Comparative chromosome studies in species of subtribe Orchidinae (Orchidaceae)

In our study, FISH mapping using 18S-5.8S-25S rDNA and 5S rDNA sequences was performed for the first time on Ophrys tenthredinifera Willdenow, 1805, Serapias vomeracea (Burman f., 1770) Briquet, 1910 and Himantoglossum hircinum (Linnaeus, 1753) Sprengel, 1826. A detailed study was also performed on O. tenthredinifera using Giemsa-staining, silver-staining, CMA fluorescence banding and fluorescence in situ hybridisation (FISH) with rDNA probes. We analysed two subspecies, i.e. O. tenthredinifera subsp. neglecta (Parlatore, 1860) E.G. Camus, 1908 and O. tenthredinifera subsp. grandiflora (Tenore, 1819) Kreutz, 2004 by the traditional Feulgen method and constructed the karyotype. The cytotaxonomic implications for both taxa are also discussed. In Himantoglossum hircinum, FISH and silver staining highlighted differences in the number of two rDNA families (35S and 5S) with respect to Barlia robertiana (Loiseleur-Deslongchamps, 1807) Greuter, 1967. In addition, fluorescence in situ hybridisation was also applied to diploid (2n = 2x = 36) and triploid (2n = 3x = 54) Anacamptis morio (Linnaeus, 1753) R.M. Bateman, Pridgeon et M.W. Chase, 1997. As far as we are aware, this is the first case of autotriploidy observed in A. morio.

Isolation and genotyping of Candida albicans involved in vaginal candidiasis among pregnant women in Sulaymaniyah and Erbil cities

Background and objective: Vaginal candidiasis is a common infection among pregnant women. The current study aimed to investigate the etiologic species of vaginal candidiasis and the genotypes of C. albicans isolated from vaginal samples among pregnant women in the Kurdistan region of Iraq. Methods: Vaginal swabs were collected from pregnant women admitted to the Maternity hospital in Sulaymaniyah and Erbil cities from March 2016 to December 2018. Candida isolates were identified on CHROMagar medium, then confirmed with PCR depending on ITS region. All C. albicans isolates were confirmed using chitin synthase gene (CHS1) and subjected to genotypic analysis based on the transposable intron in 25S rDNA with using CA25S and CA-INT primers. DNA sequencing of 25S rDNA region was done by using CA and CA-INT primers. Results: Among 340 women tested, 114 (33.53%) were positive for vaginal candidiasis. Five Candida species were identified, where they are C. albicans (56.14%), C. glabrata (24.56%), C. kefyr (11.40%), C. tropicalis (5.27%) and C. krusei (2.63%). It has been found that C. albicans significantly (P <0.01) higher than non-albicans species. The genotypes A (450 bp), B (840 bp), and C (450 and 840 bp) of C. albicans were detected. The Genotype A (54.69%) was the most prevalent, followed by Genotype B (34.38%) and Genotype C (10.94%). In regards to genetic variation, genotypes A and B were more similar compared to genotype C. Conclusion: The current study revealed a high prevalence of vaginal candidiasis with different genotypes of C. albicans among pregnant women. Therefore, it is worth considering a vaginal swab culture with clinical symptoms during the diagnosis of vaginal candidiasis. Keywords: Candida; Genotypes; Vulvovaginitis; Candidiasis; Prevalence.

Genotyping of Candida albicans Strains Obtained from Oropharyngeal Candidiasis Patients Based on ABC and RPS Typing Systems

Background: Candida albicanss has been introduced as one of the most common causes of nosocomial infections. Molecular typing methods are powerful tools in epidemiology to investigate the infection source of candidiasis, identify the transmission routes, and control the measures. Objectives: This study aimed for genotyping C. albicans species isolated from oral cavities of the non-HIV patients who suffer from oropharyngeal candidiasis via combined ABC and repeat sequences (RPS) typing systems. Methods: In this study, 31 DNA samples of clinical isolates of C. albicans were evaluated in terms of 25s ribosomal DNA region sequence or ABC typing, and ALT repeats numbers within RPS. DNA was amplified in two separate reactions, and the PCR products were electrophoresed to identify the genotypes of the isolates. Based on the band's pattern, phylogenetic analysis was conducted by UPGMA, and the discriminatory power of ABC and RPS typing was measured by Simpson’s index of diversity. Results: Genotype A with (14 isolates, 45.2%) were the most frequent and followed by genotype B (10 isolates, 32.3%) and Genotype C (7 isolates, 22.6%), respectively. In addition, genotype 3 with 25 isolates (80.6%) were the most prevalent, followed by genotype 2/3 (4 isolates, 12.9%) and genotype 3/4 (2 isolates, 6.5%) respectively. No significant relationship was found between the obtained genotypes and drug-resistant isolates (P < 0.05). Conclusions: This study showed that 25s rDNA and RPS typing is a quick, simple, and cost-effective method with average discriminatory power and good reproducibility for C. albicans genotyping. It can be used for the epidemiology of C. albicans infections.

Diversity of chromosomal composition in top onion (Allium × proliferum (Moench) Schrad. ex Willd.) accessions from the VIR in vitro collection

Background. Top onion, Allium × proliferum (Moench) Schrad. ex Willd., 1809 (2n=2x=16), is a species that is characterized by vegetative propagation by air or underground bulbs only. Accessions of this species have been shown to be hybrids of Allium cepa and Allium fistulosum (Fiskesjo, 1975; Vosa, 1976; Schubert et al., 1983; Puizina and Papes, 1999). Accessions of Allium × proliferum were obtained from various sources and conserved in the in vitro collection of VIR. However, their pedigree was unknown, therefore there was a need to determine the ploidy level and genomic composition of these accessions.Materials and Methods. Thirteen Allium × proliferum accessions from the VIR in vitro collection were studied. To characterize the ploidy level and genomic composition of the accessions, the research employed FISH with chromosome-specific markers (5S and 18S/25S rDNA) and GISH with differentially labeled DNA of the putative parent species, i.e., A. cepa and A. fistulosum.Results. According to GISH, all the studied accessions were hybrids of A. cepa and A. fistulosum. Most (10 out of 13) accessions were determined as diploid hybrids with eight A. cepa and eight A. fistulosum chromosomes. The accession К 3206 turned out to be a diploid 16-chromosome hybrid with eight A. cepa, seven A. fistulosum chromosomes and one rearranged chromosome. Accessions К 3205 and К 3202 were found to be polyploids. The A. × proliferum accession К 3202 contained seven A. cepa and 16 A. fistulosum chromosomes. The accession К 3205 is characterized by the presence of 16 chromosomes hybridizing with A. cepa DNA and 13 chromosomes hybridizing with A. fistulosum DNA. Only one chromosome of A. fistulosum in this accession was revealed to have a 5s rDNA locus.Conclusions. The above shows that the collection contains top onion accessions with karyotypic differences.

Genotyping and Antifungal Vulnerability of Candida albicans Isolated from Cancer Patients in Dewaniyeh Governorate.

Background: Candida infections are one of the major causes of morbidity in cancer patients. Aim: To determine the genotype and antifungal susceptibility of C. albicans isolated from cancer patients and compared with that from healthy individuals. Materials and methods: Oral swabs collected from cancer patients and healthy subjects were screened for the occurrence of C. albicans. Isolates were identified to species level by the conventional mycological methods. Genotypes were determined with use of 25S rDNA PCR analysis. Susceptibility testing was performed using HiCombo MIC technique. Results: Phenotypic examination showed that oral C. albicans was detected in 90% of cancer patients and 50% of healthy controls. PCR shows that the isolates of A, B, C and T genotypes, among which genotype A C. albicans was the predominant genotype. Genotype A C. albicans recognized the entirely isolates in healthy group. Isolates were most sensitive to amphotericin B. Occurrence of resistance to amphotericin B was the lowest followed by itraconazole. However, the isolates demonstrated a high rate of resistance to ketoconazole and fluconazole. Conclusion: Based on these results, 25S rDNA have been shown to be a useful criterion for distinguishing among various isolates of C. albicans. Amphotericin B is effective antifungal agents that can be used against isolates.