Phospholipase Production Research Articles

Efficient production of a novel alkaline cold-active phospholipase C from Aspergillus oryzae by molecular chaperon co-expression for crude oil degumming

A novel alkaline cold-active phospholipase C (PLC) gene (AoPC) from Aspergillus oryzae was cloned. AoPC exhibited the highest sequence similarity of 32.5% with that of a PLC from Arabidopsis thaliana. The gene was co-expressed in Pichia pastoris with molecular chaperone PDI (protein disulfide isomerases), and the highest PLC activity of 82, 782 U mL−1 was achieved in a 5-L fermentor. The recombinant enzyme (AoPC) was most active at pH 8.0 and 25 °C, respectively, and it was stable over a broad pH range of 4.5–9.0 and up to 40 °C. It is the first fungal alkaline PLC. The application of AoPC (with 25% citric acid, w/w) in oil degumming process significantly reduced the phosphorus of crude soybean oil by 93.3% to a commercially acceptable level (<10 mg kg−1). Therefore, the relatively high yield and excellent properties of AoPC may possess it great potential in crude oil refining industry.

Back to Nature: Combating Candida albicans Biofilm, Phospholipase and Hemolysin Using Plant Essential Oils.

Candida albicans is the causative agent of fatal systemic candidiasis. Due to limitations of antifungals, new drugs are needed. The anti-virulence effect of plant essential oils (EOs) was evaluated against clinical C. albicans isolates including cinnamon, clove, jasmine and rosemary oils. Biofilm, phospholipase and hemolysin were assessed phenotypically. EOs were evaluated for their anti-virulence activity using phenotypic methods as well as scanning electron microscopy (SEM) and atomic force microscopy (AFM). Among the C. albicans isolates, biofilm, phospholipase and hemolysins were detected in 40.4, 86.5 and 78.8% of isolates, respectively. Jasmine oil showed the highest anti-biofilm activity followed by cinnamon, clove and rosemary oils. SEM and AFM analysis showed reduced adherence and roughness in the presence of EOs. For phospholipase, rosemary oil was the most inhibitory, followed by jasmine, cinnamon and clove oils, and for hemolysins, cinnamon had the highest inhibition followed by jasmine, rosemary and clove oils. A molecular docking study revealed major EO constituents as promising inhibitors of the Als3 adhesive protein, with the highest binding for eugenol, followed by 1,8-cineole, 2-phenylthiolane and cinnamaldehyde. In conclusion, EOs have a promising inhibitory impact on Candida biofilm, phospholipase and hemolysin production, hence EOs could be used as potential antifungals that impact virulence factors.

Action of disinfectant solutions on adaptive capacity and virulence factors of the Candida spp. biofilms formed on acrylic resin

Understanding the behavior of Candida spp. when exposed to denture disinfectants is essential to optimize their effectiveness. Changes in the virulence factors may cause increased resistance of Candida spp. to disinfectant agents. Objective To evaluate the microbial load, cellular metabolism, hydrolytic enzyme production, hyphae formation, live cell and biofilm quantification of Candida albicans, Candida tropicalis and Candida glabrata after exposure to disinfectant solutions.Methodology Simple biofilms were grown on heat-polymerized acrylic resin specimens, and divided into groups according to solutions/strains: distilled water (control); 0.25% sodium hypochlorite (NaOCl 0.25% ); 10% Ricinus communis (RC 10%); and 0.5% Chloramine T (CT 0.5%). The virulence factors were evaluated using the CFU count (microbial load), XTT method (cell metabolism), epifluorescence microscopy (biofilm removal and live or dead cells adhered), protease and phospholipase production and hyphae formation. Data were analyzed (α=0.05) by one-way ANOVA/ Tukey post hoc test, Kruskal-Wallis test and Wilcoxon test.Results NaOCl 0.25% was the most effective solution. CT 0.5% reduced the number of CFUs more than RC 10% and the control. RC 10% was effective only against C. glabrata. RC 10% and CT 0.5% decreased the cellular metabolism of C. albicans and C. glabrata. Enzyme production was not affected. Hyphal growth in the RC 10% and CT 0.5% groups was similar to that of the control. CT 0.5% was better than RC 10% against C. albicans and C. tropicalis when measuring the total amount of biofilm and number of living cells. For C. glabrata, CT 0.5% was equal to RC 10% in the maintenance of living cells; RC 10% was superior for biofilm removal.Conclusions The CT 0.5% achieved better results than those of Ricinus communis at 10%, favoring the creation of specific products for dentures. Adjustments in the formulations of RC 10% are necessary due to efficacy against C. glabrata. The NaOCl 0.25% is the most effective and could be suitable for use as a positive control.

Yeast microbiota of free-ranging amphibians and reptiles from Caatinga biome in Ceará State, Northeast Brazil: High pathogenic potential of Candida famata

ABSTRACT: Studies on the fungal microbiota of reptiles and amphibians are necessary to better understand of host-microbe interactions and the establishment of fungal disease in these animals. However, these studies are limited. The present researchidentified yeasts from free-ranging reptiles and amphibians from the Caatinga biome andevaluated the virulence factors production, the antifungal susceptibility in planktonic and biofilm growth and the pathogenicity of Candida famata isolates. Twenty-nine isolates of the genera Candida, Cryptococcus and Rhodotorula were identified by phenotypic and/or molecular methods and production of hydrolytic enzymes in vitro by these genera of fungi was evaluated. In addition, susceptibility of planktonic cells and biofilms to azoles and amphotericin B was evaluated. The pathogenicity of C. famata, the most prevalent yeast species isolated, was evaluated using Caenorhabditis elegans model. C. famata was the most prevalent yeast in amphibian and reptilian microbiota. Phospholipase and protease production was observed in 18/29 and 11/29 of the yeast isolates, respectively, while 100% formed biofilms. Itraconazole presented high minimal inhibitory concentrations against C. famata and C. tropicalis. Amphotericin B reduced the biomass and metabolic activity of biofilms. C. famata induced the mortality of C. elegans. In conclusion, reptiles and amphibians are colonized by yeasts capable of producing important virulence factors, especially by Candida spp. that present low susceptibility to azoles which may result from imbalances in ecosystem. Finally, C. famata isolated from these animals presented high pathogenicity, showing the importance of the study of reptile and amphibians fungal microbiota.

The genotypes and virulence attributes of C. albicans isolates from oral leukoplakia.

Background There is a debate as to whether some types of oral leucoplakias (OL) are caused by Candida species, and whether they contribute to the malignant transformation, associated with a minority of such lesions. As no detailed population analysis of yeast isolates from OL is available, we evaluated the virulence attributes, and genotypes of 35 C. albicans from OL, and compared their genotypes with 18 oral isolates from healthy individuals.Material and Methods The virulence traits evaluated were esterase, phospholipase, proteinase, haemolysin and coagulase production, and phenotypic switching activity, and yeast adherence and biofilm formation. DNA from OL and control yeasts were evaluated for A, B or C genotype status.Results Phospholipase, proteinase, and coagulase activity and biofilm formation was observed in 80%, 66%, 97 % and 77 % of the isolates, respectively. Phenotypic switching was detected in 8.6%, while heamolytic, and esterase activity and adherence were noted in all isolates.Conclusions The genotype A was predominant amongst both the OL and control groups. Due to the small sample size of our study a larger investigation to define the role of candidal virulent attributes in the pathogenicity of OL is warranted, and the current data should serve as a basis until then. Key words:C. albicans, oral cavity, leukoplakia, virulence factors, genotyping.

Chitinase involved in immune regulation by mediated the toll pathway of crustacea Procambarus clarkii

Chitinase can degrade chitin and play an essential role in animal immunity and plant defense. The immune functions of Chitinase in Procambarus clarkii (P. clarkii) remain to elucidate. Here, we identified PcChitinase 2 gene sequence from P. clarkii and studied its spatial and temporal expression profiles. The PcChitinase 2 transcribed unequally in different tissues; however, its expression was highest in those of stomach, gut, and hepatopancreas. The challenge with lipolysaccharide or peptidoglycan significantly up-regulated the expression of PcChitinase 2 in hepatopancreas. The knockdown of the PcChitinase 2 gene by double-stranded RNA suppressed most of the Toll-pathway-related immune genes (phospholipase, lectin, sptazle Cactus, serine proteikinase, anti-lipopolysaccharide factor, and Toll) production were significantly increased. Our results suggest PcChitinase 2 may be involved in the innate immune responses of P. clarkii by modulating the toll pathway.

Prevalence of virulence factors and antifungal susceptibility testing of Candida species isolated from clinical specimens

Introduction: The main aim of our study is to isolate and identify Candida species from clinical samples and to detect the virulence markers and also to determine their susceptibilities to various antifungal agents. Materials and Methods: Eighty Candida isolates from respiratory samples were included in the study. Detection of various virulence markers of candida spp such as Phospholipase, Proteinase, Hemolysin and Biofilm production was carried out by Phenotypic methods. Antifungal susceptibility testing of the isolated yeasts for Amphotericin B, Itraconazole, Fluconazole, Caspofungin and Voriconazole was determined by E-test methods. Results: In our study, most common spp isolated was Candida albicans(65%) followed by Candida tropicalis(15%) and Candida glabrata(10%).Prevalence percentage of virulence factors such as Hemolysin, Proteinase, Biofilm and Phospholipase production in Candida spp were 85%, 77.5%, 68.8% and 45% respectively. The susceptibility pattern of Candida isolates to antifungal agents showed highest resistant rate of 33.8% for Itraconazole and lowest resistant rate of 2.5% for Caspofungin. Conclusion: Presence of virulence factors in Candida species might indicate invasiveness and its relation with infection. Hence the significance of virulence determination and antifungal susceptibility testing should be adopted as a routine procedure in the laboratory. Keywords: Candida species, E-test, Hemolysin Biofilm formation, Proteinase, phospholipase activity.

A comparative study on production of extracellular hydrolytic enzymes of Candida species isolated from patients with surgical site infection and from healthy individuals and their co-relation with antifungal drug resistance

BackgroundSurgical site infection (SSI) is a crucial dilemma of surgery. Patients with SSIs not only face difficulty in treatment but also bear extra cost with high mortality rate. Resistant strains of Candida have emerged as an important nosocomial pathogen. Proteinase and phospholipase are exo- enzymes of Candida species, have importance with respect to their contribution in diseases. This study focused on prevalence of Candida species in surgical wound, their resistance to antifungal drugs, co-relation of these resistance with virulence potential of Candida species and comparison of production level of exo-enzymes of Candida species isolated from patients with SSIs and healthy individuals to highlights their role in SSIs.ResultsA total of (n = 555) swab samples were investigated. (n = 450) samples were collected from patients with SSIs and (n = 105) were collected from healthy individuals. Samples were subjected for the identification of Candida species which were subsequently investigated for antifungal susceptibility, MICs and enzymatic activity of Candida species. Out of 128 strains of Candida spp. isolated from SSIs, 54(42.18%) were identified as C. albicans followed by C. glabrata 32(25%), C. parapsilosis 17(13.28%), C. krusei 13(10.16%) and C. tropicalis 12(9.38%). C. albicans isolates showed 100% susceptibility to voriconazole and amphotericin B followed by itraconazole 98% and fluconazole 89%. Out of 6 fluconazole resistant C. albicans 5(83.33%) were able to produce phospholipase while out of 48 fluconazole-susceptible strains 17(35.42%) were found to be phospholipase producer. Out of 54 C. albicans isolated from surgical wound 46(85.18%) and 49(90.74%) were found to be phospholipase and proteinase producer respectively, whereas out of 20 C. albicans isolates from healthy subjects 14(70%) produce proteinase and 12(60%) produce phospholipase. There were significant statistical differences found between the level of enzyme production by C. albicans, in relation to both sites (P = 0.014).ConclusionStudy revealed that prevalence of Candida species is high in SSIs. Phospholipase and proteinase activity were more pronounced in Candida Species from surgical wound in contrast to species from healthy individuals suggests these enzymes may have been responsible for the severity of infection in surgical wound patients.

Prevalence, virulence and antifungal activity of C. albicans isolated from infected root canals

BackgroundThere is limited data on the prevalence of Candida species in infected root canal systems of human teeth. We attempted to investigate the prevalence, genotype, virulence and the antifungal susceptibility of Candida albicans isolated from infected root canals of patients with primary and post-treatment infections in a UAE population.MethodsMicrobiological samples from 71 subjects with infected root canals were aseptically collected, and cultured on Sabouraud dextrose agar, and C. albicans was identified using multiplex polymerase chain reaction, and the isolates were further subtyped using ABC genotyping system. Their relative virulence was compared using further four archival samples of endodontic origin from another geographical region, and four more salivary isolates, as controls. The virulence attributes compared were biofilm formation, and production of phospholipase and haemolysin, and the susceptibility to nystatin, amphotericin B, ketoconazole, and fluoconazole was also tested.Results4 out of 71 samples (5.6%) yielded Candida species. On analysis of variance among the groups, the intracanal isolates, mainly Genotype A, possessed a high degree of phospholipase and haemolysin activity (p < 0.05). The UAE and Finland isolates were stronger biofilm formers and had superior phospholipase production capacity compared with the salivary isolates. All isolates were sensitive to the antifungal chemicals used. The salivary isolates were more sensitive to fluoconazole compared to other groups (p < 0.05).ConclusionThe prevalence of C. albicans in infected root canals of patients attending a dental hospital in UAE is low. The strains isolated were good biofilm formers, possessed strong phospholipase and haemolysin activity and were mostly of the genotype A. The fact that the root canal isolates possessed significant hydrolase activity, imply that they are equipped with virulence attributes necessary for survival within a harsh intracanal ecosystem.

Invasive Factors Recognition in Aspergillus Section Nigri Isolates from Patient and Environmental Samples in the Centre Region, Cameroon

Background: Aspergillus section Nigri species are invasive opportunistic pathogens, seen in individuals with various immune disorders. The invasive capacity involves the production of varieties of enzymes such as lipases, phospholipase, proteases and hyaluronidase. The determination of proteinase, phospholipase, esterase and biofilm production in patient and environmental isolates approve the pathogenic strength of the species.&#x0D; Aims: To evaluate the invasive capacity of Aspergillus section Nigri isolates from patients and environmental samples.&#x0D; Methods: Our study is cross sectional and experimental, performed at the outpatient clinic of the otorhinolaryngology department of Central and University teaching hospital during a period of 12 months from March 2018 to February 2019. To determine the invasive capacity of Aspergillus section Nigri species, 400 samples were evaluated in the study (that is; 250 from patients and 150 samples from environment). Patient samples and hospital environment samples were evaluated by standard phenotypic methods for detecting of Proteinase, phospolipase, esterase and biofilm. The variables were statistically analyzed using Chi-square test of independent and SPSS (Version 16.0).&#x0D; Results: The isolates recovered from the patient sample shows maximum invasive capacity as compared to the environmental isolates, that is for 44 isolates; 42 isolates showed proteinase activity and biofilm production, followed by phospholipase activity 36, and then esterase 32.The isolates recovered from the hospital environment also showed the production of the various invasive factors, that is for 16 isolates; 15 isolates showed biofilm production, followed by proteinase activity 6, phospholipase 5 and esterase 4.The disparities of the invasive capacity in patient and environment isolates virulence were statistically significant for proteinase, phospholipase and esterase ( that is; p-value &lt;0.05). Majority the isolates recovered from patients and the environment were potential producers of biofilm.&#x0D; Conclusion: The isolates recovered from patients sample showed high invasive capacity as compare to the isolates recovered from the environment. This highlights the implications of phospholipase enzyme, proteinase enzyme, esterase enzyme and biofilm used by Aspergillus section Nigri isolates as means of survival in the host system.

Enzyme production by Candida albicans and Candida dubliniensis in periodontal HIV-positive patients receiving and not receiving antiretroviral therapy.

Candida dubliniensis (Cd) and Candida albicans (Ca) are the most frequently isolated yeasts in HIV+ patients. Some of the enzymes produced by these yeasts are considered virulence factors since they contribute to pathogenicity of Candida spp. The aim of the present study was to compare production of enzymes such as phospholipase (Ph), proteinase (P), and hemolysin (H) by Cd and Ca strains isolated from periodontal HIV-positive patients receiving and not receiving highly active antiretroviral therapy (HAART). Subgingival biofilm samples were obtained using paper points, and a sample of oral mucosa was taken using a swab. Phenotypic and molecular methods were used to isolate 39 strains of Candida, including 25 strains of Cd and 14 strains of Ca, obtained from 33 periodontal pocket samples and 6 oral mucosa samples collected from 15 HIV+ patients (8 receiving and 7 not receiving HAART). Malt egg-yolk agar, albumin agar and blood agar were used to evaluate pH, P and H production respectively. The strains were inoculated in duplicate and incubated at 37 ºC. Colony and halo diameters were measured. A greater proportion of Ca was observed in patients not receiving HAART, and a higher proportion of Cd was observed in those under HAART, Chi2 p&lt; 0.001. Phospholipase production was observed in 92.9% percent of isolated Ca strains but in none of the isolated Cd strains. Proteinase production was high in Ca and Cd strains isolated from patients not receiving HAART. Hemolysin production was observed in all the studied strains, though it was significantly higher (p=0.04) in Ca and Cd strains isolated from patients not receiving HAART. To sum up, the proportion of Candida dubliniensis strains was highest in the subgingival biofilm of patients receiving HAART, and Cd strains were found to express fewer virulence factors than Ca strains.

Assessment of different Enzymatic Virulence Traits contributing to pathogenicity of Candida Albicans

Background: Candidiasis is regarded as one of the frequently occurring fungal infections of humans elicited by yeast that belongs to the genus Candida. Among the different species of Candida, Candida albicans is the most pathogenic specie. The pathogenicity of this specie depends on various virulence factors. This study was designed to determine the activity of some important enzymatic virulence factors such as phospholipase, proteinase and hemolysin produced by Candida albicans strains.&#x0D; Methodology: A total of 300 clinical isolates of C. albicans were obtained and identified by conventional standard diagnostic criteria. For the evaluation of phospholipase production, the egg yolk agar medium was employed. The minimum agar medium was used for proteinase activity and for hemolysin production, blood plate assay was used. The results were statistically analyzed by SPSS 16.0 version software.&#x0D; Results: Of a total of 300 isolates reviewed, the majority of the isolates were recovered from females, i.e. 234 (78%); however, the number of isolates obtained from males were 66 (22%). Amongst all the strains of C. albicans, phospholipase and hemolysin enzymes were produced primarily by 89.3% of the total C. albicans strains. While proteinase enzyme was produced by 86% of the total C. albicans isolates. A p-value &lt; 0.05 was measured as statistically significant.&#x0D; Conclusion: From the existing study, it can be determined that these enzymes are produced by most of the pathogenic strains of C. albicans. The detection of these virulence enzymes is tremendously significant in designing of drugs to combat these opportunistic infections in future.

Prevalence of virulent Candida spp. in complicated urinary tract infection of nephrolithiatic patients from surgical units of tertiary care hospitals Islamabad

Candida species are the commensal organisms of human mucosa and opportunistically cause the diseases in susceptible persons. This study aimed to determine the prevalence and virulence of different Candida spp. among nephrolithiatic patients and their association with complicated UTI (cUTI). A total of 164 urine samples were collected from surgical units of two tertiary care hospitals (Poly Clinic and Pakistan Institute of Medical Sciences Hospital, Islamabad). From 74 kidney stone patients, 77 isolates of Candida spp. were confirmed through standard microbiological and molecular characterization. C. albicans was the predominant species with 51 isolates (66.2%) followed by 26 (33.8%) of C. non-albicans. The nephrolithiatic patients suffering from cUTI were more prone to be infected with Candida (P=0.047). Among all isolates, 83% (64) of the Candida isolates were biofilm formers, 80% (60) showed the esterase production and 64.9% (50) showed phospholipase production. Candida isolates positive for various virulence factors were more prevalently isolated from both catheterized and recurrent UTI patients. Among Candida spp., 16.9% (13) isolates showed resistance to fluconazole and 19.5% (15) against voriconazole and 11 isolates were resistant for both tested antifungals. Candida isolated from cUTI cases showed comparatively enhanced virulence attributes and antifungal resistance, suggesting that these factors might have role in development of cUTI in nephrolithiatic patients. Hence, this work highlights the high prevalence of both C. albicans and non albicans spp. in nephrolithiatic patients. So, there is need to administer evidence based antifungal therapy rather than empirical therapy to reduce the cUTI in nephrolithiatic patients.

Mezoneuron benthamianum inhibits cell adherence, hyphae formation, and phospholipase production in Candida albicans.

The aim of this study was to evaluate the phytochemical constituents, antioxidant, antifungal, and anti-virulence activities of traditionally used Mezoneuron benthamianum leaves. Extracts were prepared using acetone and methanol, and the preliminary phytochemical screening was performed. The antioxidant activity was studied using the DPPH method. Anti-Candida albicans activity was established and the effect on the germ tube and phospholipase production, as well as on the host cell adherence was assessed. The extracts showed the presence of anthraquinones, cardiac glycosides, flavonoids, reducing sugars, saponins, steroids, tannins, and terpenoids. Gallic acid and trans-resveratrol were among the predominant phytochemicals found in M. benthamianum. The crude extracts presented significantly higher antioxidant activity than the ascorbic acid standard. At 0.39mg/mL, acetone extract inhibited the growth of Candida albicans. At lower concentrations (200-50µg/mL), it significantly inhibited the adherence ability (up to 51%), formation of hyphae (up to 65%), and the production of phospholipase. In conclusion, at high concentrations, M. benthamianum kills C. albicans, and at lower concentrations, it can inhibit the virulence properties of this pathogen. This study on crude extract validates the traditional use of this plant. However, further research is required to establish the anti-virulence activity of the two compounds and their therapeutic potential.

Phenotypic and molecular detection of virulence factor genes SAP4 and PLB in Candida albicans isolates from the Western part of India

BackgroundSince the last two decades, substantial increase in fungal infections has been observed in immunocompromised hosts. Virulence factors of Candida albicans play a role in adherence, haemolytic activity, phenotypic switching and production of hydrolytic enzymes. The secreted aspartyl proteinases (SAP family) contribute to adhesion, tissue damage and invasion, while phospholipase (PLB) supports the hydrolysis of phospholipids. Very few studies showed the correlation of phenotypic activity and detection of genes contributing to the similar enzyme activity. Therefore, our study aimed at demonstrating correlation between in vitro phenotypic production of phospholipase and proteinase enzymes with genetic level detection of SAP and PLB genes. MethodsThe present study was carried out on a total of 799 samples over a period of one year. Culturing was carried out on Sabouraud's dextrose agar. Confirmation and speciation of Candida spp. was carried out using cornmeal agar and CHROMagar and by the germ tube test, urease test and automated identification system Vitek-2, and antifungal susceptibility testing was performed on Candida isolates. Phenotypic phospholipase and proteinase activity was analysed, and molecular detection of SAP4 and PLB genes was carried out. ResultsIn our study, we have screened 799 samples for mycological protocol; of which, 269 (33.6%) were Candida species, 44% (119) of which were C. albicans. Proteinase activity was exhibited by 77 (64.7%) and phospholipase activity was exhibited by 73 (61.34%) isolates, while 46.2% exhibited both activities among the C. albicans species. The PLB gene was detected in 97.3% isolates, while SAP4 was detected in 94.7% of C. albicans isolates. ConclusionThe study of in vitro expression of virulence factors and gene detection of C. albicans will help to improve the prognosis despite the nature of the sample.

Isolation and Diagnosis of Candida albicans Yeast From Patients Infected With Oral Candidiasis in Mosul City and Study its Activity in Production of Phospholipase and Hemolycin

In this study, 63 oral swabs were collected . Thirty six yeast isolates were obtained (57.14%) grown on sabouraud dextrose agar medium (SDA). Symbols were given to isolates from RA1 to RA36 in order to differentiate them. The isolates were identified by growth on HiCrome Candida differential agar M1297A and identification by Vitek 2 Compact system. The identification results showed 5 species of the isolated yeasts belonging to the genus Candida, 29 isolates belonging to the species C. albicans (80.55%) and one species from C. famata, C. glabrata, C. kefyr and C. lusitaniae (2.77% for each species). Moreover one isolate was obtained from each of the following yeasts: Saccharomyces serevisiae, Malassezia furfur and Rhodotorula glutinis (2.77% for each one). This study included the evaluation of two virulence factors for all C. albicans isolates , phospholipase enzyme production on egg yolk agar and hemolytic activity on sugar-enriched sheep blood agar. . Phospholipase production test showed differences in phospholipase production among the isolates , twenty three isolates (79.31%) were producers and precipitation zone (Opaque zone) was observed around the grown colonies with different Pz values and the isolate RA13 exhibited highest activity (Pz value = 0.53). The results showed also that all isolates were producers of hemolysin with different hemolytic indexes (Hi). Isolate RA7 represented the highest one in activity (Hi = 2.16). Hemolysis of all isolates involved beta hemolysis.

Study of Some Virulence Factors of Candida Albicans Causing Intestinal Infection

The incidence of systemic candidiasis which is caused by Candida albicans has increased so the present study sheds light on detecting some virulence factors of Candida albicans like production of phospholipases and proteinases, besides the adherence on epithelial cells (from human mouth) and drug tolerance were concerned. Proteinase enzyme was produced by 20 C.albicans isolates (47.62%) out of 42 isolates. On the other hand, 18 out of 42 (42.86%) isolates of C.albicans could produce the phospholipase enzyme. The ability of the isolates for attachment onto the surface of epithelial cells was studied and found that 39 isolates of C.albicans could adherence epithelial cells and 20 isolates recorded percentage 95- 98%. As the resistance to antifungals concerned, we found that 32 isolates of Candidawere resistant to nystatin. Amphotericin B was the most effective against all isolates of Candida. Microscopic examination results of histological section taken from rats’ intestine tissues that artificially infected with C.albicans which included degeneration, necrosis, hemorrhage.

Study of Some Virulence Factors of &lt;i&gt;Candida albicans&lt;/i&gt; Causing Intestinal Infection

The incidence of systemic candidiasis which is caused by Candida albicans has increased so the present study sheds light on detecting some virulence factors of Candida albicans like production of phospholipases and proteinases, besides the adherence on epithelial cells (from human mouth) and drug tolerance were concerned. Proteinase enzyme was produced by 20 C. albicans isolates (47.62%) out of 42 isolates. On the other hand, 18 out of 42 (42.86%) isolates of C. albicans could produce the phospholipase enzyme. The ability of the isolates for attachment onto the surface of epithelial cells was studied and found that 39 isolates of C. albicans could adherence epithelial cells and 20 isolates recorded percentage 95- 98%. As the resistance to antifungals concerned, we found that 32 isolates of Candida were resistant to nystatin. Amphotericin B was the most effective against all isolates of Candida . Microscopic examination results of histological section taken from rats’ intestine tissues that artificially infected with C. albicans which included degeneration, necrosis, hemorrhage.

Relationship between antifungal susceptibility profile and virulence factors in Candida albicans isolated from nail specimens.

INTRODUCTION: The aim of this study was to evaluate some virulence factors in Candida albicans isolates from patients with onychomycosis and determine the correlation between these factors and the antifungal resistance profile. METHODS: Seventy species of C. albicans were confirmed using polymerase chain reaction amplification of the HWP1 gene. According to the Clinical & Laboratory Standards Institute guidelines, the susceptibility profile of four antifungal agents was investigated, and the production of aspartyl protease, phospholipase, haemolysin, and biofilm was determined. The correlation between these profiles was also investigated.RESULTS: The isolates indicated different levels of resistance and production of virulence factors. Significant correlations were observed between the minimum inhibitory concentration (MIC) of fluconazole/itraconazole and biofilm production, between phospholipase production and fluconazole/itraconazole MIC, and between fluconazole MIC and hemolytic activity in C. albicans isolates. The results also showed significant correlations between phospholipase activity and biofilm production.CONCLUSIONS: Our findings will contribute to a better understanding of the pathogenesis of C. albicans and characterize the relationship between virulence factors and antifungal resistance, which may suggest new therapeutic strategies considering the possible involvement of the virulence mechanism in the effectiveness of treatment.

Newly Isolated Lipolytic and Oleaginous Fungal Strain, Production, Optimization and Biochemical Characterization of the Extracellular (phospho)lipase

The present study investigates the biochemical characterization of an extracellular (phospho)lipase from a wood fungus Peziza sp. (medium optimization, inducer concentration and substrate specificity measurements). The strain was identified on the basis of ITS1/ITS4 primers. A 604 bp fragment was amplified by PCR and the obtained nucleotide sequence, showed 99% identity with the ITS region of isolates named as Peziza sp. Interestingly, Peziza sp. has both lipase and phospholipase activities with the same level which require both the presence of Ca2+ and bile salts. Our result shows that the lipase hydrolyzes preferably the olive oil at 45 °C, pH 8. Whereas, the phospholipase activity was detected on pure PC at 45 °C, pH 9. Lipid extraction from dry biomass using chloroform/methanol (2/1) and quantitative measurement using electron microscope showed that intracellular triglycerides content was significantly high and reaches 20.88%. Gas chromatography analysis shows a majority of C18:1 (76, 98%) and C18:2 (9, 33%). Whereas, saturated fatty acids ranging C16–C20 represent only 11.5% of total lipids composition. Lipid accumulation test in Pezizales sp under fluorescence microscope