Genetic diversity and seroprevalence of Toxoplasma gondii in COVID­19 patients; a first case-control study in Iran.

BACKGROUND: Toxoplasmosis is a serious or life-threatening disease in immunosuppressed patients and pregnant women. This study examined the likely association between Toxoplasma gondii infection and COVID-19 patients with moderate illness. METHODS: Seventy blood samples were collected from patients at the Health Reference Laboratory of Tabriz, Northwest Iran from April 2021 to September 2021. In addition, 70 healthy subjects of the same age (37 ± 15 years) and sex distribution were ethnically matched. Sera samples were examined for the detection of anti-Toxoplasma antibodies using ELISA. Nested-PCR targets were amplified based on the B1 and GRA6 genes. GRA6 amplicons were subjected to sequencing and phylogenetic analysis. RESULTS: The seroprevalence of toxoplasmosis based on IgG titer was 35.7% in the COVID­19 patients and 27.1% in the control group, representing not to be associated with the Toxoplasma seropositivity in COVID­19 patients (P = 0.18) compared to healthy subjects. Anti-T. gondii IgM was not found in any of the patients and healthy individuals. According to PCR amplification of the B1 and GRA6 genes, the frequency of T. gondii in COVID-19 patients was 14.2% (10/70). However, no T. gondii infection was detected in the healthy group. The CD4+T cell count was relatively lower in toxoplasmosis-infected patients (430-450 cells/mm3) than in control group (500-1500 cells/mm3). High genetic diversity (Hd: 0.710) of the type I strain of T. gondii was characterized in the patients. Present results showed that consumption of raw vegetables and close contact with stray cats can increase the transmission of T. gondii to COVID-19 patients (P < 0.01). CONCLUSIONS: The current study revealed that T. gondii type I infection is unequivocally circulating among the COVID-19 patients in Tabriz; However, no significant association was observed between the occurrence of Toxoplasma and the severity of COVID-19. To make more accurate health decisions, multicenter investigations with a larger sample size of different ethnic groups of the Iranian population are needed.

Prevalence of toxoplasmosis in patients infected with tuberculosis; a sero-molecular case-control study in northwest Iran.

In this study, we investigated the possible association between TB and Toxoplasma gondii infection. One hundred confirmed TB individuals living in northwest Iran were classified into three subgroups; newly diagnosed patients (NTB), old diagnosed patients (OTB) and multidrug resistance patients (MDR-TB). One hundred healthy subjects in the same age and sex distribution were ethnically matched. Sera samples were screened for anti-Toxoplasma antibodies. Nested-PCR was performed by targeting the B1 and GRA6 genes. The frequency of Toxoplasma infection based on IgG titer was 71.1% in the OTB subgroup and 33% in the control group, indicating significant association between Toxoplasma seropositivity and OTB (P = 0.001). According to phylogenetic network, the type I strain of Toxoplasma was identified in the OTB subgroup (10.1%). We concluded that patients with OTB subgroup are at high risk for acquisition of Toxoplasma infection which could reactivate the latent toxoplasmosis.

Global assessment of genetic paradigms of Pvmdr1 mutations in chloroquine-resistant Plasmodium vivax isolates.

BACKGROUND: Chloroquine (CQ) is generally prescribed as the front-line antimalarial drug of choice to treat Plasmodium vivax infections; however, some clinical CQ-resistant P. vivax isolates have been indigenously reported around the world during the last decade. METHODS: In this study, P. vivax isolates (n=52) were obtained from autochthonous samples in southeast Iran during 2015-2017. The genomic DNA of samples was extracted, amplified (nested PCR) and sequenced by targeting the multidrug-resistance 1 gene. To verify the global genetic diversity of CQ-resistant P. vivax strains, the sequences of Pvmdr1 originating from Asia and the Americas were retrieved. RESULTS: A total of 46 haplotypes were grouped into three distinct geographical haplogroups. The haplotype diversity and occurrence rates of Pvmdr1 976F/1076L mutations indicate that the efficacy of CQ is being compromised in Mexico, China, Nicaragua, Thailand, Brazil (2016), Ethiopia, Mauritania (2012) and southwest India in the near future. The cladistic phylogenetic tree showed that Pvmdr1 sequences isolated from the southeast Asian clade has a partial sister relationship with the American clade. CONCLUSIONS: The current findings will serve as a basis to develop appropriate malaria control strategies and public health policies in symptomatic imported malaria cases or plausible CQ-resistant P. vivax strains.

Molecular evaluation of pvdhfr and pvmdr-1 mutants in Plasmodium vivax isolates after treatment with sulfadoxine/pyrimethamine and chloroquine in Iran during 2001-2016.

The rising use of sulfadoxine/pyrimethamine (SP) in the treatment of chloroquine (CQ)-resistant Plasmodium falciparum has resulted in increased exposure to P. vivax isolates in Iran, where both species are being circulated. In this investigation, the frequency of pvdhfr and pvmdr-1 mutants was assessed in P. vivax strains during 2001-2016 after the introduction of SP/CQ in malarious areas of Iran. The P. vivax isolates (n, 52) were obtained from autochthonous samples in Southeast Iran during 2015-2016. The genomic DNA was extracted and examined using nested polymerase chain reaction-(PCR) and sequencing. Mutations were detected in pvdhfr codons P33L (21.2%), T61 M (25%), S93H (3.9%), and S117 T (1.9%) and 5 isolates showed double mutations (33 L/61 M, 7.7%; 33 L/117 T, 1.9%). No mutation was identified in pvdhfr codons F57 and S58. The pvmdr-1 1076 L mutation was detected in 93.3% of P. vivax isolates. The findings indicated that the frequency of three codons of pvdhfr F57/S58/S117 has decreased from 2001 (1.05%/7.0%/16.9%) to 2016 (0%/0%/1.9%). Genomic analysis of pvmdr-1 showed that the frequency of 1076 L has gradually increased from 2013 (93%) to 2016 (93.3%) (P > .05). The results demonstrated that P. vivax isolates are probably being exited under SP pressure, which reflects the appropriate level of training for field microscopists, as established by Iranian policymakers. Emergent pvdhfr codons 33L, 61M, and 93H should be noticed in plausible drug tolerance and treatment plans. The high prevalence of pvmdr-1 1076L mutation shows that efficacy of CQ combination with primaquine may be in danger of being compromised, however further investigations are needed to evaluate the clinical importance of CQ-resistant P. vivax isolates.

Epidemiology of malaria in East Azerbaijan province, Iran, from 2001 to 2013.

Malaria is one of the most important parasitic diseases worldwide, which is characterized by high morbidity and mortality in tropical and subtropical regions. The aim of this study was to evaluate epidemiology of malaria in East Azerbaijan province, Iran, from 2001 to 2013. During 13 years, blood samples were taken from all suspected malaria cases using lancet and then peripheral blood smear was prepared using one blood drop. The smears were stained by Giemsa's stain and were examined under a light microscope with 1000X of magnification. All demographic variables and epidemiological recorded data were obtained from Health Center and were analyzed by SPSS v. 16 software using descriptive statistical tests. Total of 133 cases were fined to be infected by malaria in 13 years that the highest rate (54.13 %) was observed in Kaleybar county. One hundred and fifteen (86.46 %) and 18 (13.54 %) out of 133 infected individuals were male and female, respectively. Mean age of the infected people was 31.57 years. The most affected age group was 30-40 years. One hundred twenty seven (95.48 %) and 6 (4.52 %) cases were infected by Plasmodium vivax and Plasmodium falciparum, respectively. Based on the findings of this study, the incidence of malaria has been declined continuously over the past decade in East Azerbaijan province, Iran.