Molecular diagnosis of Trichomonas vaginalis in liquid-based Papanicolaou samples in Shiraz, southern Iran.

BACKGROUND: Trichomoniasis is a parasitic infection of the urinary and genital tract, caused by Trichomonas vaginalis. This study aimed to investigate the molecular diagnosis of T. vaginalis infection in liquid-based Papanicolaou samples in Shiraz, southern Iran. MATERIALS AND METHODS: In this cross-sectional study, 534 liquid-based Papanicolaou samples were collected from women referring to the laboratory of Motahari Clinic of Shiraz University of Medical Sciences in 2021. Genomic DNA were extracted from the samples and examined for evidence of T. vaginalis using polymerase chain reaction (PCR) using TVK3 and TVK7 specific primers. RESULTS: The mean age of participants was 39.28 ± 9.89 with a maximum age of 65 and a minimum age of 19 years. T. vaginalis DNA fragments were detected in 4.86% (26/534) of the cases. There was significantly higher prevalence in the age groups of 21 to 30 and 41 to 50 years (46.15%, p = 0.001 and 38.46%, p = 0.015, respectively). Furthermore, the results showed an association between a history of foamy discharge and Trichomonas positivity (p = 0.001). CONCLUSION: T. vaginalis infection is common in liquid-based Papanicolaou samples of women who attended regular health check-ups in the study area. Screening for trichomoniasis in populations, particularly if using highly sensitive methods such as PCR, may lead to increased detection and treatment.

A machine learning-based system for detecting leishmaniasis in microscopic images.

BACKGROUND: Leishmaniasis, a disease caused by a protozoan, causes numerous deaths in humans each year. After malaria, leishmaniasis is known to be the deadliest parasitic disease globally. Direct visual detection of leishmania parasite through microscopy is the frequent method for diagnosis of this disease. However, this method is time-consuming and subject to errors. This study was aimed to develop an artificial intelligence-based algorithm for automatic diagnosis of leishmaniasis. METHODS: We used the Viola-Jones algorithm to develop a leishmania parasite detection system. The algorithm includes three procedures: feature extraction, integral image creation, and classification. Haar-like features are used as features. An integral image was used to represent an abstract of the image that significantly speeds up the algorithm. The adaBoost technique was used to select the discriminate features and to train the classifier. RESULTS: A 65% recall and 50% precision was concluded in the detection of macrophages infected with the leishmania parasite. Also, these numbers were 52% and 71%, respectively, related to amastigotes outside of macrophages. CONCLUSION: The developed system is accurate, fast, easy to use, and cost-effective. Therefore, artificial intelligence might be used as an alternative for the current leishmanial diagnosis methods.

In vitro evaluation of CRISPR PX-LmGP63 vector effect on pathogenicity of Leishmania major as a primary step to control leishmaniasis.

Cutaneous leishmaniasis (CL) is caused by intracellular obligate parasites (Leishmania spp.) carried by the blood-sucking of female sandflies and transmitted between mammalian hosts. Despite the high incidence and prevalence of Leishmania cases in many countries, it has been a neglected tropical disease. The current treatment approaches are limited by the complications such as loss of fertility and drug resistance. It is, therefore, essential to find new medicines to treat leishmaniasis. CRISPR/Cas9 as a powerful genome-editing tool provides the opportunity to create precise genetic manipulation to investigate the molecular basis of different leishmaniasis cases. Therefore, our main goal was to evaluate the CRISPR PX-LmGP63 vector effect on pathogenicity of Leishmania majorin vitroto challenge for using CRISPR/Cas9 as a therapeutic CL through the reduction of L. major pathogenicity by manipulating the GP63 gene. In this study, L. major parasites were transfected with CRISPR/Cas9 vectors constructed by electroporation and then added to macrophage cells on RPMI. The effect of CRISPR/Cas9 constructs on GP63 mutation, viability, and status of L. major was investigated by counting phagocytic parasites into macrophages and DNA sequence analysis. Our data validate that the use of CRISPR/Cas9 in L. major creates a new stop codon and disrupts the frame sheet of the gene by creating a new insertion (thymine), which prevents its expression. In addition, the parasite count was significantly different in the case and control of infected macrophages (P < 0.05). This study shows the successfully targeted manipulation of the L. major GP63 gene via the adaptation of the CRISPR/Cas9 editing tool. The manipulation of GP63 revealed a reduction in the infection load compared to wild-type parasite infection. Therefore, more studies are necessary for this field to help achieve a new method for the prevention and treatment of CL disease.

Seroprevalence and risk factors for Toxocara infection among children in a rural community in Fars province, southern Iran.

The current study was performed to find out the seroprevalence of Toxocara infection in children living in a rural community in Fars province, southern Iran. Venous blood was taken from 617 children and evaluated for anti-Toxocara antibodies, using an ELISA system. Of the 617 studied children, 318 (51.5%) were boys and 299 (48.5%) were girls. Mean age of the participants was 9.2 (±10.7) years. Most cases of the recruited subjects (37.4%) were in the age group of 0-5 years. Anti-Toxocara antibodies were detected in sera of 39 (6.3%) of children. From these, 23 (62.2%) were boys and 14 (37.8%) were girls. The infection rate was almost the same in different age groups. No statistically significant differences were seen between seropositivity to Toxocara infection and gender or age of the participants. Dogs or cats ownership was not associated with Toxocara seropositivity. The adjusted associations of the study variables suggested that the visceral leishmaniasis (VL) seropositivity is significantly associated with the seroprevalence of Toxocara infection (P < 0.001). Findings of the current study revealed that Toxocara infection is a common infection among children in the studied rural community in Iran. Preventive measurements are necessary to minimize the rate of Toxocara infection in children in such communities.

Preparation and characterization of artemether-loaded niosomes in Leishmania major-induced cutaneous leishmaniasis.

Cutaneous leishmaniasis is the most prevalent form of leishmaniasis worldwide. Although various anti-leishmanial regimens have been considered, due to the lack of efficacy or occurrence of adverse reactions, design and development of novel topical delivery systems would be essential. This study aimed to prepare artemether (ART)-loaded niosomes and evaluate their anti-leishmanial effects against Leishmania major. ART-loaded niosomes were prepared through the thin-film hydration technique and characterized in terms of particle size, zeta potential, morphology, differential scanning calorimetry, drug loading, and drug release. Furthermore, anti-leishmanial effect of the preparation was assessed in vitro and in vivo. The prepared ART-loaded niosomes were spherical with an average diameter of about 100 and 300 nm with high encapsulation efficiencies of > 99%. The results of in vitro cytotoxicity revealed that ART-loaded niosomes had significantly higher anti-leishmanial activity, lower general toxicity, and higher selectivity index (SI). Half-maximal inhibitory concentration (IC50) values of ART, ART-loaded niosomes, and liposomal amphotericin B were 39.09, 15.12, and 20 µg/mL, respectively. Also, according to the in vivo study results, ART-loaded niosomes with an average size of 300 nm showed the highest anti-leishmanial effects in animal studies. ART-loaded niosomes would be promising topical drug delivery system for the management of cutaneous leishmaniasis.

Association between Toxocara Spp. Exposure and Schizophrenia: A Case-Control Study.

Background: Toxocariasis is a zoonotic disease caused by the larvae of Toxocara canis or Toxocara cati. Patients with schizophrenia may be at higher risk of infection, possibly due to their cognitive and personal self-care impairments. We aimed to assess the association between Toxocara spp. exposure and schizophrenia. Methods: This case-control study was conducted on 109 patients with schizophrenia admitted to Ibn Sina Hospital, Shiraz, Iran, and 104 age- and gender-matched healthy controls from May to September 2021. A questionnaire was obtained and serum samples were tested for IgG antibodies to Toxocara excretory/secretory (TES) antigens using an enzyme-linked immunosorbent assay (ELISA). Results: Anti-Toxocara IgG was detected in 12 schizophrenic patients and 10 control subjects, giving respective seroprevalences of 11.0% (95% confidence interval [95% CI]=5.8-18.4%) and 9.6% (95% CI=4.7-17.0%). Univariate logistic analyses estimated an odds ratio (OR) of 1.16 (95% CI=0.44-3.16); however, it was not statistically significant (P=0.915). Individuals with a history of eating unwashed vegetables or fruits (23.1%, 95% CI=9.0-43.6, crude odds ratio [COR]=3.21, 95% CI=1.13-9.13) and rural residency (19.5%, 95% CI=8.8-34.9, COR=2.74, 95% CI=1.06-7.05) had significantly higher rates of seropositivity using the univariate logistic analyses. After multivariate logistic analyses, the differences were not statistically significant. Conclusion: The toxocariasis seroprevalence among schizophrenic and healthy participants was not significantly different (11% vs. 9.6%). Since the disease severity, onset, and cognitive sequelae are not the same among schizophrenic patients, clinically matched studies with larger samples are required to address the current inconsistency between the studies.

Anti-Toxoplasma In Vitro and In Vivo Activity of Pyrus boissieriana Arbutin-Rich Fraction.

PURPOSE: Pyrus boissieriana is a rich source of arbutin and has been used in herbal medicine to treat infectious diseases. This study aimed to investigate the effect of the arbutin-rich fraction of Pyrus boissieriana aerial parts on Toxoplasma gondii In Vitro and In Vivo. METHODS: An arbutin-rich fraction of P. boissieriana was prepared beforehand. Flow cytometry was used to evaluate the effect of different concentrations (1-512 µg/ml) of the P. boissieriana arbutin-rich fraction on Toxoplasma tachyzoites (RH strain). The cytotoxicity of the concentrations on the macrophage J774 cell line was also investigated by MTT assay. For In Vivo investigation, 4-6-week-old female mice infected with the RH strain of T. gondii were treated with different doses (16, 32, 64, 256, and 512 mg/kg) of the fraction using gavage. RESULTS: The highest and lowest lethality of the tachyzoites were 89.6% and 25.9% related to the concentrations of 512 µg/ml and 1 µg/ml, respectively, with an IC50 value of 18.1 µg/ml ± 0.37. The cytotoxicity test showed an IC50 value of 984.3 µg/ml ± 0.76 after 48 h incubation. The mean survival of mice at the lowest treated dose (16 mg/kg) was 6.6 days, and it was 15 days at the highest dose (512 mg/kg). The concentrations of 512, 256, 128, and 64 mg/kg of the fraction compared to the negative control (6.2 days mean survival) significantly increased the survival time of mice (P < 0.001, P = 0.009, P = 0.018, and P = 0.021, respectively). CONCLUSION: The results showed that the arbutin-rich fraction of P. boissieriana is effective against T. gondii In Vitro and In Vivo and may be a reliable alternative to conventional treatment for toxoplasmosis, although further studies are necessary.

In vitro and in vivo anti-parasitic activity of curcumin nanoemulsion on Leishmania major (MRHO/IR/75/ER).

The present study aimed to assess the anti-leishmanial effects of curcumin nanoemulsion (CUR-NE) against Leishmania major (MRHO/IR/75/ER) in both in vitro and in vivo experiments. CUR-NE was successfully prepared via the spontaneous emulsification method. The in vitro effect of various concentrations of CUR-NE against L. major promastigotes was assessed using the flow cytometry method. In vivo experiments were carried out in BALB/c mice inoculated subcutaneously with 2 × 106 L. major promastigotes. Mice were treated with topical CUR-NE (2.5 mg/ml), intra-lesion injection of CUR-NE (2.5 mg/ml), topical CUR suspension (CUR-S, 2.5 mg/ml), topical NE without CUR (NE-no CUR), amphotericin B as the positive control group, and infected untreated mice as the negative control group. In vitro exposure of promastigotes to CUR-NE showed a dose-dependent anti-leishmanial effect, with a 67.52 ± 0.35% mortality rate at a concentration of 1250 µg/ml and an IC50 of 643.56 µg/ml. In vivo experiments showed that topical CUR-NE and CUR-S significantly decreased the mean lesion size in mice after four weeks from 4.73 ± 1.28 to 2.78 ± 1.28 mm and 4.45 ± 0.88 to 3.23 ± 0.59 mm, respectively (p = 0.001). Furthermore, CUR-NE significantly decreased the parasite load in treated mice compared with the negative control group (p = 0.001). Results from the current study demonstrated the promising activity of CUR-NE against L. major in both in vitro and in vivo experiments. Moreover, CUR-NE was more efficient than CUR-S in healing and reducing parasite burden in mouse models. Future studies should aim to identify molecular mechanisms as well as the pharmacologic and pharmacokinetic aspects of CUR-NE.

Toxoplasma gondii infection in patients with brain tumors in Southern Iran: a case-control study.

Considerable evidence points to a dominant role of inflammation in tumor pathology. The biological response of the immune system can be triggered by Toxoplasma gondii as a common brain-tropic parasite. The aim of this study was to investigate an association between Toxoplasma infection and brain tumors. This case-control study was performed on sera of brain tumor patients (n = 124) and age- and sex-matched control subjects (n = 124) in Southern Iran. Data related to tumor site and type were collected during sample collection. Anti-Toxoplasma IgG was assessed by enzyme-linked immunosorbent assay (ELISA). Seroprevalence anti-Toxoplasma IgG was significantly higher in brain tumor patients 30.6% (38/124) compared with 12.1% (15/124) of the healthy controls (OR 3.211; 95% CI 1.658 to 6.219; p = 0.001). The highest seroprevalence was detected in patients with ependymoma (100%), followed by glioblastoma (83%), pituitary adenoma (47.3%), astrocytoma (27.2%), schwannoma (23%), and meningioma (22.6%). The parasite infection was correlated to brain tumor's location i.e., the patients with frontal lobe and sella region tumors had higher seropositivity compared with others (P < 0.05). The higher prevalence of Toxoplasma infection among patients with brain tumor compared with the control group indicates a probable association between the infection and brain tumors.

Protoscolicidal effects of curcumin nanoemulsion against protoscoleces of Echinococcus granulosus.

BACKGROUND: The aim of the present study was to assess in vitro protoscolicidal effects of curcumin nanoemulsion (CUR-NE) against protoscoleces of cystic echinococcosis (CE)/hydatid cysts. METHODS: The CUR-NE was prepared via spontaneous emulsification of soybean as the oil phase, a mixture of Tween 80 and Tween 85 as the surfactant, ethanol as the co-surfactant and distilled water. Various concentrations of CUR-NE (156, 312, 625 and 1250 µg/ml) were exposed to collected protoscoleces of infected sheep liver hydatid cysts for 10, 20, 30, 60 and 120 min. Viability of the protoscoleces were assessed using eosin exclusion test. Morphological changes of the protoscoleces were observed using differential interference contrast (DIC) microscopy. RESULTS: The mean particle size and zeta potential of CUR-NE included 60.4 ± 14.8 nm and - 16.1 ± 1.1 mV, respectively. Results showed that the viability of the protoscoleces decreased significantly with increases in CUR-NE concentrations (p < 0.001). The mortality rates of protoscoleces with exposure to concentrations of 1250 and 625 µg/ml of CUR-NE for 60 min were 94 and 73.33%, respectively. Mortality of the protoscoleces was 100% after 120 min of exposure to 1250 and 625 µg/ml concentrations of CUR-NE. Using NIC microscopy, extensively altered tegumental surface protoscoleces was observed after protoscoleces exposure to CUR-NE. CONCLUSION: The findings of the present study revealed the in vitro protoscolicidal potential of CUR-NE. Therefore, CUR-NEs are addressed as novel protoscolicidal agents, which can be used as an alternative natural medicine to kill the protoscoleces, owing to their low toxicity and significant inhibition potency. However, further studies are necessary to investigate pharmacologic and pharmacokinetics of CUR-NEs.

Synthesis and anti-Toxoplasma activity of indole-triazole compounds on tachyzoites of RH strain.

BACKGROUND: Conventional treatment for toxoplasmosis have severe side effects and the inability to completely eradicate the disease. Therefore, the acquisition of new anti-Toxoplasma drugs has always been of interest among researchers. In the present study, we prepare a new indole-triazole derivatives and evaluated their potential anti-parasitic activity against tachyzoites of Toxoplasma RH strain. MATERIALS AND METHODS: In this study, after synthesis of the two new compounds of indole-triazole, the effect of their different concentrations (2-1024 µg/ml) were determined on Toxoplasma tachyzoites using flow cytometry. Furthermore, tachyzoites were exposed to different concentrations of compounds (4, 16, 64, 265, 1024 µg/ml) for 1.5 h and their infectivity were evaluated in BALB/c mice. RESULTS: The flow cytometry results indicated the benzyl derivative of indole-triazole in various concentrations had a lethal effect on tachyzoites between 11.93% and 89.66%, while the naphthalene derivative had a lethality of 26.63%-66.82%. The infectivity analysis showed that the survival time of mice at concentrations of 1024 µg/ml and 512 µg/ml of benzyl derivatives was significantly increased (P = 0.008 and P = 0.016, respectively), compared to that in the negative control group. Furthermore, survival time of mice was statistically significant at the concentration of 1024 µg/ml for naphthyl derivative (P = 0.012). CONCLUSION: Findings of the current study suggested indole triazole compounds, based on their structure and enzymes targeting, have a considerable effect on tachyzoites of T. gondii RH strain and can be considered as a new anti-Toxoplasma agent.

The Relationship of Latent Toxoplasmosis and Cigarette Smoking: Seroprevalence, Risk Factor, and Case-Control Study in Fars Province, Southern Iran.

Toxoplasmosis is a parasitic disease with worldwide prevalence. Despite the relatively similar effects of toxoplasmosis and smoking on alteration in neurotransmitters, especially dopamine, little is known about the relation of Toxoplasma gondii infection and addiction to cigarette smoking. Therefore, the main objective of this study was to assess the relationship between latent toxoplasmosis and smoking. Through a case-control study, 216 regular cigarette smokers and 324 nonsmoker age- and gender-matched subjects were evaluated for anti-T.gondii IgG antibodies with enzyme-linked immunosorbent assay (ELISA). During the sampling, a structured questionnaire was used to obtain the demographic information of participants and the risk factors of acquired Toxoplasma. The median ages of case and control groups were 51.04 ± 18.1 (22-97 years) and 51.03 ± 16.5 (21-89 years), respectively (p = 0.99). Anti-T.gondii IgG antibodies were detected in 44 (20.37%) cases and in 135 (41.67%) controls. There was a statistically significant difference for the positivity rate between the smokers and the control group (OR = 0.35; 95%CI: 0.19-0.65; and p = 0.001). The overall prevalence was 33.14%. This study indicated the inverse association between seropositivity to Toxoplasma infection and cigarette smoking. This relationship could be due to the changes that latent toxoplasmosis has on the neurotransmitters, especially dopamine, which needs more research.

Cytotoxic screening and in vitro effect of sodium chlorite against Leishmania major promastigotes.

Cutaneous leishmaniasis (CL) is one of the most important parasitic diseases in the world. Despite the existence of many therapeutic strategies, the treatment of this infection still faces problems. Sodium chlorite as an antimicrobial agent has been shown to have acceptable tissue regenerative and wound healing properties. Therefore, the present study aimed to analyze the in vitro effects of different concentrations of sodium chlorite on Leishmania major promastigotes and macrophage cells. The inhibitory and toxicity effect of various concentrations (0.0035, - 1.8 mg/ml) of sodium chlorite on the standard Iranian strain of L. major promastigotes were evaluated via counting the cells and flow cytometry. Furthermore, cytotoxicity on promastigotes and J774 macrophage cell line were performed by MTT assay. The results of the inhibitory test demonstrated that sodium chlorite had dose-dependent, anti-leishmanial activities. The half-maximal inhibitory concentration (IC50) for promastigotes and J774 cells by cytotoxicity test was detected at 0.17 mg/ml and 0.08 mg/ml after 48 h respectively. Flow cytometry results showed that 27.34% death of promastigotes was observed in 0.0035 mg/ml of sodium chlorite and 78.12% in 1.8 mg/ml. The results of the present study showed that sodium chlorite could be used as an effective treatment for CL, especially in cases resistant to treatment with pentavalent compounds. However, the toxicity of this substance in high concentrations should be considered in clinical setting.

Acute toxoplasmosis can increase serum dopamine level.

Toxoplasmosis is a globally parasitic zoonotic disease transmitted by Toxoplasma gondii protozoa. This infection in its chronic form can cause a change in its host's specific behavior and is also associated with developing neuropsychological symptoms in humans. Changes in neurotransmitters' levels, especially dopamine, have been identified as a behavior change factor in the infected host. This study aimed to evaluate serum dopamine levels in acute murine toxoplasmosis. In this study, 50 mice infected with Toxoplasma were studied in 5 separate groups, and ten healthy mice were considered as negative control. For five consecutive days after parasite injection, blood sampling and serum isolation were performed daily from one of the groups. Serum dopamine levels were measured by HPLC method. Statistical studies showed that serum dopamine on the first to the fourth day after parasite inoculation was the same as the negative control, but the fifth day began to increase. The present study results indicate that dopamine production in mice infected with Toxoplasma gondii increases from day five after infection. This result suggests that in acute toxoplasmosis, dopamine production is low, and the trend of chronic disease increases dopamine production.

Toxoplasma Reduces Complications of Parkinson's Disease: An Experimental Study in BALB/c Mice.

Background: Parkinson's disease (PD) has been described in dopamine brain level reductions. Conversely, several studies have shown that Toxoplasma parasite can increase the level of dopamine in an infected host. This study was conducted to assess the serum, cerebral dopamine levels, and downregulation of Parkinson's disease manifestations in mice with chronic toxoplasmosis. Methods: PD induction was done by oral inoculation of rotenone into BALB/c mice. To induce the chronic infection, cysts of T. gondii Prugniaud strain (genotype II) were injected intraperitoneally into the mice. The rotarod test was used for the evaluation of functional motor disorders in experimental mice. The serum and cerebral dopamine levels of the mice were also measured by using high-performance liquid chromatography (HPLC) on consecutive periods (10 days). Results: Findings of the rotarod test showed the highest and lowest average of running duration belonged to the uninfected mice and PD mice, respectively. Remarkably, the running duration of infected mice with PD was higher than PD mice. As well, the level of serum and cerebral dopamine increased in mice with PD and toxoplasmosis in comparison with PD mice. Conclusion: Increasing the serum and cerebral dopamine levels in mice infected with toxoplasmosis is related to the presence of the parasite. Moreover, the dopamine upregulation due to the infection is effective in the reduction of PD complications.

The relationship between Toxoplasma gondii infection in mothers and neonate's gender.

BACKGROUND: The secondary sex ratio can be affected by various factors such as stress, immunosuppression, and age of parents in addition to mother infectious disease (Maternal infections). Toxoplasmosis is one of the critical maternal parasitic infections during pregnancy. Besides the complications of the acute form of the disease, hormonal shifts, and even alterations in the secondary sex ratio can be induced by the manipulative activity of the chronic form of the disease. Therefore, this study aimed to evaluate the correlation between Toxoplasma gondii infection in mothers and neonate's gender. METHODS: In this case-control study, 137 seropositive mothers to Anti-Toxoplasma IgG(case) was compared to 137 age-matched subject Toxoplasma-seronegative mothers(control) in terms of their neonate's gender. These individuals were randomly selected based on exclusions and inclusions criteria of the study from among 2014 mothers who had been tested for Toxoplasma infection during pregnancy from 2015 to 2018 in Shiraz, Iran. RESULTS: From a total of 2014 studied pregnant mothers, 326 (16.2%) mothers were seropositive to anti-Toxoplasma IgG, and 1688 (83.8%) were negative for IgG. It was found that the numbers of female and male neonates were 136 (45.48%) and 163 (54.51%) in the control group whereas, they were 165 (49.84%) and 166 (50.15%) in the case group, respectively. The sex ratio was 1.006:1 in Toxoplasma-seropositive and 1.2:1 in Toxoplasma-seronegative mothers. The number of male and females offsprings indicated a significant difference in Toxoplasma-seronegative mothers (54.5%, p = .015). Moreover, comparing the number of males and females between the two randomly selected groups showed that female gender is significantly more than male gender in seropositive mothers to Toxoplasma (54.8%, p = .014), which means that of 301 females, 165 offspring were born to seropositive mothers. No significant difference was observed for the sex ratio of aborted fetuses between groups. However, in the Toxoplasma-seropositive group, the sex ratio of aborted fetuses showed that the aborted male fetuses were significantly higher in number. (31 male vs 13 female, p < .001). CONCLUSION: Comprehensively, a significant relationship was found between chronic Toxoplasma infection and secondary sex ratio. However, it is suggested that this relationship be investigated in further studies as well as an animal study.

Prevalence and Predictors of Toxoplasma gondii Infection in Psychiatric Inpatients in Fars Province, Southern Iran.

Background: Psychiatric patients are at increased risk of exposure to Toxoplasma gondii infection, which may be linked to their living facilities and behaviors. Limited knowledge on the prevalence of T. gondii infection and its associated risk factors in psychiatric patients are available to the international medical communities. Thus, the aim of the current study was to assess seroprevalence of T. gondii and its associated risk factors in psychiatric inpatients in Fars Province, southern Iran. Methods: This cross-sectional study was carried out on psychiatric patients hospitalized in Ibn Sina Hospital affiliated to Shiraz University of Medical Sciences, Fars Province, southern Iran, March to July 2021. Blood samples were collected from 318 psychiatric patients and assessed for the detection of IgG against T. gondii using enzyme-linked immunosorbent assay (ELISA). Moreover, structured questionnaires were completed for the participants at the time of sampling. Logistic regression analysis was used to assess possible associations between the latent toxoplasmosis and the variables. Results: The overall seroprevalence of anti-T. gondii IgG in psychiatric inpatients was 22.3% (71/318; 95% CI = 17.9-27.3). Multivariate analyses revealed that age > 30 years [adjusted odds ratio (AOR) = 2.24, 95% CI = 1.10-4.60, p = 0.03], contact with cats (AOR = 2.52, 95% CI = 1.14-5.58, p = 0.03), raw vegetable consumption (AOR = 3.65, 95% CI = 1.74-7.65, p = 0.001), raw/undercooked meat consumption (AOR = 4.30, 95% CI = 1.47-12.63, p = 0.008), suicide attempt (AOR = 3.77, 95% CI = 1.58-8.97, p = 0.003) and cigarette smoking history (AOR = 0.38, 95% CI = 0.17-0.83, p = 0.02) were independent risk factors for T. gondii infection. Conclusion: The current results demonstrated that contact with cats, raw vegetable consumption and raw/undercooked meat consumption were independent risk factors for T. gondii seropositivity. Moreover, the current study showed significant associations between seropositivity of T. gondii and suicide attempts as well as negative associations between seropositivity of T. gondii and cigarette smoking in psychiatric inpatients using multivariate logistic regression.

Mining the Proteome of Toxoplasma Parasites Seeking Vaccine and Diagnostic Candidates.

Toxoplasma gondii is a pathogenic protozoan parasite that infects the nucleated cells of warm-blooded hosts leading to an infectious zoonotic disease known as toxoplasmosis. The infection outcomes might be severe and fatal in patients with immunodeficiency, diabetes, and pregnant women and infants. The One Health approach to toxoplasmosis highlights that the health of humans is closely related to the health of animals and our common environment. The presence of drug resistance and side effects, the further improvement of sensitivity and specificity of serodiagnostic tools and the potentiality of vaccine candidates to induce the host immune response are considered as justifiable reasons for the identification of novel targets for the better management of toxoplasmosis. Thus, the identification of new critical proteins in the proteome of Toxoplasma parasites can also be helpful in designing and test more effective drugs, vaccines, and diagnostic tools. Accordingly, in this study we present important proteins found in the proteome of the life cycle-specific stages of Toxoplasma parasites that are potential diagnostic or vaccine candidates. The current study might help to understand the complexity of these parasites and provide a possible source of strategies and biomolecules that can be further evaluated in the pathobiology of Toxoplasma parasites and for diagnostics and vaccine trials against this disease.

Evaluation of the Tyrosine and Dopamine Serum Levels in Experimental Infected BALB/c Mice with Chronic Toxoplasmosis.

BACKGROUND: Toxoplasma parasite alters the transduction of neurotransmitter signals and leads to changes in the level of brain neurotransmitters including tyrosine and dopamine, so behavior changes can occur in infected hosts. Based on this concept, this study was conducted for evaluation of the tyrosine and dopamine serum levels in infected mice with chronic toxoplasmosis. MATERIALS AND METHODS: Toxoplasma gondii (Prugniaud strain II) was injected intraperitoneally into BALB/c mice to induce chronic toxoplasmosis. Modified agglutination test (MAT), polymerase chain reaction (PCR), and microscopic methods were conducted to confirm the induction of chronic toxoplasmosis. The infected mouse sera were separated at days 40, 50, 60, 70, and 80 for evaluation of tyrosine and dopamine serum levels using high-performance liquid chromatography (HPLC). RESULTS: Microscopic methods confirmed the formation of the Toxoplasma cysts in mouse tissues. Inducing chronic toxoplasmosis is also confirmed by MAT, PCR, and histological methods. HPLC results indicated a decrease in serum tyrosine level at day 40 in infected mice in comparison to control, and the levels were too low to be measured at other times. However, a significantly high serum dopamine level was observed that gradually increased after parasite inoculation. CONCLUSIONS: No detection of tyrosine level in most of the sample groups is probably related to the very low concentration of tyrosine in sera. However, low concentration of tyrosine at day 40 and increase of dopamine in most of the sample groups suggest the production of dopamine from tyrosine due to the presence of Toxoplasma in infected mice.

Introduction of protocols for mass production of Toxoplasma gondii tachyzoites of the genotype II PRU strain.

Background: Few investigations of genotype II of Toxoplasma gondii, the most prevalent form of the Toxoplasma parasite in humans, have been carried out on due to the rapid conversion of tachyzoites to bradyzoites in its life cycle. The current study aimed to create animal and in vitro models for production of the tachyzoites of the Prugniaud (PRU) genotype II strain. Methods: To develop an immunocompromised model and obtain tachyzoites of the PRU strain, BALB/c mice were orally treated with dexamethasone (10 mg/kg), cyclophosphamide (36 mg/kg), and cyclosporine (18 mg/kg) from 5 days prior to inoculation. Then, 10-15 tissue cysts of PRU strain were inoculated intraperitoneally into the mice. The tachyzoites obtained from mice were then cultivated in a HeLa cell culture. The resulting yield of tachyzoites was cryopreserved in 92% fetal calf serum, 8% dimethyl sulfoxide. The infectivity of these tachyzoites was evaluated using in vivo and in vitro examinations. Results: Numerous tachyzoites were observed in the peritoneal fluid of the immunosuppressed mice within 10-15 days after inoculation, and many tachyzoites were harvested from the HeLa cell culture. Trypan Blue staining showed 80% viability of the tachyzoites recovered from cryopreservation and this was confirmed by HeLa cell culture. In addition, mice infected intraperitoneally with the recovered tachyzoites presented with cysts in the brain after 2 months. Conclusion: We have developed an animal model for mass production of T. gondii tachyzoites of the PRU strain. This method can provide fresh viable tachyzoites of Toxoplasma gondii for use as and when required in future investigations.