Identification Of Fungal Isolates Research Articles

Aspergillus fumigatus identification and antibiotic sensitivity in Iraqi patients

Pulmonary aspergillosis is a severe infection in immunocompromised caused by Aspergillus fungi, with Aspergillus fumigatus being the most frequently cultured of this genus, although A. flavus, A. niger, A. terrus also include If left untreated, invasive aspergillosis can have mortality approaching 100%. A total of 105 samples were collected from TB patients with symptom of lung infection . These sputum samples were obtained from patients treated in Public health department Chest and Respiratory diseases clinic, from February 2023 to April 2023. All samples were transferred to the laboratory for culturing to isolation and identification fungi after labeling each sample with questionnaire form. Lacto phenol cotton blue staining method was used in identification of fungal isolates, fungi isolated by inoculating sample on culture media such as sabouraud dextrose agar (SDA), and disc diffusion. The sample mean ± standard deviasion (SD) of age was 48.5±14.2 years (ranged 9-75 years). Male to female ratio was 2:1.5, age groups number and percentage were 32(30.5%), 13(12.4%), 14(13.3%), 14(13.3%), 16(15.2%) and 16(15.3%) in the age groups <=29, 30-39, 40-49, 50-59, 60-69 and >=70. Laboratory investigations indicated that 40 (38.1%) of the total samples diagnosed with a fungus species, 16 (15.3%) of them present with Candida species followed by 14 (13.3%) with A.fumigatus, 5 (4.8%) with A. niger, 3 (2.8%) with A.terreus and 2 (1.9%) A.flavus, while negative results were indicated in 65 (61.9%) of collected samples. A.fumigatus. isolates were subjected to susceptibility testing using different antifungals. The antifungal drugs NYS, KEA, CMZ, ICZ and PCZ were the most effect. Resistant outcomes were reported in FCZ, while intermediate results were showed in MIC and AMB. The results showed an increase in the prevalence of Aspergillus fumigatus in Iraqi patients. This prevalence may be alarming on public health.

Hygienic Evaluation of Electrolyzed or Ozonated Water-Washed Apples Stored under Different Temperatures and Relative Humidity Conditions

The presence of spoilage and pathogenic microorganisms is a concern for the safety of apples. In the present study, we evaluated the hygienic status of electrolyzed water (EW)- or ozonated water (OW)-washed apples, which were stored over 2 weeks under the combination of 2 different temperatures (25 and 30°C) and 2 different relative humidity (RH) conditions (85 and 90%). The average numbers of bacteria or fungi from unwashed and washed apples (EW or OW) did not show statistically significant differences at storage for 0, 1, or 2 weeks and had an increased tendency as the storage temperature, RH, and period increased. Identification of fungal isolates from apples revealed 3 main genera (Fusarium sp., Trichoderma sp., and Alternaria sp.) together with 8 minor genera (Meyerozyma sp., Aspergillus sp., Glomerella sp., Neofusicoccum sp., Penicillium sp., Hypoxylon sp., Talaromyces sp., and Coprinellus sp.). Moreover, sensory tests using EW- or OW-washed apples showed that OW did not significantly affect 5 quality characteristics (appearance, taste, flavor, texture, and overall acceptability). Our data suggest that EW or OW washing did not significantly reduce the levels of microorganisms on apples relative to the unwashed and that EW or OW washing did not deteriorate the quality of washed apples.

Screening of biohydrogen producing endophytic fungi from biodiesel plants

ABSTRACTBiohydrogen from endophytic fungi constitutes an alternative biofuel resource with considerable advantages over biodiesel and ethanol extracted from existing food crops. Endophytes common in plant tissues mimic host physiological behavior and produce primary and secondary metabolites like oils and fats. Therefore, endophytes from 7 selected biodiesel plants, and 30 endophytic fungi isolated from 170 different plant parts were screened. The methodology included staining, lipid analysis, DNA isolation, molecular characterization, identification of fungal isolates by ITS sequences, biodiesel analysis, characterization of volatile organic compounds (VOCs), and statistical analysis. The fungal endophyte isolate SPSRJ28 was the most significant with an accumulation capacity of more than 20% of the total lipid content of its dry biomass, validating its oleaginous nature. In addition, the endophytes investigated in the present study yielded various VOCs that were validated by chromatography for both standards and isolates. When the VOCs were subsequently analyzed for their prospective usage as myco‐diesel, the presence of methane in the isolate SPSRJ28 was found to be highly significant. These findings remarkably support the investigation of endophytic fungi and their relationship with oleaginous plants as alternative sources of bioactive metabolites from microbial cell factories for myco‐diesel generation.

Identification of Fungal Isolates in Patients With Pulmonary Tuberculosis Treated at a Tertiary Care Hospital.

Introduction Pulmonary tuberculosis (TB) has long been associated with opportunistic fungal infections and could prove lethal if these fungal infections are not detected in the early stages of tuberculosis. TB patients are mostly immunocompromised, and an association with a fungal infection fuels each other, thus decreasing host immunity and making it difficult to treat. Extensive use of antibiotics and steroids has resulted in increasing trends of these fungal infections globally. Material and methods This observational, retrospective hospital-based medical record review study was conducted in the Department of Microbiology at the Indira Gandhi Institute of Medical Sciences (IGIMS), Patna, Bihar, India. A total of 200 medical records of pulmonary tuberculosis patients diagnosed by using sputum as clinical samples were evaluated and analysed for two years, from January 2020 to December 2021. This study was started after approval from the institutional ethical committee. Data were obtained from the mycology test records from the Department of Microbiology and from the data files of the medical records section over a period of two years. Results Our study included the medical records of 200 pulmonary tuberculosis patients who underwent treatment at IGIMS Patna. Out of 200 patient records, 124 (62%) were males and 76 (38%) were females. The male-to-female ratio was 1.6:1. After analysis and evaluation of 200 medical records of pulmonary tuberculosis patients, fungal species were detected in 16 (8%) sputum samples. Among 16 culture-positive sputum samples, 10 (8.06%) and six (7.1%) were diagnosed in male and female patients, respectively. Fisher's exact test showed a non-significant two-sided p-value of 1.000 with a relative risk of 0.9982. The prevalence, or positivity rate, was 8% in two years. The age group of 31-45 years had the most fungal co-infection at 37.5%. Among the fungal isolates, 5/16 (31.25%) were yeasts, and the remaining 11/16 (68.75%) isolates were mycelial fungi. Conclusion According to the findings of the present study, pulmonary fungal infections co-exist in tuberculosis patients, although the prevalence rates of all the coinfections are low and statistically not significant. Being chronic in nature and with confusing clinical and radiological findings, these fungal infections are misdiagnosed as reactivation of tuberculosis. Hence, the increasing rate of morbidity and mortality can decrease if adequate measures are taken for the diagnosis at an early stage and appropriate treatment of these fungal mycoses with antifungal therapy is instituted.

Identification of Intestinal Fungal Microflora and Bacterial Pathogens in the Collected Adult Ixodes ricinus from the Northern Provinces of Iran.

Ticks are vectors of many pathogens that involve various important diseases in humans and animals, they have several diverse hosts consequently can retain a diverse group of indigenous microbes, from bacteria to fungi. Little is known about the prevalence and diversity of tick microflora colonizing the midgut and their effects on ticks and their interaction. This information is important for development of vector control strategies. This study was carried out in northern Iran during autumn 2019. Ticks, Ixodes ricinus caught alive on the bodies of domestic animals in the fall. The tick homogenate was prepared. The identification of fungal isolates was carried out according to a combination of macro and microscopic morphology and molecular sequencing. Pathogenic bacteria of the family Borreliaceae, Francisella tularensis, Borrelia burgdorferi and Coxiella burnetii were tested by real-time PCR. A total of 133 mature I. ricinus ticks were collected from domestic animals, including 71.5% cattle and 28.5% sheep. The tick frequency rates were 87.21% for Mazandaran, 8.28% for Golestan and 4.51% for Gilan Provinces. Total prevalence of fungal tick contamination was 53.4% (75/133) of which Trichoderma harzianum (57%) was the most prevalent species followed by Aspergillus spp. (42%), Mortierella alpine (19%) and Penicillium polonicum (14%). All tick samples were negative for three pathogenic bacteria including Francisella tularensis, Coxiella burnetii, and Borrelia burgdorferi by real-time PCR analysis. These results show a first picture of the microbial diversity of ticks and highlight the importance of microbiota and their role in host-pathogen interaction.

MICROBIAL PROFILE AND PREDISPOSING FACTORS OF MYCOTIC KERATITIS IN A TEACHING HOSPITAL, CENTRAL INDIA

Introduction: Mycotic keratitis is one of the commonest causes of eye infections especially in male which works mainly in outdoor. Filamentous fungi (mould-like fungi) were the common cause of Mycotic keratitis and common presentations seen in developing countries. The aim of present study assess predisposing factors and microbial profile of Mycotic keratitis in central India. Methods: The study was conducted in the collaboration of Department of Microbiology and Ophthalmology in G. R. Medical College, Gwalior, India. Samples were collected from corneal scraping by sterile cotton swab and inoculated on culture media Sabouraud dextrose agar (SDA) for fungal culture. Material obtained from corneal scrapping also performed 10% KOH mount preparation, gram’s staining and Lactophenol cotton blue (LPCB) mount. Identification of fungal isolates was done by colony morphology, growth characteristics and relevant standard tests. Results: A total of 237 out of 740 (32%) fungal isolates was obtained from suspected patients of Mycotic keratitis, out of which Fusarium species (33.2%) was the most predominant followed by Curvularia species (21.9%) and Aspergillus species (16.7%), Ocular trauma was the key predisposing factors. Conclusion: Trauma was the key predisposing factor of keratitis because of filamentous fungi. The commonestcausative agent of fungal keratitis in current study was Fusarium species followed by Curvularia species. Early proper diagnosis and management of fungal keratitis reduces the ophthalmic morbidity and mortality

Study of Molds in Post COVID -19 Patients: An experience from Tertiary Care Centre

Introduction: Coronavirus disease 2019 (COVID-19) pandemic has emerged as a global health crisis in the century with a heavy toll on human life. The second wave of COVID-19 was lethal for the patients as they had to combat the virus along with the opportunistic fungal and bacterial infections triggered by it. Mucormycosis gained attention for its widespread existence during this period of the second wave of COVID-19. Material & Methods: Various clinical samples like biopsy tissues from paranasal sinuses, deep nasal swabs, bronchoalveolar lavage, sputum, etc. were studied for the presence of fungal elements by KOH examination and culture on Sabouraud’s dextrose agar. Identification of fungal isolate was done by growth characteristics, LPCB wet mount, and slide culture. Result: During the study period, a total of 214 samples were received of which 17 (7.9%) were positive for fungal growth. Rhizopus arrhizus was most frequently isolated. Diabetes mellitus and the use of steroid during COVID-19 hospitalization was observed to be common risk factors. Rhino-cerebral mucormycosis was found to be the commonest type. Conclusion: Invasive mould infections can lead to fatal outcomes, if not detected timely. Accurate identification of the fungus is important for the administration of appropriate antifungal therapy.

IDENTIFICATION OF FUNGAL ISOLATES ASSOCIATED WITH ANTHRACNOSE DISEASE ON CHILLI AND ITS PATHOGENIC LEVEL

Many chilli producers in Malaysia are facing a huge problem of anthracnose disease that infect the chilli fruits either at pre- or post-harvest stage. Due to this infection, the chilli fruits become rotten and could be colonized by grey conidial masses which is unfit for human consumption and cannot be sold. This study was aimed to isolate the fungal species associated with anthracnose symptoms collected from several mini-markets near Gong Badak, Terengganu. Pathogenicity test was conducted for all these isolates to evaluate their pathogenic level. Based on the results, five fungal isolates were successfully isolated and morphologically identified as Colletotrichum spp. All these isolates were labelled as UMTT46C, UMTT48C, UMTT50C, UMTT52C and UMTT54C. From these, fungal isolate (UMTT54C) showed the highest pathogenic level with disease severity (DS), 68.9%. Other Colletotrichum spp. recorded percentage of DS ranged from 53.3% to 60%. Identification of fungal isolates and their pathogenic level from the infected chilli is very important to assist in control strategy which can further provide a good protection measure for many chilli producers.

Antifungal Susceptibility Pattern in Otomycosis among Patients Attending a Tertiary Healthcare Institution in Port Harcourt, Nigeria

Antifungal resistance is one of the factors considered to hamper effective treatment of otomycosis. Consequently, this study was aimed at isolating etiological agent for otomycosis among patients that visited University of Port Harcourt Teaching Hospital from June 2017 to June 2018 and carry out antifungal susceptibility testing with the isolates. Ear swab from 120 patients and 120 control group was aseptically collected. Nystatin, voriconazole and fluconazole were used for antifungal susceptibility testing by agar diffusion method. Isolation and identification of fungal isolates was by Standard microbiological methods. From our result, fungi were isolated from ear swab of 34 patients and 5 from the control. The isolates and their frequency of occurrence are Candida sp. (61.8%), Aspergillus sp. (23.5%) and Penicillium sp. (8.8%), while Candida albicans was the only isolate seen among the control group. Susceptibility result revealed that Candida sp. and Penicillium sp. as being sensitive to voriconazole. (76.9%) and fluconazole (66.7%) respectively. Findings from this study underscore the importance of antifungal susceptibility testing before treatment of otomycosis in order to achieve high success rate thereby reduce cases of antifungal resistance.

Fungal Diversity of Food Supplements Sold on the Markets of Abidjan (Côte d'Ivoire): Case of Spirulina (<i>Arthrospira platensis</i>) and Moringa (<i>Moringa oleifera</i>) Powders

The consumption of food supplements such as Spirulina and Moringa is increasing in developing countries. However, these foods, due to certain processing conditions, are likely to be contaminated by moulds potentially producing dangerous mycotoxins. The objective of this study is to determine the level of contamination and the diversity of fungal flora found in Spirulina (<i>Arthrospira platensis</i>) and Moringa (<i>Moringa oleifera</i>) powder produced and marketed in Abidjan. A total of 360 samples of powder, including 144 of Spirulina and 216 of Moringa, were collected from different sales outlets. The identification of fungal isolates was carried out on the basis of classical mycology criteria. The prevalence of fungal strains in the analyzed products was 85.83%, which 77.99% in Spirulina and 22.01% in Moringa. The predominant species were <i>Aspergillus flavus</i> (24.6%), <i>Aspergillus niger</i> (21.68%), <i>Penicillium sp</i> (20.71%), and <i>Aspergillus fumigatus</i> (12.62%). The study shows that Spirulina powder and Moringa powder sold in markets are contaminated with moulds, some of which are potentially mycotoxin producers posing a health risk to consumers. Strict hygiene measures must be observed during the production and marketing of Moringa and Spirulina powders in order to prevent any poisoning among consumers.

INCIDENCE OF TOXIGENIC MOULDS IN FOOD SPICES SOLD IN OPEN MARKETS IN BENIN CITY, NIGERIA

Spices are parts of plants or plant substances primarily used for flavouring and preservation of food. Spices can be exposed to microbial contamination and toxic substances such as mycotoxins during processing storage and distribution. The aim of the study was to determine the incidence of toxigenic moulds in food spices sold in open markets in Benin City. A total of 35 samples were obtained from three markets. Potato dextrose agar (PDA) and Aspergillus flavus and parasiticus agar (AFPA) were employed for the isolation, characterization and identification of fungal isolates using standard microbiological procedures. The pH was determined using an electronic pH meter, titratable acidity and moisture content were also determined using appropriate methods. Fungi isolated in this study included; Fusarium sp., Alternaria sp., Rhizopus sp., Geotrichum sp., Cladosporium sp., Aspergillus niger and Penicillium sp. The most occurring fungus was Fusarium sp. (26%) while Alternaria sp. (2%) was the least occurring fungus. The fungal load ranged from 0.33 - 28.67 x 103 sfu/g and the pH values of samples ranged from 4.83 – 5.67. The titratable acidity ranged from 0.037 – 0.521 mg/l, while the moisture content ranged from 2.0 – 28.0%. The study revealed the presence of toxigenic moulds such as Fusarium and Penicillium species in food spices sold in open markets. This may have resulted from improper storage, handling and poor sanitary conditions in the open markets.

FUNGAL COMPOSITION AND PROXIMATE ANALYSIS OF POULTRY FEEDS SOLD IN BENIN CITY, NIGERIA

Poultry farming contributes significantly to poverty alleviation by providing employment opportunities to Nigerians. Feed quality is the most prominent challenge of the poultry sector. This study was aimed at assessing the fungal and proximate qualities of poultry feeds sold in Benin City, Nigeria. Poultry feed samples (starter, grower and finisher) were obtained from five stores in Benin City. Enumeration and isolation of fungi was carried out using the spread plate method on potato dextrose agar and Aspergillus flavus and parasiticus agar (AFPA). Identification of fungal isolates was based on cultural and morphological characteristics. Proximate composition was determined by standard methods. The total fungal counts of starter, grower and finisher feeds ranged from 0.10 – 8.50 x 105 sfu/g. Fungal species isolated were Aspergillus flavus, Aspergillus niger, Penicillium spp., Saccharomyces spp., Trichoderma spp., Rhizopus spp. and Mucor spp. The results from proximate analysis were as follows: 36.50±0.92 - 70.92±0.51% (protein), 3.53±0.04 - 8.45±0.16% (fibre), 11.61±0.72 - 13.91±0.27% (fat), 2.70 - 8.60% (moisture content) and 3.65 - 37.83% (carbohydrate). The temperature and relative humidity of storage ranged from 33.90oC - 35.20oC and 69% - 75% respectively. Results showed that poultry feeds sampled have diverse fungi present in them. Control measures should be adhered to in poultry industries and stores to prevent microbial contamination.

Demographic profile of fungal rhinosinusitis at a tertiary care centre

&lt;p class="abstract"&gt;&lt;strong&gt;Background: &lt;/strong&gt;Fungal rhino sinusitis is rampant in the North Indian state of Punjab. The demographic profile, presentation and comorbidities have been analysed.&lt;/p&gt;&lt;p class="abstract"&gt;&lt;strong&gt;Methods: &lt;/strong&gt;110 samples from the nasal cavity were collected in Oto-rhino-Laryngology services and processed in the Microbiology services of Dayanand Medical College and Hospital, Ludhiana. Nasal secretions and surgically excised tissue were processed and subjected to direct microscopy by potassium hydroxide (KOH) preparation as well as inoculated on Sabrouraud’s dextrose agar. Identification of fungal isolates was done as per standard procedures.&lt;/p&gt;&lt;p class="abstract"&gt;&lt;strong&gt;Results:&lt;/strong&gt; Fungal sinusitis was most common followed by acute invasive sinusitis, chronic invasive, granuloma (fungal ball) and granulomatous invasive fungal sinusitis. Males were affected more than females and the majority of the patients were from urban background. Allergic form of disease was commonly seen in younger age group as compared to the invasive form of the disease which was seen more in older patients. Nasal discharge was common presentation followed by nasal obstruction. Bronchial asthma and diabetes mellitus were co-morbidities. KOH positivity was 44.5% and fungal culture positivity was 33.6%. The common fungal isolate in this group was Aspergillus flavus (22.7%). The most common fungal isolate in the invasive group was Rhizopus spp. (4.5%) and Mucor spp. (3.6%).&lt;/p&gt;&lt;p class="abstract"&gt;&lt;strong&gt;Conclusions: &lt;/strong&gt;Elderly diabetic and asthmatic males are more susceptible to invasive fungal rhino sinusitis while the young males are vulnerable to the allergic fungal rhino sinusitis. Thereby control of diabetic status and appropriate allergy testing and desensitization is emphasised.&lt;/p&gt;

Identification of Fungal Isolates from Soils of Rice and Napier Grass

Microbial diversity of cultivated soil is the key resource for maintaining the functional capacity of agricultural and natural ecosystem. The research aimed to study the prevalence of soil fungi from the soils of rice and napier grass fields. Soil samples were collected from the fields of rice in flooded condition and napier grass in the upland condition at the University farm. Soil fungi were cultured on RB media through the serial dilution method. A total of 13 isolates were cultured, in which 11 from the soil collected from rice cultivated field and 2 from napier grass field. Seven out of 11 isolates collected from rice and 2 from napier grass were selected and identified through the morphological characters such as colony color, mycelium diameter, color and segmentaiton, spore size and color. Mycelium growths were examined on the PDA and RB media. Fungal isolates collected in this study were morphologically identified/ characterized as Aspergillus niger, A. flavus, A. terreus, A. carbonarius, A. nidulans, Alternaria sp., Penicillium and Talaromyces. Fungal species differed among the soils were discussed.

Cross-pathogenicity of Colletotrichum isolates on tropical fruit species in Brazil

Due to their perishable nature, tropical fruits are prone to pre-and post-harvest loss attributed to biotic and abiotic factors. Although the production of tropical fruits such as banana (Musa acuminata), guava (Psidium guajava), mango (Mangifera indica), papaya (Carica papaya), and passion fruit (Passiflora edulis) is in the increasing trend, the post-harvest loss remains a challenge for the production of excess tropical fruits worldwide. Severity and cross-pathogenicity of different Colletotrichum species attributed to the reasonable loss of such fruits, hence molecular detection and discrimination of such fungal species are indispensable for post-harvest disease management. In the present study, the rDNA ITS was used for molecular identification of fungal isolates from five tropical fruits at the post-harvesting stage. Most of the identified fungi belong to the genus Colletotrichum. Furthermore, the cross-pathogenicity and disease symptom severity of the isolates across these fruit samples was also investigated. The result showed that most isolates are host-specific, passion fruit being the ideal host for all the discriminated isolates. The symptoms caused by Colletotrichum isolates showed similar patterns as revealed by lesion diameter analysis. The work provided important data on the cross-pathogenicity and disease symptom characteristics of Colletotrichum pathogens known to affect the five tropical fruits investigated.

Oral Prevalence of Candida Species in Patients Undergoing Systemic Glucocorticoid Therapy and the Antifungal Sensitivity of the Isolates.

BackgroundCandida species are commonly detected as colonizers of the oral cavity; candidiasis or candidemia can develop in patients who are immunocompromised. Use of topical or inhaled glucocorticoids can alter the spectrum of Candida species and can promote oral candidiasis. The present study aims to evaluate the diversity of Candida species in the oral cavity and their susceptibility to antifungal agents in patients undergoing treatment with systemic glucocorticoids (SGCs) compared with non-users.MethodsWe conducted a descriptive, analytical, cross-sectional study that enrolled 120 patients with oral problems who were undergoing treatment with SGCs and who were admitted to the hospital of the First Affiliated Hospital, College of Medicine, Zhejiang University and Zhejiang Hospital, Hangzhou, China, between February 2019 and September 2019. One hundred and twenty age-and sex-matched patients were recruited as the SGC non-user control group. Demographic data included oral complaints and underlying diseases; symptoms of oral candidiasis were identified on physical examination. Candida species were collected using a concentrated oral rinse. Identification of fungal isolates was based on conventional phenotypic methods assisted by DNA sequence analysis of the internal transcribed spacer (ITS) rDNA gene region. Antifungal activities of anidulafungin, amphotericin B, micafungin, caspofungin, 5-flucytosine, posaconazole, voriconazole, itraconazole, and fluconazole were evaluated using the Sensititre YeastOneTM YO10 panel supplemented by the CLSI-M27-A3 protocol.ResultsFifty-two (43.33%) out of the 120 patients undergoing with SGCs were diagnosed with oral candidiasis, compared with 14 (11.67%) of the non-users (P < 0.05). Likewise, we collected 88 strains from 73.33% of the SGC users compared with only 48 (40%) from non-users (P < 0.05). Candida albicans was detected most frequently in both groups (45.45% vs 66.67%, respectively; P = 0.033); the overall frequency of non-Candida albicans (NCA) strains isolated from patients treated with SGCs were significantly higher than that identified among non-users (51.14% vs 33.33%, respectively; P = 0.046), although there were no significant differences concerning any single species of NCA. Resistance of C. albicans to itraconazole (P = 0.004) and fluconazole (P = 0.001) was significantly higher in patients treated with SGCs than in non-users; however, echinocandins, amphotericin B, voriconazole, and posaconazole were all active against strains from both participant groups with no significant differences detected.ConclusionTaken together, our findings indicate that SGC therapy may result in an increased prevalence of oral candidiasis as reflected by the clinical presentations and strains isolated; these findings were also associated with an increased frequency of NCA strains. SGC therapy was also associated with an increased frequency of C. albicans strains that were resistant to both itraconazole and fluconazole. The impact of SGC therapy on Candida species in the oral cavity requires further study.

Fungal neonatal and infantile sepsis in Egypt: Risk factors and identification of fungal isolates

Background : Invasive fungal diseases (IFDs) are opportunistic infections associated with significant mortality in paediatric patients, especially in those with compromised immune system and neonates with very low birth weight (VLBW). The objectives of this study are to determine the prevalence, clinical features and fungi isolates of neonatal sepsis in three hospitals in Egypt. Methodology : The study is a cross sectional survey of 176 neonates with clinical sepsis admitted to the neonatal intensive care units (NICU) of the three hospitals over a period of one year (February 2015 to January 2016). A minimum of two blood samples (collected within 24 hours) from each neonate were cultured for bacteria in automated BacT/AlerT and conventional culture bottles, while Saboraud-Brain Heart Infusion broth was inoculated for fungi culture. Positive growths from the broth were sub-cultured on Sabouraud Dextrose Agar (SDA) plates for aerobic incubation at 25oC and 37oC for 2 weeks. Identification of fungi colonies on SDA was by conventional morphology and confirmation on chromogenic agar media. Phylogenetic analysis of representative fungi isolates was done by partial nucleotide sequencing of D1-D2 domain of the large subunit rRNA gene. Results : Of the 176 neonates, blood culture was positive for pathogens in 55 (31.3 %) samples and fungi were isolated in 26 (14.8 %); yeast (25) and mould (1). The commonly isolated yeasts were Candida albicans, Candida tropicalis , and Candida krusei representing 34.6%, 30.8% and 23.1%, respectively of the total fungi isolated. The phylogenetic analysis in comparison to Genbank data showed defined clades for Candida tropicalis, Candida parapsilosis, Candida albicans and Pichia kudriavzevii Conclusion : This current study highlights the changing pattern of neonatal infections in Egypt caused by Candida , with increasing incidence of infections caused by non-albicans Candida species. Key words : fungal infection, neonatal, risk factors, PCR, yeast

Dimeric naphtho-γ-pyrones and further diverse bioactive metabolites from the marine-derived Aspergillus flavus Af/MMA 2018

Background and objective Sponge-associated fungi are known for their production of structurally diverse secondary metabolites, many of which exhibit different pharmacological activities. Materials and methods Isolation and identification of fungal isolate from the marine sponge Echinodictyum flabelliforme collected from the Red Sea coast of Hurghada, Egypt, was done. Working up and purification with the ethyl acetate extract produced by the marine-derived Aspergillus flavus Af/MMA 2018 afforded nine bioactive compounds. Structure of the isolated compounds was determined on the basis of NMR (1D and 2D) and mass (EI, ESI, HRESI MS) spectra and by comparison with the corresponding literature studies. Biologically, the antimicrobial, antioxidant, and antitumor activities (using Ehrlich cells) of compounds were studied in comparison with the original extract. Results and conclusion Working up and purification of the ethyl acetate-extracted residue produced by the marine-derived A. flavus Af/MMA 2018 afforded nine diverse bioactive compounds: five dimeric naphtho-γ-pyrones, that is, aurasperone A (1), aurasperone B (2), aurasperone D (3), aurasperone F (4), and aurasperone E (5), along with β-sitosterol glucoside (6), cerebroside C (7), glyceryl linoleate (8), and linoleic acid (9). Cerebroside C showed strong antimicrobial activity against different test organisms, whereas aurasperone E showed maximum DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) scavenging activity (67.41%) after 1 h, and by using different concentrations, giving 98.99% at 1000 μg/ml. The maximum antitumor activity against Ehrlich ascites carcinoma cells (70.9%) was attributed to the dimeric naphtho-γ-pyrone aurasperone E.

Ribosomal subunit protein typing using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the identification and discrimination of Aspergillus species

BackgroundAccurate identification of Aspergillus species is a very important subject. Mass spectral fingerprinting using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is generally employed for the rapid identification of fungal isolates. However, the results are based on simple mass spectral pattern-matching, with no peak assignment and no taxonomic input. We propose here a ribosomal subunit protein (RSP) typing technique using MALDI-TOF MS for the identification and discrimination of Aspergillus species. The results are concluded to be phylogenetic in that they reflect the molecular evolution of housekeeping RSPs.ResultsThe amino acid sequences of RSPs of genome-sequenced strains of Aspergillus species were first verified and compared to compile a reliable biomarker list for the identification of Aspergillus species. In this process, we revealed that many amino acid sequences of RSPs (about 10–60%, depending on strain) registered in the public protein databases needed to be corrected or newly added. The verified RSPs were allocated to RSP types based on their mass. Peak assignments of RSPs of each sample strain as observed by MALDI-TOF MS were then performed to set RSP type profiles, which were then further processed by means of cluster analysis. The resulting dendrogram based on RSP types showed a relatively good concordance with the tree based on β-tubulin gene sequences. RSP typing was able to further discriminate the strains belonging to Aspergillus section Fumigati.ConclusionsThe RSP typing method could be applied to identify Aspergillus species, even for species within section Fumigati. The discrimination power of RSP typing appears to be comparable to conventional β-tubulin gene analysis. This method would therefore be suitable for species identification and discrimination at the strain to species level. Because RSP typing can characterize the strains within section Fumigati, this method has potential as a powerful and reliable tool in the field of clinical microbiology.

Epidemio-Clinico-Microbiological Study of Mycotic Keratitis in North-West Region of Rajasthan

Mycotic keratitis, with its diverse clinical presentation and difficulties in treatment, makes it a challenging task for clinicians and an important object of study. The aim of present study was to determine the frequency of occurrence and epidemiological association with identification of fungal isolates from mycotic keratitis cases. This was a prospective and observational study conducted in Mycology Laboratory, Department of Microbiology, S.P. Medical College, Bikaner, on corneal scrapings and swabs of a total of 480 patients attending the Ophthalmology OPD, P.B.M. Hospital, Bikaner, during July 2005 to June 2012. Out of 480 suspected cases, 180 were found to be positive for fungus by smear/culture examination. Increased incidence was seen in the months of May to September with Aspergillus fumigatus being the most common isolate. Mycotic keratitis, though an age-old disease, still presented with challenging aspects of diagnosis and treatment. The study showed fungal keratitis is prevalent in rural parts of north-west Rajasthan, mainly found in males (age group 21-40years) with low socio-economic status (farm or factory workers). The most common cause of fungal keratitis was found to be species of Aspergillus.