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.

Case Report: Toxocariasis Manifesting as Eosinophilic Pleural Effusion.

Toxocariasis is a prevalent zoonosis caused by infection with the larvae of Toxocara canis or Toxocara cati. It ranges in severity from mundane to life-threatening, depending on organ involvement. The lungs are often affected, manifesting as coughing, wheezing, and chest pain. However, pleural effusions rarely occur in patients with pulmonary toxocariasis. We report the case of a 74-year-old man with highly suspected toxocariasis who presented with an eosinophilic pleural effusion and eosinophilia. He developed dyspnea and a right-sided pleural effusion. Thoracentesis revealed an exudative effusion containing numerous eosinophils. The pleural effusion continued to increase, and the eosinophilia rapidly progressed. Although the patient had not recently had contact with animals or known exposure to contaminated food, water, or soil, toxocariasis was confirmed by positive serological test results for anti-Toxocara antibodies in the serum and pleural effusion. The patient was cured with albendazole treatment for 28 days. The pleural effusion and eosinophilia resolved and did not recur. Clinicians should consider toxocariasis in the differential diagnosis of patients presenting with eosinophilic pleural effusions.

Parasitic contamination of fresh vegetables and fruits sold in open-air markets in peri-urban areas of Jimma City, Oromia, Ethiopia: A community-based cross-sectional study.

BACKGROUND: Consuming contaminated raw vegetables and fruits is one of the primary means of parasite transmission to humans. Periodic monitoring of parasitic contamination in these food items is a crucial step in preventing the spread of parasitic disease in the community. This study was aimed at detecting intestinal parasitic contamination caused by consuming raw vegetables and fruits sold in three open-air markets and its associated factors in peri-urban areas of Jimma City, Oromia, Ethiopia. METHODS: A cross-sectional study was conducted on fruits and vegetables collected from three peri-urban open-aired markets (namely; Hora Gibe, Bore and Jiren markets) in peri-urban areas of Jimma City between July and September 2021. A total of 187 fresh vegetable samples and 188 fruits were collected and examined for intestinal parasite contamination. About 200g of fruit and vegetable samples were processed and examined microscopically for parasite contamination, utilizing direct wet mount and modified Zeihl-Neelson staining methods in accordance with standard protocols. A structured questionnaire was used to collect data on the socio-demographic characteristics of vendors and risk factors for fruit and vegetable contamination. All data were analyzed using SPSS version 20.0. RESULT: Of the 187 fresh vegetable samples and 188 fresh vegetable samples, 105 (56.1%) and 68/188 (36.2%) of vegetables and fruit samples, respectively, were found contaminated with one or more intestinal parasites. Remarkably, high level of contamination in fresh vegetable samples was recorded both in Carrot (Daucus carota) 63.8% (30/46) and Lettuce (Lactuca sativa) 63.1% (29/46) while Green pepper (Capsicum spp.) is the least contaminated. In fruit samples, Avokado (Persea americana) 42.6% (20/47) and Banana (Musa acuminata) 14.9% (7/47) were the most and the least commonly contaminated items respectively. The identified helminthes and protozoans were Ascaris lumbricoides, Strongyloides stercoralis, Hymenolepis nana, Entamoeba histolytica/dispar, Giardia lamblia, Cryptosporidium spp., Toxocara spp. And Fasciola spp. The most predominant parasite encountered was A. lumbricoides 46(12.3%) whereas both Toxocara spp. 12(6.9) and Fasciola spp. 2(0.5) were the least detected parasites. It is worth-mentioned that the rate of contamination in Bore market (38.15%) was higher compared with Jiren market (34.7%) and Hora Gibe market (27%). However, the rate of contamination in vegetables and fruit obtained from the three district was non-significant (p = 0.19). Contamination was more common in vegetables than fruits (AOR = 5.78, p<0.001). It was also observed that decreased parasitic contamination was significantly associated with washing the products before displaying it for selling (p < 0.001). CONCLUSION: The study has identified a high rate of raw vegetables and fruits contaminated with intestinal helminthes and protozoan. Contaminated fresh vegetables and fruits in open-aired peri-urban markets of Jimma city, Ethiopia may play a significant role in transmission of intestinal parasitic infections to humans, particularly A. lumbricoides infection. Therefore, it is urgently needed for health authorities to educate the public on the proper handling of vegetables and fruits prior to consumption.

Serosurvey of Toxoplasma gondii and Toxocara spp. co-infection in pregnant women in low-income areas of Brazil.

Introduction: Despite human toxoplasmosis and toxocariasis having been listed among the top six most neglected parasitic zoonoses worldwide, presenting similar associated risk factors and transmission routes, few studies have been conducted in pregnant women and the consequences of concurrent infection remain to be fully established. Accordingly, the present study has serologically assessed the co-infection and associated risk factors for Toxoplasma gondii and Toxocara spp. in pregnant women, assisted by the public Unified National Health System (SUS) in southeastern Brazil. Materials and Methods: Blood samples were collected and tested for IgG antibodies against Toxoplasma gondii by chemiluminescence immunoassay and against Toxocara spp. by enzyme-linked immunosorbent assay (ELISA). An epidemiological questionnaire was applied to gather socioeconomic information to assess the risk factors associated with seropositivity to toxocariasis/toxoplasmosis by univariate analysis followed by logistic regression. Results: Overall, seropositivity was 69/280 (24.6, 95% CI: 19.96-30.01) for T. gondii and 56/280 (20.0, 95% CI: 15.73-25.08) for Toxocara spp. Co-infection was observed in 25/280 (8.9, 95% CI: 6.12-12.85) pregnant women, with increased odds (OR: 3.3, CI 95%: 1.77-6.14, p = 0.0002). Logistic regression revealed that a higher educational level (high school or college) significantly reduced the likelihood of co-infection seropositivity, owning cats increased the odds of toxocariasis, and older pregnant women presented significantly higher T. gondii seropositivity. Conclusion: Co-infection herein highlights the importance of educational programs in the prevention of toxocariasis and toxoplasmosis in pregnant women and other high-risk populations.

Toxocara canis infection in multiple types of animals: ophthalmological and pathological observations.

Human ocular toxocariasis (OT), caused by pet roundworm Toxocara canis (Nematoda Ascaridoidea), is a worldwide ocular parasitic infection that poses a severe threat to eyesight, especially in school-aged children. However, the infection process and pathological mechanism of Toxocara are difficult to study in the human body. This study was designed to explore long-term ocular manifestations in different rodents infected with Toxocara canis, uncovering the specific pathological mechanism and migration pathway of larvae after infection. The three types of experimental animals we selected were C57BL/6 mice, Mongolian gerbils and Brown Norway rats. Mice were randomly divided into five groups and infected orally with 1000, 2000, 4000, 8000 and 10,000 T. canis eggs; gerbils were randomly divided into four groups and infected orally with 1000, 2000, 4000 and 10,000 T. canis eggs; rats were randomly divided into three groups and infected orally with 2000, 6000 and 10,000 T. canis eggs. Their ocular changes were closely observed and recorded for at least 2 months. We also enucleated the eyeballs of some animals to perform pathological sectioning and hematoxylin-eosin staining. After 3 dpi (days post-infection), hemorrhagic lesions, mechanical injury of the retina and larval migration could be observed in some infected animals. The ocular infection and mortality rates tended to be stable at 7 dpi. Larval tissue, structure disorder and inflammation could be observed in the pathological sections. In conclusion, the mice infected with 2000 T. canis eggs and gerbils infected with 1000, 2000 and 4000 T. canis eggs showing obvious ocular lesions and lower mortality rates could provide a basis for long-term observation.

Seroprevalence and Potential Risk Factors of Toxocariasis among General Population in Southwest Iran: Implications on the One Health Approach.

Toxocariasis is one of the most common zoonotic diseases distributed worldwide. This study aimed to estimate the seroprevalence of anti-Toxocara immunoglobulin G (IgG) antibodies and the associated risk factors among general populations living in urban and rural areas of Abadan and Khorramshahr cities in Khuzestan Province, Southwest Iran. This cross-sectional study was conducted between March and September 2022. There were 363 participants (190 females and 173 males) aged from <20 to ≥60 years old. Anti-Toxocara IgG antibodies in serum samples were measured using a commercially available enzyme-linked immunosorbent assay (ELISA). A structured questionnaire was employed to collect information regarding sociodemographic status and probable risk factors associated with toxocariasis. It was found that the seroprevalence rate in males (15.0%, 95% CI = 10.47-21.11) was higher than in females (10.5%, 95% CI = 6.92-15.70). Moreover, we observed that the seroprevalence was higher in participants at younger ages compared to other age ranges (COR = 2.55, 95% CI = 0.92-7.12, p =0.073). The findings of the univariate analysis revealed that residency in rural areas (p < 0.001), using unpurified water (p < 0.001), contact with dog (p =0.002), contact with soil (p < 0.001), consumption of improperly washed vegetables (p < 0.001), and history of drinking untreated water (p < 0.001) were risk factors associated with toxocariasis. Further comprehensive studies with a focus on humans and animals should be designed in different areas of the Province. The data represented by the current study are useful to health policymakers to consider precise surveillance and effective prevention measures to control this zoonotic infection among general populations.

Toxascaris leonina in dogs - A nematode species of high prevalence in some regions of Eurasia.

A recent meta-analysis of studies from around the world had shown a global prevalence of Toxascaris leonina in stray dogs of about 7%. However, studies from Eurasian regions, where higher percentages of positive dogs were often found, were under-represented in this meta-analysis. This prompted the present study. Its main objective was to examine free-roaming dogs from the capital city of Kazakhstan to obtain information on the current occurrence of T. leonina egg shedding compared to Toxocara spp. Faecal samples from 500 free-roaming dogs from the city of Astana and its suburbs were collected 1-2 days after admission to the municipal animal shelter during three time periods (May to October 2019, September 2020, and April 2021). Samples were examined by a saline flotation method. Nematode egg-positive samples were subjected to a modified McMaster method to determine the number of eggs per gram of faeces (EPG). Toxascaris leonina eggs were found in 53.8% and Toxocara spp. eggs in 14.8% of the dogs examined. The prevalence of T. leonina egg shedding was significantly associated with age class but not with sex: dogs aged between 6 months and 2 years were more likely to be T. leonina egg positive than puppies. The intensity of T. leonina egg shedding was also age dependent: dogs aged between 6 months and 2 years had significantly higher EPGs compared to younger or older animals. In contrast, the prevalence of Toxocara spp. egg shedding was associated with both age class and sex: dogs older than 2 years were less likely to shed Toxocara spp. eggs than puppies, and females were less likely to be Toxocara spp. egg positive compared to males. The present results confirm those of other studies in Eurasian regions. It is therefore reasonable to assume that the global prevalence of T. leonina infection has been underestimated and should be revised accordingly.

Migratory pattern of zoonotic Toxocara cati and T. canis in experimentally infected pigs.

BACKGROUND: Over a billion people are infected with Toxocara canis or T. cati, the roundworms of dogs and cats. Historically, T. canis has been considered the main species responsible for human toxocarosis, but as serodiagnosis cannot discriminate between the two species, this remains unresolved. We used pigs as a relevant large animal model for human infection to assess the migratory pattern of T. cati and T. canis. METHODS: Pigs were inoculated with T. cati or T. canis eggs or PBS (negative controls) and necropsied 14 or 31 days later. Different organs and tissues were examined for parasites and pathological changes. RESULTS: Overall, the two parasite species had a similar migration pattern reaching multiple organs and tissues, including the mesenteric lymph nodes, liver, lungs, and diaphragm. We recovered larvae of both species in the brain, suggesting that T. cati also can cause neurological toxocarosis in humans. Both species induced systemic eosinophilia and histopathological changes in the lungs, livers, and mesenteric lymph nodes. CONCLUSION: This study emphasises the importance of T. cati as a zoonotic agent and the need to develop diagnostic methods that can differentiate between sources of infection in humans.

Epidemiological profile of human toxocariasis in patients examined at Evandro Chagas Institute (IEC/SVSA/MS) between 2014 and 2019.

INTRODUCTION: Toxocariasis is caused by nematodes of Toxocara genus, which infest dogs and cats, with humans serving as paratenic hosts. METHODS: The epidemiological profile of patients examined for toxocariasis between October 2014 and October 2019 at Evandro Chagas Institute (IEC) was outlined. The frequency of anti-T. canis IgG antibodies were evaluated using the Enzyme Linked Immunosorbent Assay (ELISA) method. RESULTS: From a total of 734 samples, 56% were from male (p < 0.05). Regarding age, the group with the most solicitations were from ≤11 years old individuals (p < 0.05). Pará state had the highest number of exams requested (92%), with the majority from residents of urban areas, accounting for 81.5% of samples (p < 0.05). The overall toxocariasis seroprevalence was 41.8%, the male sex being the most frequent with 60.9% (p < 0.05). The most affected age group was ≤11 years old, with a total of 67.8% of positive samples (p < 0.05). CONCLUSION: The high rates obtained emphasize the need for complementary studies on toxocariasis in Brazil, especially in Pará state, contributing to epidemiological surveillance actions in the control of this infection. Besides, health campaigns for domestic and stray animals, also can contribute to a more effective surveillance in controlling parasitic infections and encourages the One Health approach.

Parasitological contamination of arable soil in selected regions of Poland - preliminary study.

INTRODUCTION AND OBJECTIVE: The hygienic status of arable soils in most developed countries has been unknown. In the presented study, a preliminary investigation was undertaken to determine the contamination with eggs of parasitic nematodes in the soil of arable fields in Poland. The aim of the study was to determine whether such contamination is common enough to constitute a significant problem and what factors may influence it. MATERIAL AND METHODS: The study was conducted in 5 Polish provinces from autumn 2021 to spring 2022. The provinces differed significantly in terms of the area of agricultural land, agricultural suitability, type of soil, scale of cattle and pig breeding, production of manure and slurry, and the use of manures and organic fertilizers for fertilization. A total of 133 soil samples were collected. Parasitological examination of soil samples was carried out using the PN-Z-19006 method [1], with confirmed high sensitivity. RESULTS: Parasite eggs were found in a total of 67 samples, of which 56 samples contained eggs of roundworms of the genus Ascaris (an average of 3.29 eggs/100 g of soil), 23 contained eggs of whipworms (an average of 1.22 eggs/100 g), and 3 contained eggs of Toxocara (1 egg/100 g). CONCLUSIONS: Differences in the percentage of positive samples were found depending on the period in which the samples were taken. The percentage of positive samples collected in autumn (53.57%) was higher than the percentage of positive samples collected in spring (48.05%). Similarly, the average number of eggs of in positive samples collected in autumn (3.43 eggs/100 g) was higher than the average number of eggs in samples collected in spring (2.90 eggs/100 g). Differences in the percentage of positive samples were also found depending on the region of origin of the samples.

Seroprevalence of Toxocara at Tra Vinh University Hospital in Vietnam.

OBJECTIVE: The study aims to assess the seroprevalence of Toxocariasis and its associated risk factors among individuals attending the outpatient department at Tra Vinh University Hospital, Vietnam, in 2022. SUBJECTS AND METHODS: A cross-sectional survey was conducted among outpatients of Tra Vinh University Hospital. Toxocariasis diagnosis was based on the Enzyme-Linked Immunosorbent Assay (ELISA) performed at the hospital's laboratory department. We assessed the seroprevalence of Toxocariasis and evaluated associated risk factors, including demographics and certain behaviors. RESULTS: Of the 249 participants surveyed, 165 tested positive for Toxocariasis, yielding a seroprevalence of 66.3% (95% CI: 60.4-72.1). Multivariate analysis revealed that age groups up to 30 and 30-60 years had higher odds of Toxocariasis infection, with adjusted odds ratios (aOR) of 2.52 (95% CI: 1.04-6.11) and 3.21 (95% CI: 1.44-7.15) respectively. Additionally, individuals residing in rural areas and those in contact with dogs or cats had increased risks, with aORs of 2.21 (95% CI: 1.21-4.01) and 2.04 (95% CI: 1.10-3.79), respectively. Notably, hand washing before eating emerged as a protective factor against Toxocariasis, presenting an aOR of 0.38 (95% CI: 0.19-0.76). CONCLUSIONS: Our findings underscore a significant seroprevalence (66.3%) of Toxocara spp. among outpatients at Tra Vinh University Hospital. Proactive measures, including hand hygiene before meals and after pet interactions, are advocated. There is a pronounced need for community-level epidemiological surveillance for human Toxocariasis.

Risk factors for toxocariasis during incarceration: the One Health intervention approach.

Despite potential exposure to soil-transmitted helminths, especially when stray dogs and cats are present, toxocariasis in inmate populations remains to be established. Accordingly, the present study assessed the seroprevalence and associated risk factors of toxocariasis at the Women's State Penitentiary of Parana, Brazil. A total of 234/370 (63.2%; 95% CI 58.2-68.0) women inmates and 28/87 (32.2%; 95% CI 23.3-42.6) correctional officers were seropositive for anti-Toxocara spp. IgG by ELISA, with inmates 2.62-fold more likely positive (p = 0.00000026). The univariate model has identified that non-white (OR = 1.58, p = 0.047) and older than 39 years (OR = 1.28, p = 0.032) inmates were associated with mild but significant odds for seropositivity. Elementary or higher educational level was considered a protective factor for seropositivity. The presence of Toxocara spp. eggs was observed in 10/15 (66.7%) collected soil samples by centrifuge-flotation in Zinc Sulfate, and molecular analysis by PCR identified only Toxocara cati in these eggs. An intervention program was established with regular trap-neuter-release, with gradual removal for adoption (donation campaigns), treatment, and euthanasia when necessary (particularly due to advanced sporotrichosis). In addition, an educational awareness agenda was proposed, aiming to reduce soil contamination and accidental intake by the incarcerated population. A total of 40 feral cats were trapped, 20 males and 20 females, mostly adults. After trapping, 36 cats were neutered, treated, and microchipped in the Veterinary Teaching Hospital (VTH) at the Federal University of Paraná. Five trapped feral cats were euthanized, four diagnosed with advanced sporotrichosis, and one already neutered cat (not herein) with complications due to feline immunodeficiency virus (FIV). Female inmates presented higher seroprevalence for Toxocara spp. antibodies when compared to correctional officers, significantly associated with age, self-declared ethnicity (non-white), and lack of formal education. Despite the non-natural scenario of a state penitentiary, the One Health approach of Toxocara spp. has highlighted the interdisciplinary nature of the study and its relevance in understanding the complex interactions between human, animal, and environmental factors, particularly impacting female inmates. Further studies should establish the rate of inmate infection over time while deprived of liberty.

Visceral larva Migrans in a Young Italian Patient: A Diagnostic Dilemma.

BACKGROUND: The association of fever, focal hepatic lesions and peripheral hyper-eosinophilia (FHLH) can be observed in both infectious and non-infectious conditions. Fascioliasis, capillariasis, toxocariasis, all causes of visceral larva migrans (VLM), represent most of the former, whilst lymphomas, eosinophilic leukemias and mastocytosis belong in the non-infectious conditions. METHODS: We prospectively followed a young patient presenting with FHLH in the Tuscany region of Italy. RESULTS: The patient was subject to serological and parasitological examination in an attempt to clarify the origin of the lesions. Serologies for both Fasciola hepatica and Toxocara spp. were positive, with the latter presenting a higher index. We opted for treatment with a prolonged course of albendazole due to the serological results and being toxocariasis more frequent in our setting. The patient was then subject to radiological follow-up. The patient responded to treatment with albendazole as shown by a decrease in eosinophils, seronegativization for Toxocara spp., clinical and radiological improvement. Toxocariasis was hence considered the most likely diagnosis. CONCLUSIONS: Parasitic infections cannot be disregarded in the presence of FHLH. Differential diagnosis between these parasitic infections can be challenging due to the presence of similar clinical presentations and serological cross-reactions, and follow-up of the patient is needed to ensure optimal treatment outcomes.

One health approach to toxocariasis in quilombola communities of southern Brazil.

BACKGROUND: Toxocariasis has been listed among the most neglected parasitic diseases worldwide, with approximately one fifth of the global population exposed, particularly those living under poverty. In Brazil, communities of descendants of enslaved blacks (quilombola) have historically had some of the highest rates of vulnerability and poverty, characterized by lack of health assistance, poor quality of life, and nutritional insecurity. METHODS: A cross-sectional sampling of quilombola individuals living in four communities of southern Brazil, as well as their dogs and the soil, was carried out from December 2021 to March 2022. Sociodemographic and other information such as water source, alimentary habits, and dog and cat ownership were gathered using a semi-structured questionnaire for assessing toxocariasis risk factors. Human serum samples were tested by ELISA for anti-Toxocara spp. IgG antibody detection was carried out on dog feces and hair, and soil samples were surveyed for presence of Toxocara spp. eggs. RESULTS: Overall, 172/208 individuals (82.7%, 95% CI = 77.0-87.2) were seropositive, the highest seroprevalence rate to date in Brazil. Male gender (P = 0.029), educational level (P = 0.026), and drinking water source (P = 0.043) were associated with seropositivity by univariate analysis. Final logistic regression revealed increased odds (P = 0.017, OR = 7.6, 95% CI = 1.5-42.7) to have seropositivity in individuals > 50 years old (< 10 years old). As expected, individuals with soil contact were more likely seropositive (P = 0.038, OR = 4.4, 95% CI = 1.1-18.8). Although retrieved in only 5/96 (5.2%) dog feces, Toxocara spp. eggs were found in 18/60 (30.0%) soil samples. CONCLUSIONS: The high vulnerability and seroprevalence observed in quilombola communities clearly demand a One Health approach for detection, monitoring, and prevention of infection by Toxocara spp. in both human and dog populations.

The Parasites of Cats in Türkiye.

Cats have an important and different place due to their close relationships with humans. Since most of the parasites they carry are zoonotic, it is important to detect them. According to the research, Dipylidium caninum, Joyeuxiella pasqualei, Toxocara spp., Toxascaris leonina, Giardia spp., Isospora spp., and Toxoplasma sp. were found to be higher in cats compared to other parasites. It has been determined that scabies and flea infestations are common as ectoparasites. This review aims to present the reported parasites and their prevalence rates in cats in Türkiye.

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.

Testing of sand from recreational areas and sandpits in Warsaw, for presence of Ascaris Lumbricoides and Toxocara spp.

INTRODUCTION: Improper maintenance of the cleanliness of sand in sandboxes can be a source of human pathogenic helminths, including Toxocara spp., Enterobius vermicularis, Ascaris lumbricoides, leading to parasitic infections. AIM OF THE STUDY: The purpose of this study was to evaluate the sanitary condition of sandboxes in playgrounds and recreational areas in Warsaw for the presence of Human roundworm (Ascaris lumbricoides) and Toxocara spp. in sand. MATERIAL AND METHODS: 450 sand samples from 90 sandboxes in the city of Warsaw were analyzed. The flotation method was used for the study, and the material was evaluated using a light microscope. RESULTS. The conducted examinations did not reveal the presence of parasite eggs, indicating that hygiene rules were observed and appropriate recommendations were followed. CONCLUSIONS: The analyzed sand samples are free of the tested parasites.

Molecular investigation of Toxocara infection from the serum of people living with HIV in Alborz, Iran.

BACKGROUND: Toxocara infection is one of the most common neglected infections of poverty and a helminthiasis of global importance. Traditional diagnostic methods such as antibodies detection in serum samples are limited due to cross-reactivity and poor sensitivity. The use of molecular base methods for diagnosis of Toxocara infection in Iran has not been fully explored. The purpose of the current study was to estimate the prevalence of Toxocara infection from serum samples of people living with HIV in Alborz province, Iran using serological and molecular methods. METHODS: Blood samples were collected from 105 people living with HIV. Epidemiological data of participant were obtained through a structured questionnaire to investigate the risk factors. Patients CD4+ T cell count were recorded. Anti-Toxocara IgG antibodies were detected by ELISA, with a cut-off point of 11. PCR was performed to detect genetic material of Toxocara species in the serum samples. RESULTS: The mean CD4+ count in HIV-infected individuals with positive toxocariasis serology was 255.1 ± 21.6 cells/µL. Seropositivity for Toxocara species was observed in 12/105 (11.4%) people living with HIV. Three samples gave positive results on PCR analysis. Based on the data, a statistically significant relationship was found between anti-Toxocara IgG antibodies seropositivity and underlying conditions (p = 0.017). No significant statistical association was observed between seropositivity for Toxocara and gender, age, exposure to domestic animals or pet keeping, education levels, and occupation (p > 0.05). The findings of PCR confirmed Toxocara DNA in 3/12 (25.0%) serum samples. CONCLUSION: These findings demonstrated for the first time that people living with HIV from Alborz province, are being exposed to this zoonosis and a relatively high seroprevalence of Toxocara in HIV/AIDS people needs comprehensive health education regarding personal hygiene and how to avoid exposure to this parasite infection, especially in people with an impaired immune system.

Zoonotic helminths of dogs and risk factors associated with polyparasitism in Grenada, West Indies.

Canine soil-transmitted helminths (STHs) cause important zoonoses in the tropics, with varying degrees of intensity of infection in humans and dogs. This study aimed to investigate the prevalence and associated risk factors for STHs in community dogs residing in Grenada, West Indies. In May 2021, 232 canine fecal samples were examined for zoonotic helminths by microscopy (following flotation), and genomic DNA from a subset of 211 of these samples were subjected to multiplex qPCR for the detection and specific identification of hookworms, Toxocara spp. and Strongyloides. Microscopic examination revealed that 46.5% (108/232, 95% CI 40­52.9), 9% (21/232, 95% CI 5.35­12.7) and 5.2% (12/232, 95% CI 2.3­8) of the samples contained eggs of Ancylostoma spp., Toxocara spp. and Trichuris vulpis, respectively. Multiplex qPCR revealed that, 42.2% (89/211, 95% CI 35.5­48.8) were positive for at least 1 zoonotic parasite. Of these, 40.8% (86/211, 95% CI 34.1­47.3) of samples tested positive for Ancylostoma spp., 36% (76/211, 95% CI 29.5­42.9) were positive for A. caninum, 13.3% (28/211, 95% CI 9­18.6) for A. ceylanicum, 5.7% for T. canis (12/211, 95% CI 2.97­8.81) and 1% (2/211, 95% CI 0­2.26) for Strongyloides spp. (identified as S. stercoralis and S. papillosus by conventional PCR-based Sanger sequencing). Using a multiple logistic regression model, a low body score and free-roaming behaviour were significant predictors of test-positivity for these parasitic nematodes in dogs (P < 0.05). Further studies of zoonotic STHs in humans should help elucidate the public health relevance of these parasites in Grenada.