Characterisation of the mitochondrial genome and phylogenetic analysis of Toxocara apodemi (Nematoda: Ascarididae).

We first sequenced and characterised the complete mitochondrial genome of Toxocara apodeme, then studied the evolutionary relationship of the species within Toxocaridae. The complete mitochondrial genome was amplified using PCR with 14 specific primers. The mitogenome length was 14303 bp in size, including 12 PCGs (encoding 3,423 amino acids), 22 tRNAs, 2 rRNAs, and 2 NCRs, with 68.38% A+T contents. The mt genomes of T. apodemi had relatively compact structures with 11 intergenic spacers and 5 overlaps. Comparative analyses of the nucleotide sequences of complete mt genomes showed that T. apodemi had higher identities with T. canis than other congeners. A sliding window analysis of 12 PCGs among 5 Toxocara species indicated that nad4 had the highest sequence divergence, and cox1 was the least variable gene. Relative synonymous codon usage showed that UUG, ACU, CCU, CGU, and UCU most frequently occurred in the complete genomes of T. apodemi. The Ka/Ks ratio showed that all Toxocara mt genes were subject to purification selection. The largest genetic distance between T. apodemi and the other 4 congeneric species was found in nad2, and the smallest was found in cox2. Phylogenetic analyses based on the concatenated amino acid sequences of 12 PCGs demonstrated that T. apodemi formed a distinct branch and was always a sister taxon to other congeneric species. The present study determined the complete mt genome sequences of T. apodemi, which provide novel genetic markers for further studies of the taxonomy, population genetics, and systematics of the Toxocaridae nematodes.

Mitogenomic phylogenies suggest the resurrection of the subfamily Porrocaecinae and provide insights into the systematics of the superfamily Ascaridoidea (Nematoda: Ascaridomorpha), with the description of a new species of Porrocaecum.

BACKGROUND: The family Toxocaridae is a group of zooparasitic nematodes of veterinary, medical and economic significance. However, the evolutionary relationship of Porrocaecum and Toxocara, both genera currently classified in Toxocaridae, and the monophyly of the Toxocaridae remain under debate. Moreover, the validity of the subgenus Laymanicaecum in the genus Porrocaecum is open to question. Due to the scarcity of an available genetic database, molecular identification of Porrocaecum nematodes is still in its infancy. METHODS: A number of Porrocaecum nematodes collected from the Eurasian marsh harrier Circus aeruginosus (Linnaeus) (Falconiformes: Accipitridae) in the Czech Republic were identified using integrated morphological methods (light and scanning electron microscopy) and molecular techniques (sequencing and analyzing the nuclear 18S, 28S and ITS regions). The complete mitochondrial genomes of the collected nematode specimens and of Porrocaecum (Laymanicaecum) reticulatum (Linstow, 1899) were sequenced and annotated for the first time. Phylogenetic analyses of ascaridoid nematodes based on the amino acid sequences of 12 protein-coding genes of mitochondrial genomes were performed using maximum likelihood and Bayesian inference. RESULTS: A new species of Porrocaecum, named P. moraveci n. sp., is described based on the morphological and genetic evidence. The mitogenomes of P. moraveci n. sp. and P. reticulatum both contain 36 genes and are 14,517 and 14,210 bp in length, respectively. Comparative mitogenomics revealed that P. moraveci n. sp. represents the first known species with three non-coding regions and that P. reticulatum has the lowest overall A + T content in the mitogenomes of ascaridoid nematodes tested to date. Phylogenetic analyses showed the representatives of Toxocara clustered together with species of the family Ascarididae rather than with Porrocaecum and that P. moraveci n. sp. is a sister to P. reticulatum. CONCLUSIONS: The characterization of the complete mitochondrial genomes of P. moraveci n. sp. and P. reticulatum is reported for the first time. Mitogenomic phylogeny analyses indicated that the family Toxocaridae is non-monophyletic and that the genera Porrocaecum and Toxocara do not have an affinity. The validity of the subgenus Laymanicaecum in Porrocaecum was also rejected. Our results suggest that: (i) Toxocaridae should be degraded to a subfamily of the Ascarididae that includes only the genus Toxocara; and (ii) the subfamily Porrocaecinae should be resurrected to include only the genus Porrocaecum. The present study enriches the database of ascaridoid mitogenomes and provides a new insight into the systematics of the superfamily Ascaridoidea.

Molecular detection and phylogenetic analysis of Toxocara vitulorum in feces and milk samples from naturally infected water buffaloes.

Toxocara vitulorum infects cattle and water buffalo, leading to mild to severe infection in calves and has wide geographic distributions, especially in tropical and subtropical countries. This work aimed to assess the prevalence, distributions, and phylogeny of T.vitulorum in water buffaloes in different parts of Sivas, one of the essential buffalo-breeding areas in Türkiye. T.vitulorum was found in 42 (8.23%) and 54 (10.58%) fecal (n:510) samples using microscopic and molecular techniques, respectively. T.vitulorum was higher in animals aged 0-6 months compared to other groups. Furthermore, when animals aged 0-6 months were grouped within themselves, the prevalence of T.vitulorum in 1-3 month-old-animals was higher than in both younger than one month and older than three months. T.vitulorum was detected in fecal samples obtained from animals older than six months. In colostrum/milk samples (n:100), T.vitulorum-larvae were found in 4% and 10% with microscopic and molecular techniques, respectively. The larvae were detected in colostrum/milk samples in the mother between the 2nd and 28th days postpartum-period. The ITS-1-gene of 11 PCR-positive samples was sequenced. The 98.99-100% nucleotide identity was determined between our T.vitulorum isolates and those present in GenBank. In conclusion, this is the first molecular survey and phylogenetic analysis of T.vitulorum in fecal and colostrum/milk samples from naturally infected water buffaloes. Data obtained in this study will help to understand the life cycle and epidemiology of the nematode. Data also revealed that veterinarians should consider older animals as well as young animals in their control program of nematode infections in farms.

Detection of Toxocara species larvae in four Iranian free-range broiler farms.

BACKGROUND: The epidemiology of Toxocara canis and Toxocara cati in food animals, associated products, and their zoonotic potential are poorly understood. A cross sectional study was designed to determine the prevalence of Toxocara spp. larvae from free-range broiler chickens in traditional farms using conventional techniques and molecular method. Eight-hundred tissue samples including liver, gizzard, lungs and heart were collected from 200 chickens belonging to different regions of Zanjan Province, Iran and were processed by conventional and molecular methods. RESULTS: Out of 800 chicken tissues, 49 samples (6.1%) were positive for nematode larvae. Polymerase chain reaction was performed to identify species-specific of Toxocara larvae. The findings showed that 10.5% (21 out of 200) chickens were infected with Toxocara species, so that 57.1% (12 out of 21) of the samples were positive for Toxocara canis and 42.9% (9 out of 21) of the samples were positive for Toxocara cati. CONCLUSION: Considering the significant contamination/infection of free-range broiler chickens with Toxocara larvae, the consumption of chicken meat and viscera, especially liver and gizzards, can play an important role in the transmission of infection to humans. Prevention and control measures focused on regular deworming of dogs and cats, increasing public awareness of Toxocara infection are recommended.

Genetic and population diversity of Toxocara cati (Schrank, 1788) Brumpt, 1927, on the basis of the internal transcribed spacer (ITS) region.

The present investigation was aimed to study the sequence, phylogenetic and haplotype analyses of Toxocara cati based on the ITS region, along with the genetic diversity, demographic history and population-genetic structure. The maximum likelihood tree based on Kimura 2-parameter model was constructed using the complete ITS region of all the nucleotide sequences (n = 57) of Toxocara spp. and other related ascarid worms available in the GenBank™. It placed all the sequences of T. cati into four major clades designated as T. cati genotypes 1-4 (TcG1-G4). A total of 66 signature nucleotides were identified in the ITS region between genotypes. The median-joining haplotype network displayed a total of 24 haplotypes, with China exhibiting the highest number of haplotypes (h = 20) followed by India (h = 4), and Japan and Russia (h = 1). It indicated a clear distinction between all the four genotypes. The pairwise FST values between all the genotypes indicated huge genetic differentiation (> 0.25) between different T. cati genotypes. Moreover, the gene flow (Nm) between T. cati genotypes was very low. Results of AMOVA revealed higher genetic variation between genotypes (92.82%) as compared to the variation within genotypes (7.18%). The neutrality indices and mismatch distributions for the G1-G4 genotypes, Indian isolates and the overall dataset of T. cati indicated either a constant population size or a slight population increase. The geographical distribution of all the genotypes of T. cati is also reported. This is the first report of genotyping of T. cati on the basis of the ITS region.

Toxocara DNA amplification in serum and tissue samples in BALB/c mice.

Toxocariasis is still a neglected parasitic disease worldwide and much about its biology and diagnosis has yet to be understood. The migration of third stage larvae via bloodstream suggests a potential use of molecular tools in diagnosis as well to deepen the knowledge about its migration behaviors. Conventional PCR was applied in serum and tissue samples from BALB/c mice infected with 5 and 500 embryonated eggs. Blood samples were collected at 15, 30, 60, 90 and 120 days post-infection. Organs were excised at 170 days post infection. There was no DNA amplification in serum samples in any group or day post-infection; contrarily, tissue samples showed DNA amplification. These results also support a continuous larval migration after and/or simultaneously with the neurotropic-myotropic phase. Thus, molecular tools might be useful as a differential diagnosis method, but do not replace immunodiagnostics techniques.

Detection of larvae of Toxocara cati and T. tanuki from the muscles of free-ranging layer farm chickens.

Although raw or undercooked livestock meat or viscera has been suggested to be a source of human toxocariasis, there have been few reports on the prevalence of Toxocara larvae in the tissue of livestock animals. To investigate the presence of Toxocara larvae in chickens, we examined 50 culled chickens from a commercial layer farm. The liver, breast meat, and thigh meat were separated individually and artificially digested to examine for the presence of larvae. Nematode larvae were detected in 2 out of 50 chickens. One larva was detected from the breast meat, and it was molecularly identified as Toxocara tanuki. The other from the thigh meat of another chicken was molecularly identified as Toxocara cati. The present study demonstrated for the first time that T. tanuki larvae do infect chickens in the natural environment. The fact that Toxocara spp. larvae were found in muscles of farm chickens suggests that consumption of raw or undercooked chicken meat may present a risk for human toxocariasis.

Comparison of the Detection of Toxocara Spp. in the Soils of Public Parks of Ahvaz (Southwest of Iran) by PCR and Loop-Mediated Isothermal Amplification (LAMP).

INTRODUCTION: The infections caused by Toxocara spp. are considered as one of the most important zoonotic diseases in the world, especially in developing countries. Human toxocariasis, particularly in children, is acquired by playing in public parks. Hence, the aim of the current study was to detect Toxocara spp. in the soils of public parks of the city of Ahvaz, southwest of Iran, using the PCR and loop-mediated isothermal amplification (LAMP) methods. METHODS: In this descriptive cross-sectional study, 260 soil samples were randomly collected from the different public parks of the city of Ahvaz. After performing zinc sulfate (ZnSO4) flotation technique, the DNA samples were extracted from the isolated Toxocara spp. eggs. Lastly, the extracted DNA was used for PCR and LAMP-based molecular detection. RESULTS: Out of 260 specimens, 57 (21.9%) samples were found positive for Toxocara spp., using the PCR method, out of that 38 (14.6%) samples were positive for T. canis and 19 (7.3%) samples were positive for T. cati. Also, out of 260 specimens, 81 (31.1%) cases were positive for Toxocara species, using the LAMP method, among them 51 (19.6%) samples were found positive for T. canis and 30 (11.5%) samples were positive for T. cati. Kappa (κ) coefficient between PCR and LAMP showed a strong agreement (0.766, P-value=0.002). CONCLUSION: The obtained data showed a relatively high outbreak of Toxocara spp. in the public parks' soils of the city, using the PCR and LAMP methods. Since the parasite can cause human toxocariasis, particularly in children; thus, the health authorities of the city of Ahvaz and other similar cities, especially in developing countries, must pay more attention to the hygiene of the public parks' soils.

Morphological and Molecular Characterization of Toxocara apodemi (Nematoda: Ascarididae) from Striped Field Mice, Apodemus agrarius, in Korea.

Adult ascarid worms from the field mice, Apodemus agrarius, were observed with a light and scanning electron microscope, and molecularly analized with 18S rRNA gene. In the scanning electron microscope, 3 prominent labia were present in the anterior end of male and female worms, but the interlabia and gubernaculum were absent. Scanning electron micrographs showed cervical alae as vestigial organs that looked like a slightly uplifted superficial sewing stitch. Total 6 pairs of post-cloacal papillae were observed on the tail of the male worms. The tail of female worms was blunt and conical shape with a spine-like structure, mucron. The eggs were sub-globular, coated with the albuminous layer and 73 by 82 µm in average size. The superficial pits of T. apodemi egg (mean 8.6×6.7 µm) are obviously bigger than those of Toxocara spp. The partial sequence of 18S rRNA showed the sequence homology of Toxocara canis (99.6%), Toxocara cati (99.4%), Toxascaris leonina (99.4%), and Toxocara vitulorum (99.2%). Conclusively, it was confirmed that ascarid nematodes, Toxocara apodemi, recovered from striped field mice in Korea are taxonomically conspecific relationship with genus Toxocara and genetic divergence from other Toxocara species.

A two-step morphology-PCR strategy for the identification of nematode larvae recovered from muscles after artificial digestion at meat inspection.

To ensure that meat from livestock and game is safe for human consumption, European legislation lays down rules for mandatory testing. Helminth larvae are a category of zoonotic foodborne pathogens that can contaminate meat. Among helminths, the only zoonotic nematode regulated in Europe regarding meat inspection is Trichinella spp.. It is precisely during Trichinella testing that other potentially zoonotic larvae can be found. Due to current lack of tools, their identification is often very complicated. Nematode larvae other than Trichinella, recovered from artificial digestions of pig and wild boar muscles from France and Germany, were subjected to a newly developed two-step identification scheme, which includes both morphological examination and molecular assays. The first step is a general orientation towards a broad taxonomic group; the second step consists of targeted identification based on the results of first step. Different parasites were identified, some of which were not zoonotic such as Metastrongylus spp. and Angiostrongylus vasorum, but others are known to be zoonotic such as Toxocara cati, Ascaris suum, and Uncinaria stenocephala. The strategy is efficient for the identification of nematode larvae recovered from muscles but could also be applied for larvae from other sources.

Molecular characterization of ascaridoid parasites from captive wild carnivores in China using ribosomal and mitochondrial sequences.

BACKGROUND: Despite the public health importance of toxocariasis/toxascariasis, only a few species of these ascaridoid parasites from wild canine and feline carnivores have been studied at the molecular level so far. Poor understanding of diversity, host distribution and the potential (zoonotic) transmission of the ascaridoid species among wild animals negatively affects their surveillance and control in natural settings. In this study, we updated previous knowledge by profiling the genetic diversity and phylogenetic relationships of ascaridoid species among eleven wild canine and feline animals on the basis of a combined analysis of the ribosomal internal transcribed spacer region (ITS) gene and the partial mitochondrial cytochrome c oxidase subunit 2 (cox2) and NADH dehydrogenase subunit 1 (nad1) genes. RESULTS: In total, three genetically distinct ascaridoid lineages were determined to be present among these wild carnivores sampled, including Toxocara canis in Alopex lagopus and Vulpes vulpes, Toxocara cati in Felis chaus, Prionailurus bengalensis and Catopuma temmincki and Toxascaris leonina in Canis lupus, Panthera tigris altaica, Panthera tigris amoyensis, Panthera tigris tigris, Panthera leo and Lynx lynx. Furthermore, it was evident that T. leonina lineage split into three well-supported subclades depending on their host species, i.e. wild felids, dogs and wolves and foxes, based on integrated genetic and phylogenetic evidence, supporting that a complex of T. leonina other than one species infecting these hosts. CONCLUSIONS: These results provide new molecular insights into classification, phylogenetic relationships and epidemiological importance of ascaridoids from wild canids and felids and also highlight the complex of the taxonomy and genetics of Toxascaris in their wild and domestic carnivorous hosts.

Toxocara "omics" and the promises it holds for medicine and veterinary medicine.

Toxocariasis is one of the most neglected worldwide zoonoses that is caused by larval nematode parasites of the genus Toxocara, Toxocara canis, and to a lesser extent, Toxocara cati, whose migration mechanism is still largely unknown. Fortunately, some advanced tools have been employed, such as genomics, transcriptomics, and proteomics, to better understand the molecular biology and regulatory mechanisms of Toxocara. Using genomics and transcriptomics, we can identify a large number of genes that participate in the development of Toxocara and the interaction of parasites and their hosts and can predict the functions of unknown genes by comparing them with other relevant species. Using proteomics, we can identify somatic proteins and excretory and secretory (ES) proteins that perform specific biological functions in tissue degradation, pathogen invasion, immune evasion or modulation. These "omics" techniques also can contribute enormously to the development of new drugs, vaccines and diagnostic tools for toxocariasis. In a word, by utilizing "omics", we can better understand the Toxocara and toxocariasis. In this review, we summarized the representative achievements in Toxocara and the interaction between Toxocara spp. and their hosts based on expressed sequence tags (ESTs), microarray gene expression, next-generation sequencing (NGS) technologies and liquid chromatography-tandem mass spectrometry (LC-MS/MS), hoping to better understand the molecular biology of Toxocara, and contribute to new progress in the application areas of new drugs, vaccines and diagnostic tool for toxocariasis in the future.

Phylogenetic relationships among Toxocara spp. and Toxascaris sp. from different regions of the world.

Toxocara and Toxascaris are parasitic nematodes that infect canids and felids although species of the genus Toxocara also infect humans. This work aimed to establish the phylogenetic and phylogeographic relationship between specimens of T. canis, T. cati, T. malaysiensis and Toxascaris leonina and to evaluate the degree of host specificity. In total, 437 samples (adults and pools of eggs) were collected from canids and felids from eight countries. Parasites were identified by morphology, PCR linked Restriction Fragment Length Polymorphism (PCR-RFLP) and partial sequencing of the mitochondrial gene cox1. Phylogenetic trees were constructed and genetic distance among isolates was estimated. Based on the molecular characterization all worms were identified in agreement with their respective hosts with the exception of three samples; two from cats and one from dogs identified as T. canis and T. cati, respectively. There was no clear geographical clustering of the samples despite this study including parasites from three continents. This is the first study, to our knowledge, to use molecular methods to identify T. canis in cats and T. cati in dogs with host specificity being the most common finding. Our developed PCR-RFLP method was found to be a facile and reliable method for identifying Toxocara species.

A systematic review and meta-analysis of canine, feline and human Toxocara infections in sub-Saharan Africa.

Toxocariasis is an emerging zoonotic disease caused by Toxocara canis and T. cati. Toxocariasis and its etiological agents are of global public health importance, whose burden appears underestimated, especially in sub-Saharan Africa (SSA). The diversity in the transmission routes of these parasites contributes to disease prevalence and often hinders disease control measures. This study aimed to review the epidemiological distribution of Toxocara infections in SSA region. A systematic review and meta-analysis were performed using PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analysis). We identified 94 relevant, peer-reviewed articles, out of which, 75 articles were found eligible based on Toxocara infections in dogs, cats and humans. Overall, 27,102 samples were examined for T. canis in dogs, T. cati in cats and Toxocara serology in humans, out of which 6142 were positive for Toxocara infection: 3717 (13.7%) in dogs (faecal, 3487; necropsy, 180; hair, 50); 266 (1%) in cats (faecal, 101; necropsy, 165); and 2159 (8%) in humans (serology). Overall mean prevalences of 19% (95% confidence interval (CI): 14-23%), 9% (95% CI: 0-28%) and 36% (95% CI: 24-49%) were recorded in dogs, cats and humans, respectively. Substantial heterogeneity was observed between studies and subgroups (I2 = 99%, P < 0.01). Findings from the review showed that studies on the epidemiology of Toxocara infections in the SSA region are limited. We strongly recommend focused, collaborative and coordinated studies to determine Toxocara spp. prevalence in various hosts, including food animals and the environment, through a 'One Health' approach across SSA countries.

Soil contamination of a public park by human and canine mastadenovirus, as well as hookworms and Toxocara spp eggs.

Public parks are leisure environments widely used by both, adults and children, often accompained by their pets. Soil contamination of these environments by enteric viruses and intestinal parasites occurs through these animals feces. The aim of this work was to detect Carnivore protoparvovirus 1 (CPV-1) and different species of Mastadenovirus in soils samples from a park located in a medium-sized city in Brazil and evaluate the presence of helminth eggs and larvae in 18 points of a public park soil samples, as well as feces found on this site during six months. Parasitological analyzes were conducted through flotation and sedimentation techniques, and polymerase chain reaction (PCR) was used for viral detection. Of the 216 soil and 16 feces samples, 49% (106/216) and 12% (2/16) were positivefor nematodes larvae, respectively, through sedimentation techniques. Toxocara spp eggs were found in one soil sample and one feces sample, Trichuris spp eggs were found in only one feces sample and Hookworms eggs were found in four soil samples. After reconstruction work in the streets near the park, 30% (64/216) of the samples were positive for Human Mastadenovirus C (HAdV-C), 1.4% (3/216) for HAdV-E and 0.4% (1/216) for Canine Mastadenovirus A (CAdV-A). The parasitic forms found in this study have demonstrated that the contamination of the park's soil pose a threat to human and animal health. This was the first study to report the presence of HAdVs and CAdVs in soil samples.

Survey of German pet owners quantifying endoparasitic infection risk and implications for deworming recommendations.

BACKGROUND: Dogs and cats can transmit zoonotic helminths to humans, e.g. Toxocara spp. and Echinococcus multilocularis. Strategic deworming may help minimize this risk. Studies in several European countries have shown that pets are dewormed less frequently against roundworms and tapeworms than recommended by the European Scientific Counsel Companion Animal Parasites (ESCCAP). The objective of this study was to identify percentages of dogs and cats falling into the different risk categories defined by the German ESCCAP guidelines and to evaluate whether deworming frequency and parasite monitoring in Germany follows these guidelines. RESULTS: According to questionnaire results from 500 dog and 500 cat owners, deworming of dogs in Germany averages 2.07 times/year while for cats this average is 1.72 times/year. In contrast, evaluation of risk factors placed only 2% (10/500) of dogs in ESCCAP category A with a recommended deworming/examination frequency of 1-2 times per year, while 4.8% (24/500) were placed in category B (4 treatments/examinations per year recommended), 30.8% (154/500) in category C (12 treatments/examinations per year against tapeworms and 4 treatments/examinations per year against roundworms recommended) and 62.4% (312/500) in category D (12 treatments/examinations per year recommended). All cats were placed either in risk group A [52.8% (264/500)] or D [47.2% (236/500)]. Generalized linear models indicated that risk group D cats were treated significantly more often against helminths than risk group A cats. There were no significant differences in deworming frequency between risk groups in dogs. The most important factor influencing deworming frequency was the frequency of veterinary visits. Dogs and cats were treated significantly more often if owners visited their veterinarian more than once yearly. CONCLUSIONS: The percentage distribution of risk groups considerably varied between dogs and cats. Nevertheless, 62% of dogs and 47% of cats were assigned to category D for which monthly treatments/examinations are recommended by the ESCCAP guidelines. Veterinarians play a key role in instructing pet owners with regard to helminthoses and their prevention, and should take the time for adequate risk assessments. The reported low deworming frequencies despite the high potential parasite infection risk suggests that pet owner advice through veterinarians needs to be improved.

Molecular characterization of Toxocara spp. eggs isolated from public parks and playgrounds in Shiraz, Iran.

Human 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.

Toxocara sero-prevalence and its relationship with allergic asthma in asthmatic patients in north-eastern Iran.

Toxocariasis is an important neglected tropical disease that has been suggested as a possible etiologic agent of asthma. The objective of the present study was to investigate possible significant association between Toxocara seroprevalence and asthma in a clinic-based case-control study. Blood samples were collected from human subjects aged 5-70 years, 50 of whom had signs of asthma and 50 of whom had no signs of asthma. Risk factors for asthma and Toxocara spp. infection were assessed by a questionnaire given to each patient. Blood samples were analysed to measure levels of anti-Toxocara spp. immunoglobulin G (IgG). Patients with bronchial asthma were observed to have higher Toxocara spp. seropositivity than that of the non-asthmatic controls (6 vs 2%, P = 0.47). The mean anti-Toxocara spp. antibody titre was not significantly higher in patients with bronchial asthma than in individuals without asthma (P = 0.395, 95% CI = 0.579-1.45). There was no significant difference in the mean age, sex, social class, exposure to smoking and presence of domestic dog or cat at home between the two groups (P ≥ 0.05). The presence of anti-Toxocara spp. IgG was statistically associated with higher blood eosinophils, but it was not associated with asthma (P ≥ 0.05). The observed relationship between exposure to Toxocara spp. infection and bronchial asthma in Iranian patients warrants further evaluation. An understanding of any potential influence on the pathogenesis of human asthma provides a potential avenue for prevention.

Morphological and Molecular Characterization of Toxocara tanuki (Nematoda: Ascaridae) from Korean Raccoon Dog, Nyctereutes procyonoides koreensis.

Present study was performed to describe the morphological and molecular characterization of Toxocara tanuki (Nematoda: Ascaridae) from Korean raccoon dog, Nyctereutes procyonoides koreensis, naturally infected in the Republic of Korea (Korea). Juvenile and adult worms of T. tanuki were recovered in 5 out of 10 raccoon dogs examined and the larval worms were detected in 15 out of 20 muscle samples (75%). Small lateral alae were observed on the cranial end of the body in male and female adults and 2 long spicules (3.0-3.5 mm) were characteristically observed in the posterior end of males. In SEM observation, 18 pairs of proximal precloacal, a precloacal median, a postcloacal median and 5 pairs of postcloacal papillae were uniquely revealed in the posterior portion of males, but the proximal papillae were not shown in the lateral ends of females. Molecular analysis on the 18S rRNA partial DNA sequences was revealed the same finding in both samples, adult worms and muscle larvae, which are closely related to T. tanuki. In conclusion, it was confirmed for the first time that T. tanuki is indigenously distributed, the Korean raccoon dog is acted as the natural definitive host of this nematode in Korea and the morphological characteristics of T. tanuki were shown in specific structure for single postcloacal median papilla in male.

Not just a walk in the park: Occurrence of intestinal parasites in dogs roaming recreational parks in Chandigarh, Northern India.

Chandigarh, a city in North-west India, has numerous parks and recreational areas where stray dogs roam freely and pet dogs are exercised. This allows for extensive human-dog interaction, which may pose a public health threat. The aim of this study was to determine the occurrence of gastrointestinal parasites of public health importance, and their seasonal variation, in canine faecal samples obtained from recreational parks in Chandigarh. A total of 212 samples were collected from 10 parks during the winter (January 2015; N = 107) and monsoon season (September 2015; N = 105), to assess the prevalence of intestinal zoonotic parasites and any seasonal variations. The samples were analysed for helminth eggs using McMaster counting chambers. Immunofluorescent antibody testing was used to analyse samples for Cryptosporidium oocysts and Giardia cysts. The Giardia-positive samples were genotyped by conventional multi-locus PCR to determine their assemblage and zoonotic potential. Among the 212 samples, strongyle-type eggs were found in 34 (16.0%), Toxocara spp. eggs were found in 6 (2.8%), taeniid eggs in 1 (0.5%), Cryptosporidium spp. oocysts in 4 (1.9%) and Giardia duodenalis cysts in 49 (23.1%). Trichuris eggs were not detected. The majority of the successfully amplified Giardia isolates belonged to canid-specific assemblages. The prevalence of Giardia cysts in faecal samples was significantly higher during winter than in the monsoon season, whereas helminth-egg prevalence unaffected by season. The prevalence of strongyle-type eggs and Giardia cysts in dog faeces was lower in more affluent areas of the city than those of less affluence. There was no significant difference in the intensity of infection between the seasons. The results indicate that faeces from dogs contaminating parks in Chandigarh do not usually contain parasite transmission stages that pose a significant risk to human health. However, the importance of minimising contamination of public parks with dog faeces is highlighted.