Evaluation of Somatic Antigens of Adult Toxocara helminthes for Detection of Human Toxocariasis.

We aimed to develop an indirect enzyme-linked immunosorbent assay (ELISA) to evaluate the presence of specific IgG against Toxocara canis and Toxocara cati somatic antigens on the serum of patients with toxocariasis. The sensitivity, specificity, positive and negative predictive values for indirect-ELISA were calculated by receiver operating characteristic curve (ROC) analysis and Youden's J using Likelihood ratio. All statistics were analysed and graphs are plotted using GraphPad Prism version 8.4.3 (Graph Pad Software, La Jolla, CA, USA), with 95% confidence interval (CI). The sensitivity, specificity, positive and negative predictive values for T. canis were 100%, 82%, 79% and 100%, respectively. The mentioned variables for T. cati were 97%, 82%, 78% and 98%, respectively. Five immune reactive bands of 38, 40, 72, 100 and 250 kDa were common in both species. Toxocara crude antigens were highly immunogenic in human sera. Immunoreactive bands against T. canis compared to T. cati somatic antigen were about two times more. Unlike Toxocara excretory-secretory antigen, that was homologue in two species, somatic antigens of T. canis and T. cati showed different immunoreactive bands in our western blot.

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.

Case series of neuroretinitis in Korea.

BACKGROUND: To present the clinical characteristics of neuroretinitis in Korea. METHODS: Twelve patients with neuroretinitis between January 2009 and September 2020 were retrospectively reviewed. Neuroretinitis was diagnosed based on fundus findings, optical coherence tomography, and fluorescein angiography. The serological findings of each patient were reviewed. RESULTS: Fifteen eyes of 12 patients (9 male and 3 female), with a mean age of 46.0 ± 10.7 years were included. Of the nine patients who underwent serological testing for Toxocara antibodies, six (66.6%) were positive. One patient had high titers of Toxoplasma immunoglobulins M and G. One patient diagnosed with dengue fever was suspected to have neuroretinitis in both eyes. There were no related abnormalities in the serological findings in four patients (33.3%) out of 12 patients. There were no suspected cases of cat-scratch disease. The six patients who were positive for Toxocara antibodies were older (mean age: 54.5 ± 9.1 years) than the others (mean age: 37.5 ± 4.4 years, p = 0.004). The four patients without any abnormal serological findings were relatively younger (mean age: 35.7 ± 3.0 years) than the other 8 patients (mean age: 51.1 ± 10.1 years, p = 0.008). CONCLUSIONS: Two-thirds of neuroretinitis patients were seropositive for Toxocara in the current cohort from Koreans. Causative factors in cases of neuroretinitis may vary according to age and region.

Synergistic effects of using sodium hypochlorite (bleach) and desiccation in surface inactivation for Toxocara spp.

Toxocara cati and T. canis are parasitic nematodes found in the intestines of cats and dogs respectively, with a cosmopolitan distribution, and the potential for anthropozoonotic transmission, resulting in human toxocariasis. Spread of Toxocara spp. is primarily through the ingestion of embryonated eggs contaminating surfaces or uncooked food, or through the ingestion of a paratenic host containing a third-stage larva. The Toxocara spp. eggshell is composed of a lipid layer providing a permeability barrier, a chitinous layer providing structural strength, and thin vitelline and uterine layers, which combined create a biologically resistant structure, making the Toxocara spp. egg very hardy, and capable of surviving for years in the natural environment. The use of sodium hypochlorite, household bleach, as a disinfectant for Toxocara spp. eggs has been reported, with results varying from ineffective to limited effectiveness depending on parameters including contact time, concentration, and temperature. Desiccation or humidity levels have also been reported to have an impact on larval development and/or survival of Toxocara spp. eggs. However, to date, after a thorough search of the literature, no relevant publications have been found that evaluated the use of sodium hypochlorite and desiccation in combination. These experiments aim to assess the effects of using a combination of desiccation and 10% bleach solution (0.6% sodium hypochlorite) on fertilized or embryonated eggs of T. cati, T. canis, and T. vitulorum. Results of these experiments highlight the synergistic effects of desiccation and bleach, and demonstrate a relatively simple method for surface inactivation, resulting in a decrease in viability or destruction of T. cati, T. canis and T. vitulorum eggs. Implications for these findings may apply to larger scale elimination of ascarid eggs from both research, veterinary, and farming facilities to mitigate transmission.

Comparison of extracellular vesicle isolation methods for the study of exosome cargo within Toxocara canis and Toxocara cati excretory secretory (TES) products.

Toxocara is a genus of nematodes, which infects a variety of hosts, principally dogs and cats, with potential zoonotic risks to humans. Toxocara spp. larvae are capable of migrating throughout the host tissues, eliciting eosinophilic and granulomatous reactions, while surviving for extended periods of time, unchanged, in the host. It is postulated that larvae are capable of altering the host's immune response through the release of excretory-secretory products, containing both proteins and extracellular vesicles (EVs). The study of EVs has increased exponentially in recent years, largely due to their potential use as a diagnostic tool, and in molecular therapy. To this end, there have been multiple isolation methods described for the study of EVs. Here, we use nanoparticle tracking to compare the yield, size distribution, and % labelling of EV samples acquired through various reported methods, from larval cultures of Toxocara canis and T. cati containing Toxocara excretory-secretory products (TES). The methods tested include ultracentrifugation, polymer precipitation, magnetic immunoprecipitation, size exclusion chromatography, and ultrafiltration. Based on these findings, ultrafiltration produces the best results in terms of yield, expected particle size, and % labelling of sample. Transmission electron microscopy confirmed the presence of EVs with characteristic cup-shaped morphology. These findings can serve as a guide for those investigating EVs, particularly those released from multicellular organisms, such as helminths, for which few comparative analyses have been performed.

The larvicidal effect of the supernatant of Lactobacillus acidophilus ATCC 4356 on Toxocara canis.

Human toxocariasis is a parasitic anthropozoonosis that is difficult to treat and control. A previous study carried out with Lactobacillus acidophilus ATCC 4356 revealed that the cell free supernatant (CFS) of this probiotic killed 100% of Toxocara canis larvae in vitro. The present study aimed to investigate the characteristics of the CFS of L. acidophilus ATCC 4356, which may be involved in its larvicidal effects on T. canis. L. acidophilus ATCC 4356 was cultured, and lactic and acetic acids present in the CFS were quantified by high performance liquid chromatography (HPLC). The levels of pH and H2O2 were also analyzed. To assess the larvicidal effect of the CFS, this was tested pure and diluted (1:2 to 1:128) on T. canis larvae. High concentrations of lactic and acetic acids were detected in the CFS. The acidity of the pure CFS was observed at pH 3.8, remaining acidic at dilutions of 1:2 to 1:16. Regarding the in vitro larvicidal effect, 100% death of T. canis larvae was observed using the pure CFS and 1:2 dilution. Based on these results, it can be inferred that the presence of higher concentrations of organic acids and low pH of the medium contributed to the larvicidal activity of the CFS of L. acidophilus ATCC 4356. In addition, the maintenance of the larvicidal effect, even after dilution, suggests a greater chance of the larvicidal effect of this CFS against T. canis in vivo.

Anthelmintic efficacy of nitazoxanide in dogs naturally infected with Toxocara canis.

Toxocara canis (T. canis) is a gastrointestinal nematode in dogs, and its larvae also infect humans, causing severe larval migratory disease. Anthelmintic drugs have become the primary means to combat T. canis. In this study, the efficacy of nitazoxanide (NTZ) was tested against all the internal stages of T. canis, including L3 larval stage in vitro experiments and gastrointestinal worm in vivo experiments. In the in vitro experiment, after treatment with NTZ at 7.81 and 62.5 µg/mL for 12 h, the larval mortality efficacy reached 90.0 and 100.0%, respectively. In the in vivo experiments, 100 mg/kg NTZ possessed good anthelmintic efficacy against T. canis, with an egg per gram (EPG) reduction of 99.19%, and 90.00% of dogs cleared with residual worms. These results were comparable to those of the positive control drug. The highest anthelmintic efficacy was observed in the group treated with 150 mg/kg NTZ. Based on faecal egg counts, the number of T. canis eggs decreased by 100.00%, and the percentage of dogs cleared with residual worms achieved 90.00% after 7 days of treatment in the 150-mg/kg NTZ treatment group. In general, NTZ showed great potential to be applied as an anthelmintic against T. canis.

A cross-sectional study to quantify the prevalence of Toxocara canis in dogs in the Central Highlands of Vietnam.

Toxocara canis is a globally distributed zoonotic parasite. The parasite has recently become a concern for public health in Vietnam. This cross-sectional study aimed to identify and quantify the risk factors associated with T. canis infection in dogs in Dak Lak province in the Central Highlands of Vietnam. The risk factors were identified using a mixed-effects logistic regression model and quantified using population attributable fractions. Examination of fecal samples collected from 1455 dogs using the sodium nitrate flotation technique showed 37.32% (95% CI: 34.83-39.86) of dogs infected with T. canis. The factors, including study location, multiple dogs living in a household, dog age, dog breed, and places keeping dogs were associated with a dog's likelihood of being T. canis infection. The household and individual dog levels contributed 17% and 82%, respectively, to the prevalence of T. canis in dogs. The adjusted population attributable fraction for confining dogs and raising an individual dog per household was 52% and 27%, respectively. The result of this study indicated that to minimize the burden of T. canis, intervention measures should target individual dogs and household levels.

Microscopic and molecular prevalence and associated risk factors with Toxocara and Blastocystis infection in dogs and cats in Mitidja, Algeria.

Domestic dogs and cats can serve as a source of environmental contamination with Toxocara spp. and Blastocystis spp., and this represents a neglected public and veterinary health problem. We assessed the microscopic and molecular prevalence of these species in a locality in Algeria and identified the associated risk factors. The faeces of 225 dogs and 78 cats were collected in Mitidja between March and July 2022. The samples were analysed by coproscopy and by polymerase chain reaction (PCR) targeting the Internal Transcribed Spacer 2 (ITS2) and Small Subunit Ribosomal (SSU-RNA) of T. canis and Blastocystis spp. respectively. The overall microscopic prevalence of Toxocara spp. in dogs and cats was 9.78 ± 1.98% and 12.82 ± 7.42%, respectively. The rate of Blastocystis spp. was 15.11 ± 2.39% and 15.38 ± 4.08% in dogs and cats, respectively while the molecular prevalence of T. canis in dogs was 4.89 ± 1.44% and in cats 1.28 ± 1.27%; the prevalence of Blastocystis spp. was 41.78 ± 3.29% and 34.62 ± 5.39% in dogs and cats, respectively. Phylogenetic and phylogeographic analyses identified the presence of the H1 subtype of T. canis in dogs, and the ST1 subtype of Blastocystis in dogs and cats. Dogs with clinical signs were more likely to be infected with T. canis (OR 6.039, P < 0.05) than healthy dogs. This study demonstrates that dogs and cats are carriers of Toxocara spp. and Blastocystis spp. and are therefore a source of environmental contamination. Veterinarians and human health professionals should work together to implement control strategies as part of a "One Health" approach to improving animal health and reducing the risk of transmission to humans.

C-type lectin 4 of Toxocara canis activates NF-ĸB and MAPK pathways by modulating NOD1/2 and RIP2 in murine macrophages in vitro.

Toxocara canis is a parasitic zoonose that is distributed worldwide and is one of the two pathogens causing toxocariasis. After infection, it causes serious public health and safety problems, which pose significant veterinary and medical challenges. To better understand the regulatory effects of T. canis infection on the host immune cells, murine macrophages (RAW264.7) were incubated with recombinant T. canis C-type lectin 4 (rTc-CTL-4) protein in vitro. The quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to analyze the nucleotide-binding oligomerization domain-containing protein 1/2 (NOD1/2), receptor-interacting protein 2 (RIP2), nuclear factor kappa-light-chain enhancer of activated B cells (NF-κB), and mitogen-activated protein kinase (MAPK) on mRNA level and protein expression level in macrophages. Our results indicated that 10 µg/mL rTc-CTL-4 protein could modulate the expression of NOD1, NOD2, and RIP2 at both the transcriptional and translational levels. The protein translation levels of NF-κB, P-p65, p38, and P-p38 in macrophages were also modulated by rTc-CTL-4 protein. Macrophages were co-incubated with rTc-CTL-4 protein after siRNA silencing of NOD1, NOD2, and RIP2. The expression levels of NF-κB, P-p65, p38, and P-p38 were significantly changed compared with the negative control groups (Neg. Ctrl.). Taken together, rTc-CTL-4 protein seemed to act on NOD1/2-RIP2-NF-κB and MAPK signaling pathways in macrophages and might activate MAPK and NF-κB signaling pathways by regulating NOD1, NOD2, and RIP2. The insights from the above studies could contribute to our understanding of immune recognition and regulatory mechanisms of T. canis infection in the host animals.

Palaeoparasitology confirms Early Lapita evidence of pig and dog at Kamgot, Bismarck Archipelago.

Little is known about helminth parasites of the Bismarck Archipelago, in either archaeological or modern contexts. This study presents a parasitological analysis of soil samples from Early Lapita habitation layers at Kamgot (3300-3000 BP). Evidence for the presence of pigs and dogs and the timing of their arrival in Early Lapita contexts have been contested in the literature. The finding of parasite eggs in samples from Kamgot supports the presence of pigs and dogs at the site. Six types of helminth eggs were identified: pig nematode Trichuris suis, dog nematode Toxocara canis, and cestode Dipylidium caninum, as well as two unknown trematodes and a possible anoplocephalid cestode, thereby indicating the local presence of other mammals or birds. This study represents the first confirmed record of ancient helminth parasites in tropical Oceania.

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.

The immunomodulatory effects of the C-type lectin protein of Toxocara canis on experimental autoimmune encephalomyelitis.

Toxocara canis is a global zoonosis infection that can cause chronic and long-term toxocariasis in their paratenic host. The excretory-secretory (ES) products of T. canis larvae are considered to be responsible for the Th2 polarization and regulatory immune responses in toxocariasis. The C-type lectin family is one of the most prominent components of ES products of T. canis infective larvae. This study aimed to investigate the ameliorative effect of a T. canis C-type lectin recombinant protein (rCTL), on experimental autoimmune encephalomyelitis (EAE) which is a T-cell-mediated autoimmune disease of the central nervous system. C57BL/6 mice were subcutaneously treated with 30 µg rCTL, three times at an interval of 1 week. EAE was induced by myelin oligodendrocyte glycoprotein 35-55 peptide (MOG35-55 peptide) immunization, and weight and clinical scores were evaluated. Real time polymerase chain reaction was performed to evaluate the expression levels of T-bet, Gata3, and Foxp3 in splenocytes. In addition, the levels of interleukin 4, interferon gamma, and tumour growth factor-ß (TGF-ß) were quantified by enzyme-linked immunosorbent assay in splenocyte culture supernatants. The results indicated that the rCTL decreased clinical disability scores and delayed the onset of EAE. Furthermore, the data showed that rCTL treatment modulated the immune response, which was associated with upregulation of the mRNA expression of the Foxp3 gene and higher production of TGF-ß in rCTL-treated mice. This study demonstrated that rCTL might be a potential agent to ameliorate EAE symptoms by stimulating anti-inflammatory responses.

A rare case of presumptive pleural toxocariasis.

Toxocariasis is a cosmopolitan helminthiasis linked to contamination with Toxocara cati or Toxocara canis. Only six isolated cases of pleural toxocariasis have been reported in the literature. We describe a case of pleurisy with isolated eosinophilia varying between 600 and 1500/mm3 likely linked o presumptive toxocariasis in a 72-year-old patient. Our patient was admitted to hospital with severe dyspnoea, asthenia and diarrhoea. Imaging studies confirmed right unilateral pleurisy without any parenchymal involvement. Serology of serum and pleural fluid was positive for anti-Toxocara antibodies by ELISA and immunoblotting. Treatment by pleural drainage and anti-parasitic medication with albendazole for 8 days resulted in the resolution of symptoms. A decrease in the levels of polynuclear eosinophils and total IgE confirmed the clinical resolution. The presence of hypereosinophilia in pleural fluid should evoke a diagnosis of pleural toxocariasis. Clinical symptoms and imaging are non-specific, but positive serology for anti-Toxocara antibodies in serum and pleural fluid can confirm the diagnosis.

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.

Toxocara tanuki larval distribution in mice and the infectivity of tissue larvae.

Toxocara tanuki is a common large roundworm in raccoon dogs. Experimental infection studies of T. tanuki in mice were conducted to clarify the distribution and infectivity of larvae in tissue. Groups of BALB/c and C57BL/6 mice (n = 5 mice/group) were each inoculated with 1000 embryonated T. tanuki eggs and necropsied at 7, 31, 91, and 182 days post inoculation (dpi). The number of larvae in the central nervous system, heart, lungs, kidneys, spleen, gastrointestinal tract, liver, and carcass was examined. Larvae obtained from the aforementioned mice on different days of the necropsy were orally inoculated into four groups of ICR mice (n = 6 mice/group) that were then necropsied at 21 dpi. Larvae were recovered from all mice. In the BALB/c and C57BL/6 mice, most of the larvae (> 88.7%) were recovered from the liver and the remainder from other tissues. The total number of larvae recovered from C57BL/6 mice was significantly higher than that from BALB/c mice, but no difference in the relative larval distribution within the viscera between the two mouse strains was observed. The mean recovery percentage of larvae from ICR mice infected with 182-day-old tissue larvae was 3.3%. Our findings showed that T. tanuki larvae migrated predominantly to the liver of mice and that the larvae maintained their infectivity for at least half a year.

Effects of metformin on parasitological, pathological changes in the brain and liver and immunological aspects during visceral toxocariasis in mice.

There are currently insufficient anthelmintic medications available for the treatment of toxocariasis. For instance, Albendazole (ABZ) is the preferred medication, but its effectiveness against tissue-dwelling parasites is limited. In addition, Metformin (MTF) is a widely used oral antidiabetic medication that is considered to be safe for treatment. This study aimed to investigate any potential effects of MTF, alone or in combination with ABZ, on mice infections caused by Toxocara canis (T. canis). The efficacy of the treatment was assessed in the acute and chronic phases of the infection by larval recovery and histopathological, immunohistochemical, and biochemical studies. The results showed that combined therapy significantly reduced larval counts in the liver, brain, and muscles and ameliorated hepatic and brain pathology. It reduced oxidative stress and TGF-ß mRNA expression and increased FGF21 levels in the liver. It decreased TNF-α levels and MMP-9 expression in the brain. In addition, it increased serum levels of IL-12 and IFN-γ and decreased serum levels of IL-4 and IL-10. In the acute and chronic phases of the infection, the combined treatment was more effective than ABZ alone. In conclusion, this study highlights the potential role of MTF as an adjuvant in the treatment of experimental T. canis infection when administered with ABZ.

The prevalence of potentially zoonotic intestinal parasites in dogs and cats in Moscow, Russia.

This study was aimed to determine the prevalence of Toxocara canis/cati, Strongyloides stercoralis, Giardia spp., and Cryptosporidium spp., which occur and are potentially zoonotic to humans in domestic dogs and cats in Moscow (Russia). The fecal flotation method and larvae detection by microscopy of a direct feces smear were performed to detect Toxocara, Giardia spp., and Cryptosporidium spp. The total parasitic prevalence in dogs was as follows: Giardia spp.: 10.2 % (226/2208), Cryptosporidium spp.: 2.7 % (60/2208), T. canis: 2 % (45/2208), S. stercoralis larvae: 1.1 % (25/2208). The younger animals under were infected more than those over 12 months of age (p<0.001). The preva lence rates were along these lines: Giardia spp. (18.2 %), Cryptosporidium spp. (5.7 %), T.canis (3 %), S. stercoralis larvae (2.3 %). The overall prevalence in cats was as follows: Giardia spp. - 5.2 % (71/1350), Cryptosporidium spp. - 4.8 % (65/1350), T. cati - 4.1 % (56/1350). Similarly to dogs, the infection rates were higher in cats under 12 months of age Giardia spp. (8.2 %), Cryptosporidium spp. (8.6 %), T. cati (7.5 %. Analysis of combined infections in dogs revealed the following combinations: Giardia spp. and Cryptosporidium spp. (35.5 %) larvae of S. stercoralis sp. and Giardia spp. (32.3 %), T.canis and Giardia spp. (22.6 %), T.canis and Cryptosporidium spp. (6.6 %), T.canis and S.stercoralis and (3.2 %), respectively. In cats, only two coinfections by Giardia spp. and Cryptosporidium spp. (58.3 %), and T.cati with Giardia spp. (41.7 %) were noticed. Further research is needed to study the spread of parasitic diseases in pet animals. The data will improve countermeasures to prevent these diseases' spread among animals and humans.

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.

Differential miRNA expression profiles in the bone marrow of Beagle dogs at different stages of Toxocara canis infection.

BACKGROUND: Toxocara canis is distributed worldwide, posing a serious threat to both human and dog health; however, the pathogenesis of T. canis infection in dogs remains unclear. In this study, the changes in microRNA (miRNA) expression profiles in the bone marrow of Beagle dogs were investigated by RNA-seq and bioinformatics analysis. RESULTS: Thirty-nine differentially expressed (DE) miRNAs (DEmiRNAs) were identified in this study. Among these, four DEmiRNAs were identified at 24 h post-infection (hpi) and all were up-regulated; eight DEmiRNAs were identified with two up-regulated miRNAs and six down-regulated miRNAs at 96 hpi; 27 DEmiRNAs were identified with 13 up-regulated miRNAs and 14 down-regulated miRNAs at 36 days post-infection (dpi). Among these DEmiRNAs, cfa-miR-193b participates in the immune response by regulating the target gene cd22 at 24 hpi. The novel_328 could participate in the inflammatory and immune responses through regulating the target genes tgfb1 and tespa1, enhancing the immune response of the host and inhibiting the infection of T. canis at 96 hpi. In addition, cfa-miR-331 and novel_129 were associated with immune response and self-protection mechanisms at 36 dpi. 20 pathways were significantly enriched by KEGG pathway analysis, most of which were related to inflammatory response, immune response and cell differentiation, such as Cell adhesion molecules (CAMs), ECM-receptor interaction and Focal adhesion. CONCLUSIONS: These findings suggested that miRNAs of Beagle dog bone marrow play important roles in the pathogenesis of T. canis infection in dogs and provided useful resources to better understand the interaction between T. canis and the hosts.