), and Panstrongylus megistus (20.54%). CONCLUSIONS: These data contribute to a better understanding of the distribution of T. cruzi infection in the Northeastern region of Brazil. Preventive measures based on vector control should be implemented in the study area in order to reduce the burden this neglected tropical disease.

Recent advances on Dirofilaria repens in dogs and humans in Europe.

Dirofilaria repens is a nematode affecting domestic and wild canids, transmitted by several species of mosquitoes. It usually causes a non-pathogenic subcutaneous infection in dogs and is the principal agent of human dirofilariosis in the Old World. In the last decades, D. repens has increased in prevalence in areas where it has already been reported and its distribution range has expanded into new areas of Europe, representing a paradigmatic example of an emergent pathogen. Despite its emergence and zoonotic impact, D. repens has received less attention by scientists compared to Dirofilaria immitis. In this review we report the recent advances of D. repens infection in dogs and humans, and transmission by vectors, and discuss possible factors that influence the spread and increase of this zoonotic parasite in Europe. There is evidence that D. repens has spread faster than D. immitis from the endemic areas of southern Europe to northern Europe. Climate change affecting mosquito vectors and the facilitation of pet travel seem to have contributed to this expansion; however, in the authors' opinion, the major factor is likely the rate of undiagnosed dogs continuing to perpetuate the life-cycle of D. repens. Many infected dogs remain undetected due to the subclinical nature of the disease, the lack of rapid and reliable diagnostic tools and the poor knowledge and still low awareness of D. repens in non-endemic areas. Improved diagnostic tools are warranted to bring D. repens diagnosis to the state of D. immitis diagnosis, as well as improved screening of imported dogs and promotion of preventative measures among veterinarians and dog owners. For vector-borne diseases involving pets, veterinarians play a significant role in prevention and should be more aware of their responsibility in reducing the impact of the zoonotic agents. In addition, they should enhance multisectorial collaboration with medical entomologists and the public health experts, under the concept and the actions of One Health-One Medicine.

Screening of Cercopithifilaria bainae and Hepatozoon canis in ticks collected from dogs of Northeastern Brazil.

Rhipicephalus sanguineus sensu lato (s.l.) is one of the most widespread ixodid ticks and is a competent vector of several vector-borne pathogens of veterinary and medical concern. For instance, this tick species transmits nematodes of the genus Cercopithifilaria and protozoa of the genus Hepatozoon to carnivores, including dogs. Here we investigated the occurrence of Cercopithifilaria spp. and Hepatozoon spp. in a population of ticks collected from naturally infested dogs living in rural areas of Northeastern Brazil. From August 2016 to June 2017, 758 tick specimens (mean ticks per month = 68.9 ± 71.4) were sampled from 75 dogs (mean ticks per dog = 10.11 ± 5.2) and dissected under a stereomicroscope in order to visualize Cercopithifilaria spp. larvae and Hepatozoon spp. oocysts and sporocysts. R. sanguineus s.l. was the only species collected, peaking in September (n = 273) and decreasing in February 2017 (n = 39). Different larval stages of Cercopithifilaria bainae were identified in 7 out of 758 (0.93%) ticks. In addition, 4 specimens (0.53%) were positive for oocysts and free sporocysts of Hepatozoon canis. The identity of both species of parasites was molecularly confirmed. These results account for the predominance of R. sanguineus (s.l.) in domestic dogs from rural locations of the study area, as well as for the presence C. bainae and H. canis in these tick populations.

Survey on helminths and protozoa of free-living Neotropical bats from Northeastern Brazil.

Bats are considered important reservoirs of pathogens of veterinary and medical relevance worldwide. However, despite the increasing attention paid towards the central role of bats in the spreading of some zoonotic infections, studies on their own parasites remain patchy especially in Neotropical regions. Understanding the relationships occurring between bats and their pathogens is a crucial step to determine the implications undermining the ecology and biology of these animals. The present study aims to assess the endoparasitic fauna of Neotropical bats from Northeastern Brazil, with a particular focus on helminths and protozoa. From October 2016 to March 2017, 54 fresh faecal samples were collected in stable shelters inhabited by three different bat species (i.e., Molossus molossus, Myotis lavali and Noctilio albiventris). Samples were analysed using four different techniques (i.e., Centrifugal Sedimentation, Flotation Technique, Spontaneous Sedimentation Technique and FLOTAC®). A total of 96.29% (52/54) samples were positive for at least one gastrointestinal parasite and 11 different families were identified. In particular, 63.6% (7/11) of the families were of helminths and 36.4% (4/11) of protozoa. Helminths were classified into two different classes (Nematoda and Cestoda) and seven families, whereas the four protozoa classes detected (i.e., Coccidia, Gregarinomorphe, Lobosea and Zoomastigophora) into four families. Eggs of Ancylostomatidae and Hymenolepididae were the most abundan. The findings of this present study demonstrate that Neotropical bats from Northeastern Brazil are parasitized by a wide number of parasites, and some of them may have important implications for Public Health.

Applicability of FLOTAC® technique in recovering equine strongyle larvae in the pasture: A comparison study.

The FLOTAC® technique represents a highly sensitive method for the isolation of oocysts, eggs, and larvae of parasites in faeces. This assay could be used for detecting free-living stages of nematodes in the pasture but no attempt has been assessed so far. Therefore, the performance of FLOTAC® technique for isolating infective larvae of nematodes in the environment was investigated and compared with the spontaneous sedimentation (SST) and centrifugal sedimentation (CST) techniques. The study was conducted in a horse farm located in northeastern Brazil, where the occurrence of strongyle larvae had been previously reported. Pasture samplings were collected monthly from January to May 2016 in a 376 m2 crop area harvested with the Guinea grass Panicum cultivar Massai. The recovery of third-stage larvae (L3) was performed using the FLOTAC®, SST and CST techniques. Values of Cohen's kappa coefficient, sensitivity, specificity, positive and negative predictive values, and accuracy of each technique were assessed. Although strongyle larvae were evenly detected, with the FLOTAC® technique yielded the highest number of positive samples (i.e., 41%, 41/100, p < .0001). The main parasites isolated belonged to the Cyathostominae and Strongylinae subfamilies. Based on these results, the FLOTAC® technique should be considered as practical and safe method for the isolation of nematode larvae in the pasture, thus opening a new potential use for this tool in the field.

Three different Hepatozoon species in domestic cats from southern Italy.

Three species of Hepatozoon, namely, Hepatozoon felis, Hepatozoon canis and Hepatozoon silvestris may affect domestic and/or wild felids. Although hepatozoonosis has been documented in a wide range of mammal species, data on cats are limited. To investigate the occurrence of these pathogens in cats, blood samples were collected from animals living in three provinces of southern Italy (Bari, Lecce, and Matera), and molecularly analysed by PCR amplification and sequencing of segments of the 18S rRNA gene. Out of 196 blood samples collected, Hepatozoon spp. DNA was amplified in ten cats (5.1%, CI: 3%-9%), with the majority of infected animals from Matera (8/34, 23.5%) and one each from the other two provinces. BLAST analysis revealed the highest nucleotide identity with sequences of H. canis, H. felis and H. silvestris deposited in GenBank. Results of this study indicate that these three species of Hepatozoon infect domestic cats in Italy. This is the first report of H. silvestris infection in a domestic cat.

Lungworms and gastrointestinal parasites of domestic cats: a European perspective.

With the exception of Aelurostrongylus abstrusus, feline lungworms have been poorly studied. Information on their distribution is patchy and mostly limited to case reports. In this study, the occurrence of feline lungworms and co-infecting gastrointestinal parasites has been investigated in 12 European countries (i.e. Austria, Belgium, Bulgaria, France, Greece, Hungary, Italy, Portugal, Romania, Spain, Switzerland and the United Kingdom). An average of 10 domestic cats, with regular outdoor access, was sampled each month for 12months, and freshly passed faeces were collected. Stools were processed using a McMaster assay and a quantitative Baermann-Wetzel method. Animals positive for lungworms and/or gastrointestinal parasites were treated with a formulation containing fipronil, (S)-methoprene, eprinomectin, and praziquantel (Broadline®, Merial), and re-sampled 28days post-treatment. The association between lungworm infection and risk factors was analysed using statistical medians/means and the efficacy of the treatment against each lungworm species was assessed. Of 1990 cats sampled, 613 (30.8%) were positive for at least one parasite, while 210 (10.6%) were infected by lungworms. The prevalence of lungworm infection varied between the sampled sites, with the highest recorded in Bulgaria (35.8%) and the lowest in Switzerland (0.8%). None of the cats from Austria or the United Kingdom were infected by lungworms. Aelurostrongylus abstrusus was the species most frequently detected (78.1%), followed by Troglostrongylus brevior (19.5%), Eucoleus aerophilus (14.8%) and Oslerus rostratus (3.8%). The overall efficacy of the treatment was 99% for A. abstrusus and 100% for T. brevior, O. rostratus and E. aerophilus. Data presented provide a comprehensive account of the diagnosis, epidemiology and treatment of feline lungworms in Europe, as well as of the occurrence of co-infections by gastrointestinal parasites.

Whence river blindness? The domestication of mammals and host-parasite co-evolution in the nematode genus Onchocerca.

The genus Onchocerca includes 34 described species and represents one of the largest genera of the filarial nematodes within the family Onchocercidae. Representative members of this genus are mainly parasites of ungulates, with some exceptions such as Onchocerca lupi and Onchocerca volvulus, infecting carnivores and/or humans. For a long time, the evolutionary relationships amongst onchocercids remained poorly studied, as the systematics of this genus was impaired by the high morphological variability of species included in the taxon. Although some molecular phylogenies were developed, these studies were mainly focused on bovine Onchocerca spp. and O. volvulus, including assessments of Wolbachia endosymbionts. In the present study, we analysed 13 Onchocerca spp. from a larger host spectrum using a panel of seven different genes. Analysis of the coxI marker supports its usefulness for the identification of species within the genus. The evolutionary history of the genus has been herein revised by multi-gene phylogenies, presenting three strongly supported clades of Onchocerca spp. Analyses of co-evolutionary scenarios between Onchocerca and their vertebrate hosts underline the effect of domestication on Onchocerca speciation. Our study indicates that a host switch event occurred between Bovidae, Canidae and humans. Cophylogenetic analyses between Onchocerca and the endosymbiotic bacterium Wolbachia indicate the strongest co-evolutionary pattern ever registered within the filarial nematodes. Finally, this dataset indicates that the clade composed by O. lupi, Onchocerca gutturosa, Onchocerca lienalis, Onchocerca ochengi and O. volvulus derived from recent speciation.