Clinical presentation and fungal species distribution in otomycosis in a tertiary care hospital

Objective: Otomycosis is a common fungal ear infection in tropical and subtropical regions worldwide. This study aimed to perform mycological analysis on fungal debris from the external auditory canals of the patients to study the most common clinical presentation and fungal species distribution in otomycosis. Materials and Methods: Patients who met the inclusion criteria for this cross-sectional study were included and evaluated after providing written informed consent. After obtaining a thorough medical history, ear swabs for culture, sensitivity, and potassium hydroxide were provided. Patients with positive culture results were studied between September 2019 and March 2021. Results: Among 103 cases observed for 18 months in the Department of ENT, Rajarajeswari Medical College, and Hospital, Bengaluru, India, we found that males (56.31%) were more affected than females (43.68%). Itching (67.96%) was the most primary and common symptom that was observed, followed by pain (20.38%), and the most common predisposing factor was the usage of earbuds (26.21%) followed by water in the ear (23.3%) and oil in the ear (16.50%). Unilateral infection was most common (96%), and the left ear was most affected (64.07%). Aspergillus niger was the most common fungal isolate (60.19%), and otomycosis was very common in postmonsoon (October–December) (58.25%). Conclusion: The most frequent fungal isolates in otomycosis are from the Aspergillus and Candida species. The left ear was typically affected by otomycosis, which frequently had a unilateral predominance. The most common clinical symptoms were itching and pain.

Expression of β-glucosidase An-bgl3 from Aspergillus niger for conversion of scopolin / 生物工程学报

Scopoletin is a coumarin compound with various biological activities including detumescence and analgesic, insecticidal, antibacterial and acaricidal effects. However, interference with scopolin and other components often leads to difficulties in purification of scopoletin with low extraction rates from plant resource. In this paper, heterologous expression of the gene encoding β-glucosidase An-bgl3 derived from Aspergillus niger were carried out. The expression product was purified and characterized with further structure-activity relationship between it and β-glucosidase analyzed. Subsequently, its ability for transforming scopolin from plant extract was studied. The results showed that the specific activity of the purified β-glucosidase An-bgl3 was 15.22 IU/mg, the apparent molecular weight was about 120 kDa. The optimum reaction temperature and pH were 55 ℃ and 4.0, respectively. Moreover, 10 mmol/L metal ions Fe2+ and Mn2+ increased the enzyme activity by 1.74-fold and 1.20-fold, respectively. A 10 mmol/L solution containing Tween-20, Tween-80 and Triton X-100 all inhibited the enzyme activity by 30%. The enzyme showed affinity towards scopolin and tolerated 10% methanol and 10% ethanol solution, respectively. The enzyme specifically hydrolyzed scopolin into scopoletin from the extract of Erycibe obtusifolia Benth with a 47.8% increase of scopoletin. This demonstrated that the β-glucosidase An-bgl3 from A. niger shows specificity on scopolin with good activities, thus providing an alternative method for increasing the extraction efficiency of scopoletin from plant material.

Evaluation of Lipid Profile and Some Renal Parameters in Some Selected Ethnic Population with Fat-mass and Obesity-associated Gene (FTO) Variants in Niger Delta, Nigeria

Aim: To evaluate lipid profile and some renal parameters in some selected ethnic population with fat-mass and obesity-associated gene (FTO) variants in Niger Delta, Nigeria. Study Design: Case-controlled observational study. Place and Duration of Study: Federal Medical Centre, Asaba, Delta State and Safety Molecular Pathology Laboratory, Enugu, Nigeria, between March 2020 and February 2022. Methodology: Changes in lipid profile and some renal parameters in FTO gene was studied in ninety-eight (98) type 2 diabetes (T2D) subjects (78 cases and 20 controls) from four different tribes in the Niger Delta region, Nigeria. Multistage sampling method was employed in the subject selection. The subjects were first separated into two groups – new cases (less than a year of diagnosis as Diabetic) and old cases (one year & above). Equal number of samples was then randomly collected from each of the cluster groups. 10mls of blood was collected into plain bottles for the assay of the above-named markers, and were assayed using spectrophotometric and ELISA methods. The data were analyzed using GraphPad Prism, version 8.0.2 and p values less than .05 were considered statistically significant. Results: The results showed that the Ijaw tribe had the highest mean total cholesterol (TCHOL), low density lipoproteins (LDL), Castelli Risk Ratio (CRR), atherogenic coefficient (AC) values (5.36 ± 0.99, 3.36 ± 0.87 mmol/l,3.76 ± 1.18 and 2.86 ± 1.16) respectively, which were significantly higher (P<.05) than those of the control group, while the Urhobo tribe had the highest mean TG and AIP values (1.47 ± 0.51 mmo/l and 0.08 ± 0.01), The control subjects had the highest mean HDL values (1.51 ± 0.49mmo/l), which were significantly higher(P<.05) than that of the control subjects. Mean creatinine level was highest in the control group (101.1 ± 21.24 µmol/L), while the Urhobo tribe had the highest mean MDRD levels (94.15 ±36.17 ml/min). Special diets did not contribute to any significant difference in the biochemical indices of the subjects apart from significant changes in the levels of triglyceride. Conclusion: From the results, we conclude that the levels of lipid profile varied with the tribe for subjects with FTO variants and control subjects and only triglyceride levels are affected by specific diets.

Environmental Effect of KPO-Fire in Niger Delta and Future Health Implication

Crude oil discovery and exploration in the Niger Delta of Nigeria which includes Delta, Bayelsa, Abia, Rivers, Cross River, Akwa-Ibom, Imo, Edo and Ondo States of Nigeria, has led to significant economic growth of the nation especially in the creation of job opportunities also the generation of revenue. Unfortunately, Oil thefts have become the idea that replaced the most crisis and agitations in the Niger Delta communities against the perceived neglect of the areas. This had resulted to widespread operation of illegal oil bunkering activities called Kpofire. Kpofire, as described by the Niger deltans, is a process of burning crude oil by illegal oil operatives or thieves at isolated locations called bunkering sites or dumpsites in order to extract refined petroleum products. The design for the extraction of the petroleum products involves the sections of steaming and collection. These sections are connected by a very long pipe. Heat is applied to the steaming section using wood from the mangroves that is called ‘Ngala’ or ‘Agala’, thereafter, crude oil is used. The products extracted are fuel, kerosene and diesel. One of the major problems of Kpofire is the introduction of soot into the environment. Soot is a product is uncontrolled and incomplete combustion or pyrolisis of materials that contain carbon. The highest constituent of soot is the Polynuclear aromatic hydrocarbon (PAH). The Niger Delta people are faced with daily environmental challenges such as inadequate housing facilities, contaminated drinking water sources, barren lands for agriculture, dead rivers for fishing activities and poor health conditions, soot, which has become a gradual but huge challenge in many Niger Delta communities, has been shown to play a huge role in respiratory, skin, and reproductive health conditions. However, vulnerability is seen more in babies and children (particularly because their respiratory organs are not matured enough), the elderly, and people with preexisting health conditions, also heart or lung diseases (such as asthma). High exposure to PAH at prenatal stages is linked to a low IQ and lead to increased behavioral problems in the early-age child and childhood asthma. A collective effort is needed to put a stop this menace called kpofire in order to avoid environmental pollution thereby endangering the health of the people of Niger Delta.

Methods of plant growth-promoting fungi application to enhance the growth and yield of wheat var. Ibaa99

Aims@#The main aim of the study was to evaluate some methods of application of Aspergillus niger AD1 and Trichoderma hamatum T-113 for enhancing the growth and yield of wheat var. Ibaa99 in pots and field conditions.@*Methodology and results@#Plant growth-promoting fungi (PGPF) loaded with peat moss were used at a rate of 100, 150 and 200 mL pot-1 or m-2 in filed soil; seed treatment (coating) with fungi suspension 19 × 107, soil treatment and combination of all the three methods was employed in the study. Wheat seeds were sown in pots and field plots during 2018-2019, and data regarding various growth and yield attributes were recorded. In both pot and field trials, the results revealed that the best treatments for the desired plant growth and yield attributes were peat moss 150 mL alone or in combination with soil and seed treatments. The soil physicochemical parameters were also improved after inoculation with selected fungal isolates in different application methods compared with un-inoculated control treatment in both pot and field conditions.@*Conclusion, significance and impact of study@#The PGPF play a vital role represented phytoremediation, phytostimulation and bio-fertilization. The isolates of PGPF, which were applied with peat moss at 150 mL to the pot and in the field alone or combined with seed treatment and soil application, were significantly the best effective method for improving wheat attributes.

An efficient marker-free genome editing method for Aspergillus niger / 生物工程学报

Aspergillus niger is an important industrial strain which has been widely used for production of enzymes and organic acids. Genome modification of A. niger is required to further improve its potential for industrial production. CRISPR/Cas9 is a widely used genome editing technique for A. niger, but its application in industrial strains modification is hampered by the need for integration of a selection marker into the genome or low gene editing efficiency. Here we report a highly efficient marker-free genome editing method for A. niger based on CRISPR/Cas9 technique. Firstly, we constructed a co-expression plasmid of sgRNA and Cas9 with a replication initiation region fragment AMA1 (autonomously maintained in Aspergillus) by using 5S rRNA promoter which improved sgRNA expression. Meanwhile, a strain deficient in non-homologous end-joining (NHEJ) was developed by knocking out the kusA gene. Finally, we took advantage of the instability of plasmid containing AMA1 fragment to cure the co-expression plasmid containing sgRNA and Cas9 through passaging on non-selective plate. With this method, the efficiency of gene editing reached 100% when using maker-free donor DNA with a short homologous arm of 20 bp. This method may facilitate investigation of gene functions and construction of cell factories for A. niger.

Profil Épidémiologique et Mycologique de l'Otomycose à l'Hôpital Général de Référence de Niamey (Niger) / Epidemiological and my cologica l profile of otomycosisat the Genera l Reference Hospital o f Niamey (Niger

But. Décrire les caractéristiques épidémiologiques et mycologiques de l'otomycose à Niamey. Méthodologie.Il s'est agi d'une étude rétrospective menée sur 3ans au service d'ORL et CCF de l'Hôpital Général de Référence, portant sur les patients chez qui le diagnostic d'otomycose a été posé. Les paramètres étudiés étaient épidémiologiques et mycologiques. Résultats. En 3 ans, nous avons enregistré 447 consultations pour otites dont 56 avaient une otomycose, soit une fréquence de 12,53% de l'ensemble des otites et 58,92% des otites externes. L'âgemoyen était de 35,75 ans (extrêmes de 2 et 70 ans). La tranche d'âge de 31 à 60 ans représentait 58,93% des patients. Nous avons retrouvéune prédominance féminine (67,85%), (p=0,0752). Les principaux facteurs favorisant l'otomycose étaient le nettoyage de l'oreille au coton tige ou à la plume de volaille : (53,57%),le port du turban ou de couvre-chef, (46,42%), la baignade (16.07%), l'étroitesse du conduit auditif externe (8,92%) et le port de prothèse auditive externe intra-conduit (5,35%,). Sur le plan mycologique, le Candida albicanset l'Aspergilus nigeront représenté respectivement 19,44% et 33,33% des espèces fongiques isolées en culture. Conclusion. L'otomycose est une infection fréquente en Orl à Niamey. Le nettoyage de l'oreille au coton tige ou à la plume de volaille venait au premier rang des facteurs favorisant l'otite fongique. L'Aspergilus nigeret le Candida albicansétaient les germes les plus fréquemment isolés.

Profile of multidrug-resistant clinical bacterial isolates at the National Hospital of Zinder (NHZ), Niger Republic in 2021

Background: Today, bacterial resistance is a public health challenge throughout the world, and infections caused by resistant bacteria are associated with increased morbidity, mortality and health care costs. The objective of this descriptive study is to determine the prevalence and distribution of multi-drug resistant (MDR) clinical bacteria isolates at the National Hospital of Zinder, Niger Republic in 2021. Methodology: We conducted a descriptive cross-sectional study of in- and out-patients from whose clinical samples' bacteria were isolated at the bacteriology unit of the laboratory. Bacteria were isolated from the clinical samples following standard aerobic cultures and identified using conventional biochemical test schemes. Antibiotic susceptibility testing (AST) was performed by the agar disk diffusion technique, and categorization of the isolates into sensitive, intermediate or resistant was done according to the recommendations of the Antibiogram Committee of the French Society of Microbiology (CA-SFM) 2020 version 1.2. MDR was defined as resistance to at least one antibiotic in three or more categories, while selected MDR bacteria such as ESBL was identified using double disk synergy test, and MRSA by cefoxitin disk diffusion test. Results: Seventy-seven (6.7%) bacterial species were isolated from 1153 clinical samples processed at the bacteriology unit of the hospital laboratory between June and December 2021, of which 65.0% (50/77) were members of the order Enterobacteriales. Escherichia coli represented 40.3% (40/77) of the isolated bacteria, Staphylococcus aureus 13.0% (10/77) and Pseudomonas aeruginosa 11.7% (9/77). The overall prevalence of MDR was 44.2% (34/77), including 61.8% (21/34) ESBL-producing Enterobacteriales (ESBL-E), 26.5% (9/34) multi-resistant P. aeruginosa and 11.7% (4/34) MRSA, with 67.6% (23/34) of the MDR isolates from outpatients. Resistance rates of the Enterobacteriales to ciprofloxacin, gentamicin, amikacin and imipenem were 62.0%, 52.0%, 38.0% and 8.0% respectively. Resistance rates of P. aeruginosa were 100.0%, 88.9%, 77.8%, 33.3%, 22.2%, and 22.2% respectively to ceftazidime, ticarcillin, imipenem, ciprofloxacin, levofloxacin, and amikacin. Resistance rates of S. aureus were 100.0%, 50.0%, 40.0%, 10.0%, 0% and 0% to penicillin G,erythromycin, cefoxitin, tetracycline, fusidic acid, and chloramphenicol respectively. ESBL-E were 47.6%,85.7% and 0% resistant to amikacin, ciprofloxacin and imipenem, and MRSA resistance rates were 75.0%, 75.0%, 50.0% and 0% to erythromycin, tetracycline, gentamicin, and chloramphenicol respectively. Conclusion: This study reports high prevalence of MDR bacteria, mainly ESBL-E, with concerning high resistance to carbapenem. Rational use of antibiotics and implementation of surveillance system for MDR bacteria must be implemented in order to limit the emergence and spread of MDR bacteria in Niger Republic.

Implementation and contribution of temperature screening and handwashing practice at points of entry for COVID-19 pandemic response in a humanitarian crisis setting

Introduction: over the last decade, insecurity in the Lake Chad Basin has triggered an unprecedented humanitarian crisis in the Niger´s Diffa Region with a significant population movement. In this humanitarian setting, we reviewed the implementation process and the contribution of temperature screening and handwashing practice at points of entry as part of non-pharmaceutical interventions against COVID-19. Methods: in Diffa, border officers were trained on the fundamentals of infection prevention and control in relation to COVID-19 readiness and response and a 14-day district response team was constituted. To examine the significance of the implementation process of temperature screening and handwashing practices at points of entry, we conducted a secondary analysis of data submitted by the six health districts of the Diffa Region between March and July 2020. Results: travellers screened for fever ranged from 10,499 (in March 2020) to 62,441 (in April 2020) with the health districts of Diffa (mean: standard error of the mean: 25,999: 9,220) and of Bosso (mean: standard error of the mean: 30.4: 19.1) accounting for the most and the least of activities during the entire period, respectively. Overall, 125/169,475 travellers presented fever and were effectively quarantined. Only the Ngourti Health District reported travellers who declined handwashing (54/169,475); this was during the first three months of the first wave of the COVID-19 pandemic. Conclusion: we have documented a successful implementation of measures related to temperature screening with some unsubstantial denial of handwashing. Given the importance of border traffic due to insecurity in the Diffa Region, maintaining temperature screening and handwashing in this humanitarian setting is necessary but requires coordinated actions of all stakeholders involved in the region.

Process optimization for enhanced production of cellulases form locally isolated fungal strain by submerged fermentation

Cellulase has myriad applications in various sectors like pharmaceuticals, textile, detergents, animal feed and bioethanol production, etc. The current study focuses on the isolation, screening and optimization of fungal strain through one factor at a time technique for enhanced cellulase production. In current study sixteen different fungal cultures were isolated and the culture which quantitatively exhibits higher titers of cellulase activity was identified both morphologically and molecularly by 18S rDNA and designated as Aspergillus niger ABT11. Different parameters like fermentation medium, volume, temperature, pH and nutritional components were optimized. The highest CMCase and FPase activities was achieved in 100ml of M5 medium in the presence of 1% lactose and sodium nitrate at 30 oC, pH5 after 72 hours. The result revealed A. niger can be a potential candidate for scale up studies.

CRISPR/Cas-based genome editing in Aspergillus niger / 生物工程学报

Aspergillus niger is a vital industrial workhouse widely used for the production of organic acids and industrial enzymes. This fungus is a crucial cell factory due to its innate tolerance to a diverse range of abiotic conditions, high production titres, robust growth during industrial scale fermentation, and status as a generally recognized as safe (GRAS) organism. Rapid development of synthetic biology and systems biology not only offer powerful approaches to unveil the molecular mechanisms of A. niger productivity, but also provide more new strategies to construct and optimize the A. niger cell factory. As a new generation of genome editing technology, the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR associated (Cas) system brings a revolutionary breakthrough in targeted genome modification for A. niger. In this review, we focus on current advances to the CRISPR/Cas genome editing toolbox, its application on gene modification and gene expression regulation in this fungal. Moreover, the future directions of CRISPR/Cas genome editing in A. niger are highlighted.

Evaluation of the antimicrobial susceptibility testing process in clinical microbiology laboratories at Niamey, Niger

Background: Risk assessment is the means of identifying and evaluating potential errors or problems that may occur in testing process. The aim of this study was to perform risk assessment of antimicrobial susceptibility testing (AST) process in clinical microbiology laboratories of Niamey, Niger Republic. Methodology: We conducted a descriptive cross-sectional study from October 1 to December 31, 2019, to evaluate AST performance in seven clinical microbiology laboratories at Niamey, the capital city of Niger republic. The evaluation focused on the determination of the criticality index (CI) of each critical point (frequency of occurrence of anomalies, severity of the process anomaly, and detectability of the anomaly during the process) in the AST process and the performance of the AST through an observation sheet using two reference strains; Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 29213. Results: The criticality index (CI) was greater than 6 for most of the critical points related to material, medium, equipment, method and labour for the AST process in all the laboratories. A range of 18-100% errors on the inhibition zone diameters of the reference strains were observed. Major and/or minor categorization (Sensitive S, Intermediate I and Resistance R) discrepancies were found at all the laboratories for either one or both reference strains. The antibiotics most affected by the S/I/R discrepancies were trimethoprim (100%), vancomycin (100%), amoxicillin (80%) and amoxicillin + clavulanic acid (70%). Conclusion: This study showed a deficiency in the control of critical control points that impacts the performance of the AST reported by the laboratories in Niger. Corrective actions are needed to improve the performance of AST in clinical microbiology laboratories in Niger

Albizia lebbeck fruit pods as a substrate for the production of lignocellulolytic enzymes by Aspergillus niger

Aspergillus niger isolated from the soil was investigated for its capability to produce various lignocellulolyticenzymes, such as LiP, endoglucanase, FPase, xylanase by solid-state fermentation, using Albizia lebbeck fruitpods as a substrate. The chemical composition of the fruit pods was studied, and the production pattern of theenzymes was examined by growing the fungi for 25 days. The LiP activity was low, whereas a good productionof endoglucanase, FPase, and xylanase enzymes was noted. A dye decolorization capacity of the A. niger wasalso studied with Congo red. Therefore, A. lebbeck fruit pods which are considered as waste and burnt off canbe utilized for the production of holocellulolytic enzymes using A. niger.

Molecular characterization of phosphate solubilizing fungi Aspergillus niger and its correlation to sustainable agriculture

Aim: The present study aimed to investigate the phosphate solubilization potential of agriculturally important fungi, i.e., Aspergillus sp. isolated from the rhizosphere of healthy plants in Abha city, Saudi Arabia.Methodology: Sixteen Aspergillus sp. isolated and tested for phosphate solubilization potential were identified by 5.8S-ITS region sequencing and characterized by 11 ISSR-PCR markers. Finally, the highest phosphate solubilization potential isolates were used in field experiments on cucumber and tomato plants. Results: All Aspergillus niger isolates showed 96–100% similarity to A. niger strains available at GenBank database, Isolate ASAB-5 was most efficient at solubilizing phosphate on Pikovskaya’s medium, with a solubilization index of 2.67, and 235.22 mg l-1 of solubilized phosphate. ISSR-PCR markers revealed is total 142 bands in all isolates, with about 32.3% showing monomorphism and 67.6% polymorphism. Based on genetic similarity and intraspecies variability, the Aspergillus isolates were grouped into two different clusters with about 67.9% genetic similarity. The results of field experiments showed no significant difference between seeds treated with culture filtrate or conidial suspension of ASAB-5; however, both differed remarkably from untreated seeds. Interpretation: The current study confirms the existence of several useful phosphate solubilizing fungi in plants, which may serve as potential biological fertilizers. They are safer than chemical fertilizers and increase the bioavailability of soil phosphates for plants

Adult nephrotic syndrome in the Niger delta sub region: a single centre study at the university of Port Harcourt teaching hospital

Background: Adult nephrotic syndrome is a common cause of chronic kidney disease globally and in Sub-Saharan Africa. In Nigeria there are only few studies on adult nephrotic syndrome, mostly from the south-western Nigeria and rarely from the Niger delta sub-region of Nigeria. This study aims at contributing to the National data on adult nephrotic syndrome, from the perspective of the Niger delta sub-region.Methods: Retrospective analysis of five-year clinical data (January 2007 to December 2011) of adult nephrotic syndrome patients in the University of Port Harcourt teaching hospital was conducted.Results: Forty-four patients, representing 1% of medical admissions and 7.3% of adult renal cases were seen during the period, with mean age of 27.7±8.5 years. There were 32 males (72.7%). Mean duration of illness was 25.3±30.3months. Peak age group was the 20-29 year age group, accounting for 52.3% of cases. Hypertension was seen in 45.5%. Mean e-GFR was 73.5±33.8 ml/min/1.73m2 with CKD 1-3 constituting 93.2% of cases. Mean 24-hour urinary protein excretion was 13.6±8.4 grams/day. Protein excretion >10grams/day was observed in 65.8% of the patients. Mean total serum protein, albumin and total cholesterol were 51.4±11.7g/dl, 22.5±9.9 g/dl and 8.1±3.0 mmol/L, respectively. Anemia was common (68%) and histology showed MCGN (52.6%), MCD (21.1%), membranous (15.8%) and FSGS (10.5%). Though response to therapy was generally poor, immunosuppressive therapy showed better outcomes.Conclusions: The prevalence and pattern of adult nephrotic syndrome in the Niger delta sub-region is similar to that in other parts of Nigeria. It is predominantly a disease of young adult males with high prevalence of hypertension, and poor histologic categories.

Improved protease activity of Pixian broad bean paste with cocultivation of Aspergillus oryzae QM-6 and Aspergillus niger QH-3

BACKGROUND: The preparation of broad bean koji is a key process in the production of Pixian broad bean paste (PBP). Protease is essential for the degradation of proteins during PBP fermentation. To obtain broad bean koji with high protease activity using the cocultivated strains of Aspergillus oryzae QM-6 (A. oryzae QM-6) and Aspergillus niger QH-3 (A. niger QH-3), the optimization of acid and neutral protease activities was carried out using Box­Behnken design with response surface methodology (RSM). RESULTS: The optimum conditions were found to be as follows: inoculation proportion (X1), 3:1 (A. oryzae QM-6: A. niger QH-3, w/w); culture temperature (X2), 33°C; inoculum size (X3), 0.5% (w/w); incubation time (X4), 5 d. The acid and neutral protease activities were 605.2 ± 12.4 U/g and 1582.9 ± 23.7 U/g, respectively, which were in good agreement with the predicted values. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles revealed that the broad bean koji extracellular proteins in the case of cocultivation were richer compared to those in the case of A. oryzae QM-6 or A. niger QH-3 strain only. In addition, the free amino acids (FAAs) in the fermentation product were 55% higher in the cocultivation process than in that involving only A. oryzae QM-6, further confirming the diversity of proteases in the fermentation products. CONCLUSIONS: The optimal conditions of koji-making in PBP were obtained using RSM. The cocultivation of A. oryzae and A. niger increases the overall enzyme activities in the culture medium and the FAAs content, which would thus have potential application in the PBP industry.

Synthesis And Biological Evaluation Of Thiosemicarbazone Analogues Of Ornidazole As Antifungal Agents

To synthesize a series of ornidazole thiosemicarbazone analogues on the basis of literature reviews of 2-Methyl-5-nitroimidazoles and thiosemicarbazones and to evaluate all the analogues in vitro for their activity against Aspergillus niger and fumigatus. Thiosemicarbazone analogues were synthesized from oxidising ornidazole with potassium dichromate and refluxing the oxidised product with substituted thiosemicarbazide using ethanol as solvent in the acidic medium overnight. All the synthesized analogues of ornidazole showed good antifungal action against fumigatus and niger except compound C-4. Unsubstituted amine analogue C-2 has shown highest percentage inhibition (96.6%, 500 μg/ml) against fumigatus while aromatic amine with or without electronegative atom analogues C-3 and C-5 has shown highest activity against Aspergillus niger which is two times than standard drug ornidazole (100%, 1000 μg/ml).

A simple and accurate method for specific quantification of biomass in mixed cultures of filamentous fungi by quantitative PCR / Un método simple y preciso para la cuantificación específica de biomasa en cultivos mixtos de hongos filamentosos por PCR cuantitativa

Abstract Production of lignocellulolytic enzymes by filamentous fungi have a great potential at industrial level due to their widespread applications. Mixed fungal cultures and particularly mixed fungal biofilms constitute a promising fermentation system for an enhanced enzyme production. However, it has not been addressed how much of this enhancement depends on the mixed biomass proportion. In this sense, the aim of this study was to develop a method to specifically and accurately quantify mixed fungal biomass. For this purpose, mixed biofilm cultures composed of Aspergillus niger and Trichoderma reesei, two filamentous fungi used industrially for cellulase production, were collected from 48 to 120 h of growth; mycelia were pulverized, and DNA was extracted for qPCR assays with specific primers for each fungus. Primers were designed from non-conserved regions of sequences of actin and β-tubulin genes of both A. niger and T. reesei. Specificity of these primers was tested in silico and experimentally. A statistically significant correlation was obtained between qPCR-calculated biomass and dry weight biomass data. By this method, it was possible to detect changes on mycelia proportions in biofilms over time, suggesting a competitive interaction between these two fungi. In conclusion, this method allows a specific and accurate quantification of mixed fungal biomass and could be also applied to different mixed culture systems for studying microbial interactions.

Resumen La producción de enzimas lignocelulolíticas por hongos filamentosos tiene un gran potencial a nivel industrial debido a sus diversas aplicaciones. Los cultivos fúngicos mixtos y particularmente las biopelículas fúngicas mixtas constituyen un sistema de fermentación prometedor para una mayor producción enzimática. Sin embargo, no se ha abordado cuánto de esta mejora depende de la proporción de biomasa mixta. En este sentido, el objetivo de este estudio fue desarrollar un método para cuantificar de forma específica y precisa la biomasa fúngica mixta. Para este propósito, se recolectaron cultivos mixtos de biopelículas de 48 a 120 h de crecimiento compuestos por Aspergillus niger y Trichoderma reesei, dos hongos filamentosos utilizados industrialmente para la producción de celulasas; el micelio se pulverizó y el ADN se extrajo para ensayos de qPCR con cebadores específicos para cada hongo. Los cebadores se diseñaron a partir de regiones no conservadas de las secuencias de los genes de actina y β-tubulina de A. niger y T. reesei. La especificidad de estos cebadores se probó in silico y experimentalmente. Se obtuvo una correlación estadísticamente significativa entre la biomasa calculada mediante qPCR y los datos de biomasa en peso seco. Mediante este método, fue posible detectar cambios en las proporciones de los micelios en las biopelículas a lo largo del tiempo, lo que sugiere una interacción competitiva entre estos dos hongos. En conclusión, este método permite una cuantificación específica y precisa de la biomasa fúngica mixta y también podría aplicarse a diferentes sistemas de cultivo mixto para estudiar interacciones microbianas.

A comparative study on enzyme production ability of microorganism producing γ-aminobutyric acid in processing of Sojae Semen Praeparatum at different pH and temperatures / 中草药

Objective: To lay a foundation for elucidating the mechanism of microbial regulation of γ-amino butyric acid (GABA) formation in the processing of Sojae Semen Praeparatum. Methods: The ability of glutamic acid decarboxylase and protease (neutral protease, alkaline protease and acid protease) production of Bacillus subtilis, Enterococcus avium, Enterococcus faecium, Bacillus amyloliquefaciens, Aspergillus niger, Aspergillus flavus, Aspergillus tamarii, Penicillium citrinum, Cladosporium tenuissimum, Phanerochaete sordida, Rhizopus oryzae, Sporidiobolus salmonicolor in different pH and temperature conditions were determined by Berthelot colorimetry and Folin phenol method. Results: The GAD and protease activities of the twelve strains were stronger at pH 5-7 and temperature 28-37 ℃. The highest GAD enzyme activity from A. flavus was 41.97 U/h, the optimum pH was 7 and the temperature was 28 ℃, followed by S. salmonicolor, A. niger, R. oryzae, and B. subtilis, respectively. The enzyme activities were 29.04, 25.78, 22.42 and 19.43 U/h. The highest neutral protease activity of B. subtilis was 24.80 U/mL, the optimum pH was 7 and the temperature was 37 ℃, followed by B. amyloliquefaciens, R. oryzae and E. avium. The enzyme activities were 16.86, 12.51 and 9.18 U/mL. The highest alkaline protease activity of B. amyloliquefaciens was 13.29 U/mL, the optimum pH was 7 and the temperature was 34 ℃, followed by B. subtilis and R. oryzae. The enzyme activities were 8.86 and 6.20 U/mL, respectively. The ability of 12 kinds of microorganisms to produce acid protease was generally poor. Conclusion: The optimal pH and temperature of the 12 kinds of microorganisms selected for this experiment are basically consistent with the natural processing of Sojae Semen Praeparatum. Among them, fungi have stronger GAD production capacity and bacteria have stronger neutral protease production capacity. The high GABA concentration of Sojae Semen Praeparatum is caused by the joint action of multiple strains.

Rice farmer's poverty and its determinants: evidence from Dogofiri village of Office du Niger zone in Mali / A pobreza dos agricultores de arroz e seus determinantes: evidências da aldeia de Dogofiri na zona do Office du Niger, Mali

ABSTRACT: The aim of this research was to contribute to a better understanding of rice farmer's poverty of Office du Niger (ON) in Mali at village-level. Data were collected through survey with 110 head family farms in the village of Dogofiri. Quantitative data were analyzed using descriptive statistics. Multiple linear regression models were used to analyze the main determinants of poverty. Results indicated that the factors of physical capital and human capital as well as government policy have a significant influence on the poverty of family farms through production, age, family size, education and health support, agricultural credit and water fees. Policies aimed to improve the family farm's income and boosting rice production to alleviate poverty ought to be based on these factors.

RESUMO: O objetivo desta pesquisa é contribuir para uma melhor compreensão da pobreza dos produtores de arroz do Office du Niger (ON), no Mali, ao nível das aldeias. Os dados foram coletados por meio de pesquisa em 110 propriedades familiares da aldeia de Dogofiri. Os dados quantitativos foram analisados por meio de estatística descritiva. Modelos de regressão linear múltipla foram utilizados para analisar os principais determinantes da pobreza. Os resultados indicaram que os fatores de capital físico e de capital humano, bem como a política governamental, influenciam significativamente na pobreza das propriedades familiares por meio da produção, idade, tamanho da família, educação e apoio à saúde, crédito agrícola e taxas de água. Políticas destinadas para melhorar a renda da agricultura familiar e aumentar a produção de arroz para atenuar a pobreza devem basear-se nesses fatores.