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Arginine kinase in Toxocara canis:Exon-intron organization,functional analysis of site-directed mutants and evaluation of putative enzyme inhibitors

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Asian Pacific Journal of Tropical Medicine 2016年第10期9卷 973-979页

作者： Susiji Wickramasinghe Lalani Yatawara Mitsuru Nagataki Takeshi Agatsuma Department of Parasitology Faculty of MedicineUniversity of Peradeniya Department of Medical Laboratory sciences Faculty of Allied Health SciencesUniversity of Peradeniya Department of Environmental Health Sciences Kochi Medical School

Objective: To determine exon/intron organization of the Toxocara canis(T. canis) AK(TCAK) and to test green and black tea and several other chemicals against the activity of recombinant TCAK in the guanidino-specific region by site-directed mutants. Methods: Amplification of genomic DNA fragments containing introns was carried out by PCRs. The open-reading frame(1 200 bp) of TCAK(wild type) was cloned into the BamH 1/SalI site of pM AL-c2X. The maltose-binding protein-TCAK fusion protein was expressed in Escherichia coli TB1 cells. The purity of the expressed enzyme was verified by SDS-PAGE. Mutations were introduced into the guanidino-specific region and other areas of pM AL/TCAK by PCR. Enzyme activity was measured with an NADH-linked assay at 25℃ for the forward reaction(phosphagen synthesis). Results: Arginine kinase in T. canis has a seven-exon/six-intron gene structure. The lengths of the introns ranged from 542 bp to 2 500 bp. All introns begin with gt and end with ag. Furthermore, we measured the enzyme activity of site-directed mutants of the recombinant TCAK. The K_m value of the mutant(Alanine to Serine) decreased indicating a higher affinity for substrate arginine than the wild-type. The K_m value of the mutant(Serine to Glycine) increased to 0.19 mM. The Km value(0.19 mM) of the double mutant(Alanine-Serine to Serine-Glycine) was slightly greater than in the wild-type(0.12 mM). In addition, several other chemicals were tested; including plant extract Azadiracta indica(A. indica), an aminoglycoside antibiotic(aminosidine), a citrus flavonoid glycoside(rutin) and a commercially available catechin mixture against TCAK. Green and black tea(1:10 dilution) produced 15% and 25% inhibition of TCAK, respectively. The extract of A. indica produced 5% inhibition of TCAK. Moreover, green and black tea produced a non-competitive type of inhibition and A. indica produced a mixed-type of inhibition on TCAK. Conclusions: Arginine kinase in T. canis has a seven-exon/six-intron gene structure. However, further studies

关键词： Toxocara canis Arginine kinase Gene structure Site directed mutagenesis Inhibition kinetics

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Multifocal granulomata in presumed Toxocara canis infection in adult

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World Journal of Ophthalmology 2013年第4期3卷 38-40页

作者： Lakshmi Kuniyal Jyotirmay Biswas Department of Vitreoretina Sankara Nethralaya Chennai TN 600006 Andhra Pradesh India Department of Uvea Sankara Nethralaya Chennai TN 600006 Andhra Pradesh India

Human infection of Toxocara canis in eye is usually an outcome of accidental ingestion of the embryonated eggs. The average age at diagnosis of ocular Toxocariasis is 7.5 years, ranging from 2 to 31 years. It constitutes 1%-2% of uveitis in children. Diagnosis is based upon the clinical features observed in a young patient and confirmed by the presence of specific Ig G in the serum or aqueous humor by Enzyme-linked immunosorbent assay test. We report a case of Presumed Toxocara infection in 45-year-old male which is unique in presentation with multifocal granulomata in retina. Our Pub Med search could not produce case with similar presentation. Probably this is the first reported case of multifocal granulomata in presumed ocular Toxocara in any age

关键词： Presumed Toxocara canis Ocular Multifocal Granulomata

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Soil-transmitted helminth egg contamination from soil of indigenous communities in selected barangays in Tigaon, Camarines Sur, Philippines

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Asian Pacific Journal of Tropical Medicine 2020年第9期13卷 409-414页

作者： James Owen C.Delaluna Mary Jane C.Flores Vicente Y.Belizario Jr. Jose Isagani B.Janairo Derick Erl P.Sumalapao Biology Department College of ScienceDe La Salle UniversityManila Philippines Biological Control Research Unit Center for Natural Sciences and Environmental ResearchDe La Salle UniversityManila Philippines Department of Parasitology College of Public HealthUniversity of the PhilippinesManila Philippines Department of Epidemiology and Biostatistics College of Public HealthUniversity of the PhilippinesManila Philippines

Objective:To provide baseline data on the prevalence and intensity of soil-transmitted helminthiasis egg contamination in the soil among indigenous communities.Methods:A total of 317 soil samples from three barangays of indigenous communities communities in Tigaon,Camarines Sur,Philippines were examined for soil-transmitted helminthiasis egg contamination using optimized sugar flotation method.Results:Of the soil samples examined,141(44.48%)were contaminated by Ascaris spp.,Toxocara spp.,and Trichuris spp.with cumulative prevalence varying across the study sites(PToxocara spp.and Trichuris spp.with a prevalence of 41.96%,7.57%,and 5.36%,respectively.Interestingly,Toxocara pp.has the highest intensity of contamination,followed by Ascaris spp.and Trichuris spp.in term of geometric mean soil-transmitted helminthiasis eggs recovered per one gram soil sample(34.25,21.45,and 11.85 respectively).Each study site harbors significant amount of soiltransmitted helminthiasis eggs and zoonotic Toxocara eggs,which present high risk of soil-transmitted helminthiasis infection,particularly among children observed to play and cohabitate with animals known to be hosts of these parasites.Conclusions:The alarming rate of soil-transmitted helminthiasis and Toxocara egg contamination reported in this study suggests that additional measures should be undertaken to control soil-transmitted helminthiasis and zoonotic intestinal infections in the country.

关键词： Indigenous peoples Soil-transmitted helminths Ascaris Toxocara Trichuris Public health

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A case report of ocular toxocariasis

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Asian Pacific Journal of Tropical Biomedicine 2011年第2期1卷 164-165页

作者： Azira NMS Zeehaida M Department of Medical Microbiology and Parasitology School of Medical SciencesUniversiti Sains Malaysia

Ocular toxocariasis is prevalent among children.The symptoms and signs may mimic other ocular pathologies such as malignancies and other infectious diseases(such as toxoplasmosis and syphilis). We presented a case of progressive blurring of vision in a single eye of a 9-year-old boy.The presence of anti-Toxocara antibody in serum samples helps to confirmation the diagnosis in our patient.Despite of treatment,the boy had lost his vision on the affected eye.

关键词： Ocular toxocariasis Ocular larva migrant Toxocara cati Vision

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Disseminated toxocariasis in an immunocompetent host

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Asian Pacific Journal of Tropical Biomedicine 2014年第10期4卷 838-840页

作者： Madan Raj Aryal Paras Karmacharya Amrit Pokharel Smith Giri Ranjan Pathak Richard Alweis Department of Medicine Reading Health SystemPA 19611USA Department of Medicine Tribhuvan University Teaching HospitalKathmanduNepal Department of Medicine University of Tennessee Health Science CenterTN 38163USA

Toxocariasis is a zoonotic infection caused by Toxocara canis,or less commonly,Toxocara cati,which is one of the most common zoonotic infections worldwide.It commonly affects the pediatric and immunocompromised population;however,it has rarely been reported in the immunocompetent adults.Two of the well-recognized syndromes in children are visceral larva migrans and ocular larva migrans.Infection in adults usually ranges from asymptomatic to nonspecific symptoms which makes the diagnosis challenging.A case of 36 year-old male was presented with disseminated toxocariasis with pulmonary and hepatic involvement and striking peripheral eosinophilia.

关键词： Toxocariasis Toxocara canis Mebendazole Albendazole

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Control of Canine Endoparasites,Especially Isospora spp.,with Procox^(■) in Naturally Infected Puppies:Parasitological,Bacteriological and Health Parameters

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Open Journal of Veterinary Medicine 2013年第2期3卷 121-130页

作者： Barbara Anna Rauscher Sabine Schafer-Somi Monika Ehling-Schulz Karin Mostl Stefanie Handl Barbara Hinney Joachim Spergser Roland Schaper Anja Joachim Institute of Parasitology Department of PathobiologyUniversity of Veterinary Medicine ViennaViennaAustria Centre for Artificial Insemination and Embryo Transfer Department for Small Animals and HorsesUniversity of Veterinary Medicine ViennaViennaAustria Functional Microbiology IBMHDepartment of PathobiologyUniversity of Veterinary Medicine ViennaViennaAustria Institute of Virology Department of PathobiologyUniversity of Veterinary Medicine ViennaViennaAustria Institute of Animal Nutrition Department for Farm Animals and Veterinary Public HealthUniversity of Veterinary Medicine ViennaViennaAustria Clinical Microbiology and Infection Biology IBMHDepartment of PathobiologyUniversity of Veterinary Medicine ViennaViennaAustria Bayer Animal Health GmbH MonheimGermany

The effect of Procox? (Bayer, emodepside/toltrazuril suspension for dogs) against natural infections with Isospora spp. was investigated. Two groups were treated either with 0.5 ml suspension/kg of body weight (0.45 mg emodepside and 9 mg toltrazuril/kg of body weight) in the 3rd, 5th and 7th week of life (w.o.l.) (Procox? group;n = 28) or with a control anthelminthic (Dewormed Control group;n = 26). Animals were surveyed weekly from the 3rd w.o.l. by coproscopy and clinical examination. Faecal samples were examined microbiologically from the 4th to the 8th w.o.l. and faecal inflammatory markers canine calprotectin and canine S100A12 were measured in the 8th w.o.l. Specific antibody titres were evaluated in serum samples from five litters before and after vaccination against canine distemper virus and canine parvovirus 2. The prevalence of Isospora-positive animals increased to 67% in the Dewormed Control group (n = 15 puppies from four parasite-positive litters), while in the Procox? group (n = 15 puppies) it was less than 34% with significantly lower excretion (p ? was easily applied and effective;adverse effects did not occur. The level of seroconversion or titre increase upon vaccination was higher in parasite-free animals (91%) compared to Procox?-treated puppies (30%) and the Control animals (10%). Animals from parasite-free litters showed significantly different excretion patterns for haemolytic Escherichia coli and Clostridium perfringens, while there was no difference between Procox?-treated and Control animals. In some animals kept under poor hygienic conditions diarrhoea was noted in association with C. perfringens, E. coli or Salmonella. Concentrations of inflammatory markers in the faeces did not significantly differ between the Procox? and the Control group. Adequate control of parasitic and bacterial infections in suckling puppies requires both antiparasitic treatment and hygiene. Even when parasites do not cause overt effects treatment is recommended in cases with a history of parasite infections.

关键词： Dog Emodepside Gut Health Isospora Toltrazuril Toxocara Vaccination

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