Validation protocol for hold time study of prepared. Comparison of aspergillus niger spore production on potato dextrose agar pda and crushed corncob medium article pdf available in the journal of general and applied microbiology 565. Extraction and purification of protease from aspergillus. Study on the use of agricultural wastes for cellulase. Various agrowastes 1, 2 have been used for of citric acid, aspergillus niger being most commonly used as microbial source 3, 4. For inoculum preparation, 100 ml of vogels medium containing 0. This inoculum is solidstate dry inoculum, and described inoculum comprises the inoculation material of freezen protective medium and predetermined amount, and wherein, described inoculum is platelike substantially. The fungus used for this study was isolated from onion left at room temperature to undergo spoilage. Isolation and characterization of citric acid producing. Research article citric acid production by aspergillus. Comparative evaluation of two different methods of. Research article citric acid production by aspergillus niger cultivated on parkia biglobosa fruit pulp helenshnadaauta, 1 khadijattoyinabidoye, 2 hauwatahir, 3 aliyudabaiibrahim, 2 andsesanabiodunaransiola 1 departmentofmicrobiology,federaluniversityoftechnology,minna,n igeria department of microbiology, usmanu danfodiyo university, sokoto. Inoculum preparation involves obtaining the organisms in an optimal state that is compatible with inoculation into cell culture, tissue culture, media, and fermentors.
Inoculum suspensions were prepared from fresh, mature 3 to 5dayold cultures grown on sabouraud agar or potato dextrose. Simultaneous saccharification and fermentation of corn. Production of acid protease by aspergillus niger using. A sterile wireloop was used to dislodge the spore clusters under sterilize conditions and then shaken thoroughly to prepare a homogenized spore suspen. The isolation of the species of the nigri section on creatine sucrose agar crea enabled to distinguish the aspergillus sp species, which was characterized by the lack of sporulation and by the. The type of sporulation medium and time of incubation had an effect on spore viability and citric acid production by mycelia grown from aspergillus niger spores. The study was conducted to investigate the potential of parkia biglobosa fruit pulp as substrate for citric acid production by aspergillus niger. Pdf inoculum standardization for antifungal susceptibility testing. Then the first reactor was inoculated with biomass prepared as indicated above. This inoculum is solidstate dry inoculum, and described inoculum comprises the inoculation material of freezen protective medium and predetermined amount, and wherein, described inoculum is. Ashour1 1botany and microbiology department, college of science, king saud university, riyadh 12824, saudi arabia.
The inoculum preparation of aspergillus niger to solidstate fermentation was carried out by inoculating the fungus in 1 l erlenmeyers flasks containing 30 ml of solidified pda medium and incubated at 30c for 5 days. Effect of increasing inoculum sizes of aspergillus hyphae. Purification and characterization of the glucoamylase from. Paper open access growth profile of aspergillus niger on red.
Inoculum standardization for antifungal susceptibility. The studies revealed that production parameters ph, inoculum size. Production and characterization of lipases by two new. The first method was adjustment of inoculum size by haemocyto. Maximal phytase activity of aspergillus niger was detected in media with 1. Some observations on the growth of aspergillus niger from. Development and temperature gradient online monitoring of. The reliability of both methods was assessed by colony counting. Nccls document m38p suggests inoculum preparation by spectrophotometric procedure.
Influence of inoculum size on phytase production and. Two methods of inoculum preparation for filamentous fungi were compared. Both strains were maintained on potatodextrose agar medium. Research article citric acid production by aspergillus niger cultivated on. The studies revealed that production parameters ph, inoculum size, substrate. The parkia biglobosa pulp powder has high sugar content and this makes it suitable for the growth of aspergillus niger. This paper discusses the effects of three aspergillus niger inoculum concentrations 0. Identification of fungi of the genus aspergillus hydrolytic enzymes like lipases and amylases 1, 26. The maximum production of cellulase was obtained after 72 h of incubation in ssf and 96 h in smf. Once the culture was prepared in growing culture form, it was subcultured on potato. Aspergillus niger and aspergillus terreus selected from the ciatej collection, were identified by molecular techniques. Citric acid production potential of aspergillus niger.
The progressive increase of invasive disease and reports of resistance among aspergillus species emphasizes the need for reproducible antifungal susceptibility testing. Pda slants preparation potato dextrose agar pda slants were prepared according to the manufacturers instructions in order to cultivate the selected a. A comparative study of the preparation and properties of ribonucleic acids of yeast. The invention discloses a kind of inoculum and preparation method thereof.
Itz was significantly affected by inoculum size of a. Inoculum preparation spore suspensions of the isolates were prepared by adding 10 ml of sterilized distilled water containing 2 drops of 0. The isolates were subcultured from the stock water suspensions on sabouraud agar and on potato dextrose agar. The isolated cultures of aspergillus niger were maintained as stock culture on potato dextrose agar slants. Pdf two methods of inoculum preparation for filamentous fungi were compared. The studies revealed that production parameters ph, inoculum size, substrate concentration. Spore suspension inoculum was prepared by washing the fungal culture grown on pda plate.
Aspergillus niger was used for cellulase production in submerged smf and solid state fermentation ssf. Development and temperature gradient online monitoring of a. Conversion of food waste to single cell protein using. Johnson agar sja and potato dextrose agar gave higher citric acid titres than maltextract agar. After that, the spores were dissolved in a solution of tween 80 0. Therefore, a vegetative mycelial inoculum was developed in a liquid medium and used as the inoculum for ssf krishna and nokes, 2000. This strain was isolated from citrus fruit and exhibits high extracellular pectinase production 19. Production of citric acid from molasses free science. Confirm inoculum numbers by plating 100l on pda plates overnight at 37c using the following dilutions. Isolation and inoculum preparation of aspergillus niger. Preliminary studies on the microbial degradation of. Lipase production by aspergillus niger using sheanut cake. A spore suspension was obtained by adding 20 ml of a 0. A total of 156 isolates belonging to 8 different species were tested.
The inoculum preparation was based on the method described by alam et al. A fast, practical and reproducible procedure for the. Mechanical cultivation of aspergillus niger spores in industrial large scale as the fermentation inoculum in citric acid plants was a difficult problem. Briefly, the inoculum 112 suspensions were prepared in. Some observations on the growth of aspergillus niger asm. Effects of aspergillus niger inoculum concentration upon the kinetics. In some cases an extended incubation is required for proper sporulation of the. The invention has been developed primarily for use as a solid, discshaped, inoculum and will be described hereinafter with reference to this application. By the spectrophotometric method for inoculum preparation, the best scenario is retrieval of an inoculum that varies between 4. Colour and size of spores which are species dependent influence the od values. Inoculum suspensions are prepared from fresh, mature 2 to 5dayold cultures grown on potato dextrose agar slants at 35c.
The chrysophyllum albidum peel was dried, sieved to remove dirt, dry milled and the powder used as substrate for citric acid production. The percent decolorization of synthetic colorants obtained by using spore inoculum of aspergillus oryzae jsa1 are shown in table 1. Isolates of all species except members of the genus fusarium were incubated at 35c. The cmcase and fpase activities recorded in ssf were 8. Inoculum suspensions were prepared from fresh, mature 3 to 5dayold cultures grown on. The nonidentity of metaphosphatase with apyrase and pyrophosphatase.
Pdf comparison of aspergillus niger spore production on. One hundred eightytwo filamentous fungi pathogenic for humans were used. Spore suspension inoculum was prepared by washing the fungal culture grown on pda plate according to procedure followed by alam et al. This was maintained on molasses agar, which contained 300 gl cane molasses ph adjusted at 6. Alternatives as spore enumeration could be more appropriate. Research article citric acid production by aspergillus niger. Among the inorganic and organic nitrogen sources, ammonium nitrate in concentration of 0. Preparation of final inoculum for particular chambers. The second method was spectrophotometric adjustment at 530 nm. Viability increased with time of incubation, but higher production.
Development of culture inoculum for scaleup production of citric acid from oil palm empty fruit bunches by aspergillus niger. Aspergillus niger is also cultured for the extraction of the enzyme, glucose oxidase, used in the design of glucose biosensors, due to its high affinity for. Inoculum development for enzyme production a fungal strain of a. A fast, practical and reproducible procedure for the standardization. The objective of this study was to develop a fast and precise method of evaluating the cell density of an aspergillus spore suspension, as an alternative to. A collection of 28 clinical isolates was tested against amphotericin b, itraconazole, voriconazole, and terbinafine. The agreement between the hematocytometer counts and the colony counts cfu per milliliter was 97. Microorganism and inoculum preparation aspergillus niger nrc1ami was obtained from the national research center culture collection nrc. This strain was isolated from citrus fruit and exhibits high extracellular pectinase production. Bioprocess optimization for pectinase production using. Sep 01, 2006 aspergillus fumigatus is most frequently isolated from clinical specimens, but other important species include aspergillus flavus, aspergillus niger and aspergillus terreus1. Kinetics of enhanced ethanol productivity using raw starch. Development of culture inoculum for scaleup production of. Inoculum size 104 cfuml versus 105 cfuml and glucose supplementation 0.
Aspergillus fumigatus is most frequently isolated from clinical specimens, but other important species include aspergillus flavus, aspergillus niger and aspergillus terreus1. Inoculating the fermentor with a preculture of pellets, instead of directly with spores, and carrying out fermentation at rev min. Identification of the fungal isolate was based on cultural and microscopy characterization following procedures from barnet and hunter 1972. The second choice, specific to the madison chamber will be a final inoculum consisting of 20 ml of 1 x 10. After 3 days, the fungus growth on the pda slants was. Preparation of inocula and fermentation procedure a. They were grown at 30 0c for 5 days and then stored at 4 c for regular sub culturing. Cn102533555b inoculum and preparation method thereof. A high nitrogen concentration increases the growth of fungi and the consumption of sugars but decreases the amount of citric acid produced because it is a limiting factor in citric acid production 11. Preparation of inoculum for inoculum preparation, aspergillus niger ipbcc. Pellet growth and citric acid yield of aspergillus niger 110. Inoculum preparation will mean to create a precise, quantified concentration of viable aspergillus fumigatus conidia in a diluent suitable for suspending and. The medium used to prepare the plates is double strength to allow for a 50% dilution once the inoculum is added. Antifungal susceptibility testing remains less well developed and utilized than antibacterial testing, the scientific support for its validity has benefited greatly by.
Morphological development of aspergillus niger in submerged. However, it will be appreciated that the invention is not limited to this particular. Spores were collected with 50 ml of sterile distilled water containing 0. Citric acid production potential of aspergillus niger using. Microorganism, maintenance, and inoculum preparation the microorganism used was aspergillus niger bk01, which was isolated from rice. Viability increased with time of incubation, but higher production of. Paper open access growth profile of aspergillus niger on. From the results obtained it showed that the combination of both aspergillus niger and pseudomonas sp. The influence of several test variables on susceptibility testing of aspergillus spp. Validation protocol for hold time study of prepared inoculum. The aspergillus niger strain is genetically modified to contain several copies of the wild type aspergillus nigerphytase gene and is referred to as aself cloned microorganism. Citric acid production by aspergillus niger cultivated on. About 810 ml of distilled water was poured each time onto the.
Briefly, the inoculum 112 suspensions were prepared in sterile saline 0. Sja also gave better germinability than the other media. Effect of different cultural conditions for phytase. Prepare a final inoculum consisting of at least ml of 1 x 10. Citric acid is a well known industrially important organic acid being utilized immensely in medicine, dairy, pharmaceutical and biochemical industries. Colony counts were done for all inoculum preparations. It can also be deduced that aspergillus niger had a greater impact on the degradation as it gave a better percentage reduction compared to pseudomonas sp. Analysis of the influence of tween concentration, inoculum. Enzymatic saccharification of pretreated rice straw by. Thirteen fungal isolates were obtained from soil samples. Pdf morphological development of aspergillus niger in.
When the culture was grown for 12 days in sterile base media containing 50 mg%, 75 mg%, 100 mg% and 200 mg% melanoidin. Inoculum preparation and verification 110 111 spore concentration was adjusted to around 106 conidiaml. Once the culture was prepared in growing culture form, it was subcultured on potato dextrose. Decolorization study on synthetic colorants by using spore. Jan 25, 2006 microorganism, inoculum preparation, medium. The present invention relates to microbial inocula and in particular to standardised inocula for producing reference cultures of microorganisms. Inoculum standardization for antifungal susceptibility testing of. Polygalacturonase by aspergillus niger using seaweed waste. Krishman ps, bajaj the polyphosphatases of aspergillus niger. Aspergillus niger strains used in the study include the local isolates which were obtained from various locations research article citric acid fermentation by aspergillus niger amal a. Microorganism and inoculum preparation aspergillus niger as02 obtained from the culture stockofmycologylaboratory,departmentofcropprotection, ahmadu bello university, zaria, nigeria was used in this study.
The agreement between the hematocytometer counts and the colony counts. This wide range of sizes of inoculum suspensions could have a significant influence on mics. A spore inoculum at a level of 10% was found best for protease production 8. Two different methods of inoculum preparation for susceptibility testing were analysed.
Inoculation methods and aeration conditions in the production of pellet cultures of aspergillus niger 110 with acceptable citric acid yields were studied. Comparative evaluation of two different methods of inoculum. Inoculum standardization is a crucial step in such procedures. The culture was maintained on potato dextrose agar pda medium slants and preserved under refrigeration at 4 c. Antifungal susceptibility testing in aspergillus spp. Aspergillus niger nrc1ami was obtained from the national research center culture collection nrc. Morphological development of aspergillus niger in submerged citric acid fermentation as a function of the spore inoculum level. Reducing sugar was estimated by 3,5dinitrosalicylic acid and citric acid was estimated spectrophotometrically using pyridineacetic anhydride methods. The effect of temperature plays an important role in the production of citric acid by aspergillus niger grewal and kalra, 1999. The production of citric acid using chrysophyllum albidum an indigenous underutilized fruit waste peel and genetically characterized strains of aspergillus niger was carried out. Production of chitosan by submerged fermentation from. Effects of aspergillus niger inoculum concentration upon the.
Decolorization study on synthetic colorants by using spore inoculum of aspergillus oryzae jsa1. The first method was adjustment of inoculum size by haemocytometer counting. Development and temperature gradient online monitoring of a vehicular rotary solid. Phytase production by aspergillus niger and aspergillus. The aspergillus niger strain selected for this study needs a vegetative inoculum for ssf, because spore production is minimal on the plate culture. Preliminary studies on the microbial degradation of plastic. Morphological development of aspergillus niger in submerged citric. Standard operating procedure for preparation of aspergillus. Influence of inoculum size on phytase production and growth.
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