Dataset Information

Comparative characterization of microRNAs from the liver flukes Fasciola gigantica and F. hepatica.

ABSTRACT: MicroRNAs (miRNAs) are key regulators of gene expression at the post-transcription level. The present study specifically explored and compared the miRNA expression profiles of F. gigantica and F. hepatica using an integrated sequencing and bioinformatics platform and quantitative real-time PCR. Nineteen and 16 miRNA candidates were identified from F. gigantica and F. hepatica, respectively. The two parasites shared 11 miRNAs, with 8 also showing similarity to miRNAs of Schistosoma japonicum. Another 8 miRNAs were identified as F. gigantica-specific and 5 as F. hepatica-specific, most of which were novel. Predicted target analysis with 11465 mRNA and EST sequences of F. hepatica and F. gigantica revealed that all of the miRNAs had more than one target, ranging from 2 to 398 with an average of 51 targets. Some functions of the predicted targets were only found in F. gigantica, such as "transcription regulator", while some others were only found in F. hepatica, such as "reproduction" and "response to stimulus", indicating the different metabolism and gene regulation patterns of the two parasites. The present study represents the first global comparative characterization of miRNA expression profiles of F. gigantica and F. hepatica, which has provided novel valuable resources for a better understanding of the two zoonotic trematodes.

SUBMITTER: Xu MJ

PROVIDER: S-EPMC3534066 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

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Comparative characterization of microRNAs from the liver flukes Fasciola gigantica and F. hepatica.

Xu Min-Jun MJ Ai Lin L Fu Jing-Hua JH Nisbet Alasdair J AJ Liu Qing-You QY Chen Mu-Xin MX Zhou Dong-Hui DH Zhu Xing-Quan XQ

PloS one 20121231 12

MicroRNAs (miRNAs) are key regulators of gene expression at the post-transcription level. The present study specifically explored and compared the miRNA expression profiles of F. gigantica and F. hepatica using an integrated sequencing and bioinformatics platform and quantitative real-time PCR. Nineteen and 16 miRNA candidates were identified from F. gigantica and F. hepatica, respectively. The two parasites shared 11 miRNAs, with 8 also showing similarity to miRNAs of Schistosoma japonicum. Ano ...[more]

PMID: 23300925

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Project description:Fasciola gigantica, responsible for the zoonotic disease fasciolosis, pose a great threat to the livestock and human health worldwide. The triclabendazole (TCBZ) has been used for decades as a broad spectrum anthelmintic to control this perilous disease but the emergence of resistance in flukes against TCBZ has prompted researchers across the world to explore for new drugs and antigenic targets. World Health Organization has strongly recommended the utilization of neurobiologically significant biomolecules as new drug/antigenic targets because of their significant role in the physiology of parasites. Monoamine Oxidase (MAO) is an important neurobiological enzyme which catabolizes aminergic neurotransmitters thus preventing prolonged excitation of neurons and in non-neuronal cells it prevents cellular toxicity due to accumulation of toxic monoamines. Owing to the important role of MAO in the survival and perpetuation of parasites, multipronged approaches were undertaken for the characterization of MAO-A in F. gigantica. The activity of MAO was found to be 1.5 times higher in the mitochondrial samples than the whole homogenate samples. The adult worms of the F. gigantica appeared to possess both the isoforms of MAO i.e., MAO-A and MAO-B. The zymographic studies revealed strong enzyme activity in its native state as assessed through prominent dark bands at 250KDa in the zymogram. The enzyme was also found to be highly immunogenic as revealed by high antibody titer at 1:6400 dilution. The immunogenicity of MAO-A enzyme was further established in the Western Blots in which a strong band of 50KDa was distinctly evident. Despite ubiquitous presence of MAO in F. gigantica some regions like tegumental surface and intestinal caecae displayed strong immunofluorescence as compared to other regions. The detection of MAO-A in the F. gigantica samples in Dot-Blot assay indicate a great potential of this molecule for the immunodiagnostics of fasciolosis, particularly in the field conditions. The enzyme activity was sensitive to the specific inhibitor clorgyline in a concentration dependant manner, particularly in the late incubation period. The zymographic results also exhibited similar trend. The strong intensity of spots in Dot-blots indicate high immunogenicity of the MAO protein. The intensity of bands/spots in the samples of worms treated with clorgyline also declined, clearly indicating that the tropical liver fluke possesses prominent MAO-A activity.

Project description:Fasciolosis is a food- and water-borne disease caused by digenean trematode species, Fasciola hepatica and F. gigantica. They are widely distributed and infect a wide range of definitive hosts, causing enormous economic loss due to reduced productivity in domestic ruminants. The two species have been previously reported to be co-endemic in KwaZulu-Natal and Mpumalanga provinces of South Africa. Hybridization between the two species has been reported elsewhere. Despite the overlap of the two species in two provinces, there has been no attempt to determine the presence of the intermediate forms or hybrids. Therefore, this study aimed at morphological and molecular characterization of Fasciola spp. collected from cattle slaughtered at abattoirs located in the two provinces of South Africa, where two species are endemic. A total of 71 fluke specimens were collected cattle from abattoirs in Enhlazeni and Nelspruit in Mpumalanga province and Pietermaritzburg in KwaZulu-Natal province of South Africa, and Zimbabwe (Bulawayo abattoir). Fasciola gigantica specimen collected from Zimbabwe where it has been confirmed as the only species occurring and this was used as control in the morphological and molecular assessment of the collected specimens. Of the 71 specimens collected, 37 were classified as F. hepatica, 12 as F. gigantica and 22 as Fasciola spp using morphological characters. Of these species, 11 of 37 F. hepatica and 6 of 22 Fasciola sp were found to be aspermic or having very scanty sperm. Fifteen flukes which were spermatic were all identified morphologically as F. gigantica whilst 5 flukes which were aspermic were identified morphologically as F. hepatica. Molecular analysis of the same 15 spermatic specimens confirmed the presence of F. hepatica (n = 9) and F. gigantica (n = 6) using the CO1 marker and as F. hepatica (n = 4), F. gigantica (n = 7) and Fasciola sp. (n = 1) for the same specimens using the ITS-1/5.8S/ITS-2 marker. The remaining 4 aspermic flukes (one did not resolve) were all identified morphologically as F. hepatica and molecular analysis confirmed them as F. hepatica (n = 4) by both CO1 and ITS-1/5.8S/ITS-2. Phylogenetic analysis based on both CO1 and ITS-1/5.8S/ITS-2 showed that F. hepatica species formed a moderately supported monophyletic clade with F. gigantica. Their ancestral history was further confirmed by haplotype network, which formed novel haplotypes that corresponded with the structure of the phylogenetic tree. Results from this study showed that morphological characters alone have limitations in identifying F. hepatica and F. gigantica in areas where the two species occur, although both methods confirmed the presence of F. gigantica occurring in Zimbabwe, F. hepatica in KwaZulu-Natal, and both species occurring in Mpumalanga province. Therefore, the use of morphological techniques, complemented by molecular techniques are recommended, especially in endemic areas where the two species are co-endemic.

Dataset Information

Comparative characterization of microRNAs from the liver flukes Fasciola gigantica and F. hepatica.

Publications

Comparative characterization of microRNAs from the liver flukes Fasciola gigantica and F. hepatica.

Similar Datasets

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