High density linkage maps are essential for precise mapping of gene loci controlling quantitative traits. In the present study, a high density linkage map of spring wheat recombinant inbred lines population derived from a cross between Yecora Rojo and No. 49 was constructed using Genotyping by Sequencing ((GBS)) method. To identify single nucleotide polymorphisms (SNPs), Sequencing of parents and recombinant inbred lines was carried out at three lanes of Illumina Hiseq 2500. In total, more than 36 thousand SNP were identified of which 804 SNPs were not aligned to any wheat chromosome. Among these SNPs, 3042, 3977 and 769 SNPs were belonged to A, B and D sub genomes, respectively. After removing SNPs with ≥ 20% missing data and minimum allele frequency ≤ 0. 05 as well as loci segregating from 1: 1 Mendelian ratio, in total 5831 polymorphic SNPs identified and assigned to 21 chromosomes of wheat. These markers spanned 3642. 14 cM of the wheat genome with an average marker density of 0. 62 (markers/cM).

Group B streptococci ((GBS)) or Streptococcus agalactiae are members of the normal flora of the female genital tract. (GBS) normally colonizes the vagina in many women asymptomatically. During labor this organism may infect the newborn, leading to neonatal sepsis and meningitis. This study aimed to investigate the prevalence of group B streptococcus in pregnant women by a rapid and easy culture method. It seems that in cases in which (GBS) carriage is not suspected until the time of labor, using such a quick and specific culture method would be valuable. A total of 330 vaginal swabs were collected from women attending delivery room at Hedayat hospital, Tehran, Iran, from April through July 2008. Cotton swabs contaminated with vaginal fluid were placed into Amies transport medium and transported to the Avicenna laboratory daily. Vaginal specimens were cultured on selective (GBS) Agar Base medium (ISLAM) for isolation and detection of group B streptococcus. The plates were incubated at 35-37°C under anaerobic condition for 24 hours. Incubated S.agalactiae developed orange/red pigmented colonies in (GBS) agar plates. Among the 330 women, the results of the culture were positive for (GBS) in 68 women (20.6%). Statistical analyses showed no significant relationship between demographics, reproductive histories and obstetric characteristics of subjects with the test results. Solely the antibiotic therapy was associated with (GBS) colonization. The results are indicating that the relatively high maternal (GBS) colonization rate in pregnant women warrants a routine screening and prophylactic treatment of the infected women. Colonization with group B streptococcus can be identified directly by (GBS) agar medium and decrease the time to detection of (GBS).

Background: Over the last two decades CONs have emerged as opportunistic pathogens, especially among immune compromised hosts and patients with implanted biomaterials. The increasing incidence of these micro-organisms in hospital- acquired infections necessitates the need for an accurate identification of staphylococcus isolates at the species level. Since tuf gene is known as the most accurate for specifying CONs, we aimed to identify species of 50 consecutive clinical CONs isolates by phenotypic characteristics and tuf gene Sequencing.Materials and Methods: A total of 50 CONs was isolated from various clinical specimens of hospitalized patients in Shahid Mohammadi hospital, Bandar-Abbass. Phenotyping was carried out by differential - biochemical tests. Genotyping was performed by Sequencing of tuf gene, followed by blast and construction of phylogenetic tree.Results: The clinical isolates consisted of 25 (50%) S. epidermidis, 22 (44%) S. saprophyticus and 3 (6%) S. hemolyticus. S. epidermidis and S. saprophyticus strains were mostly isolated from blood and urine cultures, respectively. Vancomycin was found to be the most effective antibiotic followed by cefalexin and of loxacin. Blast searches and phylogenetic tree inferred from the neighbor-joining method of 8 isolates revealed that 5 isolates had atuf sequence 100 % identical totuf gene of Staphylococcus epidermidis strain SeMCV45, and 3 isolates was 99% similar to that of Staphylococcus haemolyticus strain ShlMCV14 isolated from media cultures.Conclusions: CONS are involved in a wide range of nosocomial infections. PCR and Sequencing of the tuf gene is a reliable and valuable approach for the Genotyping and identification of CONS species in epidemiological studies.

Background: Group B Streptococci ((GBS)) or Streptococcus agalactiae is one of the most common causes of sepsis and meningitis in neonates and of invasive diseases in pregnant women. It can also cause infectious disease among adults with underlying medical conditions like immunocompromised individuals. Polysaccharide capsule is an important virulence factor. Nine (GBS) serotypes (Ia, Ib, II to VIII) based on capsular polysaccharide antigens have been described. Distribution of capsular serotypes varies over time and by geographic location. The aim of this study was to detect the capsular serotype distribution in (GBS) clinical isolates based on Genotyping of cps-gene cluster and to determine the predominant serotypes of (GBS).Methods: In this cross sectional study a total of 50 (GBS) strains were isolated from various clinical sources including: urine, vagina, semen and urethral secretions. (GBS) was identified by Gram stain, catalase test, CAMP test and also resistance to 0.04 U Bacitracin and SXT disks. DNA was extracted from all the isolates using the wizard SV Genomic DNA Purification system, Promega, USA. The capsular serotype of the isolates was assigned by using a specific-two Multiplex PCR assay. For statistical analysis, Chi-square method was used. SPSS V.13 was also used.Results: In the 50 (GBS) isolates, the predominant serotypes were III with 25 isolates (50%) and serotype V with 8 isolates (16%). Seven isolates (14%) belonged to serotype Ia and 7 isolates (14%) belonged to serotype II, respectively. Serotypes Ib, IV, VI, VII and VIII were not found and 3 strains were classified as nontypeable.Conclusion: Based on the results of this study, serotypes III and V were the predominant serotypes in (GBS) clinical isolates.

Background: Cutaneous leishmaniasis (CL) is one of the most common parasitic diseases in many regions of Iran. It has a major role in deprived societies. We aimed to identify Leishmania species based on molecular as ITS1-rDNA-PCR internal transcribed spacer 1 (ITS1) region, microscopy, and culture techniques in diagnosing cutaneous leishmaniasis. Methods: From April 2018 to May 2020, we conducted a cross-sectional study involving 32 patients with suspected CL lesions in Sistan and Baluchistan Province, located in southeast Iran. Samples were subjected to microscopic examination, culture, and PCR amplification targeting the internal transcribed spacer 1 (ITS1) region. DNA Sequencing was performed on PCR-positive samples for species identification and phylogenetic analysis. Results: PCR demonstrated superior sensitivity (93.75%, 30/32) compared to culture (56.25%, 18/32) and microscopic examination (53.1%, 17/32). Molecular analysis revealed that L. major was the predominant causative agent of CL in the study area, with L. tropica occurring less frequently. Sequencing and phylogenetic analysis of the ITS1 region showed high intraspecies similarity among L. tropica isolates, while L. major isolates exhibited greater genetic diversity. Conclusion: This study shows the co-existence of L. major and L. tropica in Mirjaveh, southeast Iran, with L. major as the primary cause. While L. tropica isolates displayed high genetic similarity, L. major samples were more diverse, indicating different epidemiological patterns. These findings highlight the importance of molecular methods for accurately identifying Leishmania species and understanding CL epidemiology in the region.

Background: The present study was aimed to investigate molecular diversity of Echino-coccus granulosus isolates collected from human clinical samples using two mitochondrial genes cox1 and nad1 in Iran.Methods: Forty seven human hydatid cysts were collected through surgery from two hospitals in Tehran during 2010-2012. To determine the fertility of protoscoleces, the cyst fluids were subjected to morphological microscopic examinations. Protoscoleces were removed from each cyst and their total genomic DNAs were extracted. PCR was performed to amplify fragments of 450 and 400 base pair (bp) for cox1 and nad1 genes, respectively. Genotype diversity and sequence variation of the strains were studied by bioinformatics software and in comparison with those mtDNA sequences already de-posited in GenBank.Results: Sixteen, (53.3%), 13 (43.3%), and 1 (3.3%) samples were related to lung, liver, and spleen, respectively. The remained 17 unfertile samples were excluded from the study. From the 29 isolates, 86.7% (n=26) and 10% (n=3) were related to G1, and G3 genotypes, respectively. The sole isolate with G6 genotype was obtained from lung sample. Analysis of concatenated sequences of cox1+nad1 indicated the presence of 11 haplotypes among our strains that were related to genotypes G1 (n=9), G3 (n=1) and G6 (n=1).Conclusion: In consistent to other reports from Iran, genotypes G1, G3, and G6 were observed in our human isolates. The rate of G3 genotype was however higher than other studies implying that human can be considered as a new appropriate host for G3 genotype. Further studies with more sample size from different geographic areas of Iran are needed for E. granulosus mapping.

Background: Dermatophytes are a group of fungi that attack keratinous tissues of the skin, hair, and nail in humans and animals, and cause infections called dermatophytosis (tinea). Since identification of pathogenic fungi at the species level is essential for the detection of the source, control and prevention, and identifying epidemiology of infection, it is necessary to use specific and sensitive diagnostic methods to identify the causes of dermatophytosis. Methods: The clinical samples (skin, nail, and hair) of patients with dermatophytosis in Mashhad City, Iran, were cultured in Mycosyl Agar culture media, and the DNA of obtained dermatophyte colonies were extracted by specific kit. The internal transcribed spacer (ITS) gene was amplified and sequenced by ITS1, ITS4 primers. Finally, the Sequencing results were analyzed using SeqMan software, and were compared with the data of the global genebank. Findings: In this study, 80 dermatophyte isolates were sequenced, which included 9 dermatophyte species as 23 (28. 8%) Trichophyton (T. ) interdigital, 18 (22. 5%) T. tunsorans, 10 (12. 5%) Epidermophyton fluccosum, 10 (12. 5%) of T. mentagrophytes, 8 (10%) Microsporum canis, 4 (5%) T. rubrum, 4 (5%) T. benhamiae, 2 (2. 5%) Nannizzia (N. ) fulvum, 1 (1. 2%) N. persicolor. Conclusion: According to report the rare species of dermatophytes in this study, the use of molecular methods such as Sequencing of the ITS gene can determine the diversity of dermatophytes in a region more precisely than morphological methods.