Serosurvey and associated risk factors of anti-Toxocara spp. antibodies in bovines from slaughterhouses of southeastern Brazil.

BACKGROUND: Toxocariasis, caused by a nematode species of the genus Toxocara, has been described as one of the most prevalent zoonotic helminthiases worldwide. Human transmission may occur by ingesting Toxocara spp. larvae from raw or undercooked meat or organs; however, no comprehensive serosurvey study has been conducted to date investigating the role of cattle as paratenic hosts. The aim of the study reported here was to assess the prevalence of anti-Toxocara spp. antibodies and associated risk factors in bovines from two slaughterhouses located in Presidente Prudente, southeastern Brazil. METHODS: Blood samples were collected and tested by indirect enzyme-linked immunosorbent assay (ELISA). Cattle farmers voluntarily responded to an epidemiologic questionnaire. RESULTS: Overall, 213 of the 553 (38.5%) bovine samples were assessed as seropositive for anti-Toxocara spp. antibodies by indirect ELISA. Multivariate analysis revealed that the source of beef cattle and the presence of dogs or cats at the farm were associated with seropositivity. The use of feedlot systems was associated with lower likelihood of seropositivity. CONCLUSIONS: These results indicate a high level of anti-Toxocara seropositivity in slaughterhouse cattle, with potentially contaminated meat posing an infection risk to humans. In addition, the presence of dogs and cats where the slaughtered beef cattle were raised was statistically associated with bovine seropositivity, probably due to the overlapping environment at the farm and the lack of pet deworming. The use of feedlot systems was a protective factor likely due to the absence of dog and cat contact, elevated feeding troughs that avoid contact with contaminated soil or grass, and younger age at slaughter of feedlot cattle. In summary, bovines may be used as environmental sentinels of Toxocara spp. contamination, and high seropositivity of slaughterhouse cattle may indicate a potential risk of human toxocariasis through the ingestion of raw or undercooked contaminated meat.

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.

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 species environmental contamination of public spaces in New York City.

Human toxocariasis has been identified as an under-diagnosed parasitic zoonosis and health disparity of significant public health importance in the United States due to its high seropositivity among socioeconomically disadvantaged groups, and possible links to cognitive and developmental delays. Through microscopy and quantitative PCR, we detected that Toxocara eggs are widespread in New York City public spaces, with evidence of significant levels of contamination in all five boroughs. The Bronx had the highest contamination rate (66.7%), while Manhattan had the lowest contamination rate (29.6%). Moreover, infective eggs were only found in the Bronx playgrounds, with over 70% of eggs recovered in embryonic form and the highest egg burden (p = 0.0365). All other boroughs had eggs in the pre-infectious, unembronyated form. Toxocara cati, the cat roundworm, was the predominant species. These results suggest that feral or untreated cats in New York City represent a significant source of environmental contamination. These findings indicate that human toxocariasis has emerged as an important health disparity in New York City, with ongoing risk of acquiring Toxocara infection in public spaces, especially in poorer neighborhoods. There is a need for reducing environmental Toxocara contamination. Additional rigorous public health interventions should explore further approaches to interrupt transmission to humans.

History of Toxocara and the associated larva migrans.

This review covers the systematics and nomenclature of the Ascaridoid genus toxocara, and more specifically the species Toxocara canis and Toxocara cati. Also discussed is the discovery of the persistence of these larvae in the tissues of paratenic hosts, and the role that other species of this genus might or could play in other such hosts; including those where the life cycle has been described, i.e., Toxocara vitulorum, Toxocara pteropodis, Toxocara mackerrasae, and Toxocara tanuki. Also examined is the work that led to the realization that the larval stage leaving the egg actually being a third rather than a second stage larva. Also discussed on the work showing that the larvae can persist in paratenic host with remarkable longevity without undergoing any morphological change for years and that these larvae can be transmitted from one paratenic host to another by ingestion. People are usually infected by the ingestion of eggs containing third-stage larvae, but infections also occur on occasions from the ingestion of uncooked paratenic hosts.

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.

Ascarid infection in wild Amur tigers (Panthera tigris altaica) in China.

BACKGROUND: Wild Amur tigers are a sparsely populated species, and the conservation of this species is of great concern worldwide, but as an important health risk factor, parasite infection in them is not fully understanding. RESULTS: In this study, sixty-two faecal samples were collected to investigate the frequency and infection intensity of Toxocara cati and Toxascaris leonina in wild Amur tigers. The T. cati and T. leonina eggs were preliminary identified by microscopy, and confirmed by molecular techniques. Infection intensity was determined by the modified McMaster technique. Phylogenetic trees demonstrated that T. cati of wild Amur tiger had a closer relationship with which of other wild felines than that of domestic cats. T. leonina of Amur tiger and other felines clustered into one clade, showing a closer relationship than canines. The average frequency of T. cati was 77.42% (48/62), and the frequency in 2016 (100%) were higher than those in 2013 (P = 0.051, < 0.1; 66.6%) and 2014 (P = 0.079, < 0.1; 72.2%). The infection intensity of T. cati ranged from 316.6 n/g to 1084.1 n/g. For T. leonina, only three samples presented eggs when the saturated sodium chloride floating method was performed, indicating that the frequency is 4.83% (3/62). Unfortunately, the egg number in faecal smears is lower than the detective limitation, so the infection intensity of T. leonina is missed. CONCLUSIONS: This study demonstrated that ascarids are broadly prevalent, and T. cati is a dominant parasite species in the wild Amur tiger population.

Presence of Toxocara spp. and Other Zoonotic parasites Ova in Children's Playground in Karaman, Turkey

Objective: Human toxocariosis (HT) is a widespread and neglected parasitic disease around the world and it is caused by Toxocara canis and Toxocara cati, a common nematode found in dogs and cats. Childiren are caught to HT after ingestion of embriyonated Toxocara spp. eggs via contaminated materials such as soil, hair and etc. The aim of this study is to investigate Toxocara spp. and other zoonotic parasites in children's playgrounds in Karaman province of Turkey. Methods: In total, 103 samples (68 sand soil, 26 soil and 9 stool) from 20 randomly selected children's playgrounds in May 2018 in Karaman province, were investigated. Samples were examined by flotation in saturated NaCl solution and parasite ova were diagnosed under the light microscope morphologically. Results: Of the 20 screened playgorunds, 11 [55%, confidence interval (CI=33.6-75.2)]and 27 analyzed sample (26.2%, CI=18.4-35.2) were positive one or more parasite species. While Toxocara spp. eggs were the most common species in total (19.4%, CI=12.6-27.8), taeniid (Taenia spp., Echinococcus spp.) eggs and Ancylostoma spp. eggs were found in seven (6.8%, CI=2.97-12.7) and one (0.97%, CI=0.05-4.21) samples respectively. Also, one soil sample was found to be contaminated with both Toxocara and taeniid eggs. Conclusion: These results demonstrate that children's playgrounds in Karaman may be a source for HT and other zoonotic infections. We advise to be fenced children's playgrounds in order to prevent pet animal's accessibility.

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.

Prevalence survey of gastrointestinal and respiratory parasites of shelter cats in northeastern Georgia, USA.

The goal of this study was to assess the prevalence of gastrointestinal and respiratory parasites of shelter cats from northeast Georgia, thus promoting a more targeted approach in parasite diagnosis and treatment. Fecal samples of cats kept in a shelter located in Lavonia, northeastern Georgia, USA, were processed for the presence of parasites using double centrifugation sugar flotation (n = 103) and Baermann techniques (n = 98). Flotation revealed eggs of Toxocara cati (17.5%), Ancylostoma sp. (11.7%), Taeniidae (3.9%), Spirometra mansonoides (2.9%), Mesocestoides sp. (1%), Dipylidium caninum (1%), and Eucoleus aerophilus (1%), and oocysts of Cystoisospora felis (16.5%), and Cystoisospora rivolta (8.7%). Baermann diagnosed Aelurostrongylus abstrusus larvae in 5 cats (5.1%), while fecal flotation alone identified only 2 of these infections. Taeniidae eggs were identified to species-level by PCR and sequencing targeting the cytochrome oxidase c subunit 1 (cox1) of the mitochondrial DNA. All isolates belong to Hydatigera taeniaeformis sensu stricto, which is the first unequivocal report of the species in North America. Overall, 45.6% of the cats were infected with at least one parasite. This prevalence of infection is much higher than what is generally reported in client owned animals, highlighting the importance of using appropriate fecal diagnostic techniques to detect gastrointestinal and respiratory parasites on newly adopted cats. Correct diagnosis may direct appropriate treatment and control strategies, which would mitigate the risk of infection of other animals in household, and human exposure to zoonotic parasites.

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.

Dog walking brings Toxocara eggs to people's homes.

To find out the transmission routes for Toxocara infection, the possibility of transfer of Toxocara eggs from the soil on the paws of animals and on the shoes of people was explored. For this purpose, a study was conducted to find helminth eggs in washings from the paws of dogs after walking, from the shoes of their owners, as well as non-dog owners. Toxocara eggs were detected in 19.4% of the dogs' paws washings and in 11.4% of washings from the shoes of their owners. The number of eggs found on the paws was about twice as high as on the shoes. The mean number of eggs in the sample was 2.9 in washings from the paws and 1.8 from the shoes. In the samples, Toxocara cati eggs prevailed both in occurrence and in abundance. Out of the total number of positive samples, the eggs of T. cati were found in 83%, and T. canis in 42%. 79% of the found eggs belonged to T. cati and 21% to T. canis. In the washings from shoes of people that do not own dogs, the eggs of parasites were not found. This study demonstrates that the helminth eggs can be transferred from contaminated soil to people's homes on the paws and shoe soles. Even animals without a patent infection may take part in propagation of infection causing risks of human toxocariasis. In dogs, in addition, the transferred on paws T. canis eggs can serve as a causative agent of permanent, cumulative subclinical infection with a deferred manifestation in posterity. It is supposed that infestation "through the paws" is one of the probable routes of transmission of toxocariasis in dogs.

Identifying anti-Toxocara IgG antibodies in horses of Mexico / Identificação de anticorpos IgG anti-Toxocara em cavalos do México

Both the presence of owned dogs and stray dogs allows the spread of Toxocara, a parasite whose eggs can be found in soil, water and food. Animals, including horses, serve as definitive and paratenic hosts. In México, where consumption of horse meat is common, Toxocara is a zoonotic parasite. The aim of this study was to identify the presence of anti-Toxocara antibodies in work horses and horses intended for human consumption by ELISA. ELISA was chosen for analysis as paratenic hosts do not shed Toxocara eggs in their feces. Blood samples were collected from a total of 188 horses, 94 of which were work horses and 94 horses from the slaughter house. Samples were analyzed by ELISA, and the general equine seroprevalence was found to be 44.6% (n = 188). Adult horses for slaughter had a 61.7% greater presence of anti-Toxocara antibodies (p = 0.006). Toxocara IgG antibodies were found in horses, confirming that horses are paratenic hosts and possible sources of infection for other animals and people.(AU)

Tanto a presença de cães com dono quanto de cães vadios permitem a disseminação de Toxocara, e o parasita está presente no solo, na água e nos alimentos. Animais, incluindo cavalos, apresentam-se como hospedeiros definitivos e paratênicos. No México, o consumo de carne de cavalo é comum, e Toxocara é um parasita zoonótico. ELISA foi escolhido para análise, já que hospedeiros paratênicos não jogam ovos de Toxocara em suas fezes. O objetivo deste estudo foi identificar a presença de anticorpos anti-Toxocara por ELISA, em cavalos de trabalho e em cavalos para o consumo humano. As amostras de sangue foram retiradas de 188 cavalos: 94 cavalos de trabalho e 94 cavalos de trabalho do matadouro. Soros dos animais foram analisados por ELISA e 44,6% dos equinos apresentaram anticorpos anti-Toxocara. Cavalos adultos para abate têm 61,7% mais elevada a presença de anticorpos anti-Toxocara (P = 0,006). Anticorpos IgG Toxocara foram encontrados em cavalos, confirmando cavalos paratênicos como hospedeiros e possíveis fontes de infecção para outros animais e pessoas.(AU)

Toxocara Seroprevalence and Associated Risk Factors in the United States.

Caused by the parasitic nematodes Toxocara canis and cati, toxocariasis in humans can result in covert toxocariasis, ocular toxocariasis, visceral larval migrans, and neurotoxocariasis. A common infection, toxocariasis exposure varies widely within and between countries, with a previous estimate of Toxocara seroprevalence using data from 1988 to 1994 in the United States of approximately 13%. Age, poverty, sex, educational attainment, ethnicity, and region have been associated with Toxocara seroprevalence. In this study, we sought to determine the seroprevalence of and factors associated with Toxocara seropositivity in the United States using data from the National Health and Nutrition Examination Survey from 2011 to 2014 to provide a more recent estimate of Toxocara seroprevalence in the United States. We found an overall Toxocara seroprevalence of 5.1%. Increasing age, male sex, low educational attainment, low income, and immigration status each was associated with Toxocara seropositivity. Mexican Americans had reduced odds of exposure. These findings show that exposure to Toxocara continues in the United States and that several demographic factors influence the risk of exposure.

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

The definitive genetic identification of Toxocara species is currently based on PCR/sequencing. The objectives of the present study were to design and conduct an in silico polymerase chain reaction-restriction fragment length polymorphism method for identification of Toxocara species. In silico analyses using the DNASIS and NEBcutter softwares were performed with rDNA internal transcribed spacers, and mitochondrial cox1 and nad1 sequences obtained in our previous studies along with relevant sequences deposited in GenBank. Consequently, RFLP profiles were designed and all isolates of T. canis and T. cati collected from dogs and cats in different geographical areas of Iran were investigated with the RFLP method using some of the identified suitable enzymes. The findings of in silico analyses predicted that on the cox1 gene only the MboII enzyme is appropriate for PCR-RFLP to reliably distinguish the two species. No suitable enzyme for PCR-RFLP on the nad1 gene was identified that yields the same pattern for all isolates of a species. DNASIS software showed that there are 241 suitable restriction enzymes for the differentiation of T. canis from T. cati based on ITS sequences. RsaI, MvaI and SalI enzymes were selected to evaluate the reliability of the in silico PCR-RFLP. The sizes of restriction fragments obtained by PCR-RFLP of all samples consistently matched the expected RFLP patterns. The ITS sequences are usually conserved and the PCR-RFLP approach targeting the ITS sequence is recommended for the molecular differentiation of Toxocara species and can provide a reliable tool for identification purposes particularly at the larval and egg stages.

Quantifying the risk of zoonotic geohelminth infections for rural household inhabitants in Central Poland.

The objective of this study was to evaluate the risk of soil transmitted zoonotic helminth infections for families with young children, inhabitants of villages in the Mazowieckie Province of Central Poland. Epidemiological survey was conducted at 33 randomly selected households with 2-3 children present. Examination of soil samples from yards surrounding the houses for the presence of geohelminth eggs was conducted, the households were inspected, and family members interviewed using a designed questionnaire. Among 55 localities examined, i.e. 33 backyards, 10 vegetable gardens and 12 sandpits, contamination was found in 2 backyards (6.1%) and 1 sandpit (8.3%) at 3 households (9.1%). Of the total 550 examined soil samples, 4 (0.7%) were found to contain Trichuris and Toxocara eggs, with an average density of 1.5 and 2.0 eggs per sample. The study showed a low level of soil contamination, which was the result of inhabitants care about the sanitation of their domiciles. However, the results of the questionnaire survey demonstrated the need to warn rural residents about the risk factors for zoonotic helmints infections. In particular, parents should be advised how to minimize the threat of parasitic diseases for children in the rural environment. The presented study showed that promotional campaigns implemented in recent years on the prevention of parasitic zoonoses have had little effect to increase the awareness of the rural community. The present results confirmed that Toxocara is the most common zoonotic agent among geohelminths in the rural environment.

Enzyme-linked immunosorbent assays for coproantigen detection of Ancylostoma caninum and Toxocara canis in dogs and Toxocara cati in cats.

We report the development and field validation of 2 ELISAs for the detection of Ancylostoma caninum or Toxocara canis coproantigens in the feces of dogs with experimental and natural infections, and evidence of cross-reactivity with respective feline counterparts. A. caninum-specific coproantigens were detected in feces of experimentally infected dogs starting at 9 d post-infection (dpi), whereas eggs were not seen until 23 dpi. T. canis-specific coproantigens were detected in 3 of 5 experimentally infected dogs by 31 dpi, and 4 of the 5 animals by 38 dpi. T. canis eggs were seen in feces of 4 of the 5 animals by 38 dpi. One dog had delayed coproantigen detection and low egg output. Additionally, 817 canine and 183 feline fecal samples from naturally infected animals tested by flotation were subjected to coproantigen ELISA testing. Of these 1,000 canine and feline samples, 13 and 23 samples, respectively, were positive for "hookworm" or "roundworm" eggs; 19 and 26 samples were ELISA positive, respectively. The T. canis ELISA detected T. cati coproantigen in cat fecal samples. Discrepant ELISA and flotation results were obtained for 16 hookworm- and 13 roundworm-positive samples. Re-examination of the egg-positive, ELISA-negative samples indicated several instances of possible misidentification or coprophagy, whereas detection of antigen in samples without egg observations is likely a reflection of true infection status with egg shedding below detection levels. There is good indication, based on accumulated field data, that these antigen tests also detect other hookworm and ascarid species.