The current study was performed to evaluate the liver function status as well as molecular characterization of the recovered worms in rats experimentally infected with F. hepatica. Sixteen male Wister rats aged 30 day...The current study was performed to evaluate the liver function status as well as molecular characterization of the recovered worms in rats experimentally infected with F. hepatica. Sixteen male Wister rats aged 30 days were randomly allocated into two groups (n = 8). The first group was infected orally with 15 viable encysted metacercaria of F. hepatica per animal. The other group was kept non-infected (control group). At zero time (before infection), the 2<sup>nd</sup>, 4<sup>th</sup>, 6<sup>th</sup>, 8<sup>th</sup>, 10<sup>th</sup>, 12<sup>th</sup> and 14<sup>th</sup> weeks post-infection (WPI), blood and serum samples were collected via puncture of retro-orbital plexus of veins from each rat. Serum enzyme level (AST and ALT) and total protein were measured, and the serum protein profile was carried out using agarose gel electrophoresis. During the period of the experiment, serum ALT and AST activities and serum total globulins significantly increased while serum total proteins and albumin markedly decreased in the infected group. On the 14<sup>th</sup> WPI, the data of the electropherogram showed that globulin fractions (α1-, β- and γ-globulin) levels were significantly increased while α2-globulin was markedly decreased in the infected group. The molecular analysis confirmed the amplification of the ITS1, ITS2 and NDI genes of F. hepatica recovered from the infected liver of rats with amplicon sizes of 630, 510 and 560 bp, respectively. Sequencing of the amplified ITS gene resulted in the determination of 3 strains (PP108836, PP108837, and PP108838). Also, analysis of the ITS2 gene resulted in obtaining 3 isolates under the accession numbers (PP109065, PP109066, and PP109067). In conclusion, fasciolosis in the rat model is suitable for routine experimental infections and caused a pronounced liver dysfunction with discharging of the Fasciola eggs in the faeces and the development of adult stages in the bile ducts. Furthermore, molecular techniques are a sensitive tool for the identification and characterisation of the Fasciola parasite.展开更多
Lymnaeid snails are key intermediate hosts for the development and survival of Fasciola spp.,the causative agent of Fascioliasis which are economically important parasites infecting humans and livestock globally.The c...Lymnaeid snails are key intermediate hosts for the development and survival of Fasciola spp.,the causative agent of Fascioliasis which are economically important parasites infecting humans and livestock globally.The current control method for treating Fascioliasis is heavily reliant on anthelmintic drugs,particularly Triclabendazole(TCBZ)which has resulted in drug-resistant parasites and poses significant risk as there are no long-term efficacious alternatives available.Sustainable control measures at the farm level could include both parasite and snail control will play an important role in Fasciola spp.control and reduce the reliance on anthelmintic drugs.Implementation of such sustainable control measures requires effective identification of snails on the property however Lymnaeid snails are small and difficult to physically locate.Snail identification using an environmental DNA approach is a recent approach in which physically locating snails are not required.Austropeplea tomentosa,is the primary intermediate snail host for F.hepatica transmission in South-East Australia and we present an in-field loop-mediated isothermal amplification and water filtering method for the detection of A.tomentosa eDNA from water samples to improve current surveillance methods.This methodology is highly sensitive with a detection limit of 5×10^(−6)ng/μL,detected in<20 minutes,with cumulative sample preparation and amplification time under 1 hour.This proposed workflow could assist in monitoring areas to determine the risk of Fascioliasis infection and implement strategies to manage snail populations to ultimately reduce the risk of infection for humans and livestock.展开更多
文摘The current study was performed to evaluate the liver function status as well as molecular characterization of the recovered worms in rats experimentally infected with F. hepatica. Sixteen male Wister rats aged 30 days were randomly allocated into two groups (n = 8). The first group was infected orally with 15 viable encysted metacercaria of F. hepatica per animal. The other group was kept non-infected (control group). At zero time (before infection), the 2<sup>nd</sup>, 4<sup>th</sup>, 6<sup>th</sup>, 8<sup>th</sup>, 10<sup>th</sup>, 12<sup>th</sup> and 14<sup>th</sup> weeks post-infection (WPI), blood and serum samples were collected via puncture of retro-orbital plexus of veins from each rat. Serum enzyme level (AST and ALT) and total protein were measured, and the serum protein profile was carried out using agarose gel electrophoresis. During the period of the experiment, serum ALT and AST activities and serum total globulins significantly increased while serum total proteins and albumin markedly decreased in the infected group. On the 14<sup>th</sup> WPI, the data of the electropherogram showed that globulin fractions (α1-, β- and γ-globulin) levels were significantly increased while α2-globulin was markedly decreased in the infected group. The molecular analysis confirmed the amplification of the ITS1, ITS2 and NDI genes of F. hepatica recovered from the infected liver of rats with amplicon sizes of 630, 510 and 560 bp, respectively. Sequencing of the amplified ITS gene resulted in the determination of 3 strains (PP108836, PP108837, and PP108838). Also, analysis of the ITS2 gene resulted in obtaining 3 isolates under the accession numbers (PP109065, PP109066, and PP109067). In conclusion, fasciolosis in the rat model is suitable for routine experimental infections and caused a pronounced liver dysfunction with discharging of the Fasciola eggs in the faeces and the development of adult stages in the bile ducts. Furthermore, molecular techniques are a sensitive tool for the identification and characterisation of the Fasciola parasite.
基金supported by Cooperative Research Centres Project(CRCP)awarded to Geneworks and La Trobe University.L.T.is supported by an Australian Research Training Program scholarship and the Tim Healy Memorial Scholarship awarded by The Department of Primary Industries South Australia(PIRSA).
文摘Lymnaeid snails are key intermediate hosts for the development and survival of Fasciola spp.,the causative agent of Fascioliasis which are economically important parasites infecting humans and livestock globally.The current control method for treating Fascioliasis is heavily reliant on anthelmintic drugs,particularly Triclabendazole(TCBZ)which has resulted in drug-resistant parasites and poses significant risk as there are no long-term efficacious alternatives available.Sustainable control measures at the farm level could include both parasite and snail control will play an important role in Fasciola spp.control and reduce the reliance on anthelmintic drugs.Implementation of such sustainable control measures requires effective identification of snails on the property however Lymnaeid snails are small and difficult to physically locate.Snail identification using an environmental DNA approach is a recent approach in which physically locating snails are not required.Austropeplea tomentosa,is the primary intermediate snail host for F.hepatica transmission in South-East Australia and we present an in-field loop-mediated isothermal amplification and water filtering method for the detection of A.tomentosa eDNA from water samples to improve current surveillance methods.This methodology is highly sensitive with a detection limit of 5×10^(−6)ng/μL,detected in<20 minutes,with cumulative sample preparation and amplification time under 1 hour.This proposed workflow could assist in monitoring areas to determine the risk of Fascioliasis infection and implement strategies to manage snail populations to ultimately reduce the risk of infection for humans and livestock.
