Differentiation of Toxocara canis and Toxocara cati based on PCR-RFLP analyses of rDNA-ITS and mitochondrial cox1 and nad1 regions

AbstractHuman toxocariasis, a worldwide parasitic disease, is caused by the larval stage of intestinal nematodes of dogs and cats, namely Toxocara canis and Toxocara cati. Human infection occurs by the accidental ingestion of embryonated eggs present in the soil, vegetables or on other contaminated surfaces, as well as via consumption of uncooked paratenic hosts, such as bird meat and giblets. The objective of this study was to evaluate the contamination of soil in public parks and playgrounds in Shiraz using microscopy and molecular methods. A total of 150 soil samples were collected from public parks and playgrounds in various areas of Shiraz, southern Iran. The samples were treated with saturated zinc sulphate solution, and Toxocara spp. eggs were detected by microscopic observation followed by nested polymerase chain reaction (PCR). To differentiate T. canis and T. cati eggs from each other, PCR restriction fragment length polymorphism (RFLP) of the internal transcribed spacer (ITS)-rDNA region by SalI endonuclease enzyme was used. PCR-sequencing was performed to confirm the results of the PCR-RFLP method. Based on the flotation results of the 150 soil samples, six (4%) were found to be positive for Toxocara spp. eggs, whereas nested-PCR showed 24 samples to be positive (16%). Based on the PCR-RFLP method and the sequence of the ITS-rDNA region, a total of 23 out of 24 isolates were confirmed as T. cati and one out of 24 as T. canis. The results showed a higher number of soil samples to be positive for Toxocara by the molecular method than microscopy, and higher T. cati infection in soil samples, which could have an important role in human infection with toxocariasis in this region.

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A Sensitive, One-Way Sequential Sieving Method to Isolate Helminths’ Eggs and Protozoal Oocysts from Lettuce for Genetic Identification

Pathogens ◽

10.3390/pathogens9080624 ◽

2020 ◽

Vol 9 (8) ◽

pp. 624

Author(s):

Annina R. Guggisberg ◽

Cristian A. Alvarez Rojas ◽

Philipp A. Kronenberg ◽

Nadia Miranda ◽

Peter Deplazes

Keyword(s):

Toxoplasma Gondii ◽

Simultaneous Detection ◽

Toxocara Canis ◽

Genetic Identification ◽

Diagnostic Strategy ◽

Toxocara Cati ◽

Test Kit ◽

Contaminated Food ◽

In Vivo Testing

Different helminths and protozoa are transmitted to humans by oral uptake of environmentally resistant parasite stages after hand-to-mouth contact or by contaminated food and water. The aim of this study was to develop and validate a method for the simultaneous detection of parasite stages from fresh produce (lettuce) by a one-way isolation test kit followed by genetic identification (PCR, sequencing). Three sentinel zoonotic agents (eggs of Toxocara canis, Echinococcus multilocularis and oocysts of Toxoplasma gondii) were used to investigate the practicability and sensitivity of the method. The detection limits (100% positive results) in the recovery experiments were four Toxocara eggs, two E. multilocularis eggs and 18 T. gondii oocysts (in 4/5 replicates). In a field study, helminth DNA was detected in 14 of 157 lettuce samples including Hydatigera taeniaeformis (Syn. Taenia taeniaeformis) (four samples), T. polyacantha (three), T. martis (one), E. multilocularis (two) and Toxocara cati (four). Toxoplasma gondii was detected in six of 100 samples. In vivo testing in mice resulted in metacestode growth in all animals injected with 40–60 E. multilocularis eggs, while infection rates were 20–40% with 2–20 eggs. The developed diagnostic strategy is highly sensitive for the isolation and genetic characterisation of a broad range of parasite stages from lettuce, whereas the sensitivity of the viability tests needs further improvement.

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Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati

Parasitology ◽

10.1017/s0031182021000342 ◽

2021 ◽

pp. 1-8

Author(s):

Héctor Gabriel Avila ◽

Marikena Guadalupe Risso ◽

Paula Ruybal ◽

Silvia Analía Repetto ◽

Marcos Javier Butti ◽

...

Keyword(s):

Low Cost ◽

Lamp Assay ◽

Toxocara Canis ◽

Toxocara Cati

Abstract

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M�glichkeiten und Grenzen einer Artdifferenzierung der Larven von Ascaris lumbricoides suis, Parascaris equorum, Toxocara canis, Toxocara cati und Toxascaris leonina in histologischen Schnitten beim nicht ad�quaten Wirt (Maus) II. Teil

Zeitschrift für Parasitenkunde Parasitology Research ◽

10.1007/bf00348668 ◽

1964 ◽

Vol 25 (1) ◽

pp. 52-67 ◽

Cited By ~ 2

Author(s):

J. Lamina

Keyword(s):

Ascaris Lumbricoides ◽

Toxocara Canis ◽

Toxocara Cati ◽

Parascaris Equorum ◽

Toxascaris Leonina

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Distribution of Toxocara infection in the environment and in definitive and paratenic hosts in Estonia

Acta Veterinaria Hungarica ◽

10.1556/avet.54.2006.3.10 ◽

2006 ◽

Vol 54 (3) ◽

pp. 399-406 ◽

Cited By ~ 16

Author(s):

Heli Talvik ◽

Epp Moks ◽

Erika Mägi ◽

T. Järvis ◽

Illa Miller

Keyword(s):

Rattus Norvegicus ◽

Risk Group ◽

Toxocara Canis ◽

Toxocara Cati ◽

Low Level ◽

Larval Infection ◽

Small Intestines ◽

Paratenic Hosts ◽

The Town ◽

Home Territory

The aim of the study was to elucidate the distribution and possible transmission routes of Toxocara spp. infection in Estonia. Out of 454 faecal and sand samples collected from park lawns and sandpits in the town of Tartu, 19 were Toxocara positive (4.2%). Out of the 45 sandpit samples 17.8% were Toxocara positive. Cat faeces was found in 21 sandpit samples. Parasitological necropsies were performed on 41 euthanised stray dogs and 27 cats in the Tallinn Dog Home. Additionally, 13 wild free-roaming brown rats (Rattus norvegicus) were captured from the Tallinn Dog Home territory, necropsied and studied for the presence of Toxocara larvae. Toxocara canis adults were found in 14.6% of the dogs and Toxocara cati (syn. mystax) adults in the small intestines of 48.2% of the cats examined. Larval infection was detected in the kidney and liver in 5 dogs (12.2%). Our study demonstrated only low-level larval Toxocara infections in adult dogs. Toxocara larvae were not found in cats and brown rats. According to the results of this study, cats more often carry Toxocara infection than dogs. Under our conditions, stray and free-roaming cats are the main contaminators of the environment with Toxocara eggs. Children playing in sandpits are the main risk group for larval toxocarosis.

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Genetic identification of bovine leukaemia virus

Foods and raw materials ◽

10.21603/2308-4057-2018-2-314-324 ◽

2018 ◽

Vol 6 (2) ◽

pp. 314-324 ◽

Cited By ~ 3

Author(s):

Irina Donnik ◽

Ramil Vafin ◽

Aram Galstyan ◽

...

Keyword(s):

Phylogenetic Analysis ◽

Research Methods ◽

Bovine Leukemia Virus ◽

Leukemia Virus ◽

Molecular Genetic ◽

Genetic Research ◽

Genetic Identification ◽

Gene Fragment ◽

Phylogenetic Classification ◽

Pcr Rflp

Molecular genetic research methods make it possible to evaluate the genetic diversity of bovine leukemia virus (BLV) and are the most informative approaches to its genetic identification. Molecular genetic research methods work well for the phylogenetic analysis of sequenced nucleotide DNA sequences of the provirus, as well as for the polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP) according to the phylogenetic classification of the pathogen. The purpose of the research was to study the scientific and methodological approaches to the genetic identification of bovine leukemia virus, integrated into the molecular monitoring of infection of cattle with BLV genotypes. The authors used PCR-RFLP-genotyping and comparative phylogenetic analysis of aligned nucleotide sequences of the env gene fragment of the BLV provirus isolates to detect the genotypic affiliation of the cattle from twenty-one livestock farms of the Republic of Tatarstan. As a result, isolates of four out of ten BLV genotypes were found in the Tatarstani cattle, namely genotypes 1, 4, 7, and 8. The research involved a comparative analysis of 505 nucleotide sequences of a fragment of the BLV env gene, including those deposited in GenBank NCBI. The analysis confirms the inconsistency of several earlier PCR-RFLP typing strategies with the current approach in assessing the genotypic diversity by phylogenetic analysis. The improved strategy of PCR-RFLP genotyping of BLV corresponds with its modern phylogenetic classification. The strategy makes it possible to identify all the known genotypes of the viral pathogen. Its validity has been proved by in silico modelling of restrictogrammes and a phylogenetic analysis of the env gene fragment of 57 reference isolates of ten BLV genotypes that generate 57 genotype-associated combinations of diagnostically significant PCR-RFLP profiles.

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PCR-RFLP ITS-based analysis of wine yeast autochthonous strains isolated from different grape cultivars in Taman subregion

E3S Web of Conferences ◽

10.1051/e3sconf/202128505020 ◽

2021 ◽

Vol 285 ◽

pp. 05020

Author(s):

Elena Lobodina ◽

Ivan Suprun ◽

Natalya Ageeva ◽

Ekaterina Al-Nakib

Keyword(s):

Genetic Analysis ◽

Restriction Analysis ◽

Its Region ◽

Wine Yeast ◽

Product Size ◽

Rdna Its ◽

Hanseniaspora Uvarum ◽

Pcr Rflp ◽

Autochthonous Yeast ◽

Rdna Its Region

The studies present the results of morphological, cultural and genetic analysis of the ITS1-ITS4 region of the autochthonous yeast strains genome by using the HaeIII restriction enzyme. On the red and white grapes varieties, based on the morphology of the cells, autochthonous strains belonging to the genus Saccharomyces prevail – 83.3%, what is confirmed by genetic analysis of rDNA ITS region. Restriction analysis showed that all strains of the genus Saccharomyces belong to the species Saccharomyces cerevisiae / S. paradoxus. The percentage of Saccharomyces isolated on the Pervenets Magaracha variety is 86.7%, Krasnostop Anapsky - 80%. The non-Saccharomyces yeast had a product size of 750 bp, presumably of the species Hanseniaspora uvarum.

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Uveitis associated with Toxocara; Clinic, Diagnosis, and Treatment

Güncel Retina Dergisi (Current Retina Journal) ◽

10.37783/crj-0172 ◽

2019 ◽

pp. 244-246

Keyword(s):

Parasitic Infection ◽

Vitreoretinal Surgery ◽

Toxocara Canis ◽

Clinical Findings ◽

Posterior Pole ◽

Larva Migrans ◽

Toxocara Cati ◽

Ocular Toxocariasis ◽

Fluid Analysis ◽

Ocular Larva Migrans

Ocular toxocariasis or ocular larva migrans is a parasitic infection via the ingestion of dog nematode Toxocara canis and cat nematode Toxocara cati larvae. It usually affects only one eye of the child under the age of sixteen. The most common clinical findings in ocular toxocariasis are peripheral granuloma, posterior pole chorioretinal eosinophilic granulomas, and endophthalmitis or pars planitis. It is diagnosed with clinical findings in developing countries, ELISA antibody tests, and, if necessary, intraocular fluid analysis can be made. Ocular toxocariasis must be distinguished from retinoblastoma and other congenital and inflammatory eye conditions of childhood. In treatment, besides anthelmintic agents, steroid use and vitreoretinal surgery may be needed.

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Toxocarosis

10.1093/med/9780198570028.003.0071 ◽

2011 ◽

Author(s):

Sheelagh Lloyd ◽

Eric R. Morgan

Keyword(s):

Immune Responses ◽

Toxocara Canis ◽

Larva Migrans ◽

Toxocara Cati ◽

Stray Dogs ◽

Baylisascaris Procyonis ◽

Wild Canids ◽

Ocular Larva Migrans ◽

Free Ranging

Toxocara canis and the syndromes of visceral and ocular larva migrans (VLM, OLM), covert toxocarosis, and neurological toxocarosis are described. Other potential agents, particularly Toxocara cati and Baylisascaris procyonis , are described. The transmission dynamics of toxocarosis to humans have never been fully elucidated, but the potential roles of pet and stray dogs, foxes, cats, and the influence of their population densities, and age demographies, are discussed in relation to contamination of the environment with eggs. Routes of infection with eggs by geophagia, poor hygiene outdoors and with dogs, and fly-borne contamination of food, and meat-borne ingestion of larvae are described. The development of prolonged in vitro culture and analyses of T. canis larval excretions/secretions (TES) and surface antigens helped explain the importance of the rapid production and shedding of TES in the prolonged course of infection and pathogenesis of disease. TES also have greatly improved serodiagnosis. However, we still have insufficient understanding of differences in the aetiology of the larvae or differences in immune responses among individuals to account for development of VLM, covert toxocarosis, or OLM in different individuals. Our understanding of the immunopathological response of the host to TES has emphasized the need for anti-inflammatory therapy in treatment; unfortunately, less information is available on the true efficacy of the anthelmintics available. The complexity of the T. canis life cycle in dogs is described and therapeutic regimens to prevent excretion of eggs by pet dogs are given. This, plus adequate control or exclusion of stray or wild canids from a property could prevent most cases of VLM. Control of infection from free-ranging stray dogs, cats and foxes, will be difficult and more data are needed to clarify the importance of these and of fly-borne and meat-borne transfer of infection to humans for control.

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PCR-based genetic identification of marlin and other billfish

Marine and Freshwater Research ◽

10.1071/mf98007 ◽

1998 ◽

Vol 49 (5) ◽

pp. 383 ◽

Cited By ~ 8

Author(s):

B. H. Innes ◽

P. M. Grewe ◽

R. D. Ward

Keyword(s):

Mitochondrial Dna ◽

Genetic Test ◽

Rflp Analysis ◽

Restriction Enzymes ◽

Genetic Identification ◽

Dna Molecule ◽

Pcr Rflp ◽

D Loop ◽

Specific Variation ◽

Species Specific

A genetic test was developed for the identification of the six species of billfish found in Australian waters (black marlin, Indo–Pacific blue marlin, striped marlin, Indo–Pacific sailfish, shortbill spearfish and broadbill swordfish). The test was based on the PCR–RFLP analysis of a 1400 bp region of the mitochondrial DNA molecule, the d-loop, using four restriction enzymes (Hinf I, Rsa I and Sau3A I andTaq I). A total of 33 composite haplotypes were observed among 160 fish; all were species-specific. Three of the species—black marlin, striped marlin and broadbill swordfish—showed sufficient intra-specific variation to be useful in population structure analyses.

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