Epidemiological aspects of the occurrence of anti-Sarcocystis spp. andanti-Toxoplasma gondii antibodies in dogs in the Northern region of Brazil / Aspectos epidemiológicos da ocorrência de anticorpos anti-Sarcocystisspp. e anti-Toxoplasma gondii em cães da região norte do Brasil

We conducted aseroepidemiological study on the occurrence of anti-Sarcocystisspp. and anti-Toxoplasma gondii antibodies in dogs from family farming properties in the municipality of Ji-Paraná, Rondônia.Blood samples were collected from apparently healthy dogs between September 2012 and November 2013. In total, 181 blood serum samples were analyzed using an indirect immunofluorescence assay, among which 57 (31.49%) and 20 (11.04%) were positive for anti-T. gondii and anti-Sarcocystis spp., respectively. Statistical analyses showed that the type of food fed to the dogs was associated with the occurrence of anti-Sarcocystisspp. antibodies. In contrast, age and access to bovine carcasses were the risk factors for anti-T. gondii.The high occurrence of seropositive dogs for Sarcocystis spp. and T. gondii evidences the wide distribution of these agents in the studied area, possibly due to human and animal exposure to these protozoan species. In addition, anti-T. gondii antibodies were directly proportional to dog age. The increase in the number of positive animals with age was statistically significant. Furthermore, high antibody titers (up to 800) against Sarcocystis spp. in dogs suggest the possibility of recent exposure, in addition to environmental contamination by oocysts/sporocysts eliminated by the feces of these animals.

Conduzimos um estudo soroepidemiológico sobre a ocorrência de anticorpos anti- Sarcocystis spp. e anti-Toxoplasma gondiiem cães de propriedades de agricultura familiar no município de Ji-Paraná, Rondônia. Amostras de sangue foram coletadas de cães aparentemente saudáveis, entre setembro de 2012 e novembro de 2013. Ao todo, foram analisados 181 soros sanguíneos por meio do ensaio de imunofluorescência indireta, sendo positivas 57 (31,49%) e 20 (11,04%) amostras para anticorpos anti-T. gondii e anti-Sarcocystis spp., respectivamente. As análises estatísticas demonstraram que o tipo de alimentação fornecida aos cães esteve associado à ocorrência de anticorpos anti-Sarcocystis spp. Em contraste a idade e o acesso à carcaça bovina foram fatores de risco para a presença de anticorpos anti-T. gondii. A alta ocorrência de cães soropositivos para Sarcocystis spp. e T. gondii evidencia a ampla distribuição desses agentes na área estudada, possivelmente devido à exposição humana e animal a essas espécies de protozoários. Além disso, o resultado dos anticorpos anti-T. gondii relacionados a idade do cão mostraram diferença estatística, com aumento significativo no número de animais positivos com a idade. Além disso, altos títulos de anticorpos (até 800) contra Sarcocystis spp. em cães sugerem a possibilidade de exposição recente, além da contaminação ambiental por oocistos/esporocistos eliminados pelas fezes desses animais.

Uso da miltefosina no tratamento clínico de cães com leishmaniose visceral: revisão de literatura / Use of miltefosin in the clinical treatment of dogs with visceral leishmaniosis / Uso de miltefosina en el tratamiento clínico de perros con leishmaniosis visceral: revisión de la literatura

A leishmaniose visceral canina é uma doença de caráter zoonótico, acometendo os seres humanos e diversas espécies de animais silvestres e domésticos. Objetivou-se com o presente estudo realizar uma revisão de literatura sobre o uso da miltefosina no tratamento clínico de cães com leishmaniose visceral. Trata- se de uma revisão de literatura, a qual foi realizada por meio de consultas à periódicos e livros presentes na biblioteca do Cesmac. Foram utilizadas bases de dados como: portal Capes, SCIELO, Google Acadêmico; pesquisa em monografias, teses e dissertações. Causada pelo protozoário Leishmania chagasi, sendo o cão doméstico o principal reservatório desse protozoário. Por representar um problema grave de saúde pública e ser considerada uma doença potencialmente fatal (quando não tratada precocemente e adequadamente), faz- se importante que o clínico esteja familiarizado com os sinais clínicos, exames complementares e principais protocolos terapêuticos, em especial a utilização da miltefosina no tratamento da leishmaniose visceral em cães. Por ser uma zoonose que causa graves problemas de saúde pública e que vem crescendo cada vez mais no Brasil, cabe aos médicos veterinários assumirem o compromisso na conscientização sobre a importância do diagnóstico precoce além de promoverem o bem-estar animal e a saúde pública.(AU)

Canine visceral leishmaniasis is a zoonotic disease, affecting humans and several species of wild and domestic animals. The objective of the present study was to carry out a literature review on the use of miltefosine in the clinical treatment of dogs with visceral leishmaniasis. This is a literature review, which was carried out through consultations with periodicals and books present in the Cesmac library. Databases such as: Capes portal, SCIELO, Google Scholar; research in monographs, theses and dissertations. Caused by the protozoan Leishmania chagasi, with the domestic dog being the main reservoir of this protozoan. As it represents a serious public health problem and is considered a potentially fatal disease (when not treated early and properly), it is important that the clinician is familiar with the clinical signs, complementary exams and main therapeutic protocols, especially the use of miltefosine in the treatment of visceral leishmaniasis in dogs. As it is a zoonosis that causes serious public health problems and that has been growing more and more in Brazil, it is up to veterinarians to make a commitment to raise awareness of the importance of early diagnosis in addition to promoting animal welfare and public health.(AU)

La leishmaniosis visceral canina es una enfermedad zoonótica que afecta a los seres humanos y a varias especies de animales salvajes y domésticos. El objetivo de este estudio fue realizar una revisión bibliográfica sobre el uso de la miltefosina en el tratamiento clínico de perros con leishmaniosis visceral. Se trata de una revisión bibliográfica, que se realizó mediante consultas a publicaciones periódicas y libros presentes en la biblioteca del Cesmac. Se utilizaron bases de datos como: portal Capes, SCIELO, Google Académico; investigación en monografías, tesis y disertaciones. Causada por el protozoo Leishmania chagasi, siendo el perro doméstico el principal reservorio de este protozoo. Dado que representa un grave problema de salud pública y se considera una enfermedad potencialmente mortal (cuando no se trata de forma temprana y adecuada), es importante que el clínico esté familiarizado con los signos clínicos, las pruebas adicionales y los principales protocolos terapéuticos, especialmente el uso de miltefosina en el tratamiento de la leishmaniosis visceral en perros. Siendo una zoonosis que causa graves problemas de salud pública y que viene creciendo cada vez más en Brasil, corresponde a los veterinarios asumir el compromiso de concienciar sobre la importancia del diagnóstico precoz y promover el bienestar animal y la salud pública.(AU)

Risk Factors Associated With Echinococcosis in the General Chinese Population: A Meta-Analysis and Systematic Review.

Background: Echinococcosis is a severe zoonotic disease that imposes a substantial burden on human life. This meta-analysis aimed to summarize available data on the prevalence of human echinococcosis and identify the key risk factors for echinococcosis in the Chinese general population. Methods: Relevant studies were comprehensively searched in the PubMed, EMBASE, Web of Science, Cochrane, Chinese National Knowledge Infrastructure (CNKI), Chongqing VIP Information (VIP), Wanfang and SinoMed databases until August 22, 2020. A random-effects model was used to estimate the pooled odds ratio (OR) and 95% confidence interval (95% CI). The I2 and Q statistics were calculated to evaluate the heterogeneity, and potential sources of heterogeneity were identified using sensitivity analysis and subgroup analysis. Publication bias was estimated by funnel plots and Egger's test. Results: A total of 1026 studies were identified through the database search, of which 26 were eligible for this meta-analysis. The pooled prevalence of AE and CE were 2.88% and 5.66%, respectively. Ethnicity (OR = 2.93, 95% CI: 1.81-4.75; I2 = 0), herdsman occupation (OR = 2.66, 95% CI: 2.25-3.14; I2 = 8.0%), not washing hands before meals (OR = 2.40, 95% CI: 1.34-4.28; I2 = 82.8%) and being female (OR = 1.45, 95% CI: 1.26-1.66; I2 = 33.9%) were risk factors for AE. The top five risk factors for CE were ethnicity (OR = 3.18, 95% CI: 1.55-6.52; I2 = 79.2%), nomadism (OR = 2.71, 95% CI: 1.65-4.47; I2 = 55.8%), drinking nonboiled water (OR = 2.47, 95% CI: 1.36-4.47; I2 = 85.7), feeding viscera to dogs (OR = 2.35, 95% CI: 1.89-2.91; I2 = 21.5%), and herdsman occupation (OR = 2.19, 95% CI: 1.67-2.86; I2 = 85.1%). Conclusions: This study generalized articles that have contributed to our current understanding of the epidemic of human echinococcosis (AE and CE) in China over the years. The results support that the ethnicity and dog-related factors are major risk factors for both CE and AE. The identification of echinococcosis risk factors may aid researchers and policymakers in improving surveillance and preventive measures aimed at reducing Echinococcus granulosus and Echinococcus multilocularis infection in humans.

Transcriptomic Profile of Canine DH82 Macrophages Infected by Leishmania infantum Promastigotes with Different Virulence Behavior.

Zoonotic visceral leishmaniosis caused by Leishmania infantum is an endemic disease in the Mediterranean Basin affecting mainly humans and dogs, the main reservoir. The leishmaniosis outbreak declared in the Community of Madrid (Spain) led to a significant increase in human disease incidence without enhancing canine leishmaniosis prevalence, suggesting a better adaptation of the outbreak's isolates by other host species. One of the isolates obtained in the focus, IPER/ES/2012/BOS1FL1 (BOS1FL1), has previously demonstrated a different phenotype than the reference strain MCAN/ES/1996/BCN150 (BCN150), characterized by a lower infectivity when interacting with canine macrophages. Nevertheless, not enough changes in the cell defensive response were found to support their different behavior. Thus, we decided to investigate the molecular mechanisms involved in the interaction of both parasites with DH82 canine macrophages by studying their transcriptomic profiles developed after infection using RNA sequencing. The results showed a common regulation induced by both parasites in the phosphoinositide-3-kinase-protein kinase B/Akt and NOD-like receptor signaling pathways. However, other pathways, such as phagocytosis and signal transduction, including tumor necrosis factor, mitogen-activated kinases and nuclear factor-κB, were only regulated after infection with BOS1FL1. These differences could contribute to the reduced infection ability of the outbreak isolates in canine cells. Our results open a new avenue to investigate the true role of adaptation of L. infantum isolates in their interaction with their different hosts.

Dermal microfilariae of dogs, jackals and cats in different regions of Iran.

BACKGROUND: Due to the complexity of retrieving skin-dwelling microfilariae, filarioids of dogs presenting dermal microfilariae (e.g. Cercopithifilaria spp., Onchocerca lupi) are relatively unknown compared to Dirofilaria spp. and Acanthocheilonema spp. whose microfilariae circulate in the blood. Although Cercopithifilaria spp. and O. lupi filarioids are distributed worldwide, there is a paucity of information on their occurrence in Iran. The aim of this study was to investigate these filarioids in a large population of dogs from different regions of Iran. METHODS: From October 2018 to September 2020, skin biopsies were obtained from dogs housed in shelters (n = 557) and privately owned dogs (n = 26) in seven provinces of Iran (Hamedan, Kermanshah, Yazd, Mazandaran, Khuzestan, Lorestan, Esfahan), as well as from three road-killed jackals (Canis aureus) and three cats (Felis catus) in Hamedan province. The skin biopsies were first soaked in saline solution at room temperature overnight, and examined for dermal microfilariae under the microscope. Positive skin specimens and sediments were tested by PCR for a 304-bp region of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene and amplicons were sequenced. RESULTS: Microfilariae of Cercopithifilaria spp. were found in skin biopsies of 32 of the 583 (5.5%) dogs tested, with infection rates of up to 25% in Kermanshah. No microfilariae were recovered from skin biopsy samples collected from dogs in Khorramabad and Ahvaz, nor from the examined jackals and cats. None of the privately owned dogs were found to be infected. Morphologic and morphometric characteristics of the microfilariae were consistent with C. bainae. Eighteen skin samples were positive for the cox1 gene, of which 15 sequences showed a nucleotide identity of 100% and three of 93.4% with the reference sequence of C. bainae available in GenBank (haplotype I; GenBank accession number: JF461457). CONCLUSIONS: The data from this study broadens current knowledge on the geographical distribution of C. bainae in dogs in Middle Eastern countries. Further studies on different wild canine species in the country (e.g. jackal, fox, wolf) could provide further information on the epidemiology of these filarioids. A particular focus should be put on zoonotic O. lupi given the reports of its presence in human patients from this country.

Otimização de novos candidatos a fármacos para leishmaniose visceral canina / Optimization of new drug candidates for canine visceral leishmaniasis

A leishmaniose visceral canina se trata de enfermidade infecciosa, zoonótica, de caráter insidioso e multissistêmica. Fatores como a ineficácia dos métodos de controle aplicados sobre os reservatórios caninos (incluindo eutanásia) e a toxicidade dos fármacos atualmente disponíveis para o tratamento humano, justificam urgência na busca de novos candidatos a fármacos. O presente estudo teve por objetivo avaliar o potencial anti-Leishmania de 25 compostos sintéticos derivados do gibilimbol. Destes, quatro apresentaram atividade em amastigotas sem citotoxicidade a células de mamíferos, com o CE50 em amastigotas variando entre 5,1 e 13,5 µM, e dois (LPM-21 e LPM-22) foram submetidos à predição drug-likeness in silico e ensaios de mecanismo de ação em promastigotas de Leishmania (L.) infantum. Ambos demonstraram alta absorção gastrointestinal, permeabilidade à barreira hematoencefálica, solubilidade moderada e alta (respectivamente), ausência de moléculas de interferência, e perfil drug-likeness favorável para futuros candidatos a fármacos. Nos ensaios de mecanismo de ação, LPM-21 induziu tardiamente a permeabilização plasmática parasitária (220 minutos), além de ter ocasionado a despolarização da membrana mitocondrial, redução nos níveis de ROS, alcalinização dos acidocalcissomos e importante elevação do Ca2+ intracelular, fatos que sugerem um mecanismo de ação tendo como principal alvo de atuação os acidocalcissomos, resultando no extravasamento do Ca2+ e subsequente despolarização da membrana mitocondrial. O composto LPM-22 não alterou a permeabilidade da membrana plasmática, não atuou sobre o potencial de membrana mitocondrial, induziu a acidificação dos acidocalcissomos, elevação do Ca2+ e redução nos níveis de ROS, podendo sugerir danos à membrana mitocondrial e consequente elevação deste íon. A espectrometria de massas por MALDI-TOF revelou aumento na intensidade dos picos em promastigotas incubadas com LPM-21 e LPM-22. Logo, este trabalho concretiza o potencial anti-Leishmania dos derivados sintéticos do gibilimbol, em especial os compostos LPM-21 e LPM-22, os colocando como futuros candidatos para estudos in vivo. (AU)

Canine visceral leishmaniasis is an infectious, zoonotic, insidious and multisystemic disease. Factors such as the ineffectiveness of the control methods applied to canine reservoirs (including euthanasia) and the toxicity of the drugs currently available for human treatment, brings the urgency in search for new drug candidates. The present study aimed to evaluate the anti-Leishmania potential of 25 synthetic compounds derived from gibilimbol, a phenolic isolate extracted from the leaves of Piper malacophyllum. Four compounds showed activity in amastigotes without cytotoxicity to mammalian cells, with EC50 in amastigotes ranging between 5.1 and 13.5 µM, and two (LPM-21 and LPM-22) were submitted to the in silico drug-likeness prediction, and mechanism of action assays in Leishmania (L.) infantum promastigotes. Both demonstrated high gastrointestinal absorption, permeability to the blood-brain barrier, moderate and high solubility (respectively), and favorable drug-likeness profile for future drug candidates, without revealing PAINS molecules. In the mechanism of action assays, LPM-21 induced parasitic plasmatic permeabilization only at later times (220 minutes), in addition to depolarization of the mitochondrial membrane, reduction in ROS levels, acidocalcisomes alkalinization and significant increase in intracellular Ca2+, suggesting a mechanism of action targetting acidocalcisomes, resulting in extravasation of Ca2+ and subsequent depolarization of the mitochondrial membrane. In contrast, LPM-22 did not alter the permeability of the plasma membrane and the mitochondrial membrane potential, resulting in acidification of acidocalcisomes, elevation of Ca2+ and reduction in ROS levels, which may suggest damage to the mitochondrial membrane and consequent calcium elevation. Mass spectrometry by MALDI-TOF revealed an increase in peak intensity in promastigotes incubated with LPM-21 and LPM-22. Finally, this work realizes the anti-Leishmania potential of synthetic gibilimbol derivatives, especially the compounds LPM-21 and LPM-22, placing them as future candidates for in vivo studies. (AU)

Ocorrência de Babesia sp, Ehrlichia canis e Hepatozoon canis em cães domiciliados, em dois municípios do estado do Espírito Santo - Brasil / Occurrence of Babesia sp, Ehrlichia canis and Hepatozoon canis in domiciled dogs in two municipalities of the state of Espírito Santo - Brazil / Ocurrencia de Babesia sp, Ehrlichia canis y Hepatozoon canis em perros domiciliados em dos municipios del estado de Espírito Santo - Brasil

As doenças transmitidas por carrapatos são afecções de grande importância na clínica médica de pequenos animais, devido à alta casuística e ampla distribuição vetorial no território brasileiro. Os principais agentes responsáveis pelas infecções em cães são Babesia sp., Ehrlichia canis e Hepatozoon canis. Os animais infectados são assintomáticos ou apresentam sinais clínicos inespecíficos, sendo necessário a utilização de testes diagnósticos para definição do agente etiológico, e diagnóstico seguro. O objetivo do presente estudo foi determinar a ocorrência desses micro-organismos em cães naturalmente infectados, domiciliados nos municípios de Vila Velha e Anchieta, Espírito Santo, utilizando diferentes testes de detecção: Reação em cadeia polimerase (PCR), sorologia para detecção de anticorpos anti Ehrlichia canis e pesquisa de hematozoários em esfregaço sanguíneo. Foram analisadas 65 amostras de sangue obtidas por venopunção de veia cefálica de cães. No teste de PCR, 4,62% dos animais foram positivos para Babesia vogeli e 1,54% para Ehrlichia canis sendo os resultados para Hepatozoon canis negativos. No teste sorológico para E. canis 90,77% dos animais foram positivos para a presença de anticorpos, e na pesquisa em lâminas de esfregaço sanguíneo 3,02% apresentavam outros hemoparasitas. Os resultados indicam a dispersão desses hemoparasitas na população canina da região de estudo, entretanto com baixa ocorrência. O teste de PCR demonstrou-se como o mais sensível no qual Babesia vogeli foi o agente mais observado.(AU)

Tick-borne diseases are diseases of great importance in the medical practice of small animals, due to the high casuistry and wide vectorial distribution in the Brazilian territory. The main agents responsible for infections in dogs are Babesia sp., Ehrlichia canis and Hepatozoon canis. Infected animals are asymptomatic or present nonspecific clinical signs, requiring the use of diagnostic tests to define the etiologic agent, and safe diagnosis. The objective of the present study was to determine the occurrence of these microorganisms in naturally infected dogs domiciled in the municipalities of Vila Velha and Anchieta, Espírito Santo, using different detection tests: polymerase chain reaction (PCR), serology to detect antibodies against Ehrlichia canis and research of hematozoa in blood smears. Sixty-five blood samples obtained by venipuncture of the cephalic vein of dogs were analyzed. In the PCR test, 4.62% of the animals were positive for Babesia vogeli and 1.54% for Ehrlichia canis, and the results for Hepatozoon canis were negative. In the serological test for E. canis, 90.77% of the animals were positive for the presence of antibodies, and in the research in blood smear slides, 3.02% presented other hemoparasites. The results indicate the dispersion of these hemoparasites in the canine population of the study region, however with low occurrence. The PCR test proved to be the most sensitive, in which Babesia vogeli was the most observed agent.(AU)

Las enfermedades transmitidas por garrapatas son enfermedades de gran importancia en la práctica médica de los pequeños animales, debido a la alta casuística y amplia distribución vectorial en el territorio brasileño. Los principales agentes responsables de las infecciones en los perros son Babesia sp., Ehrlichia canis y Hepatozoon canis. Los animales infectados son asintomáticos o presentan signos clínicos inespecíficos, siendo necesario el uso de pruebas diagnósticas para la definición del agente etiológico, y el diagnóstico seguro. El objetivo del presente estudio fue determinar la ocurrencia de estos microorganismos en perros infectados naturalmente, domiciliados en los municipios de Vila Velha y Anchieta, Espírito Santo, utilizando diferentes pruebas de detección: reacción en cadena de la polimerasa (PCR), serología para detectar anticuerpos anti Ehrlichia canis e investigación de hematozoos en frotis de sangre. Se analizaron sesenta y cinco muestras de sangre obtenidas por venopunción de la vena cefálica de los perros. En la prueba PCR, el 4,62% de los animales fueron positivos para Babesia vogeli y el 1,54% para Ehrlichia canis, y los resultados para Hepatozoon canis fueron negativos. En la prueba serológica para E. canis, el 90,77% de los animales fueron positivos a la presencia de anticuerpos, y en la investigación en láminas de frotis de sangre el 3,02% presentaron otros hemoparásitos. Los resultados indican la dispersión de estos hemoparásitos en la población canina de la región de estudio, aunque con una baja presencia. La prueba PCR resultó ser la más sensible, en la que Babesia vogeli fue el agente más observado.(AU)

[Establishment and preliminary application of a recombinase-aided isothermal amplification assay-based multiplex nucleic acid assay for detection of three Echinococcus species].

OBJECTIVE: To establish a multiplex nucleic acid assay for rapid detection of Echinococcus multilocularis, E. granulosus and E. shiquicus based on the recombinase-aided isothermal amplification assay (RAA) and to preliminarily assess its diagnostic efficiency. METHODS: The mitochondrial genomic sequences of E. multilocularis (GenBank accession number: NC_000928), E. granulosus (GenBank accession number: NC_044548) and E. shiquicus (GenBank accession number: NC_009460) were used as target sequences, and three pairs of primers were designed based on the RAA primer design principle and synthesized for the subsequent multiple RAA amplification. The genomic DNA of E. multilocularis, E. granulosus and E. shiquicus at different concentrations and the recombinant plasmids containing the target gene at various concentrations were amplified to evaluate the diagnostic sensitivity of the multiplex RAA assay, and the genomic DNA of E. multilocularis, E. granulosus, E. shiquicus, Taenia multiceps, T. saginata, T. asiatica, Dipylidium caninum, T. hydatigena, Toxocara canis, Fasciola hepatica, T. pisiformis, Mesocestoides lineatus and Cryptosporidiumn canis was detected using the multiplex RAA assay to evaluate its specificity. In addition, the reaction condition of the multiplex RAA assay was optimized, and was then employed to detect the tissues with echinococcosis lesions, simulated canine fecal samples and field captured fox fecal samples to examine its application values. RESULTS: The multiplex RAA assay was effective to specifically amplify the mitochondrial gene fragments of E. multilocularis, E. granulosus and E. shiquicus within 40 min at 39 °C, with sequence lengths of 540, 430 bp and 200 bp, respectively. This multiplex RAA assay showed the minimum detection limits of 2.0, 2.5 pg/µL and 3.1 pg/µL for detection of the genomic DNA of E. multilocularis, E. granulosus and E. shiquicus, and presented the minimum detection limit of 200 copies/µL for detection of the recombinant plasmids containing E. multilocularis, E. granulosus and E. shiquicus target genes. This multiplex RAA assay was effective to simultaneously detect single and multiple infections with E. multilocularis, E. granulosus and E. shiquicus, but failed to amplify the genomic DNA of T. multiceps, T. saginata, T. asiatica, D. caninum, T. hydatigena, T. canis, F. hepatica, T. pisiformis, M. lineatus and C. canis. In addition, the optimized multiplex RAA assay was effective to detect all positive samples from the tissue samples with echinococcosis lesions, simulated canine fecal samples and field captured fox fecal samples, which was fully consistent with the detection of the single PCR assay. CONCLUSIONS: A sensitive and specific multiplex nucleic acid assay for rapid detection of E. multilocularis, E. granulosus and E. shiquicus has been successfully established.

Prevalence and risk factors of Taenia hydatigena in dogs, pigs, and cattle in the Central Highlands of Vietnam.

Taenia hydatigena is a globally distributed canine tapeworm. The canine tapeworm results in economic impacts for farmers owing to organ condemnation. T. hydatigena utilizes dogs and other carnivores as definitive hosts while swine and ruminants serve as intermediate hosts. T. hydatigena is endemic in Vietnam; however, information on the prevalence and risk factors associated with infection is scarce. This cross-sectional study aims to identify the prevalence of T. hydatigena taeniasis in dogs and T. hydatigena cysticercosis in pigs and cattle. The risk factors associated with taeniasis in dogs were identified using a fixed effects logistic regression model and quantified using population attributable fractions. The prevalence of T. hydatigena taeniasis in dogs and T. hydatigena cysticercosis in pigs and cattle was 10.31% (95% CI 8.21 to 12.84%), 7.60% (95% CI 6.34 to 9.08%), and 11.11% (95% CI 7.63 to 11.81%), respectively; these levels were lower than those reported in other regions of Vietnam. The population attributable fraction of risk factors associated with T. hydatigena taeniasis in dogs for dogs living in proximity of an abattoir, those having access to raw pork or beef viscera, and those living in multi-dog households were 80%, 19%, and 7%, respectively. This current study identified and quantified the risk factors for taeniasis in dogs thus advocating for targeted community intervention programs to break the lifecycle of T. hydatigena in Dak Lak province.

Esophageal spirocercosis with pulmonary egg deposition and secondary hypertrophic osteopathy in a dog from Costa Rica.

Spirocerca lupi is the etiologic agent of spirocercosis in canids from the tropical and subtropical regions of the world. A 1-year-old dog was presented with weight loss and diarrhea. Upon examination, thickened limbs, a mass in the lung's right middle lobe and an esophageal mass were found. The pulmonary mass aspirate revealed S. lupi eggs surrounded by a purulent infiltrate. Limb bone biopsies showed irregular mineralization and cartilaginous metaplasia. Treatment with doramectin resolved the initial clinical signs. However, regurgitation was reported 5 months later. Therefore, the dog received additional doramectin and later vomited a worm morphologically consistent with a S. lupi adult. Herein we report the esophageal migration of S. lupi with egg deposition to the lung parenchyma, accompanied by secondary hypertrophic osteopathy. This report highlights the presence of S. lupi in Costa Rica and the importance of including this parasite as a differential diagnosis when compatible signs are identified.

Investigation of possible alternate animal reservoir hosts of Opisthorchis viverrini.

Chronic opisthorchiasis caused by Opisthorchis viverrini (O. viverrini) adversely affects human health and is associated with a fatal bile duct cancer (cholangiocarcinoma). Although cats and dogs are known animal reservoir hosts of opisthorchiasis, there is limited information about whether other fish-eating animals are fluke reservoirs. Wetlands along Chi River of Thailand have abundant intermediate host snails and fish for O. viverrini and diverse other animal species. This led to our investigation into whether other fish-eating animals can also become infected and be alternate reservoir hosts for human opisthorchiasis. Our preliminary study focused on the O. viverrini infection status of crab-eating or long-tailed macaques roaming in Kosumpi National Forest Park in Chi River Basin, Kosumpisai District of Mahasarakam Province, and rodents, small residential mammals and fish-eating birds living in Lawa wetland complex of Khon Kaen Province. Fecal samples of each animal were collected and modified formalin ether concentration technique was applied to identify infections. Additionally, participatory epidemiology was used to retrieve information from local communities on opisthorchiasis transmission in these animals. No O. viverrini infection was found in any fecal samples suggesting that monkeys, rodents, small residential mammals and birds in these two wetlands probably do not serve as alternate reservoir hosts of O. viverrini.

Leishmaniose visceral em um Pastor Belga: relato de caso / Visceral leishmaniasis in a Belgian Pastor: case report

O presente trabalho tem como objetivo relatar um caso de um Pastor Belga, do município de Ponta Porã, Mato Grosso do Sul, positivo para Leishmaniose Visceral, atendido em 2017 em uma clínica veterinária localizada em Pedro Juan Caballero, Paraguai. O diagnóstico foi confirmado através dos sinais clínicos característicos, e dos exames ELISA e PCR positivos. O animal foi submetido ao tratamento clínico para melhora dos sintomas, cujo tratamento antiparasitário inicial foi realizado com a associação de estibogluconato de sódio 75 mg/kg e alopurinol 100 mg seguido de aloputinol 100mg de uso contínuo e uso da coleira antileishmaniose. Tratamento esse considerado eficiente, com melhora clínica do animal. Após 24 meses o animal foi diagnosticado com tumor de mama e lesão da bolsa escrotal, sendo submetido a tratamento clínico e cirúrgico. Com 30 e 36 meses do diagnóstico inicial repetiu-se os exames ELISA (positivo) e PCR (negativo), e então o animal foi considerado curado clinicamente devido à ausência de sinais clínicos. Tendo em vista a complexidade dos fatores no ciclo de transmissão, conclui-se que as medidas em saúde ainda são insuficientes para o controle efetivo da doença. É importante o papel do Médico Veterinário na saúde pública, devido a obrigatoriedade de notificação de casos de Leishmaniose Visceral Canina, sendo necessários esforços nas diferentes áreas da saúde animal, humana e do meio ambiente, visando medidas de vigilância e controle da doença no país.

The present work aims to report a case of a Belgian Shepherd, from the municipality of Ponta Porã, Mato Grosso do Sul, positive for Visceral Leishmaniasis, treated in 2017 in a veterinary clinic located in Pedro Juan Caballero, Paraguay. The diagnosis was confirmed through the characteristic clinical signs, and the positive ELISA and PCR tests. The animal was submitted to clinical treatment for improvement of symptoms, whose initial antiparasitic treatment was performed with the association of sodium stibogluconate 75 mg/kg and allopurinol 100 mg followed by alloputinol 100mg of continuous use and use of the antileishmaniasis collar. This treatment was considered efficient, with clinical improvement of the animal. After 24 months the animal was diagnosed with a breast tumor and scrotum injury, and was submitted to clinical and surgical treatment. At 30 and 36 months from the initial diagnosis, the ELISA tests (positive) and PCR (negative) were repeated, and then the animal was considered clinically cured due to the absence of clinical signs. Considering the complexity of the factors in the transmission cycle, it is concluded that the health measures are still insufficient for the effective control of the disease. The role of the veterinarian in public health is important, due to the obligatory notification of cases of Canine Visceral Leishmaniasis, being necessary efforts in the different areas of animal health, human and environment, aiming at measures of surveillance and control of the disease in the country.

Occurrence and molecular characterization of Giardia duodenalis from naturally infected dogs in the municipality of Santa Maria, Rio Grande do Sul, Brazil / Ocorrência e caracterização molecular de Giardia duodenalis em cães naturalmente infectados no município de Santa Maria, Rio Grande do Sul, Brasil

Giardiasis is an important and prevalent zoonosis in dogs and humans caused by Giardia spp. The close relationship between pets and humans has physical, emotional and social benefits. The dogs have an important role in Giardia duodenalis cycle and transmission. This study aimed to verify the occurrence of the parasite in dogs from Central Region, in Santa Maria, Rio Grande do Sul State, Brazil, from April to October 2018. Dog feces (230) were submitted to Faust coproparasitological and molecular analyses. The positive samples in the nested-PCR (β-giardin gene) were sent for DNA sequencing and phylogenetic analyses (Neighbor-Joining). The occurrence of G. duodenalis, was 5.6% (13/230) and 4.3% (10/230) detected by coproparasitological technique and nested-PCR, respectively. There was no difference in the sensitivity of the tests used. From the faecal samples analyzed, there were no differences among the variables: diagnostic techniques, local, sex, and age of the animals (p>0.05). Only in the stool examination methodology a difference was observed between the ages (p<0.05). G. duodenalis assemblages were C and D, frequently reported in dogs. The close relationship between dogs and people may allow co-infections of circulating parasites in the population, including Giardia spp. and increasing the risk of transmission of zoonotic agents.(AU)

A giardíase é uma zoonose importante e prevalente em cães e humanos, sendo causada por Giardia spp. A estreita relação entre animais de estimação e seres humanos traz benefícios físicos, emocionais e sociais. Os cães têm um papel importante no ciclo e transmissão de Giardia duodenalis. Este estudo teve como objetivo verificar a ocorrência do parasita em cães da Região Central, em Santa Maria, RS, Brasil, de abril a outubro de 2018. As fezes de cães (230) foram submetidas a técnica coproparasitológica de Faust e análises moleculares. As amostras positivas no nested-PCR (gene β-giardin) foram enviadas para sequenciamento de DNA e posterior análise filogenética (Neighbor-Joining). A ocorrência de G. duodenalis foi de 5,6% (13/230) e 4,3% (10/230) detectados pela técnica coproparasitológica e nested-PCR, respectivamente. Não houve diferença na sensibilidade dos testes utilizados. Das amostras fecais analisadas, não houve diferenças entre as variáveis: técnicas de diagnóstico, local, sexo e idade dos animais (p>0,05). Somente na metodologia de exame de fezes observou-se diferença entre as idades (p<0,05). As assemblages de G. duodenalis encontradas foram C e D, frequentemente relatadas em cães. A estreita relação entre cães e pessoas pode permitir co-infecções de parasitas circulantes na população, incluindo Giardia spp. e aumentando o risco de transmissão de agentes zoonóticos.(AU)

Prurito anal en niños cubanos con dipylidiosis / Anal pruritus in Cuban children with dipylidiasis

Introducción: La dipylidiosis es una zoonosis parasitaria producida por Dipylidium caninum. La infección es más frecuente en niños que en adultos debido al estrecho contacto de los primeros con mascotas (perros y gatos). La dipylidiosis es una enfermedad benigna y autolimitada, generalmente cursa de manera asintomática, aunque en ocasiones pueden aparecer algunos síntomas. Objetivo: Exponer el caso de niños cubanos con prurito anal, forma clínica poco frecuente de presentación de la dipylidiosis. Presentación de casos: Se trata de cuatro niños de La Habana, Cuba, atendidos en la atención primaria y secundaria de salud por presentar prurito anal, acompañado de la expulsión de elementos blanquecinos móviles en las deposiciones. Se les realizó estudios de las heces y técnica de Graham, con resultados negativos. La falta de diagnóstico etiológico de una posible parasitosis, propició la remisión de los casos al Laboratorio Nacional de Referencia de Parasitología del Instituto de Medicina Tropical Pedro Kouri donde se interrogó a las madres y se indicaron exámenes de las heces, que identificaron D. caninum. Conclusión: Ante niños con prurito anal, forma clínica de presentación poco frecuente de la dipylidiosis, es necesario hacer el diagnóstico diferencial de esta taeniosis zoonótica y solicitar el diagnóstico parasitológico para confirmar la parasitosis. Los resultados negativos obtenidos en la atención primaria y secundaria de salud, hacen sospechar el desconocimiento del diagnóstico de esta zoonosis parasitaria en la red nacional de salud, que, además, produjo demora de dicho diagnóstico y su confirmación en el nivel terciario de salud(AU)

Introduction: Dipylidiasis is a parasitic zoonosis produced by Dipylidium caninum. The infection is more frequent in children than in adults due to the close contact of children with pets (dogs and cats). Dipylidiasis is a benign and self-limiting disease, usually occurs asymptomatically, although sometimes some symptoms may appear. Objective: Present the case of Cuban children with anal pruritus, a rare clinical form of presentation for dipylidiasis. Presentation of cases: Four children from Havana, Cuba, treated in primary and secondary health care services after presenting anal pruritus, accompanied by the withdraw of mobile whitish elements in the stools. Stool studies and Graham's technique were performed, with negative results. The lack of etiological diagnosis of a possible parasitosis, led to the referral of the cases to the National Reference Laboratory of Parasitology of Pedro Kouri Institute of Tropical Medicine where the mothers were questioned and stool tests were indicated, which identified D. caninum. Conclusion: In children with anal pruritus, a rare clinical form of presentation of dipylidiasis, it is necessary to make the differential diagnosis of this zoonotic taeniosis and request the parasitological diagnosis to confirm this parasitosis. The negative results obtained in primary and secondary health care services make suspect the ignorance of the diagnosis of this zoonotic taeniosis in the national health network, which, in addition, produced delay in the diagnosis and its confirmation at the tertiary level of health(AU)

Report of rare case of intense parasitism by 34 specimens of Dioctophyme renale in a dog.

Dioctophymosis is caused by Dioctophyme renale, a nematode that usually affects the right kidney of carnivores. The aim of this study was to report on a case of a dog with progressive weight loss and swollen abdomen that was diagnosed as presenting dioctophymosis. The patient underwent surgical treatment through which 34 nematodes were found, of which 18 were female and 16 were male, with a maximum length of 74 centimeters. The parasites were free in the abdominal cavity and inside the right kidney, and had caused peritonitis, free fluid, severe adherences between the abdominal organs and renal perforation. Parasitic diseases with a high number of specimens of this species are uncommon in dogs. The disease progresses with an inflammatory reaction and consequent formation of adherences and granulomatous tissue. This surrounds the eggs that were eliminated in the abdominal cavity by the free parasites. This disease occurs frequently in the city of Pelotas, Rio Grande do Sul, Brazil, where the patient of this report was living. To the best of authors' knowledge, this is the case with the largest number of specimens of D. renale removed from a single dog in vivo.

Assessment of the exposure to Echinococcus multilocularis associated with carnivore faeces using real-time quantitative PCR and flotation technique assays.

The eggs of Echinococcus multilocularis, the infectious stage, are spread into the environment through wild and domestic carnivore faeces. The spatial location of the faeces containing infective E. multilocularis eggs is a key parameter for studying areas of exposure and understanding the transmission processes to the intermediate hosts and humans. Echinococcus multilocularis faecal prevalence is often assessed by detecting E. multilocularis DNA, not necessarily eggs. This work aimed to determine the percentage of faeces containing E. multilocularis eggs in a rural town and its surroundings and whether this level of precision is relevant in assessing exposure to E. multilocularis. For this purpose, we developed a combined molecular and microscopic approach to investigate the E. multilocularis exposure of potential hosts in the environment from field-collected carnivore faeces. Carnivore defecation patterns were then spatialized to study the spatial distribution of E. multilocularis. Faeces were screened for E. multilocularis DNA using a specific real-time quantitative PCR (qPCR). Echinococcus multilocularis eggs were morphologically identified from E. multilocularis-specific qPCR-positive faeces after sucrose flotation and individually confirmed through specific PCR and sequencing. The spatial distribution of E. multilocularis was studied using Kulldorff statistics. Echinococcus multilocularis eggs were identified mostly in fox faeces positive for E. multilocularis DNA by qPCR (n = 27/70) and only from 1 of 15 copro-samples from dogs and 1 of 5 from cats. The faecal prevalence of E. multilocularis DNA and eggs was overdispersed, with the same geographical patterns. These data suggest that E. multilocularis DNA and/or egg detection in carnivore faeces, mainly that of foxes, is appropriate in ecological studies of E. multilocularis transmission.

Case Report: Autochthonous Case of Human Visceral Leishmaniasis in the West Bank, Palestine.

Human visceral leishmaniasis (HVL) is a parasitic disease infecting children in the Mediterranean region. Here, we portray a case of a 2-year-old child with an epidemiological description of the situation surrounding the case. The patient was suffering from recurrent fever, weakness, and abdominal discomfort associated with loss of appetite. Routine blood investigations showed pancytopenia, whereas examination revealed hepatomegaly. A diagnosis of HVL was made by demonstrating amastigotes in a Giemsa-stained smear from a bone marrow aspirate followed by genotyping by PCR and sequencing. In conclusion, early detection of VL infection followed by appropriate treatment protocols is essential to saving the patient.

Diversity of Taenia and Hydatigera (Cestoda: Taeniidae) in domestic dogs in Kenya.

Taenia species of domestic dogs can cause cysticercosis and coenurosis in a wide range of intermediate hosts including humans. Most taeniids of dogs are globally distributed, but some wildlife-transmitted species can be specific for certain regions. Generally, little information exists on the species composition and frequency in most regions of the world, which impairs risk assessment and control strategies. This study determined the range of taeniid species in dogs in four widely spaced areas of Kenya by genetic identification of eggs in faeces collected from the environment. Individual taeniid eggs were characterised by nested polymerase chain reaction of NADH dehydrogenase subunit 1 and cytochrome C oxidase 1 genes, restriction fragment length polymorphism and partial sequencing. Overall 79/1621 (4.9%) faecal samples contained eggs of Taenia or Hydatigera (8.0% in Turkana, 4.8% in Isiolo, 3.8% in Maasai Mara and 1.3% in Meru). Taenia hydatigena and T. multiceps were the most frequent, found in 36 and 15 samples, respectively. Other eggs found in the faeces belonged to T. serialis (sensu lato), T. madoquae (the first record in domestic dogs), T. ovis, T. saginata and Hydatigera taeniaeformis. Polymorphism of nad1 sequences revealed 22 and 8 haplotypes of T. hydatigena and T. multiceps, respectively. The results show the involvement of dogs in both domestic and sylvatic transmission cycles. In addition to the species range, this study provides data on the intraspecific diversity of T. hydatigena and T. multiceps in Kenya, which will serve as baseline information for further studies into cysticercosis and coenurosis in livestock and humans in the region.

The fox tapeworm, Echinococcus multilocularis, in grey wolves and dogs in Slovakia: epidemiology and genetic analysis.

Echinococcus multilocularis, the causative agent of human alveolar echinococcosis, is an important emerging parasite in the northern hemisphere. In epidemiological studies, the highest attention is being paid to foxes as the main reservoir hosts responsible for geographic expansion from multiple focal populations and the invasion of urban habitats, but little information is available on the parasite distribution in other carnivores. Hence, the study was designed to obtain updated information about the occurrence and genetic diversity of E. multilocularis in grey wolves and dogs in Slovakia. Faecal samples of wolves were collected from three locations under a certain level of environmental protection in the central and eastern parts of the country, and the presence of the parasite DNA was detected in 35.7% of 112 samples, with the highest rate (51.2%) recorded in the Poloniny National Park in north-eastern Slovakia. Among 110 faecal dog samples, E. multilocularis was detected in three faeces from segregated Roma settlements in the eastern part of the country, which accounted for an overall positivity of 2.7%. Sequence analysis of two mitochondrial genes, 12S rRNA and NADH dehydrogenase subunit 1, revealed four haplotypes in 13 isolates from wolves and dogs originating from four sites in eastern and central Slovakia, with all samples bearing a European-type pattern of E. multilocularis. The more than one-third positivity rate of E. multilocularis in wolf faecal samples dispersed over a large part of the country has corroborated the extensive circulation of the parasite in wildlife and confirmed the need to improve intervention control strategies.

Rhodnius prolixus Colonization and Trypanosoma cruzi Transmission in Oil Palm (Elaeis guineensis) Plantations in the Orinoco Basin, Colombia.

Trypanosoma cruzi is the etiological agent of Chagas disease that infects more than seven million people in Latin America. The parasite is transmitted by triatomine insects, of which some species are often associated with palms. The establishment of oil palm plantations (Elaeis guineensis) in the Orinoco region (Colombia) has been rapidly growing, possibly constituting a new environment for the establishment and increase in triatomine populations. In this study, the potential of Rhodnius prolixus to colonize E. guineensis plantations and maintain T. cruzi transmission was assessed. Fieldwork was conducted in two areas located in the department of Casanare for sampling E. guineensis and Attalea butyracea palms, sampling for triatomines to determine their abundance and prevalence of T. cruzi infection. To assess T. cruzi transmission potential in the area, sylvatic and domestic mammals were sampled. Results showed that palm infestation with triatomines was higher in A. butyracea than in E. guineensis palms and T. cruzi infection in triatomines varied between habitats for one study area, but was constant in the other site. Trypanosoma cruzi-infected mammals in the E. guineensis plantations were mainly generalist rodents, suggesting that these mammals could have an important role in T. cruzi transmission in plantations. In conclusion, E. guineensis plantations in the Orinoco region are suitable habitats for R. prolixus and T. cruzi transmission.