Biochemical Liver Functions and Molecular Identification of Fasciola hepatica from Experimentally Infected Rat Model

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 2nd, 4th, 6th, 8th, 10th, 12th and 14th 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 14th 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.

Changes of the Levels of Blood NO and TNF-αConcentrations in Water Buffaloes and Goats Infected with Fasciola hepatica

Changes of concentrations were studied in water buffaloes and goats infected with Fasciola he-patica on blood NO(nitric oxide) and TNF-α(tumor necrosis factor-α). Twenty healthy male castrated water buffaloes of 2 - 3 years old and weighing 300 - 500 kg as well as six goats were confirmed free of fasciolosis by fecal examination and Dot-ELISA. Two studies were conducted using the water buffaloes. In the first experiment, 8 water buffaloes were randomly divided into control group (n = 3) and infection group (n = 5). Each buffalo in the infected group received orally 60 metacercariae of F. hepatica per day for 20 days (total 1 200 metacercariae) to produce a chronic infection. In the second experiment, 12 water buffaloes were randomly divided into infected (n=9) and control group (n = 3). Each buffalo in the infected group was given a single oral dose of 1 600 metacercariae to produce an acute infection. The 6 goats were randomly divided into two infected groups and a control group. The sheep in two infections received a single oral dose of 200 and 500 metacercariae respectively, the control group remained uninfected. Blood NO and TNF-a concentrations of the test animals were measured by a reductive enzyme assay and RIA, respectively. Blood NO concentration in both acutely and chronically infected water buffaloes progressively increased from week 3 post-infection and was significantly greater than that of the control group (P<0. 05) at the 5th week (acute infection) and 7th week (chronic infection), and remaining at higher concentration for the remaining period of the studies. Blood TNF-αconcentrations in both chronically and acutely infected water buffaloes also increased after infection. In the goat experiment, plasma NO concentrations in both infection groups increased from week 3 after infection, and remained higher than that of the control group until the end of the experiment. TNF-α concentrations in goats in infection group 1 and 2 gradually increased after infection and were significantly greater than those of the control group from the 9th to 11th week and from the 11th to 15th week respectively.

Clinical presentation and management of Fasciola hepatica infection:Single-center experience

AIM:To identify the characteristic clinical,laboratory and radiological findings and response to treatment in patients with fascioliasis.METHODS:Patients who were diagnosed with Fasciola hepatica infection were included in this prospective study.Initial clinical,laboratory and radiological findings were recorded.All patients were followed until a complete response was achieved or for 6 mo after treatment discontinuation.RESULTS:Fasciola hepatica infection was diagnosed in 30 patients(24 females;mean age:42.6 years) between January 2008 and February 2011.Twentytwo(73%) patients had hepatic phase fascioliasis,5 patients had biliary phase,and 3 patients had biliary phase associated with acute pancreatitis.Of the 8 patients with biliary phase fascioliasis,2 patients displayed features that overlapped with both hepatic and biliary phase.Abdominal pain and right upper abdominal tenderness were the most prominent signs and symptoms in all patients.Eosinophilia was the most prominent laboratory abnormality in both patients with hepatic and biliary phase(100% and 50%,respectively).Multiple nodular lesions like micro-abscesses on abdominal computerized tomography were the main radiological findings in patients with hepatic phase.Small linear filling defects in the distal choledochus were the main endoscopic retrograde cholangiopancreatography(ERCP) findings in patients with biliary phase.Patients with hepatic phase were treated with triclabendazole alone,and patients with biliary phase were treated with triclabendazole and had live Fasciola hepatica extracted from the bile ducts during ERCP.CONCLUSION:Fasciola hepatica infection should be considered in the differential diagnosis of patients with hepatic or biliary disease and/or acute pancreatitis associated with eosinophilia.

Antifungal,molluscicidal and larvicidal assessment of anemonin and Clematis flammula L.extracts against mollusc Galba truncatula,intermediate host of Fasciola hepatica in Tunisia

Objective: To investigate the potential of anemonin and Clematis flammula(C.flammula) extracts against infective organisms.Methods: The molluscicidal activities of anemonin and C.flammula extracts against Galba truncatula Mull.(Lymnaeidae) and Fasciola hepatica larval stages contaminating this snail in Tunisia were assessed by testing six groups of snails in 250 mL of extracts and aqueous dechlorinated solutions with different concentrations(ranging from 2.5 to 20.0 mg/L) for 48 h.Besides, the antifungal potential of C.flammula leaves and flowers was evaluated by using the diffusion agar and broth dilution methods against four fungal strains: Aspergillus niger, Pythium catenulatum, Rhizoctonia solani and Fusarium phyllophilum.Results: As a result, hexane and ethyl acetate flower extracts exhibited significant molluscicidal activities with LC_(50) median lethal concentrations values of 11.87 and 11.65 mg/L, respectively while LC_(50) value of anemonin was 9.64 mg/L after 48 h exposure.The flower extracts showed a larvicidal effect with a deterioration rate exceeding 35.39% where flower ethyl acetate residue gave a deterioration rate of cercariae close to 97%.Moreover, C.flammula extracts were not noxious to the associated fauna survival.All extracts inhibited the growth of P.catenulatum, the leaves and flowers methanolic extracts had the more important fungicide action with minimum inhibitory concentrations of 1.56 and 3.12 mg/mL together with minimum fungistatic concentrations of 3.12 and 6.25 mg/mL respectively.Only flower extracts were active against Rhizoctonia solani with minimum inhibitory concentrations varying between 0.70 and 1.56 mg/mL and 6.25 mg/mL of minimum fungistatic concentration.Phytochemical tests showed that the antifungal activity may be attributed to the presence of the flavonoids/saponins in the methanolic extracts and the molluscicide effects could be due to the richness of hexane and ethyl acetate extracts on sterols and triterpenoids.Conclusions: This study emphasizes the important molluscicidal and antiparasitic effects of flower ethyl acetate extracts and anemonin compound as well as the considerable antifungal activities of methanolic extracts.These results improve the therapeutic virtues of C.flammula aerial part extracts.

Obtaining the Minimum Lethal Dose against <i>Fasciola hepatica in Vitro</i>Using Plant Extract Hexanes with Fasciolicide Activity and Toxicity Evaluation on CD1 Male Mice

Fascioliasis is a parasitic disease of worldwide distribution affecting mainly cattle and sheep. Its importance lies in the economic losses it produces in the livestock industry. Its control is carried out by using a chemical fasciolicide showing resistance problems and environmental contamination. Looking for an alternative control for this disease the present study was aimed at determining the hexane anti-Fasciola hepatica in the in vitro effect of some plant extracts and the minimum lethal dose of the mentioned extracts. All selected plants were tested in vitro at concentrations of 500, 250, 125 and 50 mg/L):Achilleamillefolium (plumajillo), Artemisiaabsinthium (wormwood), Artemisia mexicana (estafiate), Castelatortuousa (chaparroamargo), Chenopodiumgraveolens (epazote de zorrillo), Gymnospermaglutinosum (popote) Justicia spicigera (muicle), Limpia critridora (cedron), Lippiagraveolens (oregano), Menthapiperita (Mint), Populus alba (alamo) and Thymusvulgaris (thyme). Subsequently proceeded to perform a toxicity study with these fractions in CD1 male mice 10-13 weeks of age, forming groups of 3-5 animals they were administered a single oral dose being (5 mg/kg, 50 mg/kg, 500 mg/kg, 2500 mg/kg and 5000 mg/kg) and were kept under observation 20 days, later were sacrificed and a kidney and liver histology was performed, finding the safety of the extracts. To perform the toxicity study with these fractions, groups of five CD1 male-mice were formed, they were treated with oral doses of 5, 50, 500, 2500 and 5000 mg/kg, administered with a cannule. All mice were kept under observation for 20 days. Finally they were sacrificed to perform histology of the kidney and liver in search of possible side effects. Results show that none of the extracts exhibited that fasciolocide activity for mice CD1 even at the highest dose thereforefinding the safety of the extracts.

Does Fasciola hepatica infection modify the response of acute hepatitis C virus infection to IFN-α treatment?

Immunologic response to acute hepatitis C is mainly a Th1 response, whereas fasciolopsiasis is associated with a diverse T-cell response. Interferon-alpha has immunomodulatory effects and enhances Th1 immune response. Fasciola infection could theoretically interfere with the Th1 immune response, even when acquired after an initial response to interferon-alpha treatment for acute hepatitis C virus （HCV） infection. We report here the case of a male patient who acquired Fasciola hepatica infection after an initial response to IFN-alpha therapy with a favorable outcome.

Fasciola hepatica infestation as a very rare cause of extrahepatic cholestasis

Fasciola hepatica,an endemic parasite in Turkey,is still a very rare cause of cholestasis worldwide.Through ingestion of contaminated water plants like watercress,humans can become the definitive host of this parasite.Cholestatic symptoms may be sudden but in some cases they may be preceeded by a long period of fever,eosinophilia and vague gastrointestinal symptoms.We report a woman with cholangitis symptoms of sudden onset which was proved to be due to Fasciola hepatica infestation by an endoscopic retrograde cholangiography.

<i>Fasciola hepatica</i>and Associated Parasite, <i>Dicrocoelium dendriticum</i>in Slaughter Houses in Anyigba, Kogi State, Nigeria

Fasciola hepatica is a parasite of clinical and veterinary importance which causes fascioliasis that leads to reduction in milk and meat production. Bile samples were centrifuged at 1500 rpm for ten (10) minutes in a centrifuge machine and viewed microscopically to check for F. hepatica eggs. A total of 300 bile samples of cattle which included 155 males and 145 females were collected from the abattoir. Results were analyzed using chi-square (p > 0.05). The prevalence of F. gigantica and Dicrocoelium dentriticum is 33.0% (99) and 39.0% (117) respectively. Age prevalence of F. hepatica revealed that 0 - 2 years (33.7%, 29 cattle) were more infected than 2 - 4 years (32.7%, 70 cattle) while for D. dentriticum age 2 - 4 years were more infected than 0 - 2 years with prevalence of 40.2% (86) and 36.0% (31) respectively. No significant difference (P > 0.05) existed in prevalence in ages of the cattle. Out of the 300 bile samples examined, 22.3% (67 cattle) were co-infected with F. hepatica and D. dendriticum. Males were more co-infected than females having a prevalence of 24.5% (38 cattle) and 20.0% (29 cattle) respectively. Based on the age, samples of age 0 - 2 years were more co-infected than those of age 2 - 4 years with a prevalence of 23.3% (20 cattle) and 22.0% (47 cattle) respectively. The findings of this present study revealed that efforts to alleviate problems of animal health and productivity are yet to make any significant impact as this poses threat on human health. Investigation on the pattern of infections in cattle slaughtered should be strengthened.

条华蜗牛破厣前后体内总糖含量的变化

【目的】了解条华蜗牛破厣前后体内总糖含量变化规律,为深入研究蜗牛破厣生化机理提供参考。【方法】用蒽酮-硫酸比色法测定了两个体质量组条华蜗牛在破厣、休眠、取食或饥饿等条件下足和内脏的总糖含量。【结果】条华蜗牛破厣后体内含糖量升高,体质量组IV蜗牛足、内脏含糖量都高于相应体质量组I。破厣前蜗牛足内含糖量与破厣后的差异显著(P<0.05),而内脏的差异不显著。休眠蜗牛体内含糖量随休眠天数增加有降低的趋势,除体质量组I蜗牛足内含糖量休眠1 d后(27.62μg/mg)显著高于5 d~15 d外,其他休眠时间之间差异都不显著。蜗牛取食后,足和内脏含糖量变化趋势基本类似,都是取食12 h~24 h后增加,然后有所下降。除体质量组IV足外,取食24 h后都显著高于取食12 h和36 h。活动蜗牛体内含糖量随饥饿时间延长有降低趋势,饥饿36 h后含糖量均显著低于饥饿12 h。【结论】条华蜗牛破厣前后体内总糖含量变化与多个因素有关,总糖量变化规律反映了蜗牛破厣的生理生化特征。

肝片吸虫检测方法概述

肝片吸虫是一种人兽共患寄生虫,是目前全世界分布最广泛的吸虫之一,能感染包括人在内的多种哺乳动物,引起肝片吸虫病,不仅严重危害人类健康,还影响畜牧养殖业的发展,造成巨大的经济损失。肝片吸虫因其复杂的生活史和种类广泛的中间宿主和终末宿主,导致其致病机制尚不明确,且肝片吸虫感染引起的动物临床症状与其他肝胆疾病类似,主要表现为急性或慢性肝炎和胆管炎,没有特征性症状,极易被误诊。因此,及时、准确的检测对防治肝片吸虫病尤为重要。目前,肝片吸虫的检测主要依赖病原学、免疫学、分子生物学等手段,论文就肝片吸虫检测方法的研究进展进行综述,分析各项技术的优点与局限性,以期为肝片吸虫病检测方法的研究和发展提供参考。

大片吸虫磷酸丙糖异构酶基因的克隆表达和免疫原性分析

磷酸丙糖异构酶(TPI)是生物体糖酵解的一种关键酶,对大片吸虫获取能量而赖以生存有着十分重要的作用。本试验根据肝片吸虫TPI(GenBank:KC164346.1)的基因序列,设计带有BamHⅠ和XhoⅠ作为酶切位点的引物,以大片吸虫的cDNA为模板,扩增大片吸虫磷酸丙糖异构酶(FgTPI)基因。结果:FgTPI基因开放阅读框为762 bp,编码253个氨基酸,理论等电点pI为8.07;构建的重组表达质粒FgTPI-pET-28a(+)成功在大肠杆菌中表达,重组蛋白的分子量约为32 kDa;重组蛋白免疫BALB/c小鼠,收取多克隆抗体,ELISA检测结果显示,rFgTPI能诱导较高的IgG抗体水平;用感染大片吸虫动物的血清检测重组蛋白的免疫原性,Western blot分析表明,重组蛋白rFgTPI具有良好的免疫原性;生物信息学预测显示,FgTPI主要以α螺旋和β折叠为主,β转角较少,有3个较长的无规则卷曲;通过α-磷酸甘油脱氢酶偶联法测定其活性,发现FgTPI酶促反应最佳pH值为7.5,最适温度为35℃。本试验结果为深入研究FgTPI的生物学功能、评价其作为抗大片吸虫病疫苗候选分子奠定了基础。

我国片形吸虫(Fasciola)线粒体细胞色素c氧化酶亚基基因(cox1)部分序列的多态性

用33P对引物进行标记,对来自国内不同地区不同宿主的片形吸虫(Fasciola)的线粒体基因组细胞色素氧化酶亚基基因(pcox1)部分序列进行PCR扩增及DNA单链构象多态性(SSCP)分析,结果76个虫体均成功地扩增出约450bp的基因片段;SSCP分析显示,不同地区或同一地区的不同样品在pcox1的SSCP带型上存在多态性;代表性样品的测序结果表明,其碱基序列存在差异。试验结果显示,我国片形吸虫pcox1序列种内变异不明显,种间变异显著,表明cox1序列可以作为片形吸虫分类鉴定中一个可靠的遗传标记。

条华蜗牛(Cathaica fasciola)翻身习性的研究

【研究目的】研究条华蜗牛翻身习性,并研究蜗牛体重、环境温度、光照强度、饥饿和取食等因素对翻身时间的影响作用。【方法】将蜗牛身体倒置后,用秒表记录蜗牛身体从倒置至恢复原状的翻身时间长短来比较不同因素对翻身时间的影响。【结果】翻身时间随体重、光照强度和饥饿时间增加而增加,而随温度和取食时间增加而减少。体重与翻身时间呈极显著正相关(P<0.0001)。三体重组(I、III、V)翻身时间在30～40℃和取食48h分别极显著短于相应10℃和12h的(P<0.01)。在最低强度的白炽灯光(37.8LX)下翻身时间都极显著短于最高光强(4310LX)的,相同光照强度不同光质对翻身时间也有一定影响。饥饿72h后翻身时间都极显著长于12h的。【结论】条华蜗牛身体倒置恢复原状的时间可受多个因素的影响,除受到自身体重的影响外,还受到如光照、温度、是否取食等多个外界因素的影响。

塔叶苔属(Schiffneria Steph.)(Hepaticae)的研究

塔叶苔属(Schiffneria Steph.)建立于1894年。作者在研究中国标本时,发现一新种———Schiffneria yunnanensis C.Gao& W.Li sp.nov.。塔叶苔属的配子体为叶状体,生殖枝具茎叶返祖现象,应置于叶苔亚纲的叶状体无组织分化类群———带叶苔目(Pallaviciniales)之中。

宁化县人体假性感染肝毛细线虫Capillaria hepatica调查

目的分析宁化县人体假性感染肝毛细线虫情况及相关因素。方法结合当地人体寄生虫感染调查,对水茜、淮土和河龙3个自然村的居民,以常规粪便改良加藤氏厚涂片法检查。共查881人,发现感染者8人,感染率为0.91%。经追踪调查,8例感染者在1周前均有吃过熏烤老鼠干史。结论证实当地人体粪便内发现的肝毛细线虫卵,是吞食含有该虫卵的熏烤鼠干所致。故8例的肝毛细线虫卵阳性是通过性的假性感染。

胆道肝片吸虫病1例报道

通过1例误诊为胆总管结石患者在接受ERCP治疗时,从胆道取出肝片吸虫的诊疗过程,增强医师对该病的认识。

新疆巴州部分地区秋季黑头羊消化道寄生虫虫卵检测及分析

为了解新疆巴州部分地区秋季黑头羊消化道寄生虫感染情况,于2023年8-9月从新疆巴州和静、和硕县11个乡镇(1~11号)共采集黑头羊新鲜粪便1143份,进行粪便检测。结果显示,1143份粪便样品经虫卵漂浮法共检出3种虫卵,分别为肝片吸虫、细颈线虫和小型艾美耳球虫。整理巴州不同地区、年龄、性别黑头羊消化道寄生虫感染情况,发现巴州地区黑头羊细颈线虫感染率最高,肝片吸虫和小型艾美耳球虫感染率均较低。母羊细颈线虫、肝片吸虫感染率比公羊高,小型艾美耳球虫感染率比公羊低。5~6岁黑头羊细颈线虫感染率最高,3~4岁黑头羊小型艾美耳球虫感染率最高,仅5~6岁黑头羊检出一例肝片吸虫感染。研究表明,巴州地区黑头羊存在肝片吸虫、细颈线虫和小型艾美耳球虫感染,且不同性别、年龄的黑头羊寄生虫感染情况存在一定差异。

牛舌菌(Fistulina hepatica)水溶性多糖FHP-2P的分离与理化特征

牛舌菌(Fistulinahepatica)子实体经热水抽提,乙醇沉淀,蛋白酶法和Sevag法去蛋白后得粗多糖FHP.FHP经DEAE-纤维素和SephadexG-200柱进一步纯化得到牛舌菌多糖纯品FHP-2P.经测定该多糖为均一组分,相对分子质量为7.6×104.红外扫描呈现出典型的多糖吸收峰,并含有β-型糖苷连接键;紫外扫描无核酸和蛋白质的特征吸收峰.纸层析和气相色谱分析结果表明FHP-2P的单糖组成为D-葡萄糖、D-甘露糖、D-半乳糖和D-木糖,其摩尔比为13.9∶2.85∶3.25∶0.48.

Capillaria hepatica in China

Capillaria hepatica(C.hepatica) is a parasitic nematode causing hepatic capillariasis in numerous mammals.Ecologic studies showed that the first hosts of C.hepatica were rodents,among which rats had relatively high infection rates,which explains why C.hepatica spreads globally.Anatomical studies showed that the liver was the principal site of colonization by these parasites and physical damage tended to occur.Although C.hepatica might lead to serious liver disorders,relevant clinical reports were rare,because of the non-specific nature of clinical symptoms,leading to misdiagnosis.This review mainly focuses on the biological characteristics and epidemiology of C.hepatica in China and histopathologic changes in the liver,with expectation of gaining a better understanding of the disease and seeking more effective treatment.

Effects of Mirazid~ and myrrh volatile oil on adult Fasciola gigantica under laboratory conditions

Objective:To evaluate the effects of Mirazid and myrrh volatile oil on adult Fasciola gigantica(F.gigantica) under laboratory conditions.Methods:The effects of oleoresin extract of myrrh(Mirazid) and myrrh volatile oil on the surface morphology of adult F.gigantica following treatment in vitro had been determined by scanning electron microscopy.The results were compared with those observed in the fluke tegument following incubation in triclabendazole sulphoxide(TCBZ-SO),active form.(Fasinex,Ciba-Geigy).Results:Observations of the efficacy of Mirazid oleoresin extract and myrrh volatile oil indicated that both products showed dosedependent anthelmintic efficacy.The anterior half of the fluke was consistently more severely affected than the posterior half.The surface changes induced by Mirazid oleoresin extract were less severe than those observed after exposure to either myrrh volatile oil or TCBZ-SO.Flukes showed swelling after these treatments,but its level and blebbing were much greater with myrrh volatile oil;in which patches of tegumental sloughing were observed in the apical cone and the posterior mid-body region of flukes.This was not observed after treatment with Mirazid oleoresin extract.Conclusions:The comparatively more disruption,observed in myrrh volatile oil exposed specimens,compared to that exposed to Mirazid oleoresin extract might suggest that the anthelmintic activity of Mirazid oleo resin extract was attributed to its content of volatile oil.So,increasing the concentration of myrrh volatile oil in Mirazid might possibly help to developing its anthelmintic activity._______________________________________________