Gastrointestinal parasites of dogs and foxes in the Zanjan province of Iran: With an emphasis on Echinococcus species.

Canids harbor many zoonotic parasites and play an important role in the spread of parasites in the human environment. Estimation of parasitic infection among canids as definitive hosts may help competent authorities design efficient control programs. This study was conducted to determine the prevalence of intestinal parasites in dogs and foxes with an emphasis on Echinococcus spp. A total of 500 fecal samples of dogs and 30 fecal samples of foxes were studied in the summer, autumn, and winter of 2021 in the Zanjan province using the formalin-ethyl acetate concentration technique, followed by multiplex PCR. The overall prevalence of gastrointestinal parasite infection was estimated to be 19.05%. The prevalence was 24.8%, 10.2%, and 26.7% in stray, shelter dogs and foxes, respectively. No parasites were found among pet and guard dog samples. PCR results on Taenidae eggs showed that 2.4% of samples were positive for Echinococcus granulosus and none contained E. multilocularis. Noteworthy is that E. granulosus was identified only in stray dog samples. The higher prevalence of E. granulosus infection in stray dogs in this province emphasizes the importance of monitoring the food sources consumed by these animals.

Molecular detection and phylogenic characterization of Neospora caninum in naturally infected sheep in Alborz and Qazvin provinces, the north of the central region of Iran.

Neospora caninum is a protozoan coccidian parasite that can act as a cause of abortion in sheep. The aim of this study was to investigate the presence of this parasitic agent and its role in causing abortion in sheep of Iran. Between June 2019 and February 2022, 100 samples [brain (n = 39), placenta (n = 8), embryonic membrane (n = 7), cotyledon (n = 7), umbilical cord (n = 2), homogenate mixture of tissues (heart, liver, spleen and digestive track) (n = 37)] that were collected following the necropsies of 39 aborted ovine fetuses from different parts of the Alborz and Qazvin provinces, the north of the central region of Iran were employed for DNA extraction. Nc-5 was selected as the target gene sequence for amplification of DNA by using four pairs of primers in two semi-nested PCR. Samples considered positive for the presence of the NC-5 gene were examined to further confirm the presence of the ITS1 gene. Sequence of NC-5 gene was detected from the 27 tissue samples of 23 aborted ovine fetuses. The ITS1 gene sequence was detected in all of the 27 tissue samples that were positive for the NC-5 gene analysis. Brain tissue was the most studied tissue, and the highest number of positive cases was observed in this tissue. The present study updated the situation of ovine neosporosis in the central region of Iran and confirmed the presence of the N. caninum among sheep flocks' abortion.

RFLP Analysis of Fragments of the 18S rRNA and Cox1 Genes to Identify Sarcocystis cruzi in Water Buffalo (Bubalus bubalis) In Guilan Province, North of Iran.

Background: Sarcocystosis is a zoonotic disease worldwide caused by Sarcocystis spp., some of these species can show clinical and subclinical manifestations, resulting in financial losses. Our study was performed for identifying Sarcocystis sp., in slaughtered buffalo by PCR-RFLP based strategy with sequencing in Guilan, North of Iran. Methods: Overall, 400 fresh muscle samples were prepared via naked-eye observation from 100 buffaloes (esophagus, diaphragm, shoulder, and thigh), followed by the digestion of samples. The PCR was done to amplify partial parts of the 18S rRNA and mitochondrial cytochrome c oxidase subunit I (Cox1) genes. Then, the PCR products were digested by endonuclease SspI, DraI, and FokI. Sequencing of all species was done to confirm the RFLP results. Results: Five macroscopic cysts (1.25%) were visible in the sample by naked-eye examination. Furthermore, 293 samples (73.25%) were found to be Sarcocystis sp. positive through tissue digestion and microscopic observation, whereas 376 samples (94%) were positive by PCR. In addition, the findings of PCR-RFLP and nucleotide sequence samples exhibited the infection of buffaloes with S. cruzi. Conclusion: Based on the data presented herein, Bovine sarcocystosis caused by S. cruzi is very common in buffalo in the Guilan region. Regarding the high prevalence of sarcocystosis, developing disease control and prevention policies for buffaloes is necessary, and a change of attitude in traditional farming is recommended.

The profile of inflammatory factors in dairy calves with Cryptosporidium infection.

The present study was conducted to investigate the detection and identification of Cryptosporidium species via molecular techniques and evaluate the serum concentrations of inflammatory factors in Cryptosporidium species. The fecal samples (n = 256) were collected from pre-weaned (≤ 2.00 months) calves and the positive samples were identified utilizing Ziehl-Neelsen staining. Nested species-specific multiplex PCR (nssm-PCR) and restriction fragment length polymorphism (RFLP) were used to identify the species and sub-species. The serum concentrations of IL-1ß, IL-6, IL-12, TNF-α, and IFN-γ were also assessed. The results revealed that 10.54% of samples were positive. The results of Nested-PCR showed that 92.59% of the samples were positive for C. parvum while 7.41% were positive for C. andersoni. The results of RFLP confirmed 92.59% of the samples for C. parvum, 3.70% for C. muris / C. andersoni, and 3.70% for C. muris. The serum concentrations of IL-1ß, IL-6, IL-12, TNF-α, and IFN-γ were significantly higher in the infected calves compared to those in healthy calves. However, the serum concentration of IFN-γ was significantly higher in the calves infected with C. parvum while the serum concentrations of TNF-α and IL-6 were significantly higher in those infected with C. andersoni . In conclusion, C. parvum was prevalent in the region and the calves demonstrated inflammatory responses to Cryptosporidium species.

Accurate and rapid detection of Fasciola hepatica copro-DNA in sheep using loop-mediated isothermal amplification (LAMP) technique.

Fascioliasis is a parasitic infection caused by Fasciola spp. in humans and animals. Despite significant advances in vaccination and new therapeutic agents, little attention has been paid to validate methods for the diagnosis of fascioliasis in animals. This study aimed to compare the loop-mediated isothermal amplification (LAMP) technique with PCR assay for the diagnosis of F. hepatica in sheep. In this cross-sectional study, 195 stool samples were collected from sheep for 3 months in Lorestan province, West of Iran. Specimens' parasitological examination was performed by using the direct wet mount and formalin-ether concentration method. After DNA extraction from the samples, molecular analysis was done using PCR and LAMP techniques based on the Fasciola ribosomal intergenic spacer (IGS) sequence. Of 195 specimens of sheep, 11 specimens were identified as F. hepatica-positive infection by using microscopic, PCR and LAMP assays. Kappa agreement test results showed that there was a significant agreement between the results of microscopic examination diagnostic tests, PCR and LAMP (Kappa = 0.51-0.72 and p < .001). According to the results of chi-square comparisons between parasite prevalence applying different techniques and variables of age, sex breed, and type of drinking water, there was no significant relationship (p ≥ .05). However, most of the infected sheep with Fasciola were 3- to 4-year-old females, of the Lori breed and consumed tap water. In many endemic areas, successful prevention and treatment of fascioliasis in animals depend on rapid and accurate diagnosis. Based on the results of the Kappa agreement, the significant agreement among the results of the microscopic examination, PCR and LAMP indicates the accuracy and reliability of these tests in the diagnosis of F. hepatica in sheep. However, molecular methods, especially the LAMP technique, are suggested because of their higher sensitivity and reliability for the diagnosis of F. hepatica even under field conditions.

DNA barcoding of Iranian radicine freshwater snails begins to untangle the taxonomy and phylogeography of intermediate hosts of schistosomiasis and fasciolosis from the Middle East and across Central Asia.

In the Middle East radicine snails are of considerable medical and veterinary importance acting as vectors of trematodes. In Iran, such snails are responsible for the transmission of the zoonotic trematodes Schistosoma turkestanicum and Fasciola gigantica. Historically, Radix gedrosiana has been incriminated as an important intermediate host for both trematodes, however, controversy remains over the snail's true taxonomic status. This species has been determined using morphological characters that has resulted in erroneous identification of species, affecting understanding of population biology, and ultimately affecting vector incrimination. In this current study DNA barcoding using cox1 and phylogenetic analyses revealed that snails identified as R. gedrosiana from Iran split into two separate species, Radix euphratica and Ampullaceana sp. The cox1 also provided useful insights into the evolutionary history of R. euphratica populations. Phylogeographic analyses indicated that R. euphratica had an Iraqi/Iranian origin approximately 3.3 MYA and exists as a large stable population across the Middle East and Central Asia, and a lack of genetic differentiation between geographical isolates. Such molecular barcoding techniques are crucial for the identification of radicine snails of Iran being invaluable for the monitoring of zoonotic flukes, understanding the distribution of infection and the accurate incrimination of snail vectors.

A review on inactivation methods of Toxoplasma gondii in foods.

Toxoplasmosis is an infection caused by Toxoplasma gondii, a widespread zoonotic protozoan which poses a great threat to human health and economic well-being worldwide. It is usually acquired by ingestion of water contaminated with oocysts from the feces of infected cats or by the ingestion of raw or undercooked foodstuff containing tissue cysts. The oocyst can contaminate irrigation water and fresh edible produce. It is estimated that approximately one-third of the human population worldwide harbor this parasite. Infection with T. gondii is an important cause of diseases of the central nervous system and the eye in immunocompromised and immunocompetent individuals. The purpose of this study was to evaluate the efficacy and applicability of thermal (heating, cooking, freezing and low temperature), non-thermal (high pressure processing, ionizing irradiation and curing) and chemical and biochemical (disinfection, essential oils and biochemical methods such as enzymes, nanoparticles, antibiotics and immune response) treatments for the inactivation, inhabitation or to kill T. gondii in foodstuff intended for public consumption and under experimental conditions.