Development of a copro-LAMP assay for detection of several species of Echinococcus granulosus sensu lato complex.

Cystic echinococcosis represents a significant problem in human and animal health and constitutes one of the most severe Neglected Tropical Diseases prioritized by the World Health Organization. The etiological agent is the complex Echinococcus granulosus sensu lato (s. l.), composed of several species/genotypes. Diagnosis in the definitive host and molecular epidemiology studies are important points for cystic echinococcosis control. Here we developed a new copro-LAMP assay, LAMP EGSL, for diagnosis in the definitive host for simultaneous detection of Echinococcus granulosus sensu stricto (s. s.), Echinococcus ortleppi, and Echinococcus canadensis species. Also, the analytical sensitivity, specificity and plausibility of performance in a rural context of a previously reported species-specific LAMP reaction, was evaluated. Both reactions showed high analytical sensitivity values (10 fg-100 fg DNA) and did not show cross reaction with DNA from host or other helminthic parasites. LAMP EGSL was performed with samples from an endemic area. In addition, the alkaline hydrolysis of one E. granulosus s. s. adult parasite followed by specific LAMP to E. granulosus s. s. was performed in a laboratory with low resources from another cystic echinococcosis endemic area. The results obtained suggest that LAMP EGSL represents a potential tool for canine diagnosis that could be useful for cystic echinococcosis control programs. In addition, we showed that LAMP reaction for E. granulous s. s., E. ortleppi and E. canadensis specific detection, could be useful for molecular epidemiology studies applicable to the definitive host. Both reactions were performed in endemic, rural areas without sophisticated equipment.

Occurrence of dioctophymosis in canines within a riparian zone of the Río de La Plata watercourse, in Ensenada, Buenos Aires Province, Argentina.

Dioctophymosis is a parasitic disease occasioned by the so-called "giant kidney worm", Dioctophyme renale, a nematode with an indirect life cycle. This parasite's definitive host is the mink, Mustela vison, though numerous wild and domestic mammals as well as man can serve as final hosts. The worms also can be in ectopic locations in the body. We surveyed 692 canines by ecography, urine sampling, surgery, necropsy, and clinical examination and diagnosed 244 cases of dioctophymosis (35.3%). Of the cases of dioctophymosis identified, 30.7% were obtained by ecography, 45.9% by urinalysis, and 17.6% by both those techniques -in addition to positive findings through surgery (2.5%), necropsy (2.5%), and the spontaneous elimination of the parasites (0.8%). Cases of dioctophymosis were observed in animals as young as 4months of age up to 15years. The frequency of D. renale diagnosis throughout the sampling period varied significantly. There was a statistically significant association between risk factors (swimming in the river, eating frogs, fish or eels, drinking ditch water) and the prevalence of infection. It was discussed the period missing after infection in canines.