Molecular analysis and dimorphism of azole-susceptible and resistant Candida albicans isolates

INTRODUCTION: Candida albicans is responsible for superficial or systemic infections known as candidiasis, which may be found in infected tissue as unicellular budding yeasts, hyphae, or pseudohyphae. In this study, the effects of both fluconazole and itraconazole antifungal agents on the hyphal formation and genotypic characterization of C. albicans isolates classified as either susceptible or resistant were investigated. METHODS: The hyphal production of five C. albicans isolates under the action of antifungal agents was investigated by culturing yeast on growth medium and on hyphal induction medium. The genotypic characterization was carried out for 13 isolates of C. albicans using the random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) method. RESULTS: The dimorphism analysis showed that the hyphal formation was higher in resistant than in the susceptible isolates to both azoles. The RAPD-PCR method identified the formation of two different groups. In group A, four resistant and two susceptible isolates were clustered, and in group B, one resistant and six susceptible isolates were clustered. CONCLUSIONS: Considering that hyphal formation was higher in resistant isolates in the presence of azole drugs, we confirmed that the hyphal production is closely related to susceptibility to azoles. These drugs may affect the morphogenesis of C. albicans depending on their susceptibility to these drugs. In relation to RAPD-PCR, most resistant isolates classified in group A and susceptible isolates in group B demonstrated that this method presented a similar standard between the two groups, suggesting that by this technique, a strong correlation between genotypes and fluconazole-resistant samples may be found.

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Bacterial, Viral and Fungal Infections in Patients after Allogeneic Stem Cell or Bone Marrow Transplantation - A Comparison between Patients with Related and Patients with HLA-Matched Non-Related Stem Cell Donors.

Blood ◽

10.1182/blood.v110.11.4978.4978 ◽

2007 ◽

Vol 110 (11) ◽

pp. 4978-4978

Author(s):

Christina T. Rieger ◽

Johanna Tischer ◽

Helmut Ostermann

Keyword(s):

Bone Marrow ◽

Candida Albicans ◽

Stem Cell ◽

Stem Cell Transplantation ◽

Myeloid Leukemia ◽

Fungal Infections ◽

Stem Cell Source ◽

Cell Source ◽

Group A ◽

Group B

Abstract Bacterial, viral and fungal pathogens frequently cause severe, life-threatening infections in immunocompromised patients after allogeneic stem cell transplantation (SCT). We investigated whether patients with related stem cell donors (group A) developed infections less frequently than patients with HLA-matched, non-related donors (group B). Fifty-nine consecutive patients treated at our transplantation unit between April 2004 and January 2005 were included into the analysis. We documented demographic and clinical characteristics at baseline, treatment, clinical course, microbiological examinations, clinical and radiological signs of infection and mortality. Of the total 59 patients analyzed, 22 received stem cells from related and 37 from HLA-matched non-related donors. Both groups were well balanced regarding age and weight. 50% of the patients in group A and 60% in group B were male. Most frequent diagnoses were acute myeloid leukemia (30 of 59 patients [50.8%]; group A: 68.2%; group B: 40.5%), multiple myeloma (15.2%), acute lymphoblastic leukemia (11.9%) and chronic myeloid leukemia (10.2%). Bone marrow was more often the stem cell source in group A (45.5%/ 10 patients) than in group B (10.8%/ 4 patients), peripheral stem cell transplantation respectively was predominant in the unrelated group (86.5%/ 32 patients) versus the family donor group (54.5%/ 12 patients), cord blood was used as unrelated stem cell source in1 patient (2.7%). Clinically documented infections occurred in 6% in group A and in 14% in group B. Pulmonary infiltrates were observed more frequently in group A (11 patients/ 50%) than in group B (16 patients/ 43.2%). The predominant findings were atypical infiltrates (total 16 patients), followed by signs of fungal (total 7 patients) and bacterial pulmonary infiltration (total 4 patients). Microbiologically documented infections were detected in all patients. The average number of pathogens was equal in both groups. Detected pathogens were HHV-6 (48 patients), coagulase-negative Staphylocci (17 patients), EBV (14 patients) and CMV (11 patients). Three fungal infections were detected by microbiological approaches in group A (2 × Candida albicans, 1 × Pitysporum ovale) compared to nine fungal infections in group B (5 × Candida albicans, 1 × Candida glabrata, 1 × Candida parapsilosis, 2 × Geotrichum capitatum). Two years after transplantation, 55.9% of patients were alive (group A: 68.2%; group B: 48.6%). Patients with AML had a two-year survival of 50% (group A: 53.3%; group B: 46.7%). In our study, we observed no clear relation between frequency of infection and donor type, yet there was a trend towards more invasive fungal infections in the unrelated group (13% group A vs. 24% group B).

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Superoxide Dismutase 2 Val16Ala Polymorphism is Associated with Amiodarone-Associated Liver Injury

Serbian Journal of Experimental and Clinical Research ◽

10.2478/sjecr-2019-0078 ◽

2020 ◽

Vol 0 (0) ◽

Author(s):

Branimir Radmanovic ◽

Jovan Jovanovic ◽

Natasa Djordjevic ◽

Dejan Baskic ◽

Jelena Cukic ◽

...

Keyword(s):

Liver Injury ◽

Control Group ◽

Single Nucleotide ◽

Clinical Center ◽

Group A ◽

Hardy Weinberg Equilibrium ◽

Pcr Method ◽

Group B ◽

And Control ◽

Superoxide Dismutase 2

AbstractAssociation of SOD2 V16A single-nucleotide polymorphism (rs4880) with drug hepatotoxicity were reported but relationships with amiodarone prescriptions remained unexplored. Research was an exploratory, controlled prospective clinical trial. Patients hospitalized and treated in Clinical Center in Kragujevac, Serbia (in year 2017) were divided into experimental (using amiodarone, having liver injury, n=29, 19 males, the mean age 66.8±10.4 years), control A (neither amiodarone use nor hepatotoxicity, n=29, 19, 66.1±10.3) and control B group (using amiodarone, not having hepatotoxicity, n=29, 19, 66.8±9.8). From blood samples, among other routine biochemistry, genotyping for SOD2 polymorphism Val16Ala was conducted using real-time PCR method with TaqMan® Genotyping Master Mix and TaqMan® DME Genotyping Assay for rs4880. Patients taking amiodarone and having liver injury were mostly carriers of Val/Val (TT) genotype (13 of 24 patients, 54.2%) while Val/Ala (TC) and Ala/Ala (CC) genotypes prevailed in control group A (19 of 40, 47.5%) and control group B (9 of 23, 39.1%), respectively (2=10.409, p=0.034). Frequency of Val (T) and Ala (C) alleles were 0.51 and 0.49, respectively in the whole study sample (Hardy Weinberg equilibrium, 2=0.56, p=0.454). Carriers of TT genotype had significantly higher ALT (437.0±1158.0 vs 81.9131.5 U/L), total bilirubin (28.320.5 vs 15.313.0 mol/L) and total bile acid concentrations (10.910.2 vs 6.45.3 mol/L) compared to carriers of TC genotype (U=2.331, p=0.020, U=3.204, p=0.001 and U=2.172, p=0.030, respectively). Higher incidence of 47T allele of SOD2 was inpatients with amiodarone-associated liver injury as compared to patients on amiodarone not experiencing hepatotoxic effects.

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Characterization of Clinical and Carrier Streptococcus agalactiae and Prophage Contribution to the Strain Variability

Viruses ◽

10.3390/v12111323 ◽

2020 ◽

Vol 12 (11) ◽

pp. 1323

Author(s):

Aneta Lichvariková ◽

Katarina Soltys ◽

Tomas Szemes ◽

Livia Slobodnikova ◽

Gabriela Bukovska ◽

...

Keyword(s):

Streptococcus Agalactiae ◽

Bacterial Infections ◽

Group B Streptococcus ◽

Host Bacterium ◽

Phage Integrase ◽

Integration Sites ◽

Group A ◽

Pcr Method ◽

Group B

Streptococcus agalactiae (group B Streptococcus, GBS) represents a leading cause of invasive bacterial infections in newborns and is also responsible for diseases in older and immunocompromised adults. Prophages represent an important factor contributing to the genome plasticity and evolution of new strains. In the present study, prophage content was analyzed in human GBS isolates. Thirty-seven prophages were identified in genomes of 20 representative sequenced strains. On the basis of the sequence comparison, we divided the prophages into eight groups named A–H. This division also corresponded to the clustering of phage integrase, even though several different integration sites were observed in some relative prophages. Next, PCR method was used for detection of the prophages in 123 GBS strains from adult hospitalized patients and from pregnancy screening. At least one prophage was present in 105 isolates (85%). The highest prevalence was observed for prophage group A (71%) and satellite prophage group B (62%). Other groups were detected infrequently (1–6%). Prophage distribution did not differ between clinical and screening strains, but it was unevenly distributed in MLST (multi locus sequence typing) sequence types. High content of full-length and satellite prophages detected in present study implies that prophages could be beneficial for the host bacterium and could contribute to evolution of more adapted strains.

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Changes in subgingival microflora after placement and removal of fixed orthodontic appliances

Srpski arhiv za celokupno lekarstvo ◽

10.2298/sarh1406301z ◽

2014 ◽

Vol 142 (5-6) ◽

pp. 301-305 ◽

Cited By ~ 4

Author(s):

Marija Zivkovic-Sandic ◽

Branka Popovic ◽

Jelena Carkic ◽

Nadja Nikolic ◽

Branislav Glisic

Keyword(s):

Statistical Significance ◽

Orthodontic Appliances ◽

Fixed Orthodontic Appliances ◽

Orthodontic Patients ◽

Group A ◽

Subgingival Microflora ◽

Fixed Appliances ◽

Pcr Method ◽

Group B ◽

The Right

Introduction. The placement of fixed orthodontic appliances may lead to increased plaque accumulation and changes in subgingival microflora. Objective. The aim of this study was to examine the changes in frequency of subgingival microflora that occur after placement and removal of fixed orthodontic appliance using polymerase chain reaction (PCR). Methods. This study included 33 orthodontic patients, who were divided into two groups. Subgingival plaque samples were collected from the right upper incisor (U1) and right upper first molar (U6). In group A, the samples were taken three times: before placement appliance (T1), after one month (T2), and after 3 months (T3). In group B the samples were also taken three times: before appliance removal (T1), after one month (T2), and after three months (T3). PCR method was used to determine the presence of P. gingivalis, A. actinomycetemcomitans, T. forsythia, and P. intermedia. Results. In group A the frequency of P. gingivalis showed statistically significant decrease at U1 (p=0.049) and U6 (p=0.008), from T1 to T2, and at U1 (p=0.048) from T1 to T3. In group B only the frequency of T. forsythia showed a statistically significant decrease, at U6 (T1 vs. T2, p=0.004; T1 vs. T3, p=0.0003). Regarding other analyzed bacteria, changes in the presence were noticed but no statistical significance was found. Conclusion. Placement of fixed appliances may have an impact on subgingival microflora, but in the first months after the placement and removal of the appliance changes were not significant, probably due to good oral hygiene.

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Evaluation and Comparison of the Effectiveness of Physical Methods of Disinfection on Heat-Polymerized Polymethyl Methacrylate – An In-vitro Study

Journal of Pharmaceutical Research International ◽

10.9734/jpri/2021/v33i48b33286 ◽

2021 ◽

pp. 285-293

Author(s):

Kapil Soni ◽

Vivek Choukse ◽

Rajeev Shrivastava ◽

Umesh Palekar ◽

Prabha Shakya Newaskar ◽

...

Keyword(s):

Candida Albicans ◽

Microwave Irradiation ◽

Uv Radiation ◽

Acrylic Resin ◽

Positive Control ◽

Mean Value ◽

Control Group ◽

Group A ◽

The Mean ◽

Group B

Introduction: The common relationship of Candida and the ample proof that acrylic resin dentures are related to denture stomatitis in the oral cavity, which can range from simple mucosal lesions to a fatal systemic form. The current study aims to assess and compare the efficacy of microwave irradiation and UV radiation in the disinfection of Candida albicans infection in heat-polymerized polymethyl methacrylate. Materials and Methods: A total of 90 specimens were prepared with heat-polymerized acrylic resin, out of which 30 were used for Microwave irradiation, and 30 were used for UV irradiation, and 30 were used for positive control. There were three groups: Group A (Positive Control group); Group B (Microwave irradiation); Group C (UV radiation). After preparing the samples they were chemically sterilized, then infected with C. Albicans, and again Group B is disinfected with Microwave irradiation, Group C is disinfected by UV light and Group A is kept as Positive control without disinfection. Turbidity was measured for each group after re-infection of every specimen in CFU/ml. Results: In the present study, the CFU/ml of Candida albicans in the control group was 2.97x107±0.6 x 107 (P= .000), for microwave irradiation, the mean value was 5.3±11.1 (P= .000), and for UV radiation, the mean value was 19.3±22.9 (P= .003). Conclusion: According to the results of the current study, microwave irradiation (6 minutes at 650W) is more effective than UV radiation (10 minutes each side at 254nm wavelength) of Candida albicans infection on the heat-polymerized acrylic resin in laboratory conditions.

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Genetic characterization of Lactococcus garvieae isolated from farmed rainbow trout by random amplified polymorphic DNA-PCR in Iran

Journal of the Hellenic Veterinary Medical Society ◽

10.12681/jhvms.22951 ◽

2020 ◽

Vol 71 (1) ◽

pp. 2023

Author(s):

F. FADAEIFARD ◽

M. RABIEI ◽

M. F. SHARIFPOUR

Keyword(s):

Rainbow Trout ◽

Bacterial Infections ◽

Random Amplified Polymorphic Dna ◽

Farmed Fish ◽

Biochemical Tests ◽

Bacterial Cultures ◽

Lactococcus Garvieae ◽

Strain Pattern ◽

Rapd Pcr ◽

Pcr Method

Lactococcosis is one of the main bacterial infections of fish around the world. Lactococcus garvieae has been a major cause of rainbow trout losses in freshwater farming. This study aimed to genotype and determine the variability of L. garvieae isolated from infected farmed rainbow trout in Iran by the RAPD-PCR method. Bacterial samples were collected from 12 farms located in the western part of Iran and suspected to carry Lactococcus infection. Two hundred bacterial cultures containing cocci shaped bacteria were cultured in Trypticase soy agar (TSA) and blood agar mediums. All bacterial cultures were tested by conventional microbiological and biochemical tests, and PCR assay to identify L. garvieae by 16S rDNA genes. The RAPD-PCR method was used to determine the genetic pattern of all isolates. The sample strain pattern of the isolates was analyzed in the NTSYS program. According to a similarity coefficient index of 70%, all L. garvieae isolates were separated into two groups with four RAPD profile types. The highest and the lowest genetic pairwise similarity among the isolates were 98% and 54%, respectively. The results of the present study revealed that RAPD-PCR is an applicable method to describe the genetic diversity of different strains of L .garvieae among farmed fish.

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Phylogenetic and Transmission Patterns of Extended-Spectrum β- Lactamase –Producing Escherichia Coli among Neonatal’s Gut Flora in Indonesia

Folia Medica Indonesiana ◽

10.20473/fmi.v57i3.22828 ◽

2021 ◽

Vol 57 (3) ◽

pp. 199

Author(s):

Rakhmatul Binti Sulistya ◽

Kuntaman Kuntaman ◽

Eko Budi Koendhori ◽

Terza Aflika Happy ◽

Wahyu Setyarini

Keyword(s):

Escherichia Coli ◽

Bacterial Isolates ◽

Gut Flora ◽

E Coli ◽

Extended Spectrum ◽

Rapd Pcr ◽

Significant Difference ◽

Group A ◽

Neonatal Group ◽

Pcr Method

This study aimed to explore the phylogenetic pattern of Extended-spectrum β-lactamase-producing Escherichia coli in the gut flora of neonatal into groups A, B1, B2, D and tracing the presence of spreading among the age group of neonatal. Multiplex PCR was conducted to classify the phylogenetic group of ESBL producing E. coli into groups of commensals (A, B1) and pathogen (B2, D). The spread of bacterial agents among the host (neonatal) was conducted by the RAPD-PCR method with two random primers. Among 34 bacterial isolates were identified among early neonatal group A (22,2%), B1 (11,1%), B2 (22,2%) D (33,3%) and postneonatal groups A (8%), B1 (4%), B2 (36%) D (40%). It was not a statistically significant difference (p=0.388). The dominance of pathogenic groups B2 and D as many as 70,6%. From 34 samples was found 34 types of RAPD indicated there were not any bacterial spread among neonatal. The ESBL producing E. coli among early neonatal and postneonatal were dominated by the pathogenic group of B2 and D (total=70,6%), but they were not statistically significant. There was not any spreading of bacterial agents among individual gut flora of neonatal.

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Differentiation of Candida species obtained from nosocomial candidemia using RAPD-PCR technique

Revista da Sociedade Brasileira de Medicina Tropical ◽

10.1590/s0037-86822006000200008 ◽

2006 ◽

Vol 39 (2) ◽

pp. 174-178 ◽

Cited By ~ 12

Author(s):

Henrique Maia Valério ◽

Rita de Cássia Botelho Weikert-Oliveira ◽

Maria Aparecida de Resende

Keyword(s):

Candida Albicans ◽

Candida Glabrata ◽

Candida Parapsilosis ◽

Random Amplified Polymorphic Dna ◽

Intraspecific Polymorphism ◽

Surgical Wounds ◽

Rapd Pcr ◽

Nosocomial Candidemia ◽

Belo Horizonte ◽

Catheter Urine

Thirteen strains of the genus Candida were isolated from catheter, urine and surgical wounds from individual patients of the Santa Casa de Misericórdia, Belo Horizonte, MG, Brazil. Ten strains were characterized as Candida albicans, two as Candida glabrata, and one as Candida parapsilosis. Isolates were evaluated for molecular relatedness by random amplified polymorphic DNA technique using 15 primers. The analysis of the genomic DNA obtained revealed a low intraspecific polymorphism and did not allow for the differentiation between strains of the same species obtained from distinct clinical sources (catheter, urine and surgical wounds). The RAPD profiles generated were able to differentiate among the species of Candida albicans, Candida parapsilosis and Candida glabrata strains isolated in this study.

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Determination of Genetic Diversity of Four Guppy Strains Using the Random Amplified Polymorphic DNA-PCR

Asian Journal of Biochemistry, Genetics and Molecular Biology ◽

10.9734/ajbgmb/2021/v8i130183 ◽

2021 ◽

pp. 1-8

Author(s):

Riva Hafidah ◽

Ayi Yustiati ◽

Yuniar Mulyani ◽

Ibnu Bangkit Bioshina Suryadi

Keyword(s):

Genetic Diversity ◽

Random Amplified Polymorphic Dna ◽

Descriptive Analysis ◽

Similarity Index ◽

Quantitative Descriptive Analysis ◽

Rapd Pcr ◽

Result Show ◽

Pcr Method ◽

Quantitative Descriptive

This research aims to determine genetic diversity of four strains guppy, respectively are japan blue double sword (JBD), japan blue tiger double sword (JBTD), blue moscow (BM), and panda guppy (PG) with RAPD-PCR method. The obtained genetic diversity data is used as guide reference for hybridization between four strains. The research was conducted in September 2020 to April 2021 with explorative methods and in qualitative and quantitative descriptive analysis. The research was carried out in biotechnology Laboratory, Fishery and Marine Sciences Faculty and Central Laboratory,Padjadjaran University, Indonesia.Strains of JBD, JBTD, BM obtained fromCilengkrangSubdistrict, Bandung and PG strain obtained from Parung market, Bogor. Primary OPA-03 (AGTCAGCCAC) is used for standard parameters to interpret genetic diversity among four strains of guppy. Based on results, amplification with OPA-03 primary visualize 25 bands that include five polymorphic bands and 20 monomorphic bands. The phylogenetic tree result show that there are two relationship groups. The first group are JBD, JBTD, and BM with similarity index in the range of 80-89%The first group consist two sub groups of relationship. The first sub group are JBD and JBTD with similarity index of 89%. The second sub group is BM with similarity index of 80%. The second group isPG with similarity index of 65.5%.

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Genetic structure and diversity among three Greek sheep breeds using Random Amplified Polymorphic DNA-PCR

Journal of the Hellenic Veterinary Medical Society ◽

10.12681/jhvms.14860 ◽

2017 ◽

Vol 62 (4) ◽

pp. 301 ◽

Cited By ~ 1

Author(s):

I. MASTRANESTASIS (Ι. ΜΑΣΤΡΑΝΕΣΤΑΣΗΣ) ◽

Ch. LIGDA (Χρ. ΛΙΓΔΑ) ◽

K. THEODOROU (Κ. ΘΕΟΔΩΡΟΥ) ◽

L. V. EKATERINIADOU (Λ.Β. ΑΙΚΑΤΕΡΙΝΙΑΔΟΥ)

Keyword(s):

Genetic Structure ◽

Genetic Differentiation ◽

Phylogenetic Relationships ◽

Random Amplified Polymorphic Dna ◽

Genetic Distances ◽

Genetic Identity ◽

Rapd Pcr ◽

Pcr Method ◽

Genetic Structure And Diversity

Genetic structure and diversity of 120 animals from three Greek local breeds were investigated by Random Amplified Polymorphic DNA (RAPD) - PCR method. Sheep samples originated from the Lesvos, Chios and Karagouniko breeds were treated with 11 random primers to estimate their genetic diversity and phylogenetic relationships. Our analysis comprised two levels of the breeds' genetic structure: i) the genetic differentiation among the three breeds and ii) the genetic differentiation among the flocks within each breed. This combined approach gave two main findings: i) the study of genetic distances and identity revealed that the Karagouniko sheep breed is genetically distinct from Chios (GD=0.1979) and Lesvos (GD=0.1691) breeds, while ii) the Chios and Lesvos breeds are genetically similar (GI=0.9631); half of the flocks of Lesvos have a relatively closer relationship with those of Chios than with the other Lesvos flocks. This is the first study that reports the close genetic relationship between the Chios and Lesvos breeds and gives strong evidence to hypotheses about their related origin. Furthermore, the study of polymorphic loci revealed particular indicators located in Karagouniko breed, as definitional datum of genetic identity or as a fingerprint of breed. Therefore, RAPD-DNA methods can be an efficient tool for the determination of phylogenetic relationships and genetic identity among the bloodstock breed of sheep.

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