ABSTRACT
Abstract Giardia duodenalis is a zoonotic parasite that infects the gut of a wide range of vertebrates, including numerous wildlife species. However, little is known about this protozoan parasite in reptiles. Fecal samples from 31 wild lizards were collected in Galicia (northwest Spain) and screened for the presence of Giardia by PCR amplification and sequencing of the ITS1-5.8S-ITS2 region in the ribosomal unit. This allowed detection of the parasite in 5 samples (16.1%), and enabled identification of G. duodenalis assemblage A2 in two samples of Iberian rock lizard (Iberolacerta monticola), G. duodenalis assemblage B in other two samples of I. monticola, and G. duodenalis assemblage E in one sample of Bocage's wall lizard (Podarcis bocagei). The results obtained after PCR amplification and sequencing of the SSU-rDNA gene confirmed the presence of G. duodenalis assemblage A in two samples of I. monticola. This is the first report of G. duodenalis in free-living lizards, although further studies are needed to distinguish between actual infection and mechanical dissemination of cysts. The detection of zoonotic and livestock-specific assemblages of G. duodenalis demonstrates the wide environmental contamination by this parasite, possibly due to human activities.
Resumo Giardia duodenalis é um parasito zoonótico que infecta o intestino delgado de uma ampla gama de vertebrados, sendo detectado em numerosas espécies selvagens. No entanto, pouco se conhece sobre a presença deste parasito protozoário em répteis. Para estudar a presença de Giardia, foram obtidas amostras fecais provenientes de 31 lagartos e coletadas em diferentes localizações de Galicia (Noroeste da Espanha). Mediante a aplicação da técnica de PCR e posterior sequenciamento da região ITS1-5.8S-ITS2 da unidade ribossômica, detectou-se Giardia em 5 amostras (16,1%), identificando-se o genótipo A2 de G. duodenalis em 2 amostras de lagartos da montanha (Iberolacerta monticola), G. duodenalis genótipo B em outras 2 amostras de I. monticola e G. duodenalis genótipo E em outra amostra de lagarto de Bocage (Podarcis bocagei). Os resultados obtidos, após amplificação e sequenciamento de um fragmento do gene SSU-rDNA, confirmam a presença de G. duodenalis genótipo A em 2 amostras de I. monticola. Esta é a primeira vez que se descreve G. duodenalis em lagartos selvagens, embora sejam necessários outros estudos complementares para confirmar se estes animais sofrem uma infecção real ou se apenas atuam como disseminadores mecânicos da contaminação ambiental. Além disso, a detecção de genótipos zoonóticos e específicos de ruminantes domésticos demonstra a contaminação do ambiente selvagem por G. duodenalis, possivelmente devido à atividade humana.
Subject(s)
Animals , Giardiasis/veterinary , Giardia lamblia/classification , Lizards/parasitology , Zoonoses/parasitology , Giardiasis/parasitology , Feces/parasitologyABSTRACT
The aim of this paper was to discover the prevalence and assemblages of Giardia intestinalis, harbored in sheep and cows on familiar farms from five states of the Mexican Republic. Stool samples from 265 sheep and 174 cows were analyzed by centrifugation and flotation in zinc sulfate to search for cysts and ova. The samples with Giardia cysts were processed in a Sheather solution in order to isolate them. Afterwards, cultures were established in TYI-S-33, each one of which was the Giardia DNA source. The DNA was obtained and used as a template to amplify a fragment of the glutamate dehydrogenase (gdh) enzyme. The 430 bp amplicons were restricted with Nla IV and Rsa I in order to identify the restriction fragments length polymorphisms (RFLP's) patterns. From the cyst analysis, Giardia cysts in nine cows (5.1%) and 30 sheep (11.3%) were found. Then 10 axenic cultures (5 from sheep and 5 from cows) were set up. From the RFLP's pattern it was found that one cow had assemblage (AI), another two had a mixture of assemblages (AI + BIII) and the other two had (E + BIII). In sheep, it was found that two sheep had assemblage (AI) and the other three had a mixture of assemblages (AI + BIII). This is the first report in which zoonotic assemblages (A-I and BIII) predominance in ruminants from five states of Mexico have been demonstrated. Therefore, it is necessary to carry out further studies aimed at discovering other Giardia genotypes and transmission patterns between animals and humans in Mexico.
Con el fin de determinar la frecuencia y genotipos de Giardia intestinalis en ovinos y bovinos de traspatio de algunos estados de la República Mexicana, en este trabajo se colectaron heces de 265 ovinos y 174 bovinos, para la búsqueda de Giardia mediante coproparasitoscópicos (CPS) de concentración flotación. De las muestras fecales que resultaron positivas se obtuvieron los quistes por el método de Sheather. Los quistes se desenquistaron in vitro y los trofozoítos se mantuvieron en cultivo TYI-S-33 axénico. El ADN de los trofozoítos se obtuvo mediante extracciones fenólicas y se amplificó un segmento de ≈ 430 pb del gen de la enzima glutamato deshidrogenasa (gdh) por medio de la reacción en cadena de la ADN polimerasa (PCR), el producto se restringió con las enzimas Nla IV y Rsa I y se obtuvieron los polimorfismos de los fragmentos de restricción (RFLP). En los CPS se encontró a Giardia en nueve bovinos (5.1%) y 30 ovinos (11.3%). Se establecieron 10 cultivos axénicos (5 de bovinos y 5 de ovinos). En un bovino se encontró el genotipo (AI), dos tuvieron mezcla de los genotipos (AI + BIII) y los otros dos fueron (E + BIII). Un ovino fue genotipo (AI) y tres tuvieron mezcla de los genotipos (AI + BIII). Éste es el primer informe que presenta predominio de genotipos zoonóticos (AI y BIII) en ovinos y bovinos de México. Es necesario investigar los genotipos de Giardia y patrones de transmisión entre animales y humanos en México.