Molecular Genotyping of Giardia duodenalis in Humans in the Yazd County, Central of Iran.

Background: We aimed to investigate the molecular genotyping of Giardia duodenalis in Humans in the Yazd County, Central, Iran. Methods: Total of 35 fecal samples were collected from patients referred to Yazd Central Laboratory, Yazd, Iran from February to July 2022. All the samples were included in this study after microscopic observation of G. duodenalis. DNA samples were extracted using related kit and were analyzed by Nano Drop. The molecular assessment was carried out using semi-nested PCR using the target gene of gdh. All amplified samples were sequenced using Sanger method. BLAST analyzed the sequences for assemblage identification. Results: Out of 35 samples, 24 (68.57%) and 11 (31.43%) were male and female, respectively. All included samples were amplified using the specific gdh primer pair. The molecular analysis showed 17 isolates (48.57%) as assemblage BIV, 8 isolates (22.86%) as assemblage BIII, 6 isolates (17.14%) as assemblage AII and 4 isolates (11.43%) as assemblage AIII (P<0.05). Conclusion: Assemblages A and B are the most prevalent in Central Iran. The molecular identification of G. duodenalis isolates from animals and implementing control programs.

Differential gene expression (DGE) analysis in persons with a history of giardiasis.

Giardiasis, which is caused by Giardia duodenalis, has clinical symptoms such as steatorrhea and can be very dangerous in children. In addition, some documents reported that this parasite is present inside the tissue of patients with cancer. In this study, we analyzed the gene expression profiles of some main genes important to apoptosis and anti-apoptosis in humans.Expression profile arrays of Genomic Spatial Event (GSE) 113666, GSE113667, and GSE113679 obtained from Gene Expression Omnibus were used for meta-analysis using R commands. Cytoscape and STRING databases used the protein-protein Interaction network. Then, the Kyoto Encyclopedia of Genes and Genomes and Gene Ontology analysis was performed. Similar genes in Homo sapiens were identified using Basic Local Alignment Search Tool analysis. The validation was performed on eight people using real-time Polymerase chain reaction. In addition to the candidate genes, the gene expression of some other genes, including Serine/Threonine Kinase 1 (AKT1), Cyclin Dependent Kinase Inhibitor 2A (CDKN2A), Kirsten Rat Sarcoma (KRAS), and Phosphatidylinositol-4,5-Bisphosphate 3-Kinase Catalytic Subunit Alpha (PIK3CA) were also examined. Analysis of the expression of serum amyloid A1 (SAA1), Regenerating Islet-Derived 3 Gamma (REG3G), and REG3A genes did not show any difference between the two groups of healthy and diseased people. Examining the mean expression of the four genes AKT1, CDKN2A, KRAS, and PIK3CA showed that three genes of AKT1, CDKN2A, and KRAS had increased expression in people with a history of giardiasis compared to healthy people. We showed that the gene expression pattern differs in apoptosis and anti-apoptosis signaling in people with a history of giardiasis. Giardia duodenalis seems to induce post-non-infectious symptoms with stimulation of human gene expression.

Expression profile in Leishmania major exposed to Staphylococcus aureus and group A beta-hemolytic Streptococcus.

BACKGROUND & OBJECTIVES: The interaction of Leishmania spp. with microbiota inside the midgut vector has significant output in pathogenesis. This study aimed to identify the profile of Leishmania majorgene expression of LACK, gp63, and hsp70after exposure to Staphylococcus aureusand group A beta-hemolytic Streptococci (GABHS). METHODS: Leishmania major (MRHO/IR/75/ER) promastigotes were exposed with S. aureus, with GABHS, and with both GABHS and S. aureus at 25°C for 72 h. The gene expression analysis of Lmgp63, Lmhsp70,and LmLACKwas assessed using SYBR Green real-time PCR by ΔΔCt. All experiments were repeated in triplicate. Statistical analysis was done using two-way ANOVA. A P-value less than 0.05 was considered significant. RESULTS: Lmgp63 was expressed in the group exposed to GABHS with 1.75-fold lower than the control group (p=0.000). The LmLACK had expression in both groups exposed with GABHS and GABHS with S. aureus with 2.8 and 1.33-fold more than the control group, respectively (p=0.000). The Lmhsp70 gene expression was reported in the group exposed with GABHS with relative quantification of 5.7-fold more than the control group. INTERPRETATION & CONCLUSION: This study showed that the important genes encoding LACK, gp63, and hsp70 changed their expression after exposure to the S. aureus and GABHS.

Bacterial and Fungal Occurrence in Hydatid Cysts from Livestock in Central Iran.

Echinococcus granulosus sensu lato causes Cystic echinococcosis. This study investigated the bacterial and fungal species in the liver and lung hydatid cysts obtained from sheep, goats, cattle, and camels slaughtered in Yazd abattoir, Central Iran. In this study, 84 hydatid cysts were obtained from 20 sheep, 13 goats, 25 cattle, and 26 camels. The fertility and viability rates were assessed using light microscopy and eosin staining, respectively. The aspirated hydatid cysts were cultured to detect the presence of any bacteria and fungi. Bacterial isolates were identified by biochemical tests. DNA was also extracted from germinal layers, and then genotyping was carried out targeting the cox 1 gene. The statistical analysis was performed by SPSS version 16.0. This study showed that 22.62% (19/84) of hydatid cysts had bacterial occurrence, and none of the samples had fungal species. Among the fertile cysts, 52.6% had bacterial occurrence, of which 40% were viable. Most bacteria detected in hydatid cysts included Staphylococcus saprophyticus, Escherichia coli, and S. epidermidis. Hydatid cysts with bacterial occurrence were identified as G1-G3, G5, and G6/G7. The bacterial species occurrence in hydatid cysts had no significant relationship with fertility and viability (P > 0.05), without any significant relation with viability (P > 0.05), animal species (P > 0.05), involved organ in animals (P > 0.05), and hydatid cyst genotypes (P > 0.05). It should also be mentioned that this is the first study to assess the relationship between hydatid cyst genotyping and the occurrence of fungal and bacterial species.

The role of ATP-binding cassette transporter genes expression in treatment failure cutaneous leishmaniasis.

Leishmaniasis is one of the common diseases transmitted by sand flies in tropical and subtropical regions of the world. Currently, antimonial derivatives are the first line of treatment. Some of the members of the ATP-binding cassette (ABC) family of Leishmania are shown to be associated with no response to treatment. In this study, we evaluated ABCI4, ABCG2, ABCC7, ABCB4, and ABCC3 genes expression in Leishmania isolated from patients with non-healing cutaneous leishmaniasis and treatment response isolates. We selected 17 clinical isolates including 8 treatment failure and 9 treatment response samples from September 2020 to March 2021. The isolates were obtained from patients of Health Center Laboratory of Varzaneh, Isfahan, Iran with cutaneous leishmaniasis. The diagnosis was performed using microscopic observation. The samples were directly collected from the lesions. The expression profiling of genes was assessed using SYBR Green real-time PCR that was analyzed with delta-delta Ct. All treatment failure clinical isolates were L. major. Gene expression analysis in treatment failure isolates showed that the ABC transported genes had a different pattern in each isolate. Treatment failure has been reported for cutaneous leishmaniasis worldwide. Knowledge of the molecular mechanisms of treatment failure could solve this problem. ABC transporter genes are considered controversial over the mechanisms of treatment failure outcomes. In this study, we showed that ABC transporter genes could be considered one of the important mechanisms.

Cystic Echinococcosis in Central Iran: G1 and G6 Genotypes in Patients.

Background: Cystic echinococcosis (CE) is caused by Echinococcus granulosus sensu lato. In Central Iran, no molecular information is available on CE in humans. Therefore, in this study, we identified the genotyping of hydatid cysts obtained from patients with CE in central Iran using mitochondrial cytochrome c oxidase subunit I (cox1) gene. Patients and Methods: Hydatid cysts were obtained from 19 patients referred to Shahid Sadoughi, Mojibian, and Mortaz Hospitals, Yazd, Iran from 2018 to 2020. Informed consent was obtained from all included patients. After DNA extraction, amplification was done using cox1 gene. Phylogenetic analysis was performed using MEGA7. Results: Of the 19 patients, 11 (57.9%) were male and eight (42.1%) were female. The mean age of the patients was 35.645 ± 2.55 years old. Regarding cyst location, of eight isolates from lung, six and two belonged to G1 and G6, respectively; and all liver cysts were G1 genotype. The spleen and neck cysts had G1 and G6 genotypes, respectively (p > 0.05). All cysts with a diameter in the range of 5-10 cm (n = 9) and large cysts (>10 cm; n = 5) were identified as G1 (p = 0.002). The maximum likelihood tree topology demonstrated the maximum similarity of G1 among Iran and worldwide (99%-100% likelihood). Conclusions: Based on our results, it seems that the sheep-dog cycle in the infection of humans by Echinococcus granulosus in this study area has the most important role compared with the other cycles such as the camel-dog one.

A comparison of different O-antigen serogroups of Escherichia coli in semen samples of fertile and infertile men.

OBJECTIVE: Male genital tract infections have been associated with infertility, and Escherichia coli has drawn increasing attention as an important bacterium in this context. This investigation aimed to characterize and compare the distributions of O-antigen serogroups of E. coli in the semen samples of fertile and infertile men. METHODS: In this case-control study, semen samples were collected from 618 fertile and 1,535 infertile men. The E. coli-positive samples were evaluated in terms of concentration, morphology, viability, and motility parameters according to the World Health Organization 2010 guidelines. Finally, different serogroups of E. coli were identified by multiplex polymerase chain reaction targeting the O-antigen variations of the bacterium. RESULTS: The prevalence of E. coli among fertile men was significantly higher than among infertile men (p<0.001). The sperm morphology, viability, and motility in the E. coli-positive fertile group were significantly higher than in the E. coli-positive infertile group (p<0.001). E. coli O6 was the most prevalent serogroup found in both groups. However, there was no significant difference in the frequency of different serogroups of E. coil between the two groups (p=0.55). CONCLUSION: Despite the higher prevalence of E. coli among fertile men, E. coli had more detrimental effects on semen parameters in infertile men. There was no significant difference in E. coli serogroups between the fertile and infertile groups.

A Meta-Analysis of the Prevalence of Toxoplasmosis in Livestock and Poultry Worldwide.

Toxoplasma gondii causes toxoplasmosis with a global prevalence in the world. A large proportion of human illness is most frequently associated with consuming raw and undercooked meat or other animal products containing infective parasitic stages of T. gondii. This systematic review and meta-analysis study evaluated the prevalence of toxoplasmosis in cattle, sheep, camels, goats, and poultry worldwide. The search was performed in databases including PubMed, WoS, Scopus, Science Direct, Google Scholar, and ISC from 2000 to 2019 in Persian and English. The main inclusion criteria were the prevalence of toxoplasmosis among livestock and poultry and the prevalence indices by sample size. During these 20 years, the overall prevalence of toxoplasmosis in livestock and poultry was 28.3% (95% confidence interval (CI) 25-31.9%) using the random-effects meta-analysis model. The highest prevalence of T. gondii in livestock and poultry animals was found in Asia in 2014 with 89.8% (95% CI 78.5-95.5%). The lowest prevalence was found in Asia in 2013 with 1.26% (95% CI 0.4-3.8%). A quarter of livestock and poultry were infected with T. gondii. Since livestock products are globally important sources of people's diet, our findings are useful for policymakers to control T. gondii infection in livestock.

Occurrence and genetic characterization of Echinococcus granulosus sensu lato from domestic animals in Central Iran.

BACKGROUND: The species complex of Echinococcus granulosus sensu lato (s.l.) causes cystic echinococcosis distributed worldwide. There is no genotype information from hydatid cysts in the intermediate hosts in Central Iran. Therefore, in this study, we analyzed the hydatid cysts in livestock slaughtered in an abattoir in this region. Six hundred fifty-seven hydatid cysts were isolated from 97 animals, including sheep, cattle, camels, and goats slaughtered in Yazd abattoir from September 2018 to January 2020. The demographic data was collected as well as cyst location, fertility, and viability. Out of 657 samples, 164 samples were genotyped. Then, phylogenetic analysis was performed using MEGAX. Statistical analyses were done using SPSS version 16.0 by chi-square with a significant difference of less than 0.05. RESULTS: Out of 164 samples, the G1-G3 complex genotype had the most frequency in samples, with 135 cases recognized. The G6/G7 was observed in 19 isolates and G5 was reported in nine samples. One sample was detected as Taenia hydatigena. CONCLUSIONS: This study showed that G1-G3 and G6/G7 genotypes were presented in all animals, but G5 was reported only in cattle, goats, and camels. It is the first molecular identification of cystic echinococcosis in Central Iran. Hence, reporting G5 in livestock in this area should be considered due to transmission to humans.

Mitogen-Activated Protein Kinase and Aquaglyceroporin Gene Expression in Treatment Failure Leishmania major.

PURPOSE: Leishmaniasis comprises various clinical forms mainly including cutaneous, muco-cutaneous, and visceral leishmaniasis; caused by Leishmania species. Antimoniate is the first-line treatment but some cases showed no response to treatment in the worldwide. In this study, mitogen-activated protein kinase (MAPK) and aquaglyceroporin 1 (AQP1) gene expressions were assessed in treatment failure clinical isolates of Leishmania major. Also, molecular and phylogenic analyses of the mentioned isolates were performed. METHODS: Samples were obtained from the patients with suspicious CL referred to the laboratory of Diagnosis Center, Gorgan Province, Iran, from October 2016 to December 2019. Detection and identification of the parasite was performed. The genes expressions of MAPK1 and AQP1 were done using SYBR Green real-time PCR. The AQP1 gene from the isolates with treatment failure was sequenced and analyzed using BLAST and multiple alignments. The phylogenic analysis was done using MEGA7. The statistical analysis was done using SPSS 16.0 by non-parametric Mann-Whitney U test. RESULTS: All clinical isolates were detected L. major. The mean AQP1 and MAPK1 gene expressions in treatment failure isolates were 58.71 and 6.139 fold less than the ones in treatment response isolates, respectively. Based on the AQP1 gene sequence, a nucleotide change of aspartic acid with asparagine at the site 234 was observed. Phylogenic tree analysis showed three groups with the minimum dissimilarity of 0.008 between TF isolates with the standard L. major strains. CONCLUSION: We showed that MAPK1 and AQP1 may have critical roles in response to antimoniate in clinical isolates L. major in this study.

Relationship between Serum Tumor-Related Markers and Genetically Modified Rice Expressing Cry1Ab Protein in Sprague-Dawley Rats.

Genetically Modified (GM) foods are among the most important achievements of biotechnology. Given the safety importance of transgenic rice, this study was conducted to investigate the effect of GM rice consumption on serum concentrations of tumor markers in rats. In this experimental intervention, we used the blood serum samples from the Biobank taken from 60 males and 60 female Sprague-Dawley (SD) rats fed on three different diets, including rat's standard food, non-GM rice, and GM rice after 90 day. Tumor markers including Carcinogenic embryonic antigen (CEA), Alpha-Fito protein (AFP), Cancer Antigen 19-9 (CA19-9), Cancer Antigen 125 (CA125), and Cancer Antigen15-3 (CA15-3) were assessed by enzyme-linked immune sorbent assay (ELISA) method. Statistical analysis was conducted via SPSS software. The results show that the concentrations of tumor markers were within the normal range in the SD rats treated with diet, and since the concentration of tumor markers was lower than the acceptable index determined, according to the kit standard in both groups, no obvious carcinogenic effect was found. However, these findings are not enough to make a final decision regarding the safety assessment of GM rice consumption.

A comparison of different O-antigen serogroups of Escherichia coli in semen samples of fertile and infertile men.

OBJECTIVE: Male genital tract infections have been associated with infertility, and Escherichia coli has drawn increasing attention as an important bacterium in this context. This investigation aimed to characterize and compare the distributions of O-antigen serogroups of E. coli in the semen samples of fertile and infertile men. METHODS: In this case-control study, semen samples were collected from 618 fertile and 1,535 infertile men. The E. coli-positive samples were evaluated in terms of concentration, morphology, viability, and motility parameters according to the World Health Organization 2010 guidelines. Finally, different serogroups of E. coli were identified by multiplex polymerase chain reaction targeting the O-antigen variations of the bacterium. RESULTS: The prevalence of E. coli among fertile men was significantly higher than among infertile men (p<0.001). The sperm morphology, viability, and motility in the E. coli-positive fertile group were significantly higher than in the E. coli-positive infertile group (p<0.001). E. coli O6 was the most prevalent serogroup found in both groups. However, there was no significant difference in the frequency of different serogroups of E. coil between the two groups (p=0.55). CONCLUSION: Despite the higher prevalence of E. coli among fertile men, E. coli had more detrimental effects on semen parameters in infertile men. There was no significant difference in E. coli serogroups between the fertile and infertile groups.

Upstream region of OprD mutations in imipenem-resistant and imipenem-sensitive Pseudomonas isolates.

The current study was aimed at investigating the prevalence of the mutations upstream of the oprD coding region and its promoters among imipenem-resistant and sensitive Pseudomonas aeruginosa isolated from educational hospitals in Yazd City, Iran. All isolates were identified by the conventional biochemical tests. Then, the antibiotic resistance of these isolates was determined using the disk diffusion method according to the CLSI guidelines. Also, the E.test was performed to determine the minimum inhibitory concentrations (MIC) of imipenem. The mutations of this gene were recognized by the amplification of this region and subsequently sequenced. Sequencing of the genomic region upstream of oprD these regions were done in the 29 clinical strains. Statistical analysis was done by the statistical software SPSS-18. Seventy (77.7%) of isolates had MIC ≥ 16 and were resistant to imipenem. Mutations of the upstream of the oprD gene and its promoters were seen in 25 (86.2%) isolates and 4 isolates had no mutation. One isolate had a base substitution A→Cat nt 25 in the coding region and this isolate had a point mutation leading to an amino acid change at positions 9 (I→L). Our study results indicated that none of the strains had mutation in Shine-Dalgarno and the point mutations were the most common mutations upstream of the oprD coding region among P. aeruginosa isolates. Mutations were observed in imipenem-resistant isolates and it seems this mechanism is effective in resistance of isolates to imipenem and this confirmed that the indiscriminate use of antibiotic should be controlled.

Molecular characteristic of treatment failure clinical isolates of Leishmania major.

BACKGROUND: Leishmaniasis is a prevalent tropical disease caused by more than 20 Leishmania species (Protozoa, Kinetoplastida and Trypanosomatidae). Among different clinical forms of the disease, cutaneous leishmaniasis is the most common form, with an annual 0.6-1 million new cases reported worldwide. This disease's standard treatment is pentavalent antimonial (SbV) that have been used successfully since the first half of the 20th century as a first-line drug. However, treatment failure is an increasing problem that is persistently reported from endemic areas. It is important to define and standardize tests for drug resistance in cutaneous leishmaniasis. SbV must be reduced to its trivalent active form (SbIII). This reduction occurs within the host macrophage, and the resultant SbIIIenters amastigotes via the aquaglyceroporin1 (AQP1) membrane carrier. Overexpression of AQP1 results in hypersensitivity of the parasites to SbIII, but resistant phenotypes accompany reduced expression, inactivation mutations, or deletion of AQP1. Hence, in this study, a phylogenetic analysis using barcode gene COXII and kDNA minicircle and expression analysis of AQP1 were performed in treatment failure isolates to assess the isolates' molecular characteristics and to verify possible association with drug response. METHODS: Samples in this study were collected from patients with cutaneous leishmaniasis referred to the Diagnosis Laboratory Center in Isfahan Province, Iran, from October 2017 to December 2019. Among them, five isolates (code numbers 1-5) were categorized as treatment failures. The PCR amplification of barcode gene COXII and kDNA minicircle were done and subsequently analyzed using MEGA (10.0.5) to perform phylogenetics analysis of Treatment failures (TF) and Treatment response (TR) samples. Relative quantification of the AQP1 gene expression of TF and TR samples was assessed by real-time PCR. RESULTS: All samples were classified as L. major. No amplification failure was observed in the cases of barcode gene COXII and kDNA minicircle amplification. Having excluded the sequences with complete homology using maximum parsimony with the Bootstrap 500 method, four major groups were detected to perform phylogenetic analysis using COXII. The phylogenetic analysis using the barcode target of minicircle showed that all five treatment failure isolates were grouped in a separate sub-clade. CONCLUSIONS: We concluded that the barcode gene COXII and the minicircle kDNA were suitable for identification, differentiation and phylogenetic analysis in treatment failure clinical isolates of Leishmania major. Also, AQP1 gene expression analyses showed that treatment failure isolates had less expression than TR isolates. The isolate with TF and overexpression of the AQP1 gene of other molecular mechanisms such as overexpression of ATP-binding cassette may be involved in the TR, such as overexpression of ATP-binding cassette which requires further research.

Toxoplasma gondii in Sheep and Goats from Central Iran.

OBJECTIVE: Toxoplasmosis, caused by Toxoplasma gondii, infects humans by consuming infected raw or undercooked meat and foods harboring mature oocysts. In this study, we assessed the prevalence of T. gondii in sheep and goats coming from central Iran. After completing the questionnaire, about one gram of liver or diaphragm tissue was taken as a sample from 90 sheep and 90 goats slaughtered in Yazd Province and stored at - 20 ºC. DNA extraction was done, and then T. gondii was detected using nested PCR. RESULTS: This study indicated that the prevalence of T. gondii in all slaughtered animals was 11.6% (21 of 180), including 14.4% (13/90) in sheep and 8.8% (8/90) in goats. The infection rates in liver and diaphragm samples were 12.2% (11/90) and 11.1% (10/90), respectively (p = 0.8163). The infection rate in animals older than one was 16.3% (15/92), and it was 6.8% (6/88) in animals under one year of age. Therefore, no significant differences were found (p = 0.475). Infection rates were 19.5% (18/92) in males and 3.4% (3/88) in females (p = 0.0007). In conclusion, the infection rates of toxoplasmosis in livestock in this area are almost high, and therefore, it is necessary to design appropriate prevention programs to control the disease.

Safety assessment of genetically modified rice expressing Cry1Ab protein in Sprague-Dawley rats.

Rice is considered one of the most important staple food crops. Genetically modified (GM) Bt rice, harbored cry1Ab gene expressing the insect-resistance protein has been developed to resistance to the insects. In this study, we assessed the safety of the GM Bt rice on Sprague-Dawley rats for 90 days. Totally, 120 rats in both sexes were used for three different diets, including 50% GM Bt rice, feeding with 50% rice, and standard feeding. Each 40 SD rats including 20 males and 20 females were considered as each diet. The clinical variables such as body weight and food consumption were measured and a range of clinical tests was examined, including hematology, serum chemistry parameters, urinalysis profile, thyroid, and sex hormone levels. Pathological assessments were also done. The results showed that the mean weekly feed utilization (%) had no significant difference among the studied groups. Also, blood biochemistry, hematological parameters, urine analysis, and hormonal levels had no significant differences among the groups. However, alanine aminotransferase was less in males versus female feeding with GM Bt rice. No histopathological changes were observed among the groups. In conclusion, this study demonstrated that GM Bt rice had no obvious adverse effects on rats' health.

The expression profile of LmTRYP, LmTRYR, and LmHSP83 genes in treatment failure clinical isolates of Leishmania major.

Cutaneous leishmaniosis (CL) is one of the important neglected tropical diseases caused by Leishmania spp. such as L. major, L. tropica in the Old World. In recent years, some reports of treatment failure in patients with CL have been reported worldwide. Therefore, in this study, we assessed LmHSP 83, LmTRYR, and LmTRYP gene expressions in treatment failure clinical isolates of L. major. After sampling from the cutaneous lesions, DNA was extracted and then genera verification and species identification was done using ITS1-PCR-RFLP method. A part of each sample was used in order to RNA extraction and cDNA synthesized. LmHSP 83, LmTRYR, and LmTRYP gene expressions were assessed using SYBR Green real-time PCR. The treatment failure clinical isolates had the mean expression of 5.55±1.67, 247.024±23.54, and 1.204±2.14 for LmHSP 83, LmTRYR, and LmTRYP , respectively less than the same genes in treatment response isolates (P=0.001). This study recommended the other mechanisms may involve in response to treatment in treatment failure clinical isolates of L. major.

Toxoplasma gondii in Milk of Human and Goat from the Desert Area in Central Iran.

BACKGROUND: Toxoplasma gondii, an obligate intracellular protozoan, causes toxoplasmosis. The aim of this study was molecular detection of T. gondii in breast and goat milk samples by the molecular method in the central Iran. METHODS: Totally, 300 human' and 200 goats' milk samples were collected randomly from different regions of central Iran in 2018. DNA extraction was performed by the salting-out method. Molecular detection of the parasite was done by nested-PCR using the specific primer pairs. Statistical analysis was performed by SPSS 23 using descriptive and Chi-square tests. RESULTS: Out of 300 human milk samples, 1 sample (0.3%) was infected with T. gondii. Out of 200 samples of goat milk, 11 samples (5.5%) showed infection with T. gondii. The frequency of infection in goat's milk samples was 4.36% in the south and west, 1.9% in northern regions, and 2% in eastern regions of the province. The statistical analysis showed no significant difference between toxoplasmosis and different geographical regions in the province. CONCLUSION: Because of the popularity of the goat milk and the transfection probability with the milk to humans, it is recommended to boil milk prior to use. Furthermore, case contamination of T. gondii in the human milk sample showed one of the important paths for infection transmission, which requires further studies.

Isolation and Molecular Identification of Leishmania spp. Agents in Patients with Cutaneous Leishmaniasis in Yazd Province, Endemic Region of Central Iran.

BACKGROUND: Cutaneous Leishmaniasis (CL) is a major health problem in many parts of Iran. Many methods have been introduced for detection and identification of Cutaneous Leishmaniasis. The purpose of this study was isolation and molecular identification of Leishmania spp. agents in patients with CL from endemic region of central Iran. In this study, one of the main loci of central Iran named Yazd will be assessed CL identification using PCR-RFLP. METHODS: For this cross-sectional study, sampling was done from 372 suspicious patients with CL who referred to Health Centers of Yazd Province from 2016 to 2017. After collection samples of patients, DNA extraction was done from samples on slides. Genus detection was done using specific primers by PCR. RFLP analysis was done for species identification. RESULTS: Out of 372 samples, 159 samples were positive using PCR based method. Out of 159 samples, 87 (54.7%) L. major and 72 (45.3%) L. tropica were identified using RFLP analysis. The number of lesions in each patient was different but 119 (74.8%) patients showed the number of 1-3 lesions, and more lesions (more than 10 lesions) was showed in 4 (2.5%) person. CONCLUSION: The CL found in Yazd province resulted from L. major and L. tropica as the agents of rural and urban types, respectively. The prevalence of L. major and L. tropica was almost the same. This indicated that control programs could be designed for treatment and vector and reservoir control.