Latent toxoplasmosis in COVID-19 patients and link with higher mortality in COVID-19 male patients.

BACKGROUND: Immunocompromised patients may be at risk for reactivating the toxoplasmosis infection; therefore, early diagnosis would be highly desirable in these individuals. This study evaluated the possible association between coronavirus disease 2019 (COVID-19) and latent Toxoplasma gondii infection in Guilan province, Iran. MATERIALS AND METHODS: The study was performed among 210 COVID-19 patients referred to Guilan University of Medical Sciences hospitals in 2022. Peripheral blood samples were taken for serum separation, collected into tubes, and kept at - 20 °C until use. Blood samples were obtained from COVID-19 patients. IgG antibody to Toxoplasma gondii was detected by a commercial ELISA kit. Accordingly, IgG absorbance levels <9 were considered harmful, 9-11 was considered borderline, and >11 was positive. RESULTS: Toxoplasma IgG antibodies were found in 73.9 % of patients with COVID-19 in male patients. The seroprevalence of Toxoplasma in dead and lived COVID-19 male patients was 83.3 % and 66.7 %, respectively, and this difference was significant. A present study found a significant correlation between the rising titer of Toxoplasma IgG and the severity of COVID-19. There was no significant difference between the hospitalization duration factor and the seropositivity rate. CONCLUSION: Regarding the significant association between the rising titer of Toxoplasma IgG and the severity of COVID-19. The findings demonstrated an association between the severity and mortality rate of COVID-19 with higher titer Anti-Toxoplasma IgG antibodies. Toxoplasmosis is currently considered a risk factor for COVID-19.

Molecular characterization of Trichomonas infections in women of Ilam City, southwestern Iran.

Trichomoniasis is a sexually transmitted infection (STI) caused by the flagellated protozoan Trichomonas vaginalis. Little information is available on the epidemiology and genetic diversity of T. vaginalis in Ilam City, southwestern Iran. A descriptive cross-sectional investigation was carried out between July 2017 and December 2018 on the suspected women patients referred to eight gynecology clinics of Ilam City for probable Trichomonas infection. They were undergone a set of clinical, parasitological, and molecular examinations. During clinical consultation, posterior vaginal fornix secretions and urine samples were gathered from the participants. For the reasons such as physical conditions and cultural and religious constraints, most of participating women, especially young girls due to their virginity, preferred to give urine samples instead of vaginal discharge. The presence of Trichomonas was diagnosed by microscopic examination and molecular detection using conventional PCR targeting ITS1-rDNA. A total of 1765 suspected individuals were examined clinically via vaginal secretions (495 specimens) and urine samples (1270 specimens). Of them, 21 (1.18%) cases, including 13 vaginal secretions and 8 urine samples, were positive for Trichomonas infection by microscopy. Slightly more than half of the patients (11/21, 52.4%) complained of vulvar itching, burning, and frequent urination. Cervical lesions, patchy erythema, and vaginal discharge were recorded in 28.6%, 23.8%, and 19% of the patients respectively. All patients with positive microscopic identification were confirmed by amplification of 450-bp fragment of ITS1-rDNA. Phylogenetic analysis revealed a high rate of genetic homogeneity in which all our isolates together with homologous sequences from China, Philippines, Austria, and USA were clustered within the same clade. A statistically significant relationship was recorded between the patients positive for trichomoniasis and the presence of chronic disease (e.g., diabetes, immune system deficiency).

Detection of potentially pathogenic free-living amoebae from the Caspian Sea and hospital ward dust of teaching hospitals in Guilan, Iran.

Free-living amoebae (FLA) thrive in diverse environmental conditions. The present study aimed to define the FLA distribution from the Caspian Sea as well as from hospital ward dust from Guilan, Iran. Seawater (20) and hospital ward dust samples (100) were collected from May to June 2018. Seawater samples were vacuum filtered through a 0.45 µm pore-size membrane. Dust was collected using sterile gauze, washed with sterile distilled water, with washings collected thereafter. Washings were similarly filtered as seawater samples. FLA from the filtered material was cultivated in non-nutrient agar. Molecular analysis was performed by PCR and sequencing using specific primers for Acanthamoeba, Naegleria, and Vermamoeba/Hartmanella. Culture and PCR returned 50 and 65% positivity, respectively, for seawater samples where sequencing revealed Acanthamoeba T2, T5 and T6 genotypes and A. palestinensis and A. lenticulata, as well as N. dobsoni and N. clarki. In addition, 30% amoebic growth and 16% PCR detection were observed from hospital ward dust samples where sequencing revealed Acanthamoeba T2, T4 and T11 genotypes and A. castellanii, A. palestinensis and A. stevensoni as well as N. clarki. For both seawater and dust samples, Acanthamoeba was the dominant isolate. The detection of potentially pathogenic FLA from seawater may pose a threat to the public, while the presence of the same in dust spells threats to both hospital staff and patients, in particular, immunocompromised individuals. Public education, awareness, improved sanitation and hygiene, and the crafting of diagnostic strategies for the early detection of FLA in humans are necessary for the mitigation and management of potential human infection cases.

In vitro bioaccessibility, phase partitioning, and health risk of potentially toxic elements in dust of an iron mining and industrial complex.

Dust emitted from mining, ore processing, and tailing dumps have direct effects on miners who work close to these operations. The Gol-E-Gohar (GEG) mining and industrial company is one of the most important iron concentrate producers in the Middle East. The objective of the present study was to estimate the distribution, fractionation, and oral bioaccessibility of potentially toxic elements (PTEs) in dust generated by the GEG mining and industrial company. Total PTE content including Al, As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Mo, Ni, Pb, V, and Zn was quantified for suspended particulate matter (PM) in PM2.5, PM10, and total suspended particulate matter (TSP). As, Cd, Co, Cu, Fe, Ni, and Pb were quantified in fallout dust samples for oral bioaccessibility using in vitro Unified BARGE (UBM) Method and modified BCR fractionation analysis. Enrichment factors (EF) were calculated for the studied elements in PM; Cu, Fe, and As were found to be extremely enriched. Oral bioaccessibility of selected PTEs in fallout dust samples ranged from 0.35% to 41.55% and 0.06-37.58% in the gastric and intestinal phases, respectively. Regression modeling revealed that the bioaccessibilities of the PTEs could mostly be explained by total concentrations in dust particles. Average daily intake (ADI) calculations revealed that the intake of PTEs did not exceed the tolerable daily intake (TDI) values and as such was not considered a significant risk to workers. Additionally, the hazard quotients (HQ) and carcinogenic risk (CR) values were lower than the acceptable level. This study can provide further risk assessment and management of PTE pollution in occupational environments.

Challenging TaqMan probe-based real-time PCR and loop-mediated isothermal amplification (LAMP): the two sensitive molecular techniques for the detection of toxoplasmosis, a potentially dangerous opportunistic infection in immunocompromised patients.

Due to defects and drawbacks of most conventional diagnostic methods including serology for the diagnosis of toxoplasmosis as a dangerous opportunistic infection in immunocompromised individuals, the accurate, rapid, and sensitive detection of infection in such patients is essential. In this study, the TaqMan probe-based real-time PCR and, a relatively new nucleic acid amplification method, the loop-mediated isothermal amplification (LAMP) technique was compared based on the repetitive elements (RE) sequence to detect Toxoplasma gondii (T. gondii) DNA in blood samples of immunocompromised individuals. During this study, 119 blood samples from immunocompromised cancer patients with renal failure, undergoing dialysis were studied. After DNA extraction from blood samples using the salt extraction method, the molecular techniques of TaqMan probe-based real-time PCR and LAMP were used to investigate the contamination of the samples with T. gondii, based on the 529 bp (RE) sequence of T. gondii. The analytical sensitivity of LAMP and real-time PCR was evaluated by duplicating the five-step serial dilutions of T. gondii tachyzoites from 0.25 to 5×105 spiked tachyzoites per milliliter of the Toxoplasma seronegative blood sample. The extracted DNA from other parasites and human chromosomal DNA were used to determine the specificity of the molecular methods. The obtained results were analyzed using Kappa statistical test and SPSS22 software. Out of 119 studied samples, 7 (5.8%) and 5 (4.2%) samples were positive for Toxoplasma by TaqMan probe-based real-time PCR and LAMP, respectively. The limits of detection of TaqMan probe-based real-time PCR and RE-LAMP in negative serum samples were one and five tachyzoites (CT 38), respectively. Both real-time PCR and LAMP methods were 100% specific for Toxoplasma detection. Positive results were obtained only with T. gondii DNA, while other DNA samples were negative. The TaqMan probe-based real-time PCR based on the RE sequence showed higher sensitivity to T. gondii DNA detection in blood samples of cancer patients and serial dilutions of parasitic tachyzoites. The results show that TaqMan probe-based real-time PRC is a sensitive and specific method for the detection of toxoplasmosis in immunocompromised individuals, as well as the LAMP assay, which can be used as a suitable alternative diagnostic method for the detection of toxoplasmosis in such patients, without need the for any expensive equipment.

Isolation and molecular identification of free-living amoebae (Naegleria spp., Acanthamoeba spp. and Vermamoeba spp.) from mineral springs in Guilan Province, northern Iran.

Free-living amoebae (FLA) include many genera which cause serious diseases such as sight-threatening keratitis, cutaneous ulcers and fatal encephalitis. This study was conducted due to the lack of research regarding genotypes Acanthamoeba, Naegleria and Vermamoeba in mineral springs of Guilan Province in northern Iran. Twenty-five water samples were collected from mineral springs in Guilan Province. After filtration through nitrocellulose membrane, samples were cultured on non-nutrient agar plates. The morphological key of Page was used to identify free-living amoebae (FLA) using an inverted microscope. Positive cultures were analyzed by polymerase chain reaction (PCR) and genotypes based on the NCBI database. Eleven (44%) samples were positive for Acanthamoeba, Naegleria and Vermamoeba. By sequencing the positive isolates, the strains were shown to belong to Acanthamoeba castellanii (three case isolates belonged to T4 genotype), three cases of Vermamoeba vermiformis, and two cases of N. australiensis, two cases of N. pagei and one case of N. gruberi. According to our research the occurrence of Acanthamoeba, Naegleria spp. and Vermamoeba spp. in mineral springs could be hazardous for high risk persons. Regular monitoring and posting warning signs of these waters by health planners could prevent free-living amoebae mediated diseases.

Intestinal protozoan and helminthic infections among hemodialysis and cancer patients.

Intestinal parasitic infections (IPIs) can be a severe threat to immunocompromised patients. This is particularly true for those undergoing chemotherapy and hemodialysis. The present research is aimed at identifying intestinal parasites that might be present in immunocompromised patients. In this cross-sectional study 1040 stool samples were collected from March to September 2017. Six hundred and forty-one stool samples from immunocompromised patients (279 samples from hemodialysis patients and 362 samples from chemotherapy patients) and 399 samples from the control group were collected in Guilan province, Iran. The samples were tested by direct, formalin-ether methods for protozoa and ova of intestinal parasites and Ziehl-Neelsen staining methods for coccidian parasites such as Cryptosporidium species. The overall parasitic infection rate was highest (15%) in hemodialysis patients and 11.3% in chemotherapy patients, whereas the lowest rate was observed (7.3%) in the control group. The infectivity rates were statistically significant (P = 0.008) when compared with the control group. The parasites found were Blastocystis hominis (8.9% of the cases), Entamoeba coli (1.6%), Iodamoeba butschlii (0.8%), Endolimax nana (0.6%), Chilomastix mesnili (0.5%), Strongyloides stercoralis (0.5%), and Taenia species (0.15%), whereas Giardia lamblia was detected only in the control group. There was not a correlation between prevalence of parasites with age or education levels of the infected individuals. Results of the present study suggest that periodic stool examinations in special parasitological laboratories should be included as part of routine and general medical care.

Modeling and forecasting the spread and death rate of coronavirus (COVID-19) in the world using time series models.

Coronaviruses are a huge family of viruses that affect neurological, gastrointestinal, hepatic and respiratory systems. The numbers of confirmed cases are increased daily in different countries, especially in Unites State America, Spain, Italy, Germany, China, Iran, South Korea and others. The spread of the COVID-19 has many dangers and needs strict special plans and policies. Therefore, to consider the plans and policies, the predicting and forecasting the future confirmed cases are critical. The time series models are useful to model data that are gathered and indexed by time. Symmetry of error's distribution is an essential condition in classical time series. But there exist cases in the real practical world that assumption of symmetric distribution of the error terms is not satisfactory. In our methodology, the distribution of the error has been considered to be two-piece scale mixtures of normal (TP-SMN). The proposed time series models works well than ordinary Gaussian and symmetry models (especially for COVID-19 datasets), and were fitted initially to the historical COVID-19 datasets. Then, the time series that has the best fit to each of the dataset is selected. Finally, the selected models are applied to predict the number of confirmed cases and the death rate of COVID-19 in the world.

Fuzzy clustering method to compare the spread rate of Covid-19 in the high risks countries.

The numbers of confirmed cases of new coronavirus (Covid-19) are increased daily in different countries. To determine the policies and plans, the study of the relations between the distributions of the spread of this virus in other countries is critical. In this work, the distributions of the spread of Covid-19 in Unites States America, Spain, Italy, Germany, United Kingdom, France, and Iran were compared and clustered using fuzzy clustering technique. At first, the time series of Covid-19 datasets in selected countries were considered. Then, the relation between spread of Covid-19 and population's size was studied using Pearson correlation. The effect of the population's size was eliminated by rescaling the Covid-19 datasets based on the population's size of USA. Finally, the rescaled Covid-19 datasets of the countries were clustered using fuzzy clustering. The results of Pearson correlation indicated that there were positive and significant between total confirmed cases, total dead cases and population's size of the countries. The clustering results indicated that the distribution of spreading in Spain and Italy was approximately similar and differed from other countries.

A new approach in adsorption modeling using random forest regression, Bayesian multiple linear regression, and multiple linear regression: 2,4-D adsorption by a green adsorbent.

The mathematical model's usage in water quality prediction has received more interest recently. In this research, the potential of random forest regression (RFR), Bayesian multiple linear regression (BMLR), and multiple linear regression (MLR) were examined to predict the amount of 2,4-dichlorophenoxy acetic acid (2,4-D) elimination by rice husk biochar from synthetic wastewater, using five input operating parameters including initial 2,4-D concentration, adsorbent dosage, pH, reaction time, and temperature. The equilibrium and kinetic adsorption data were fitted best to the Freundlich and pseudo-first-order models. The thermodynamic parameters also indicated the exothermic and spontaneous nature of adsorption. The modeling results indicated an R2 of 0.994, 0.992, and 0.945 and RMSE of 1.92, 6.17, and 2.10 for the relationship between the model-estimated and measured values of 2,4-D removal for RFR, BMLR, and MLR, respectively. Overall performances indicated more proficiency of RFR than the BMLR and MLR models due to its capability in capturing the non-linear relationships between input data and their associated removal capacities. The sensitivity analysis demonstrated that the 2,4-D adsorption process is more sensitive to initial 2,4-D concentration and adsorbent dosage. Thus, it is possible to permanently monitor waters more cost-effectively with the suggested model application.

Genetic diversity of an avian nasal schistosome causing cercarial dermatitis in the Black Sea-Mediterranean migratory route.

This study is part of an effort to document the diversity of avian schistosomes in ducks and snails in Northern Iran, a major flyway (Black Sea/Mediterranean) for migratory birds and where cercarial dermatitis (CD) is prevalent in rice growing areas. CD is an allergic skin reaction from schistosome trematodes that emerge from aquatic snails. Most CD cases are reported from recreational swimmers or aquaculture farmers. Much of the work on the epidemiology of CD has focused in recreational waters in the Americas and Europe, with fewer studies in aquaculture, particularly in Iran. The artificial environment at aquaculture sites support dense populations of snails that are hosts to schistosomes, as well as domestic ducks. Thus, are domestic ducks reservoir hosts of species of Trichobilharzia, one of the main etiological agents of CD in Northern Iran? This study focused on a survey of domestic ducks for the presence of the nasal schistosome, T. regenti, that has been reported widely in Europe. Trichobilharzia regenti were found in domestic ducks in the Guilan Province of Iran based on morphological and molecular analyses. The presence of this species in Northern Iran indicates that the domestic duck can serve as a reservoir host for this species and that one of the local snail species is likely the intermediate host. The continued study and surveillance of this species is important because it is a neuropathic schistosome that can use a diversity of bird definitive hosts and Radix snails that are widespread across Eurasia.

Modeling caffeine adsorption by multi-walled carbon nanotubes using multiple polynomial regression with interaction effects.

Permanent monitoring of environmental issues demands efficient, accurate, and user-friendly pollutant prediction methods, particularly from operating variables. In this research, the efficiency of multiple polynomial regression in predicting the adsorption capacity of caffeine (q) from an experimental batch mode by multi-walled carbon nanotubes (MWCNTs) was investigated. The MWCNTs were specified by scanning electron microscope, Fourier transform infrared spectroscopy and point of zero charge. The results confirmed that the MWCNTs have a high capacity to uptake caffeine from the wastewater. Five parameters including pH, reaction time (t), adsorbent mass (M), temperature (T) and initial pollutant concentration (C) were selected as input model data and q as the output. The results indicated that multiple polynomial regression which employed C, M and t was the best model (normalized root mean square error = 0.0916 and R2 = 0.996). The sensitivity analysis indicated that the predicted q is more sensitive to the C, followed by M, and t. The results indicated that the pH and temperature have no significant effect on the adsorption capacity of caffeine in batch mode experiments. The results displayed that estimations are slightly overestimated. This study demonstrated that the multiple polynomial regression could be an accurate and faster alternative to available difficult and time-consuming models for q prediction.

Contribution of Blastocystishominis subtypes and associated inflammatory factors in development of irritable bowel syndrome.

Blastocystis hominis with worldwide distribution is a human intestinal protozoa found in all countries. There have been differences in the severity of the pathogenesis of various Blastocystis spp. and a concomitant variation in the plasma concentration of the cytokines in patients with irritable bowel syndrome. In the present study, we aimed to demonstrate the contribution of B. hominis subtypes in the development of irritable bowel syndrome. Stool samples were collected from patients with gastrointestinal disorders. All samples were evaluated through native-lugol method. Total DNA was extracted. A PCR protocol was developed to amplify a specific region of the SSU ribosomal DNA (rDNA) gene. Serum levels of IL-6 and TNF-α were determined by immunoassay methods. The ClustalW algorithm was applied to align and blast the nucleotide sequences of the amplified region of the SSU rDNA gene. To evaluate the phylogenetic and molecular evolutionary of the nucleotide sequences, we used the MEGA software. In this study, we found 26 haplotypes of B. hominis in the studied samples which were collectively belong to five subtypes (ST1, ST2 in patients without irritable bowel syndrome vs. ST3 and two unknown subtypes in patients with irritable bowel syndrome). Result of ELISA showed a high level of IL-6 and TNF-α in the serum of patients with irritable bowel syndrome. The genetic heterogeneity of B. hominis and the existence of different subtypes of the protozoan in patients with IBS may shed light to the fact that some subtypes of parasites may involve in the pathogenesis of IBS.

Occurrence and molecular characterization of free-living amoeba species (Acanthamoeba, Hartmannella, and Saccamoeba limax) in various surface water resources of Iran.

This study was conducted to determine the presence and molecular identity of Acanthamoeba species in the surface water resources of four provinces in Iran, namely Guilan, Mazandaran (North of Iran), Alborz, and Tehran (capital city), using culture- and molecular-based methods. During March to November 2014, 49 surface water samples were collected from environmental water sources-the distinct surface waters of Guilan, Mazandaran, Alborz, and Tehran provinces, in Iran. For the isolation of Acanthamoeba species, approximately 500 ml of the water samples were filtered through a cellulose nitrate membrane with a pore size of 0.45 µ. The filter was transferred onto non-nutrient agar plates seeded with Gram-negative bacteria (Escherichia coli) as a food source. The presence of Acanthamoeba was confirmed by the genus-specific primer pair JDP1 and 2, and/or NA primers were used to identify Acanthamoeba and certain other free-living amoebae. In total, 38 out of 49 samples were positive by culture and/or PCR for Acanthamoeba and other free-living amoebae from all three provinces. By sequencing the positive isolates, the strains were shown to belong to Acanthamoeba (16 isolates belonged to T4 and 2 isolates belonged to T5), Hartmannella vermiformis (3/24), and Saccamoeba limax (2/24). The T4 and T5 genotypes were detected in Guilan and Mazandaran provinces. Two isolates from Guilan and Tehran provinces belonged to S. limax, and H. vermiformis was detected in Guilan province. The results of this study highlight the need to pay more attention to free-living amoebae, as human activity was observed in all of the localities from which these samples were taken. These surface waters can be potential sources for the distribution and transmission of pathogenic Acanthamoeba in the study areas, and free-living amoebas (FLA) (particularly the Acanthamoeba species) can serve as hosts for and vehicles of various microorganisms.

Genotyping of Giardia lamblia and Entamoeba spp from river waters in Iran.

In this study, DNA from 55 surface and river water samples, which were collected from some water sources of Tehran and the Guilan Province, Iran, were extracted and examined for Entamoeba spp. and Giardia lamblia by PCR and genotyping. Twenty-seven samples, which were concentrated using the immunomagnetic separation technology (IMS) method, were examined for Giardia alone. Twenty-eight samples, which were concentrated using the sucrose flotation (SF) method, were examined for both Giardia and Entamoeba species. The results showed that 27/55 (17/27 and 10/28) (49 %), 4 /28 (14.28 %) and 3/28 (10.7 %) of the samples were positive for Giardia lamblia, Entamoeba spp and mixed infections (Entamoeba spp. and Giardia spp.), respectively. Sixteen out of 55 samples were negative. Entamoeba genus-specific PCR primers in single-round PCR were used to differentiate between the Entamoeba spp. (E. histolytica, E. dispar and E. moshkovskii). With respect to the 7 samples that were positive for Entamoeba, (14.28 %) 4 out of 28 were positive for E. moshkovskii, (7.14 %), 2 out of 28 were positive for E. histolytica and (3.57 %) 1 out of 28 was positive for E. dispar. Genus-specific PCR primers in a semi-nested PCR assay was performed to genotype Giardia species. Of the 27 samples that were positive for Giardia, 10 samples were sequences. All 10 successfully sequenced samples contained assemblage B of Giardia lamblia.This is first study to investigate the G. lamblia genotypes in the water supply of the Tehran and Guilan provinces, and it is the first study to investigate Entamoeba species in the water supplies of Iran. The investigated river water supplies, which are used for agriculture, camping and animal farming, were heavily contaminated by the human pathogenic Entamoeba and Giardia parasites. There is a potential risk of waterborne outbreaks in humans and animals.

Corrigendum to "Molecular detection of Toxoplasma gondii in chicken hearts from markets and retail stores in Northern Iran" [Food and Waterborne Parasitology 27 (2022) e00166].

[This corrects the article DOI: 10.1016/j.fawpar.2022.e00166.].

Isolation of Acanthamoeba species in surface waters of Gilan province-north of Iran.

We analyzed water samples to determine the prevalence of free-living Acanthamoeba in water sources from Gilan, greater area, Iran. A total of 27 surface water samples were collected from environmental sources, including natural (rivers, lakes, springs, and lagoon) and freshwater source. The samples were filtrated and transferred to non-nutrient agar plates seeded with Escherichia coli and incubated for 2 to 7 days at 30°C or 42°C. The plates were examined by microscopy to morphologically identify Acanthamoeba species. Following DNA extraction, PCR was used to confirm the microscopically identification. A total of 19 out of 27 samples (70.3%) were positive for Acanthamoeba species based on the morphological criteria, and 14 (73.7%) were confirmed by PCR method. The high frequency of Acanthamoeba spp. in different environmental water sources of Gilan is an alert for the public health related to water sources in Iran.

Modeling the stochastic mechanism of sensor using a hybrid method based on seasonal autoregressive integrated moving average time series and generalized estimating equations.

Sensor, which is one of the main components of control system, plays its vital role in measuring state and output system variables and highlights the importance of having desired statistical information about sensor output signals because it can be monitored, stored, or used as the primary input signal in other devices. However, these signals display noises (i.e. system noise and measurement noise) and even if the effects of system noises are faded away or removed from measured data, there is still some stochastic noise remained in the measurements. Even though SARIMA has been effective in modeling the stochastic noise in the sensor, the present study has found out the necessity of designing a novel approach including a combination of seasonal autoregressive integrated moving average (SARIMA) and polynomial generalized estimating equations (PGEE), to evaluate the stochastic behavior of sensors. Finally, the study tried to employ the proposed approach in real load-cell sensor data to examine its effectiveness.

Molecular detection of Toxoplasma gondii in chicken hearts from markets and retail stores in Northern Iran.

Detection of Toxoplasma gondii in chicken products indicates risk of transmission to consumers. The objective of the current study was to investigate the molecular prevalence of T. gondii in free-ranging and industrial chickens in Guilan province, Northern Iran. A total of 150 chicken heart samples including 75 free-range and 75 industrial chickens were collected from farmers' markets and chicken retailers in Guilan, Northern Iran, between October 2017 and August 2018. Genomic DNA were extracted from samples and examined for evidence of T. gondii using polymerase chain reaction (PCR) targeting the B1 gene. The B1-positive samples were further analyzed by nested-PCR for SAG1 gene. Of the 150 samples, T. gondii DNA fragments were detected in 59 (39.3%), including 30 (40%) free-range and 29 (38.7%) industrial chicken. No significant differences of T. gondii DNA detection was observed between the free-range and industrial chicken samples (p = 0.73). Four selected positive samples were used for amplifying and sequencing of the SAG1 gene. The results revealed that all four sequences of SAG1 had 100% similarity with T. gondii sequences previously isolated from an AIDS/HIV patient in Mazandaran province, Northern Iran. Furthermore, the phylogenetic analysis demonstrated that all four sequences were closely related to Type I of T. gondii. However, our Type I identification is preliminary and needs to be confirmed by further multilocus sequence typing (MLST) analysis. The findings of the present study provide new data about the presence of T. gondii DNA in chicken hearts in the study area. These results confirm that chicken can be used as sentinels for environment contamination; however, further studies are needed to determine the viability of T. gondii in chicken hearts from Iran for risk assessment.

Seroepidemiological Study of Visceral Leishmaniasis (Kala-Azar) in Children under 12 Years Old in North of Iran: An Observational Study in 2019-2020.

Background: Visceral leishmaniasis (VL) caused by the Leishmania donovani complex that is transmitted by the bites of female sandflies. Mediterranean type of VL caused by L. infantum. While, Roudbar County of Guilan Province has been introduced as a suspected cutaneous leishmaniasis focus; there are no published data on the seroprevalence of VL in Guilan Province. We aimed to investigate the sero-prevalence of this disease in Roudbar County. Methods: This descriptive cross-sectional study was carried out in 2019-2020 among children less than 12 years of age to determine the seroprevalence of VL by direct agglutination test (DAT). Blood samples were randomly collected from 918 children under 12 years of age refers to the public health center in the clusters. Results: Out of 918 children, 14 (1.52%) showed anti-Leishmania antibodies, with 4 (0.43%), 2 (0.21%), 8 (0.87%) anti-L. infantum antibodies at titers 1:800, 1:1600 and ≥1: 3200 respectively. All children with anti-Leishmania antibody titers of ≥1:800 were evaluated by a physician. Clinical manifestation of VL including fever, anemia and hepatosplenomegaly observed in a 6-year-old boy from Defraz village with anti-Leishmania antibody of titers ≥102400.This patient was admitted to the pediatric hospital in Rasht, capital of Guilan province, Iran and was successfully treated. Conclusion: VL is being circulated with low prevalence in children up to 12 years old in Roudbar, northern part of Iran. Accordingly, it is critical to improve the awareness of physicians and public health supervisors about the importance of this fatal disease in Guilan province and especially in Roudbar area.