Seromolecular study on the prevalence and risk factors of Toxoplasma gondii infection in pregnant women referred to a gynecology hospital in Urmia, northwest part of Iran in 2022.

Toxoplasmosis is a frequent infection among the human population. The infection can cause devastating complications for the fetus during pregnancy. The present study aimed to determine the serological and molecular prevalence of the infection and molecular characterization of Toxoplasma gondii isolates among pregnant women referred to Kowsar Hospital, Urmia, Iran. In a cross-sectional study, 340 blood samples were collected from pregnant women referred to Kowsar Hospital, Urmia, Iran from May to July 2022. Anti-T. gondii IgG and IgM seropositivity were determined by enzyme-linked immunosorbent assay. PCR was carried out by targeting the GRA6 gene of the parasite on all patients' buffy coats. Anti-T. gondii IgG and IgM antibodies were positive in two (0.6%) women, and 101 (29.7%) women had anti-T. gondii IgG and 70.3% were seronegative. PCR was positive in two IgM-positive women, and both isolates belonged to T. gondii carrying the GRA6 allele of lineage I. The risk of infection was significantly higher in women who had constant contact with cats and soil, and who were residents of rural areas. The two IgM-positive women were asymptomatic regarding acute toxoplasmosis. According to the results of the present study, the prevalence of toxoplasmosis in pregnant women in Urmia is similar to its prevalence in other areas in northwestern Iran, and despite the low prevalence of acute infection, it should not be ignored.

Prevalence and molecular analysis of Cryptosporidium spp. collected from surface water.

Cryptosporidium is an obligate intracellular parasite reported from all over the world. This protozoan infects a wide range of animals as well as humans. Cryptosporidium parvum and Cryptosporidium hominis are the most prevalent infecting species with mild and self-limiting infection in healthy people. The protozoan oocyst is resistant to common water purifiers. Based on emerging evidence, Cryptosporidium is one of waterborne parasites considered a major public health problem in developing and developed countries. In this study, 42 samples were collected from 14 rivers in the catchment area of Lake Urmia in northwest of Iran. Moreover, amplification of SSU rRNA gene was performed, and polymerase chain reaction products were sequenced. The results of sequencing and comparing the sequences with those in the GenBank revealed that all the 17 positive samples were C. parvum, a zoonotic species and one of the most frequent human-infecting species. Considering these data, it is highly important to inhibit the spread of this protozoan by treating livestock and preventing human and animal effluents from entering the water.

Evaluation of Adjuvant Effectiveness of Alum-Propranolol Mixture on the Immunogenicity of Excreted/Secreted Antigens of Toxoplasma gondii RH Strain.

Purpose: The introduction of novel adjuvants is an important step in attempts to develop a safe and more efficient vaccine. The present study was performed to determine whether the use of a mixed beta-adrenergic receptor antagonist propranolol (PRP) and aluminum (alum), as an adjuvant, have efficacy for Toxoplasma gondii vaccine to induce protective immunity in a mouse model. Methods: Female BALB/c mice divided into five groups were immunized with excretorys-ecretory antigens (ESA) vaccine, alum-ESA vaccine, PRP-ESA vaccine, and alum-PRP ESA vaccine, as well as with phosphate buffered saline (PBS), as a negative control group. The immune responses were evaluated by lymphocyte proliferation assay for measuring delayedtype hypersensitivity (DTH) response and by cytokine assay for evaluating IFN-γ and IL-5 levels. The survival rate of mice in all groups was assessed during a three-week monitoring period after an intraperitoneal challenge with T. gondii tachyzoites. Results: The results showed that mice immunized with PRP, as an adjuvant, could secret a higher level of IFN-γ, which was significant in comparison to other groups. However, mice vaccinated with alum-precipitated ESA antigen had ability to produce an elevated level of IL-5 compared to other mouse groups (P ≤ 0.05). Moreover, alum-PRP co-administration together with ESA vaccine resulted in the longer survival of mice. Conclusion: The findings of this study revealed that the combination of alum-PRP adjuvants and ESA vaccine of T. gondii elicits both humoral and cellular immune responses, which are comparable to either alum or PRP alone.

Prevalence and genetic characterization of Cryptosporidium in pre-weaned cattle in Urmia (Northwestern Iran).

INTRODUCTION: Cryptosporidiosis is a zoonotic disease causing digestive problems in pre-weaned calves. Considering the zoonosis of the parasite and its importance in veterinary medicine, we evaluated the prevalence and genotyping of Cryptosporidium spp. in diarrheic pre-weaned calves in the northwest of Iran. METHODOLOGY: A total of 100 stool samples of the infant calves with diarrhea were collected from industrial and conventional livestock farms in Urmia City. All the samples were tested with acid-fast staining, ELISA, and PCR. Positive samples of the PCR method were sequenced to determine the Cryptosporidium species. The obtained results were compared for the mentioned methods based on statistical factors, sensitivity, specificity, positive and negative predictive values, as well as duration of the experiment and the costs of testing. RESULTS: The results of this study showed that the prevalence of Cryptosporidium spp. in diarrheic infant calves in Urmia city was 5%, and C. parvum species of Cryptosporidium was detected in all the sequenced samples. According to the findings of the current study, the most appropriate method for the detection of the parasite is the ELISA that has a higher sensitivity and predictive value than acid-fast staining method and should be used in veterinary laboratories. CONCLUSIONS: In the current investigation, C. parvum was identified as the only infectious agent in the region and could be the main cause of human infection. More studies are needed to find the source of infection for establishing the control measures.

Sero-epidemiology of Hydatidosis Among General Population of Jolfa County, Northwestern Iran Using IHA, ELISA and Western Blot (2017-2018).

BACKGROUND: Human hydatidosis is mostly a latent and neglected disease with known endemicity in Iran. AIMS: Due to the importance of this infection in the country and its latent nature, we aimed to evaluate the serological status of hydatid cyst in northwestern Iran. OBJECTIVES: Herein, we evaluated the serological status of hydatid cyst in urban and rural inhabitants of Jolfa county, northwestern Iran during 2017-2018. METHODS: In total, 1296 blood samples were obtained from human individuals and the presence of anti-E. granulosus antibodies was investigated using IHA, ELISA and WB. RESULTS: Based on results, 25 IHA positive person were detected in the examined population, however ELISA test showed 14 of 25 IHA positive patients as negative. Also, 269 IHA negative fellows were shown as negative by ELISA. WB analysis of sera from 25 IHA positive subjects revealed consistent results with the ELISA test, and the most reactive SHCF Ag was a 37 KDa protein. The age-standardized seroprevalence of hydatidosis among Jolfa's general population was 1.12% with 95%CI: 1.02-1.20. Moreover, there existed a significant association between keeping/- contact with dogs (P = 0.022) as well as vegetable consumption (P < 0.001) with ELISA positive test results. CONCLUSION: Along with such serological evidence in this region, we highly suggest physical examination and applying imaging techniques for suspected cases in the area for a better understanding of CE.

Evaluation of Toll-Like Receptor 11 Agonist Adjuvant Activity in Immunization of BALB/c Mice with Total Lysate Antigens of Toxoplasma gondii RH Strain.

BACKGROUND: In this study, the effect of total lysate antigen (TLA) of Toxoplasma gondii on spleen lymphocyte prolifration, secretion of IL5, INF-γ, and mice survival time was evaluated using agonist of toll-like receptor (TLR) 11, as an adjuvant. METHODS: This study was done in the Department of Parasitology and Mycology of Urmia University of Medical Sciences, Urmia, Iran in 2018. First, different groups of BALB/c mice were immunized with TLA alone and also TLA + TLR11 agonist. Subsequently, factors such as spleen lymphocyte prolifration, IL-5, and INF-γ secretion and mice survival time in different groups were compared. RESULTS: Mice immunized with TLA + adjuvant showed higher immunization index than the two other groups and combination of TLR11 (as an adjuvant) and TLA significantly elevated the effect of TLA by increasing the production of INF-γ and IL-5 and by the shift of the immune system to Th1. In addition, the combination of TLA and TLR11 adjuvant increased the proliferation of lymphocytes and survival time in mice against T. gondii. CONCLUSION: Profilin (as an adjuvant) in combination with TLA could be a potent vaccine candidate that evokes a powerful specific immune response and significantly improves the efficacy of TLA vaccine by increasing the induction of INF-γ production and by shifting the immune responses to Th1 profile through increasing the INF-γ/IL-5 ratio. It causes significant protection against T. gondii after i.p. injection.

Designing and standardizing Toxocara serologic diagnostic kit and determining anti-Toxocara antibodies frequency in patients referred to health care centers in Urmia (northwest of Iran).

Toxocariasis is a helminthic zoonosis caused by larval stages of the roundworm of dog, Toxocara canis, and less frequently by T. cati, the roundworm of cats. Eosinophilia in peripheral blood may be indicative of a disease; however, it does not necessarily represent toxocariasis. Therefore, it is necessary to investigate the relationship between eosinophilia and toxocariasis in a region. The aims of this study were the diagnosis of hyperosinophilia patients using traditional ELISA kit and also by a handmade ELISA kit produced by T. canis excretory-secretory (TCES) antigens as well as the determination of the abundance of anti-Toxocara antibodies among people referred to Urmia, northwest of Iran care centers. Traditional ELISA kit was used to determine anti-TCES-specific IgG antibodies on 180 hypereosinophilic samples. These antibodies were evaluated in 1002 samples, including 180 hypereosinophilic samples and 822 random samples without eosinophilia by a handmade ELISA kit produced by TCES antigens. A Western-blot confirmatory test was performed on ELISA-positive samples. Our results showed a 17.22% prevalence rate of Toxocara antibodies among hypereosinophilic samples with traditional ELISA kit, and this rate was 3.89% in the 1002 study population with random sampling (with or without eosinophilia). Also, there was a good match between the results of handmade ELISA with those of traditional kit. The positive results in the ELISA method were confirmed by the Western-blot analysis. Our findings show that although the high eosinophil count is not necessarily a sign of toxocariasis, in Urmia distric, about one-fifth of eosinophilia cases have anti-toxocariasis antibodies. In addition, the abundance of anti-Toxocara antibodies in this area was 3.89%.