A Case-Control Seroprevalence Survey of Toxoplasmosis in Hemodialysis Patients and Healthy Subjects in Kazeroon and Jahrom Districts in Fars Province, Southern Iran.

Opportunistic parasites such as Toxoplasma gondii (T. gondii) are capable of causing neurological and ocular manifestations in patients undergoing hemodialysis. By designing a matched case-control study, we conducted a seromolecular survey of T. gondii in hemodialysis patients compared to a healthy group from Jahrom and Kazeroon cities in Fars Province, Iran. For this purpose, 75 hemodialysis patients from Kazeroon city, 75 hemodialysis patients from Jahrom city, and 75 healthy individuals were recruited for the study. The serum levels of specific immunoglobulins (IgG/IgM) in the case and control groups were evaluated using the enzyme-linked immunosorbent assay (ELISA) method. Also, buffy coat samples were used to extract genomic DNA. Then, Polymerase Chain Reaction (PCR) was accomplished using the RE and GRA6 genes of T. gondii. A standard questionnaire containing demographic factors was administered. Although the seroprevalence of the anti-T. gondii IgG antibody in hemodialysis patients from Kazeroon (18.66% (14/75)) and Jahrom (25.33% (19/75)) was higher than that in the control group (13.33% (10/75)), no statistically significant difference was observed between the case and control groups (P value = 0.373 from Kazeroon and P value = 0.354 from Jahrom). Among the studied variables, only residence (urban) was significantly associated with the anti-T. gondii IgG antibody in the case group from Kazeroon. Also, no IgM antibody titers and DNA of T. gondii were detected in the case and control groups from both cities. Although high seroprevalence of anti-T. gondii IgG antibody was seen in hemodialysis patients, further epidemiological studies with larger samples need to be done in Jahrom and Kazeroon cities and in other parts of Iran. It is also necessary for health officials to establish programs for the prevention and control of T. gondii infection in hemodialysis patients.

First molecular detection of SARS-CoV-2 virus in cockroaches.

Coronavirus is one of the main pathogens that primarily targets the human respiratory system. There are several ways to transmit this virus, such as direct contact or droplets spread by coughing or sneezing, and direct contact with fomites and surfaces is another way. This cross-sectional study was conducted in Shiraz, southern Iran, in 2021. 5 locations, including 3 hospitals and 2 dormitories, were selected for the survey. The cockroaches were collected from selected locations and transferred to the Laboratory of Medical Entomology at Shiraz University of Medical Sciences. All specimens were identified morphologically. The external and gastrointestinal washouts of collected samples with sterile phosphate-buffered saline separately were used for molecular analysis. An RT-qPCR assay, which suggests the possible insect­borne transmission, was used. External and gastrointestinal washout of B. germanica from Dastgheyb Dormitory and P. americana from Ali-Asghar Hospital were positive for contamination with the SARS-CoV-2. Cockroaches spread the virus in the environment and contaminate human food and various surfaces of buildings. Their role will be more important in crowded places such as hotels, lodging houses, restaurants, and hospitals; vector control programs should be carried out with more accuracy in such places.

Global prevalence and subtype distribution of Blastocystis sp. in rodents, birds, and water supplies: A systematic review and meta-analysis.

Animals such as rodents and birds may play a key role in the distribution of Blastocystis sp., either as introducers of the parasite into the water or as receptors of an infection already established in the water. Hence, we aimed to evaluate the prevalence and subtype distributions of Blastocystis sp. in rodents, birds, and water supplies at a global scale through a systematic review and meta-analysis approach. The standard protocol of preferred reporting items for systematic reviews and meta-analyses (PRISMA) statements were applied. Eligible prevalence studies on Blastocystis sp. in rodents, birds, and water supplies, published between 1 January 2000 and 20 January 2022 were collected using a systematic literature search in online databases (Scopus and Web of Science) and search engines (PubMed and Google Scholar). Inclusion and exclusion criteria were followed. The point estimates and 95% confidence intervals (CI) were calculated using a random-effects model. The variances between studies (heterogeneity) were computed by I2 index. In total, 20 articles (24 datasets) for birds, 18 articles (18 datasets) for rodents, and 10 articles (12 datasets) for water supply were included for the final meta-analysis. The pooled prevalence of Blastocystis sp. in birds, rodents, and water was estimated to be 29% (95% CI 12-47%), 18% (95% CI 12-23%), and 10% (95% CI 6%-15%), respectively. Considering the subtypes, Blastocystis sp. with subtypes ST7 in birds, ST4 in rodents, and ST1 in water supplies were the highest reported subtypes. The present results highlight the role of birds, rodents, and water as a reservoir for human-infected Blastocystis sp. Therefore, this global estimate could be beneficial for preventive and control measures.

Isolation and identification of potentially pathogenic free-living amoeba in dental-unit water samples.

Although the presence of free-living amoebae (FLAs) in various water sources has been reported, few studies have been surveyed on their abundance in medical-unit and dental-unit water samples. The current study aimed to identify morphological and molecular characteristics of FLA isolates in the water samples of the dental unit in Iran. A total of 232 water samples were collected from 17 dental units. Then, filtration and cultivation were conducted on a non-nutrient agar (NNA) medium. Also, polymerase chain reaction (PCR) assay and sequencing were performed by using the genus/species-specific primers plus a common primer set on positive samples. One hundred and sixty-six samples were positive for FLA by the microscopic method, whereas 114 samples were positive by the molecular method with a common primer set. Considering the PCR assay with genus/species-specific primers, 23.27% (54/232) samples were identified as Acanthamoeba spp. (belonging to T4 genotype), 36.63% (85/232) as Vermamoeba vermiformis, and 1.72% (4/232) as Vahlkampfiidae family (Naegleria lovaniensis). These results highlight a need to improve filtration systems in dental units and periodic screenings for FLA in dental-unit water.

Sero-molecular survey on Toxoplasma gondii infection among drug addicted and non-addicted individuals: a case-control study.

BACKGROUND: Up to now, epidemiological studies on the prevalence of Toxoplasma gondii infection among drug addicted individuals have been rarely performed. By designing an age and sex matched case-control study, we sought to determine the prevalence and associated factors with T. gondii infection in these population using serological and molecular techniques. METHODS: One hundred and thirty-seven drug addicted individuals and 141 healthy subjects were enrolled in this study. Informed consent as well as a standard questionnaire were obtained from all subjects participating. Blood samples were collected from each participant and the serum was screened for anti-Toxoplasma antibodies (IgG and IgM). PCR assay was performed using the primer pair targeting the RE and GRA6 genes of T. gondii. Then, PCR products were sequenced to determine genotype. RESULTS: The seroprevalence of T. gondii infection based on IgG titer was 34.3% in case and 9.9% in the control groups, revealing a statistically significant difference (OR = 4.37; 95% CI = 2.46-9.12; P = 0.001). After analyzing the variables studied through the questionnaire, age was the only significantly factor associated with the anti-T. gondii IgG antibody in case group. Considering PCR assays with RE genomic target, the prevalence of T. gondii infection was 5.1% in the case and 3.5% in control groups which the difference was no statistically significant (OR = 1.46; 95% CI = 0.45-4.73; P = 0.521). Subsequently, all sequenced samples were genotype #1 using the GRA6 genomic target. CONCLUSIONS: T. gondii exposure is relatively high among drug addicted individuals in Iran, and there is a need for health policymakers and researchers to establish enlightenment and prevention programs for these population at risk of infection.

Bionomics of Phlebotomine Sand Flies in Different Climates of Leishmaniasis in Fars Province, Southern Iran.

Background: Phlebotomus and Sergentomya are distributed in the old-world regions and transmit leishmaniases through mammalian and reptile hosts. Cutaneous leishmaniasis (CL) is one of the most important diseases in Iran. Iranian sand flies belong to three Oriental, Palearctic, and Afrotropical Regions. Fars Province is located in Palearctic, but southern parts are affected Oriental region situations on phglebotomine population variety. Therefore, a comprehensive study was required on the vectors of the disease in this Province. Methods: Regarding the approved role of the environmental factors in creating the fauna and distribution of living creatures, the "de martonne climate method" was used, and the climate was noticed as an important environmental factor for the determination of vector distribution. Accordingly, 14 sampling sites were selected from 10 foci in different climates of Fars Province. 19648 sand flies were collected from the studied areas in this Province during 2016. Results: Phlebotomus papatasi and Se. antennata were the most frequent species, which were caught from in/outdoor areas. Phlebotomus sergenti and Ph. alexandri were caught from both Palearctic and Oriental zones but were more prevalent in the cold semi-arid climate of the Palearctic zone. Moreover, they were not caught from the hot desert and summer Mediterranean climates of the Oriental zone. Conclusions: It seems that Ph. papatasi as the main vector of CL could be well distributed in different climates in Fars. Moreover, some species like Ph. sergenti preferred especial climates in the Palearctic zone. Therefore, these data could be helpful to control leishmaniases more efficiently.

Isolation and identification of potentially pathogenic free-living amoeba in dialysis fluid samples of hydraulic systems in hemodialysis units.

BACKGROUND: Free-living amoeba (FLA), including Acanthamoeba, Naegleria, Balamuthia and Vermamoeba, have been isolated from water, sand, soil, dust and air. Numerous studies considered that FLA are a significant cause of neurological and ocular complications in high-risk groups, including immunocompromised individuals. The present study aimed to identify morphological and molecular characteristics of FLA isolates in dialysis fluid samples of hydraulic systems in hemodialysis units in Iran. METHODS: A total of 328 dialysis fluid samples were collected from 16 dialysis machines, including 164 samples before hemodialysis sessions (after cleaning) and 164 samples after hemodialysis sessions (before cleaning). Filtration and cultivation were performed on non-nutrient agar medium. Also, PCR and sequencing were applied by using the genus-specific primers along with a common primer set on positive samples. RESULTS: Both morphology and molecular investigations showed that 22.5% (74/328) of dialysis fluid samples were positive for FLA. There was a positive relationship between the high frequency of FLA after hemodialysis sessions (before cleaning) compared with before hemodialysis sessions (after cleaning) (OR=2.86; 95% CI 1.5 to 5.45). Considering the PCR assay, 16.46% (54/328) samples were identified as Acanthamoeba spp. (belonging to T3 and T4 genotypes), 5.18% (17/328) as Vermamoeba vermiformis and 0.91% (3/328) as Vahlkampfiidae family (Naegleria australiensis, Naegleria pagei and Allovahlkampfia). CONCLUSION: The present results support a need to improve filtration and purification methods for dialysis fluid of hydraulic systems in hemodialysis units. They also highlight the relevance of periodic screenings for FLA-related diseases in hemodialysis patients.

Genotyping of Toxoplasma gondii Strains from Goats in Jahrom District, Southern Iran.

PURPOSE: Toxoplasma gondii is transmitted congenitally or acquired by consumption of food and water contaminated with cysts or oocysts. This study aimed at genotyping T. gondii strains from slaughtered goats in Jahrom. METHODS: A total of 561 specimens (heart, diaphragm, and tongue) from 187 slaughtered goats were collected from Jahrom slaughterhouse. After DNA extraction, the T. gondii strains were genotyped by the nested PCR-RFLP based on GRA6 and 3', and 5' ends of the SAG2 gene. RESULTS: T. gondii infection was present in 18.2% of cases. Among the examined organs, the diaphragm was more disposed to the infection (10.2%). Furthermore, infection rates of the heart and tongue were 8.6% and 3.7%, respectively. Concurrent infection in the heart and diaphragm, tongue and diaphragm, and heart and tongue were 3.2%, 0.5%, and 0.5%, respectively. In genotyping experiments, genotype I was the most frequent genotype of T. gondii (58.8%), followed by type II (23.5%), type III (11.8%), and a combination of type I and II (5.9%). CONCLUSIONS: The results of this study showed the presence of different genotypes of T. gondii in goats including three major and mixed genotypes. These results can be useful in toxoplasmosis control and prevention.

Genetic characterization of Toxoplasma gondii isolates from human spontaneous aborted fetuses in Jahrom, southern Iran.

Toxoplasma gondii (T. gondii) is an intracellular protozoan that infects the fetus through the placenta and leads to severe complications in the fetus. One of the complications of congenital toxoplasmosis is spontaneous abortion. The prevalence of toxoplasmosis infection was investigated among spontaneously aborted fetuses (SAFs), and the genotypes of parasite isolates were determined in the present study. Placentas from 330 samples of SAFs were collected in Jahrom (Fars province) from February to September 2018. DNA was extracted from each placental tissue. The T. gondii infection was detected using nested polymerase chain reaction (Nested-PCR) assay based on a 529 bp repeat element (RE) gene. Afterward, Toxoplasma was genotyped using PCR-restriction fragment length polymorphism (PCR-RFLP) based on the GRA6 gene. The frequency of T. gondii infection was found to be 14.5% (48 out of 330 samples). Genotyping of nine T. gondii isolates revealed that all belonged to genotype II. Statistically, the prevalence of T. gondii infection was significantly correlated with the education levels of the mothers and the age of the fetus (P < 0.05). The lowest prevalence of Toxoplasma infection belonged to mothers with university education and the highest frequency of infection was observed among the fetuses in the age group of 8-9 weeks. The findings of the present study suggest a significant role for toxoplasmosis in SAFs in Jahrom city.

High occurrence of Acanthamoeba spp. in the water samples of public swimming pools from Kerman Province, Iran.

Acanthamoeba spp. is a free-living amoeba that can cause major infections in humans, including keratitis and granulomatous encephalitis. Thus, water resources play an important role in transmitting Acanthamoeba spp. infection to humans. The purpose of this study was to investigate the presence of Acanthamoeba spp. in public swimming pools from three cities of Kerman Province, southeastern Iran. Eighty water samples of 20 public indoor swimming pools were taken from Kerman, Jiroft, and Kahnauj cities. Water temperature (°C), pH, and free chlorine concentration (ppm) were measured. Filtration and cultivation were applied on non-nutrient agar medium. The polymerase chain reaction was applied by using the genus-specific primers (JDP1 and JDP2) on positive samples; these primers can amplify the 423-551 bp fragment. Eighteen of the 20 swimming pools (including 32/80; 40% samples) were contaminated with Acanthamoeba spp. All swimming pools of Jiroft and Kahnauj and 88.2% of swimming pools in Kerman were contaminated. As such, all 32 Acanthamoeba isolates were amplified using the JDP primer pairs. Two genotypes, T3 and T4, were also identified. The present research is the first to report Acanthamoeba spp. in public swimming pools from Kerman Province. Due to high occurrence of this protozoan, it is recommended to use warning signs around swimming pools to create awareness of this infection.

PCR-based detection of Toxoplasma gondii from cattle in southern Iran.

OBJECTIVE: Toxoplasma gondii is a protozoan parasite that is widely prevalent in most warm-blooded vertebrates. Humans mainly become infected by eating raw or undercooked meat. This study was designed to investigate the infection of cattle with T. gondii in Jahrom, southern Iran. METHODS: Tissue samples consisting of heart, diaphragm, and tongue were collected from 125 slaughtered cattle. DNA samples were extracted from the homogenized tissues. T. gondii was detected and genotyped using nested-polymerase chain reaction (Nested-PCR) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) based on GRA6 and SAG2 (3', 5' terminal regions) genes, respectively. RESULTS: The prevalence of T. gondii DNA was 56% in cattle. The most infected tissue was the diaphragm (54.4%) followed by the heart (48.8%) and tongue (43.2%). Type II was the most prevalent genotype (70%) among T. gondii isolates. CONCLUSION: In this study, the high prevalence of T. gondii infection in cattle meat indicates the important role of cattle in the transmission of infection to humans. Therefore, incorporating the correct method of consuming meat in health education programs is crucial to prevent human infection.

Seroprevalence of Toxoplasma gondii among HIV Patients in Jahrom, Southern Iran.

Toxoplasma gondii is an important opportunistic agent especially in immunocompromised hosts and can cause significant morbidity and mortality. Hence, detection and monitoring of anti-Toxoplasma antibodies are of a great interest in HIV-infected patients. A study on the prevalence of toxoplasmosis and associated risk factors was carried out among HIV-infected patients in Jahrom, southern Iran. The prevalence of anti-Toxoplasma IgG antibodies was 21.1% in HIV-infected patients by ELISA. PCR was performed on all of the samples, and 1 of the blood samples was positively detected. Among the HIV patients, anti-Toxoplasma IgG antibodies were significantly higher in age group of 30-39 years old (P=0.05). The seroprevalence of toxoplasmosis in patients with CD4+<100 cells/µl was 33.3% that was significantly higher than the other groups (P=0.042) with or without IgG antibodies. The CD4+ count mean of seropositive patients was lower than that of seronegative patients. The seroprevalence of toxoplasmosis in patients with highly active antiretroviral therapy was significantly less than patients without therapy (P=0.02). In conclusion, this study showed low seroprevalence of latent toxoplasmosis among HIV-infected patients in the region and confirmed the need for intensifying prevention efforts among this high-risk population and also the risk of toxoplasmosis reactivation which could be important among this population.