Candida Famata Research Articles

Indigenous Yeast, Lactic Acid Bacteria, and Acetic Acid Bacteria from Cocoa Bean Fermentation in Indonesia Can Inhibit Fungal-Growth-Producing Mycotoxins

Cocoa bean fermentation is an important process in the manufacturing of cocoa products. It involves microbes, such as lactic acid bacteria, yeast, and acetic acid bacteria. The presence of mold in cocoa bean fermentation is undesired, as it reduces the quality and may produce mycotoxins, which can cause poisoning and death. Aspergillus niger is a fungus that produces ochratoxin A, which is often found in dried agricultural products such as seeds and cereals. In this study, we applied indigenous Candida famata HY-37, Lactobacillus plantarum HL-15, and Acetobacter spp. HA-37 as starter cultures for cocoa bean fermentation. We found that the use of L. plantarum HL-15 individually or in combination Candida famata HY-37, Lactobacillus plantarum HL-15, and Acetobacter spp. HA-37 as a starter for cocoa bean fermentation can inhibit the growth of A. niger YAC-9 and the synthesis of ochratoxin A during fermentation and drying. With biological methods that use indigenous Lactobacillus plantarum HL-15 individually or in combination with Candida famata HY-37 and Acetobacter spp. HA-37, we successfully inhibited contamination by ochratoxin-A-producing fungi. Thus, the three indigenous microbes should be used in cocoa bean fermentation to inhibit the growth of fungi that produce mycotoxins and thus improve the quality.

Isolation and Identification of Yeasts in White Cheese

In this study, firstly yeast was isolated from 50 white cheese samples produced by traditional methods from raw milk. The yeast species were isolated from 50 traditional white cheese samples and identified with the VITEK2 Compact system. There were obtained from white cheese samples 90 isolates 35.6% Candida sake (32), 13.3% Candida zeylanoides (12), 8.9% Candida famata (8) and 8.9% Candida kefyr (8) and the remaining 33.3% Candida sphaerica (4), Candida colliculasa (2), Candida boidinii (2), Candida lusitaniae (4), Candida parapsilosis (2), Candida sphaerica (4) Cryptococcus laurentii (4), Candida krusei (4) and Saccharomyces cerevisiae (4). Yeasts are known as a saprophyte found in many foods, but that some of them positively contribute to the fermentation. More over some yeasts are could used of starter culture during the manufacture of certain dairy product for maturation by supporting the functions. some yeasts species were isolated in this study, could be used as support culture.

Comparative Study of Antimicrobial Activity and Cytotoxicity of Saponin Extracted from Six Egyptian Sea-cucumber Species, Licorice, and Ginseng

Background: The study aimed to compare the antimicrobial activity and cytotoxicity of saponin extracted from six sea-cucumber species, licorice, and ginseng to determine the most potent one. Methods: Antimicrobial activity assessed by well diffusion and agar dilution methods. The inhibitory effects of the tested extracts on cell growth of Liver (HePG-2) and breast (MCF-7) cell lines assessed by MTT [3- (4, 5- dimethyl thiazol-2yl) - 2, 5- diphenyl tetrazolium bromide] assay. Results: The results revealed that licorice extract presented higher antibacterial activity against Gram-positive bacterial strains than sea-cucumber extracts; but has comparable results for Gramnegative (Salmonella tyhimurium, Klebsiella spp, and Acinetobacter baumanii). Holothuria atra body wall and Bohadschia tubules have the highest antifungal effect against Candida albicans, Candida krusei, Candida famata, Candida glabrata, and Candida tropicalis. Sea-cucumbers are more potent against fungal strains than licorice and ginseng extract. Licorice and ginseng presented higher cytotoxicity against HePG2 and MCF-7 cell lines than sea-cucumbers. Conclusion: This study promotes the use of licorice extract as a better antibacterial substance than sea-cucumber extracts; however, the use of sea-cucumber extracts as antifungal agents than herbal extracts. Also, we support the utilization of licorice and ginseng as a cytotoxic agent instead of seacucumbers to stop the fishing of animals and consequently save it from extinction.

The effect of Candida famata and Lactobacillus plantarum on the number of coliforms and the antibiotic resistance and virulence of Escherichia coli in the gut of broilers

This study was undertaken to determine the effect of a yeast (Candida famata) and a bacterium (Lactobacillus plantarum), administered alone or in combination in the drinking water, on the population of yeast, Lactobacillus sp. and coliforms, and the prevalence of antimicrobial resistance (AMR) and virulence genes in Escherichia coli (E. coli) isolated from digesta samples taken throughout the life of broiler chickens. Male (Ross 308) day-old chicks (220) were used. C. famata (isolated from a chicken) and L. plantarum (isolated from a pig) were administered via the drinking water. Water was provided either untreated or with C. famata (CF; 108/ml), L. plantarum (LP; 105–108/ml), or a combination of CF and LP (106–108/ml) in water hoppers on 2 days each week for 35 days. Administering probiotics did not affect the growth performance in broiler chickens. No significant interactions were observed between main effects, and neither CF nor LP had any effect on the population size of Lactobacillus sp. or coliforms. The administration of C. famata increased the population density of yeasts in the small intestine at these ages. The population density of coliforms, Lactobacillus sp. and yeast decreased with age (P < 0.001). There was no significant effect of probiotics on the prevalence of phenotypic AMR and virulence genes in these studies. The prevalence of E. coli that was resistant to ampicillin and tetracycline, as well as carrying ≥3 virulence-associated genes, was greatest at the end of the starter phase (around 8 days old), before declining through the grower and finisher phases. There was only limited evidence that administering either CF or LP affected either the AMR or the virulence of E. coli in the bird. However, tetracycline resistance in E. coli was associated (P < 0.001, P < 0.01, P < 0.05, and P < 0.05) with the carriage of the iron uptake systems of E. coli D, iron-repressible protein, increased serum survival and temperature-sensitive haemagglutinin genes respectively, suggesting that the accumulation of iron and the genetic element conferring tetracycline resistance may be intertwined.

Identification of yeasts in different rubber leaf (Hevea brasiliensis) clones and their effects on the physical properties of fermented glutinous rice tapai

Tapai is one of the most popular traditional desserts in Malaysia and other Asian countries. Traditionally, tapai is wrapped in a rubber leaf to enhance the smell and increase its palatability. The study focused on identifying the yeasts present before and after the production of glutinous rice tapai wrapped in different rubber leaves clones, namely RRIM 2025, RRIM 2002, PB 260 and PB 350. The identification of the yeast was carried out using API 20C AUX test strips for all rubber leaves clones, glutinous rice tapai wrapped in RRIM 2025, RRIM 2002, PB 260, PB 350 and in a container (control). The results showed that Crytococcus laurentii, Rhodotorula mucilaginosa 2, Candida famata, Rhodotorula minuta were present in rubber leaf clones. While the yeasts that had been identified in tapai wrapped in rubber leaf were Candida guilliermondii, Rhodotorula mucilaginosa 2, Candida parapsilosis and Trichosporon mucoides and only C. guilliermondii was found in the container. The physical properties of the tapai that are wrapped in rubber leaves have a difference in texture, pH value and total soluble solids content compared to the control sample. The tapai sample wrapped in RRIM 2025 and RRIM 2002 had a high total soluble solid content of 45.8±0.14% and 45.78±0.16% °Bx, respectively. Meanwhile, the control sample has the highest pH value and the hardest rice kernels, which were 4.71±0.05 and 218.19±25.39 N, respectively. The results showed that the different yeasts present in the rubber leaf may cause changes in the physical properties of glutinous rice tapai.

BIOFILM PRODUCTION AND ANTIMICROBIAL RESISTANCE IN VENTILATOR ASSOCIATED PNEUMONIA (VAP) PATHOGENS OF ICUs OF TERTIARY CARE CENTER OF CENTRAL GUJARAT.

Background: In critical care units, Ventilator-associated pneumonia (VAP) is a common device-associated infection in mechanically ventilated patients. Problem gets worst of associated with biofilm producing organism with higher antimicrobial resistance. The current study was carried out to observe the pattern of antimicrobial resistance, biofilm forming capacity of isolates causing Ventilator-associated pneumonia and other risk factors associated with VAP patients in intensive care units of Shree Krishna Hospital, Karamsad.&#x0D; Methodology: 97 total tracheal aspirate culture isolates recovered from 83 mechanically ventilated patients diagnosed to be suffering from VAP as per NHSN definition, admitted in various ICUs of Shree Krishna Hospital, Karamsad during the study duration were included in the study. Relevant clinical history of the patients and other details taken for various patient variable factors like age, gender, co-morbid conditions, indoor days, ventilator days, final patient outcome and other lab based investigations done as indicator of active pneumonia or sepsis from the electronic hospital database available on hospital information system. The tracheal aspirate culture isolates were then tested for antimicrobial susceptibility testing by Vitek2compact and in-vitro biofilm production assay using microtitre plate method. Objective of the present study was to determine the incidence of antimicrobial resistance, biofilm forming capacity of VAP pathogens, to determine risk factors associated and final outcome in VAP patients infected with biofilm forming pathogens. Chi-square test was used to check the relation between the categorical variables while t test was applied in case of continuous variables. A p value less than 0.05 was considered as statistically significant.&#x0D; Results: Out of total 83 patients of VAP, 97 isolates recovered in tracheal aspirate culture. Out of total 83 patients, 42 patients (49 isolates) were found Biofilm producer (BFP) and 41 patients (48 isolates) were found Biofilm non-producer (BFNP). Out of 97 culture total isolates, the most common organisms grew were Klebsiella pneumoniae (29 isolates), Acinetobacter baumani (28 isolates) and Pseudomonas aeruginosa (19 isolates) apart from them lesser number of isolates of Staphylococcus aureus (6), Escherichia coli (5), Pantoea spp. (2), Serretia marcescens (2), Pseudomonas putida (1), Sphingomonas paucimobilis (1), Stenotrophomonas maltophila (1), Enterococcus faecium (1), Candida famata (1) and Candida tropicalis (1). The antimicrobial resistance was compared in three major pathogen between BFP and BFNP isolates, i.e. Klebsiella pneumoniae, Acinetobacter baumani and Pseudomonas aeruginosa, which was found to be statistically insignificant. Mortality was recorded higher in BFP patients (16.67%) compared to BFNP patients (7.3%) of VAP, but statistically it was not found to be significant (p value &gt; 0.05).&#x0D; Conclusions: Incidence of BFP and BFNP associated VAP seen 50.51% and 49.49% respectively out of total 97 isolates. Biofilm forming pathogen causing VAP may not influence the outcome of the patient but, biofilm producer pathogens continue to be associated with pathogens causing VAP in significant amount of total cases. Typical hospital acquired strains like Klebsiella pneumoniae, Acinetobacter baumani and Pseudomonas aeruginosa is recorded frequently compared to other pathogens.&#x0D; Key words: Intensive Care Unit, anti-microbial resistance, VAP, Bio film, Health care associated infection, Indwelling device associated infection.

Raw bovine milk as a reservoir of yeast with virulence factors and decreased susceptibility to antifungal agents.

We identified 66 yeast isolates, including 26 different yeast species from 910 individual milk samples. Our results indicate that individual milk samples may serve as a source of yeasts with the potential to trigger infection and may have reduced sensitivity to tested antifungal agents.

Strategies to Increase the Production of Biosynthetic Riboflavin.

Riboflavin is widely regarded as an essential nutrient that is involved in biological oxidation in vivo. In addition to preventing and treating acyl-CoA dehydrogenase deficiency in patients with keratitis, stomatitis, and glossitis, riboflavin is also closely related to the treatment of radiation mucositis and cardiovascular disease. Chemical synthesis has been the dominant method for producing riboflavin for approximately 50years. Nevertheless, due to the intricate synthesis process, relatively high cost, and high risk of pollution, alternative methods of chemical syntheses, such as the fermentation method, began to develop and eventually became the main methods for producing riboflavin. At present, there are three types of strains used in industrial riboflavin production: Ashbya gossypii, Candida famata, and Bacillus subtilis. Additionally, many recent studies have been conducted on Escherichia coli and Lactobacillus. Fermentation increases the yield of riboflavin using genetic engineering technology to modify and induce riboflavin production in the strain, as well as to regulate the metabolic flux of the purine pathway and pentose phosphate pathway (PP pathway), thereby optimizing the culture process. This article briefly introduces recent progress in the fermentation of riboflavin.

Phytochemical screening and anti-candida activities of Crocus cancellatus herb. Ethanol extract

The antifungal, antioxidant activity, total tannin estimation, and phytochemical screening of Crocus Cancellatus extract were investigated. The best yield was obtained with ethanol solvent (36.1%). A significant amount of tannin was obtained in the plant (11.1%). In Antifungal testing against Candida spp. Results were obtained between 8.22 and 13.16 mm. The minimum and maximum results were obtained against Candida Krusei (8.22 mm) and Candida glabatra (13.16 mm) respectively. The minimum inhibitory concentration (MIC) value obtained against all candida strains resulted in the range of 12.5-25 μg/ml. The minimum inhibitory zone 0 mm of FLU/25 was obtained against Candida famata While the highest inhibitory zone of FLU/25 38.40 mm was obtained against Candida guilliermondii. In the antioxidant study, the maximum value (98.68%) was obtained at 0.2 mL, while the minimum value was obtained at 0.1 mL (96.60%). 10 compounds were determined in the content determination study with LC-MS/MS. Gallic acid (46965.14 µg/g), Quinic acid (1935.71 µg/g), and malic acid (3831.37µg/g) was obtained at the highest levels. Vanillic acid (138.2 µg/g), Protocatechuic acid (116.87µg/g), tr-Ferulic acid (51.17µg/g), Quercetin (20.56 µg/g), p-coumaric acid (12.8 µg/g), Chlorogenic acid (2.51 µg/g) and Salicylic acid (1.56 µg/g) were obtained quantitatively less. As a result of all these results, it was determined that the antioxidant activity of the plant is effective on pathogenic species thanks to its phenolic compounds.

PROPOLIS: ANTIMICROBIAL ACTIVITY AND CHEMICAL COMPOSITION ANALYSIS

Over the last few years, propolis has been the object of many studies conducted around the world, and its biological properties and chemical composition have been widely investigated. The present study focuses on the evaluation of the antimicrobial activity as well as an examination of the chemical composition of two samples of propolis from Eastern Algeria coming from the commune of El Mechrouha and Ouled Driss in the wilaya of Souk-Ahras. The two samples are tested for their antimicrobial power by undertaking the agar diffusion technique on eight pathogenic microbial strains (six bacterial strains and two fungal strains) which are: Escherichia coli, Klebsiella pneumonia, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus thoraltensis, , , Candida famata and Aspergillus niger. The results obtained clearly show the impact of propolis on the microbial susceptibility of Gram-positive bacteria (S. agalactiae and S. aureus), as well as on fungal species (C. famata and A. niger). The analysis of the chemical composition of the ethanolic extracts of the two propolis by UV-visible absorption spectrometry and thin layer chromatography showed that Algerian propolis is wealthy in phenolic compounds, and high performance liquid chromatography allowed the identification of four polyphenols (Gallic acid, Caffeic acid, Quercetin and Catechin). These outcomes permitted a first assessment of the two propolis which present comparable components in their chemical compositions.

Study of Yeasts Characteristics Isolated from the Fermented Peelings of Yams: Research of New Sources of Fermentative Strains

Yeast strains were isolated in this study from the perspective of fermentation technology. For this study, fermented peelings of three varieties of yam namely “Bete-bete”, “Kponan” and “Krengle” were analyzed. The charge of yeasts were 124000 UFC, 3200 CFU, and 118000 CFU respectively for the fermented peelings of the varieties “Bete bete”, “Kponan” and “Krengle”. Five species were identified by VITEK® 2 Systems method. Indeed, strains of Candida ciferrii, Candida famata, Candida lusitaniea, Cryptococcus laurentii and Trichosporon mucoides were isolated and identified. However, the predominance species were Candida ciferrii, and Candida famata. The strains of all the yeasts species had positive assimilation for the majority of carbon and nitrogen compounds tested. These yeasts showed also activities mainly for leucine-arylamidase, beta-N-acetyl-glucosaminidase, gamma-glutamyl-transferase, PNP-N-acetyl-BD-galactosaminidase 1 and alpha-glucosidase. The characteristics of these yeasts show thus great perspective of fermentation technology. Our work is the first ever on yeast diversity from fermented peelings of yams.

Fermentative Potential of Native Yeast Candida famata for Prokupac Grape Must Fermentation

The fermentative potential of native Candida famata isolates from wild and cultivated blackberries was evaluated for potential application in Prokupac grape must fermentation. 5 isolates, out of a total 22 isolated yeasts, were identified as C. famata. After the initial screening of fermentative performances, microfermentation was performed in a sterile grape must. Produced samples were analyzed using the HPLC technique. All isolates showed an ability to grow at lower temperatures, good tolerance to 7% ethanol and 300 ppm of SO2. C. famata isolates WB-1, WB-2 and W-5 had similar fermentation performance, but WB-1 isolate was chosen for validation at a laboratory-scale level according to a pleasant, fruity aroma, highest fermentative vigor and power, good organic acid profile and the highest level of ethanol and glycerol produced in micro-vinification experiments. Good enological performance of selected C. famata WB-1 isolate is confirmed by higher level of glycerol, lower level of ethanol and acetic acid in wine samples produced in pure and sequential fermentation, when compared to the control sample. Throughout the selection of C. famata yeasts with good enological potential, this work gives a contribution in the area of precision enology, aiming to find a perfect match between non-exploited yeasts and “autochthonous” grape cultivar Prokupac.

MALDI-TOF MS Overcomes Misidentification of the Uncommon Human Pathogen Candida famata by Routine Phenotypic Identification Methods.

Candida famata has been associated with the identifiable Candida infections that takes place in human and the identification error of this species possibly will result in misinterpretation of antifungal susceptibility and improper diagnosis; which will have a major effect on the prognosis and therapy of patients. Our objective is to correctly identify Candida spp. collected from patients at the intensive care units, New Cairo University teaching hospital in Cairo-Egypt using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). Hundred clinically isolated yeast strains were identified using API 20C AUX obtained from patients receiving care at intensive care units. ATB FUNGUS 3strips were used to detect the minimum inhibitory concentration. Thirty-three non duplicate strains identified as C. famata were subjected to re-identification by MALDI-TOF MS. Our results revealed that isolates were initially identified as C. famata 33%, C. tropicalis 15%, C. albicans12% and C. parapsillosis 10% using the phenotypic techniques. MALDI-TOF MS analyses results showed that the 33 C. famata isolates are C. tropicalis (n = 29), Trichosporon asahii (n = 2), C. parapsilosis (n = 1), and Aeromonas sobria (n = 1). Antifungal resistance was low in the Candida species, except for reduced susceptibility to itraconazole among C. krusei strains. This report shows that misidentification of C. famata is frequent when using conventional phenotypic methods of identification which result in challenges in treating fungal infections. MALDI-TOF MS is an accurate convenient substitute to classical approaches for fungal identification. In general, antifungal multidrug resistance is uncommon in our studied Candida species and yeast isolates.

CONTRIBUTION OF THE MYCOLOGY LABORATORY IN THE MANAGEMENT OF ORAL CANDIDOSIS IN IMMUNODEPRESSED PATIENTS

The authors report a series of 49 cases of oral candidiasis including 20 symptomatic, listed on 100 immunocompromised patients collected at the Mohammed V Military Hospital in Rabat over a 12-month period. The objective of this work was to define the risk factors that lay the foundation for fungal proliferation in the mouth, through early detection in asymptomatic or non-asymptomatic patients. Mycological analysis in the laboratory was based on direct examination and culture on Sabouraud chloramphenicol medium with and without cycloheximide, then identification of the fungal species by API 20 C AUX galleries and the VITEK 2 compact® . The prevalence of oral candidiasis was 49%. The mean age of the patients was 54 years with a sex ratio M / F of 1.04. The contributing factors identified were hyposialia (p = 0.0337), corticosteroid therapy (p = 0.025 and dental removable prostheses (p = 0.000791). The fungal species identified were Candida albicans (79%), Candida dubliniensis (7%), Candida ciferrii (4%), Candida famata (4%), Candida glabrata (4%) and Candida lusitaniae (2%). Conclusion: The oral localization of candidiasis remains very frequent in immunocompromised subjects. Their treatment involves first of all the search for contributing factors and early detection in the presence of asymptomatic forms that only mycological analysis can identify the variety.

Distribution of clinical Candida species and their susceptibility to antifungal agents

Background: The incidence of fungal infections, especially by Candida species, has increased in recent years. This study was designed to isolate and identify Candida species from various clinical samples, and to examine their susceptibility to antifungal agents. Methods: A total of 175 Candida species were isolated from different clinical samples, and were identified using germ tube test, Cornmeal agar, and API C 20 AUX and VITEK 2 Compact Systems. Antifungal susceptibility of these isolates was determined using ATB Fungus 3 strip and VITEK 2 antifungal susceptibility cards. The results of these two methods were comparatively assessed. Results: A total of 175 Candida strains were isolated from 114 (65.1%) urine, 42 (24%) blood, eight (4.6%) sputum, seven (4%) endotracheal, and four (2.3%) wound samples. Of isolated Candida species, 102 (58.2%) were Candida albicans, 31 (17.6%) Candida tropicalis, 17 (9.6%) Candida parapsilosis, 13 (7.4%) Candida glabrata, three (1.8%) Candida kefyr, three (1.8%) Candida krusei, two (1.2%) Candida lusitaniae, and one (0.6%) Candida famata. By VITEK 2 antifungal susceptibility cards, the overall resistance rates were 0.6% to each of amphotericin B and 5-flucytosine, and 1.8% to fluconazole. In ATB Fungus 3 strip, the resistance (1.8%) was observed only to fulconazole. All isolates were uniformly susceptible to voriconazole in both methods. Conclusion: Valuable information and data on distribution and susceptibility of Candida strains were obtained. These data may be valuable from epidemiological point of view as well as for proper and optimal therapy of Candida infections in our region (Şanliurfa, Turkey).

Kandidemi Etkenlerinin Tür Dağılımı ve Duyarlılıkları: Hastanemizde Ampirik Antifungal Tedavi Politikası Değiştirilmeli mi?

Objective: In recent years, the incidence of invasive Candida infections have increased due to the increased use of broad-spectrum antibiotics, the number of patients with immunosuppressive therapy and in the intensive care unit with the impaired condition. Knowing the species distribution and antifungal susceptibilities in Candida infections are important in managing the treatment of these infections with high mortality and morbidity. In this study, it was aimed to determine the distribution and antifungal sensitivities of Candida species that isolated from blood cultures taken from patients hospitalized in various clinics in our hospital and evaluation of the antifungal treatment policy applied in our hospital according to these results. Method: Using gradient test strips (BioMérieux E test, France) of Candida spp. grown in blood cultures (BacT/Alert 3D, BioMérieux, France), and identified by conventional methods and automated identification systems [API ID 32C, (BioMérieux, France) and MaldiTOF MS, (Bruker Daltonik GmbH, Bremen, Germany)] between July 2017 and March 2019, and their susceptibilities to antifungal agents were determined). Results: Among the 175 Candida strains isolated, 84 (48%) were Candida parapsilosis, other identified strains were as follows; Candida albicans (n=57; (32.6%), Candida glabrata (n=15; 8.6%), Candida tropicalis (n=12; 6.9%), Candida kefyr (n=3; 1.7%), one Candida dubliniensis (n=1; 0.6%), Candida famata (n=1; 0.6%), Candida lusitaniae (n=1; 0.6%) and Candida spp. (n=1; 0.6%). Antimicrobial resistance rates of isolated C. parapsilosis strains against fluconazole (54.8%), voriconazole (44.1%), posaconazole (7.1%), itraconazole (25%) and anidulafungin (1.2%), and of C. albicans against fluconazole (8.8%), voriconazole (7%), itraconazole (15.8%) were determined as indicated. Conclusion: As a result, it is noteworthy that C. parapsilosis is the most frequently isolated infection agent in the blood cultures of patients hospitalized in our hospital and that especially C. parapsilosis species show high fluconazole resistance.

Recent Advances in Construction of the Efficient Producers of Riboflavin and Flavin Nucleotides (FMN, FAD) in the Yeast Candida famata.

The approaches used by the authors to design the Candida famata strains capable to overproduce riboflavin, flavin mononucleotide (FMN), and flavin adenine dinucleotide (FAD) are described. The metabolic engineering approaches include overexpression of SEF1 gene encoding positive regulator of riboflavin biosynthesis, IMH3 (coding for IMP dehydrogenase) orthologs from another species of flavinogenic yeast Debaryomyces hansenii, and the homologous genes RIB1 and RIB7 encoding GTP cyclohydrolase II and riboflavin synthase, the first and the last enzymes of riboflavin biosynthesis pathway, respectively. Overexpression of the abovementioned genes in the genetically stable riboflavin overproducer AF-4 obtained by classical selection resulted in fourfold increase of riboflavin production in shake flask experiments.Overexpression of engineered enzymes phosphoribosyl pyrophosphate synthetase and phosphoribosyl pyrophosphate amidotransferase catalyzing the initial steps of purine nucleotide biosynthesis enhances riboflavin synthesis in the flavinogenic yeast C. famata even more.Recombinant strains of C. famata containing FMN1 gene from D. hansenii encoding riboflavin kinase under control of the strong constitutive TEF1 promoter were constructed. Overexpression of the FMN1 gene in the riboflavin-producing mutant led to the 30-fold increase of the riboflavin kinase activity and 400-fold increase of FMN production in the resulting recombinant strains which reached maximally 318.2mg/L.FAD overproducing strains of C. famata were also constructed. This was achieved by overexpression of FAD1 gene from D. hansenii in C. famata FMN overproducing strain. The 7- to 15-fold increase in FAD synthetase activity as compared to the wild-type strain and FAD accumulation into cultural medium were observed. The maximal FAD titer 451.5mg/L was achieved.

Candida parapsilosis: A rare cause of arthritis and its successful treatment with oral fluconazole

Fungal arthritis is a rare condition, often missed and underreported. Among all Candida species, Candida parapsilosis is a rare cause. A 66-year-old male with a history of arthritis on and off for the past 3 years and treated symptomatically presented with acute pain and swelling of the left knee joint for 1 week. Synovial fluid examination revealed yeast-like structures and fungal culture showed growth of Candida spp. VITEK 2 compact identified it as Candida famata but matrix-assisted laser desorption ionization–time of flight mass spectrometry assay confirmed it as C. parapsilosis. This is a rare case of nonalbicans candida arthritis, especially in this part of the country.

Prevalence, Species Distribution and Antifungal Susceptibility Profile of &amp;lt;i&amp;gt;Candida&amp;lt;/i&amp;gt; Species Isolated from Bloodstream of Critical Care Unit Patients in a Tertiary Care Hospital in Kenya

The upsurge of candidemia in the past years has been an immense encumbrance on public health and the number of deaths caused by candidemia particularly in critical care unit patients is devastating. Candida species harbor a 30% - 60% mortality rate and compared to stable people or those with less serious illnesses, this ranges from 60% to 80% of those who are chronically ill patients. Grounded on a recent report from a tertiary care hospital in Kenya showing the emergence of previously unobserved species: Candida auris, this study aimed to determine the prevalence, species distribution, and antifungal susceptibility profile of candidemia in critical care unit patients of the hospital. 378 Critical Care Unit patients were enrolled for the study from January 2019 to January 2020. Positive archived isolates were sub-cultured using Saboraud Dextrose Agar. Candida species were identified utilizing API20C AUX and Vitek-2. Antifungal susceptibility testing was conducted using the Liofilchem MIC Test strip. Out of 378 patients, thirty-one presented a positive culture for Candida species. The prevalence of Candidemia was 8.2% with 9 (29.03%) Candida auris, 8 (25.81%) Candida albicans, 6 (19.35%) Candida parapsilosis, 3 (9.68%) Candida famata, 3 (9.68%) Candida tropicalis, 1 (3.23%) Candida duobushaemolumonii, and 1 (3.23%) Candida lusitaniae. A resistance pattern to Fluconazole was observed among Candida auris and Candida parapsilosis, and resistance to Flucytosine was observed in Candida tropicalis, whereas susceptible MIC values were obtained for the other drugs. There is an increase in candidemia among critical care unit patients in the health facility posing a public health challenge. Moreover, the onset of new species Candida auris which is unprecedented in Kenya warrants enhanced infection control, and the uniform resistance of Candida auris, Candida parapsilosis, and Candida tropicalis towards Fluconazole and Flucytosine necessitate constant drug monitoring for empirical treatment regime. In contrast, the high potency of Echinocandins and Amphotericin-B demonstrate them as the drug of choice.

Overexpression of Riboflavin Excretase Enhances Riboflavin Production in the Yeast Candida famata.

Many microorganisms are capable of riboflavin oversynthesis and accumulation in a medium, suggesting that they efficiently excrete riboflavin. The mechanisms of riboflavin efflux in microorganisms remain elusive. Candida famata are representatives of a group of so-called flavinogenic yeast species that overproduce riboflavin (vitamin B2) in response to iron limitation. The riboflavin overproducers of this yeast species have been obtained by classical mutagenesis and metabolic engineering. Overproduced riboflavin accumulates in the cultural medium rather than in the cells suggesting existence of the special mechanisms involved in riboflavin excretion. The appropriate protein and gene have not been identified in yeasts till recently. At the same time, the gene BCRP (breast cancer resistance protein) has been identified in mammal mammary glands. Several homologs of the mammal BCRP gene encoding putative riboflavin efflux protein (excretase) were identified in the flavinogenic yeasts Debaryomyces hansenii and C. famata. Here we evaluate the yeast homologs of BCRP with respect to improvement of a riboflavin production by C. famata. The closest homologs from D. hansenii or C. famata were expressed under the control of TEF1 promoter of these yeasts in the wild-type and riboflavin-overproducing strains of C. famata. Resulted transformants overexpressed the corresponding genes (designated as DhRFE and CfRFE) and produced 1.4- to 6-fold more riboflavin as compared to the corresponding parental strains. They also were characterized by overexpression of RIB1 and RIB6 genes which encode the first and the last structural enzymes of riboflavin synthesis and exhibited elevated specific activity of GTP cyclohydrolase II. Thus, overexpression of yeast homolog of mammal gene BCRP may be useful to increase the riboflavin yield in a riboflavin production process using a recombinant overproducing C. famata strain or other flavinogenic microorganisms.