Sensitivity comparison for the Leishmania spp. detection in different canine tissues using PCR-HRM

ABSTRACT Background: Leishmaniasis is a vector-borne disease caused by a parasite protozoon from the genus Leishmania. Among the molecular techniques applied for detecting these parasites, real-time PCR with High Resolution Melting (PCR-HRM) proved advantageous since it simultaneously determines both the presence and species of the pathogen in one step, through amplification and later analysis of curves generated by melting temperature. Methods: Based on this molecular technique, the goal of this study was to estimate the PCR-HRM sensitivity for Leishmania spp. detection in different canine tissues by evaluating biological samples obtained from popliteal, submandibular, and pre-scapular lymph nodes, from bone marrow and ear pinnae of 28 stray dogs captured in the metropolitan area of Asunción (Paraguay). Results: The rk39 immunochromatographic test showed that 25/28 tested dogs (89%) presented antibodies against L. infantum. In 20/25 dogs that tested positive for rk39 (80%), it was possible to detect Leishmania spp. by PCR-HRM and determine that the species corresponded entirely to L. infantum. Regarding the analysis of different tissues, the parasite was detected in all popliteal lymph node samples, followed by high detection in submandibular (at 95%) and pre-scapular lymph nodes (at 90%), bone marrow (at 85%), and ear pinnae (at 85%). Conclusions: This study demonstrated that the use of real-time PCR-HRM using the molecular marker hsp70 was a highly sensitive method for simultaneously detecting and identifying Leishmania species in different tissues taken from infected dogs. In addition, the usefulness of ear pinnae as easily accessible tissue for molecular diagnosis was emphasized.

Prevalence of intestinal parasite infections in stray and farm dogs from Spain / Prevalência de infecções por parasitos intestinais em cães errantes e de fazenda na Espanha

Abstract Dogs play a potential role as reservoirs for zoonotic parasites, being especially problematic uncontrolled dog populations such as stray and farm dogs with access to populated areas. In order to investigate the prevalence of canine intestinal parasites in at-risk dog populations, we tested a total of 233 faecal samples shed by stray and dairy farm dogs from northern Spain. Telemann method was used to detect the presence of eggs and (oo)cysts of common dog intestinal parasites and Cryptosporidium was detected by PCR. One hundred and forty eight out of 233 samples (63.5%) were positive for at least one intestinal parasite, being Ancylostomidae (35.6%; 83/233) and Trichuris (35.2%; 82/233) the parasites most frequently identified. Cryptosporidium DNA was not detected in any of the faecal samples analysed. The overall prevalence was significantly higher in stray dogs than in farm dogs (72.5% vs 58.8%). Specifically, stray dogs had a significantly higher prevalence of Ancylostomatidae, Toxocara, Toxascaris and Taenidae. These dog populations are an important source of environmental contamination with intestinal parasite forms, which could be of significance to animal and human health.

Resumo Os cães desempenham um importante papel como reservatório de parasitos zoonóticos, sendo especialmente problemáticas as populações descontroladas, como a de cães errantes e de fazenda, com acesso às áreas povoadas. Para investigar a prevalência de parasitos intestinais em populações caninas de risco, foram analisadas 233 amostras fecais provenientes de cães de fazendas leiteiras e errantes do norte da Espanha. O método Telemann foi utilizado para detectar ovos, cistos e oocistos dos parasitos caninos mais comuns e para a detecção de Cryptosporidium foi utilizada a técnica da PCR. Cento e quarenta e oito de 233 amostras analisadas (63,5%) foram positivas para pelo menos um parasito intestinal, sendo Ancyostomatidae (35,6%; 83/233) e Trichuris sp. (35,2%; 82/233) os parasitos identificados com maior frequência. O DNA de Cryptosporidium sp. não foi detectado em nenhuma das amostras fecais analisadas. A prevalência geral foi significativamente maior em cães errantes do que em cães de fazenda (72,5% vs 58,8%). Especificamente, os cães errantes tiveram prevalência maior para Ancylostomatidae, Toxocara, Toxascaris e Taenidae. Essas populações de cães são importantes fontes de contaminação ambiental, pois eliminam formas de vida desses parasitos, que podem ter impacto na saúde animal e humana.

Increasing number of stray dogs- awaited human rabies disaster in Chamba town of Himachal Pradesh

Chamba is an   ancient small town of Himachal Pradesh. The town is located at an altitude of 996 metres (3,268 ft) on the bank of the Ravi River.The population  town is 19930 [Census 2011]. Since it is headquarter of district, the offices of various departments are situated here. There are large numbers of temples, palaces and because of its scenic beauty there is continuous visit of tourists to this place. All these conditions lead to heavy crowd of people in the town round the year.

Stray dogs in urban fragments: relation between population's perception of their presence and socio-demographic factors / Cães errantes em fragmentos urbanos: relação entre o relato de visualização de cães errantes e fatores sociodemográficos da população local

Stray dogs are associated with environment and public health impacts. São Paulo is a city with approximately 2.5 million owned dogs and until the time of writing no studies describing the distribution of the stray dog population have been conducted in this city. Therefore, no scientific support for intervention plans is available. The objective of this study was to evaluate the association of local sociodemographic and environmental factors with the population perception of presence of stray dogs in urban fragments of São Paulo city. A convenience sample of six urban fragments was selected. In-depth interviews were performe, using a systematic random sample of households in each study area, between October/2010 and January/2011 to identify socio-demographic factors of interest and determine the population's perception of stray dogs. Additionally, the presence of stray dogs was estimated by photographic mark-recapture method. The degree of human-dog proximity and environmental factors such as waste handling were associated to the sighting of stray dogs. Stray dogs were observed in two of the six study areas, agreeing with the greater population perception of this issue on those areas. Intervention in these factors and encouragement of responsible ownership are two potential solutions for management and reduction of problems associated with the presence of stray dogs.(AU)

A presença de cães errantes está associada a impactos no meio ambiente e na saúde pública. São Paulo é uma cidade com aproximadamente 2.5 milhões de cães domiciliados e até o presente momento, não foram desenvolvidos estudos que descrevam a distribuição da população errante na cidade. Desta forma, não há suporte científico para planejar intervenções. O objetivo deste estudo foi avaliar a associação entre os fatores sociodemográficos e ambientais da população local com a frequência com que relataram a visualização de cães errantes em fragmentos urbanos da cidade de São Paulo. Foi definida uma amostra de conveniência composta por seis fragmentos urbanos, onde um questionário foi aplicado em uma amostra sistemática dos domicílios de cada área de estudo. Este questionário, aplicado entre outubro de 2010 e janeiro de 2011, determinou os fatores socio-econômicos de interesse e a percepção da presença de cães errantes pela população. Também foi estimada a presença de cães pelo método de marcação e recaptura fotográfica. Foi possível estabelecer uma relação entre a visualização de cães errantes pela população local e o grau de proximidade com cães e o manejo de resíduos do domicílio. Cães errantes foram observados em duas das seis áreas trabalhadas, em concordância com a maior percepção da sua presença pela população local. Intervenções nestes fatores e incentivo a guarda responsável são duas ações com potencial de contribuir para redução dos problemas causados pela presença de cães errantes.(AU)

Echinococcosis and other parasitic infections in domestic dogs from urban areas of an argentinean Patagonian city

In urban populations of South America, dogs with free access to public areas represent a public health concern. The primary consequence of roaming dogs on human health is the transmission of infectious and parasitic diseases mainly through feces contamination. The main diseases likely to be transmitted are hydatidosis or echinococcosis, larva migrans, and giardiasis. In Argentina, hydatidosis ranks among the most prevalent zoonosis. Although it is considered a rural disease, the circulation of this parasite in urban areas has been documented. The aim of this work was to survey intestinal parasites in canine feces from two low-income urban neighborhoods of Bariloche city, Argentina, and to assess their seasonal variation. During 2016, 188 fresh dog feces were collected from sidewalks in 40 randomly selected blocks from the neighborhoods. Each sample was processed by Sheater flotation and tested for a coproantigen (CAg) by ELISA. The percentage of parasitized feces was 65.3% (95% CI: 55.9%-73.8%). Eleven parasite species were found, 3 protozoan, 3 cestodes, and 5 nematodes. Echinococcus sp. was present in 9.3% of the samples (95% CI: 4.7%-16.1%). Canine echinococcosis rates resulted similar to rates found previously in other neighborhoods of the city. The life cycle of Echinococcus sp. is sustained in urban areas by the entry of parasitized livestock, domiciliary slaughtering, and inadequate deposition of offal. The risk of Echinococcus sp. transmission to people in these neighborhoods is very high, due to high density of free-roaming dogs and high percentages of infected feces, similar to percentages observed in rural areas.

En las poblaciones urbanas de América del Sur, los perros con acceso libre a áreas públicas representan un problema de salud pública. La principal consecuencia es la transmisión de enfermedades infecciosas y parasitarias a través de la contaminación por heces. Las principales enfermedades que pueden transmitirse son hidatidosis, larva migrans y giardiasis. En Argentina, la hidatidosis es una de las zoonosis más prevalentes y aunque es considerada una enfermedad rural, algunos estudios muestran la circulación de este parásito en zonas urbanas. El objetivo fue registrar los parásitos intestinales en heces caninas de dos barrios de bajos ingresos de la ciudad de Bariloche, Argentina, y evaluar su variación estacional. Durante 2016, se recolectaron 188 heces frescas de perros en 40 manzanas seleccionados aleatoriamente. Las heces se procesaron mediante flotación de Sheater y una prueba ELISA de coproantigeno (CAg). El porcentaje de heces parasitadas fue del 65.3% (IC 95%: 55.9%-73.8%). Se encontraron 11 especies de parásitos, 3 protozoos, 3 cestodes y 5 nematodes. Echinococcus sp. estuvo presente en el 9.3% de las heces (IC 95%: 4.7% -16.1%). La equinococosis canina mostró valores similares a estudios previos en otros barrios de la ciudad. El ciclo de vida Echinococcus sp. se mantiene en las zonas urbanas por entrada de ganado parasitado, faena domiciliaria y deposición inadecuada de vísceras. El riesgo de transmisión de Echinococcus sp. en estos barrios es alto, debido a la alta densidad de perros sueltos y al alto porcentaje de heces infectadas, similar al de las zonas rurales.

Geographic analysis on the occurrence of human and canine leptospirosis in the city of Maringá, state of Paraná, Brazil / Análise geográfica da ocorrência da leptospirose em humanos e em cães na cidade de Maringá, Paraná, Brasil

INTRODUCTION: Leptospirosis is a zoonotic disease, the primary hosts of which are wild, synanthropic, and household animals. Humans behave as terminal and accidental hosts. The prevalence of leptospirosis depends on carrier animals that disseminate the agent, on the environmental survival of this agent, and on the contact of susceptible individuals. Each serovar has one or more hosts with different adaptation levels. The focuses of leptospirosis are infected, sick, and asymptomatic animals, which are considered to be sources of environmental infection. This study aimed to determine the risk areas for leptospiral infection in stray dogs and patients diagnosed with leptospirosis from 2006 to 2008 in Maringá, State of Paraná, Brazil. METHODS: Three hundred and thirty-five stray dogs and 25 patients were studied. Serum from both animals and patients was examined by the microscopic serum agglutination test to study anti-leptospiral antibodies. To determine the risk areas and the spatial distribution of the disease, thematic maps were designed. RESULTS: Forty-one (12.2 percent) dogs positive for one or more leptospire serovars were observed, the most frequent serovars being Pyrogenes (43.9 percent), Canícola (21.9 percent), and Copennhageni (19.5 percent). Among the humans, 2 (8 percent) were positive for serovars Pyrogenes and Hardjo Prajitno and for Pyrogenes and Cynopteri. CONCLUSIONS: Spatial analysis showed that the risk for dogs and humans in the City of Maringá to become infected with leptospires exists in both the central and the peripheral areas, a fact that reinforces the relevance of this study and of continuous epidemiological and environmental surveillance actions to control the disease in animals and in humans.

INTRODUÇÃO: Leptospirose é uma zoonose que tem como hospedeiros primários os animais silvestres, sinantrópicos e domésticos. Os humanos comportam-se como hospedeiros terminais e acidentais. Sua prevalência depende dos animais portadores que disseminam o agente, de sua sobrevivência ambiental e do contato de pessoas susceptíveis. Cada sorovar tem um ou mais hospedeiros com diferentes níveis de adaptação. Os focos de leptospirose devem-se aos animais infectados, doentes e assintomáticos, considerados como fontes de infecção ambiental. O objetivo deste estudo foi determinar áreas de risco da infecção leptospírica em cães errantes e pacientes com diagnóstico de leptospirose nos anos de 2006 a 2008, em Maringá, Estado do Paraná, Brasil. MÉTODOS: Foram estudados 335 cães errantes e 25 pacientes. Os soros, tanto dos animais como dos pacientes, foram examinados pela prova de soroaglutinação microscópica (SAM), para pesquisa de anticorpos antileptospíricos. Para determinar áreas de risco e a distribuição espacial da doença foram elaborados mapas temáticos. RESULTADOS: Foram observados 41(12,2 por cento) cães positivos para um ou mais sorovares de leptospiras, e os mais frequentes foram: Pyrogenes (43,9 por cento), Canícola (21,9 por cento) e Copennhageni (19,5 por cento). Nos humanos, a positividade foi de 2 (8 por cento) para os sorovares, Pyrogenes e Hardjo Prajitno e, Pyrogenes e Cynopteri. CONCLUSÕES: A análise espacial revelou que o risco de cães e humanos, no município de Maringá, se infectar com leptospiras está presente tanto em áreas centrais como periféricas, fato que reforça a relevância deste estudo e de ações contínuas de vigilância epidemiológica e ambiental para o controle da doença tanto nos animais como no homem.

Espécies e prevalência das infestações por carrapatos em cães de rua da cidade de Porto Alegre, RS, Brasil / Species and prevalence of ticks infestations on stray dogs in Porto Alegre city, RS, Brazil

O objetivo deste trabalho foi identificar as espécies de carrapatos que parasitam os cães da área urbana de Porto Alegre e conhecer suas prevalências nesta população. Para isso, foram examinados 450 cães de rua de Porto Alegre, colhidos os carrapatos nos animais infestados e estes identificados ao estereomicroscópio por espécie e por estágio. Dentre os animais examinados, 52,44 por cento albergavam carrapatos e destes, 93,22 por cento estavam parasitados por Rhipicephalus sanguineus, 2,97 por cento por Amblyomma aureolatum e 3,81 por cento pelas duas espécies simultaneamente. Do total de carrapatos colhidos, a espécie mais abundante nos animais foi R. sanguineus representando 99,43 por cento desse total; foram encontrados ínstares de todos os estágios parasitários de R. sanguineus e só do estágio adulto de A.aureolatum.

The purpose of this study was to identify and to know the prevalence of ticks recovered from dogs of urban area of Porto Alegre, RS. Four hundred and fifty stray dogs were examined and the ticks found were identified according to the species and parasitic stage. A percentage of 52.44 percent of those dogs were infested with ticks, among these, 93.22 percent were parasitized by Rhipicephalus sanguineus, 2.97 percent by Amblyomma aureolatum and 3.81 percent by both species. R. sanguineus was the more prevalent species among infested animals representing 99.43 percent of the total number of ticks sampled. All parasitic stages of R. sanguineus were present on the stray dogs but only the adult stage of A. aureolatum was found.