Microorganism, maintenance, and inoculum preparation the microorganism used was aspergillus niger bk01, which was isolated from rice. Paper open access growth profile of aspergillus niger on red. This strain was isolated from citrus fruit and exhibits high extracellular pectinase production. The present invention relates to microbial inocula and in particular to standardised inocula for producing reference cultures of microorganisms. Inoculum suspensions are prepared from fresh, mature 2 to 5dayold cultures grown on potato dextrose agar slants at 35c. Inoculum standardization for antifungal susceptibility. Reducing sugar was estimated by 3,5dinitrosalicylic acid and citric acid was estimated spectrophotometrically using pyridineacetic anhydride methods.
Inoculum standardization for antifungal susceptibility testing of. Research article citric acid production by aspergillus niger cultivated on. 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. Aspergillus niger nrc1ami was obtained from the national research center culture collection nrc. Citric acid production by aspergillus niger cultivated on. Influence of inoculum size on phytase production and.
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. Citric acid production potential of aspergillus niger. Confirm inoculum numbers by plating 100l on pda plates overnight at 37c using the following dilutions. 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 preparation will mean to create a precise, quantified concentration of viable aspergillus fumigatus conidia in a diluent suitable for suspending and. The progressive increase of invasive disease and reports of resistance among aspergillus species emphasizes the need for reproducible antifungal susceptibility testing. The first method was adjustment of inoculum size by haemocytometer counting. Ashour1 1botany and microbiology department, college of science, king saud university, riyadh 12824, saudi arabia. The invention has been developed primarily for use as a solid, discshaped, inoculum and will be described hereinafter with reference to this application. 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. Research article citric acid production by aspergillus niger. The inoculum preparation was based on the method described by alam et al. Isolation and inoculum preparation of aspergillus niger. Aspergillus niger was used for cellulase production in submerged smf and solid state fermentation ssf.
Cn102533555b inoculum and preparation method thereof. Inoculum preparation involves obtaining the organisms in an optimal state that is compatible with inoculation into cell culture, tissue culture, media, and fermentors. The maximum production of cellulase was obtained after 72 h of incubation in ssf and 96 h in smf. The percent decolorization of synthetic colorants obtained by using spore inoculum of aspergillus oryzae jsa1 are shown in table 1. Morphological development of aspergillus niger in submerged citric acid fermentation as a function of the spore inoculum level. Johnson agar sja and potato dextrose agar gave higher citric acid titres than maltextract agar. The effect of temperature plays an important role in the production of citric acid by aspergillus niger grewal and kalra, 1999. Comparative evaluation of two different methods of. Inoculating the fermentor with a preculture of pellets, instead of directly with spores, and carrying out fermentation at rev min. 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. A collection of 28 clinical isolates was tested against amphotericin b, itraconazole, voriconazole, and terbinafine. The cmcase and fpase activities recorded in ssf were 8. Microorganism and inoculum preparation aspergillus niger as02 obtained from the culture stockofmycologylaboratory,departmentofcropprotection, ahmadu bello university, zaria, nigeria was used in this study.
From the results obtained it showed that the combination of both aspergillus niger and pseudomonas sp. The chrysophyllum albidum peel was dried, sieved to remove dirt, dry milled and the powder used as substrate for citric acid production. Isolation and characterization of citric acid producing. Inoculum preparation spore suspensions of the isolates were prepared by adding 10 ml of sterilized distilled water containing 2 drops of 0.
However, it will be appreciated that the invention is not limited to this particular. This was maintained on molasses agar, which contained 300 gl cane molasses ph adjusted at 6. Effects of aspergillus niger inoculum concentration upon the kinetics. Preliminary studies on the microbial degradation of plastic. After 3 days, the fungus growth on the pda slants was. Development of culture inoculum for scaleup production of citric acid from oil palm empty fruit bunches by aspergillus niger. Morphological development of aspergillus niger in submerged citric. Preliminary studies on the microbial degradation of. Jan 25, 2006 microorganism, inoculum preparation, medium. Itz was significantly affected by inoculum size of a. The parkia biglobosa pulp powder has high sugar content and this makes it suitable for the growth of aspergillus niger. Anand kishore 2008 has also reported that maximum citric acid production was recorded at 30c using aspergillus niger ncim 705. Inoculation methods and aeration conditions in the production of pellet cultures of aspergillus niger 110 with acceptable citric acid yields were studied. Briefly, the inoculum 112 suspensions were prepared in.
The second method was spectrophotometric adjustment at 530 nm. A spore suspension was obtained by adding 20 ml of a 0. The studies revealed that production parameters ph, inoculum size, substrate concentration. Inoculum development for enzyme production a fungal strain of a. Paper open access growth profile of aspergillus niger on. Kinetics of enhanced ethanol productivity using raw starch. The studies revealed that production parameters ph, inoculum size. They were grown at 30 0c for 5 days and then stored at 4 c for regular sub culturing. Sja also gave better germinability than the other media. Lipase production by aspergillus niger using sheanut cake. Preparation of inocula and fermentation procedure a.
Analysis of the influence of tween concentration, inoculum. Development of culture inoculum for scaleup production of. 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. 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. Preparation of final inoculum for particular chambers. The production of citric acid using chrysophyllum albidum an indigenous underutilized fruit waste peel and genetically characterized strains of aspergillus niger was carried out. Spore suspension inoculum was prepared by washing the fungal culture grown on pda plate according to procedure followed by alam et al. Microorganism and inoculum preparation aspergillus niger nrc1ami was obtained from the national research center culture collection nrc. Effects of aspergillus niger inoculum concentration upon the. Development and temperature gradient online monitoring of a vehicular rotary solid. The influence of several test variables on susceptibility testing of aspergillus spp. 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. Pdf two methods of inoculum preparation for filamentous fungi were compared. About 810 ml of distilled water was poured each time onto the.
Then the first reactor was inoculated with biomass prepared as indicated above. Once the culture was prepared in growing culture form, it was subcultured on potato dextrose. Inoculum size 104 cfuml versus 105 cfuml and glucose supplementation 0. Conversion of food waste to single cell protein using. Production of citric acid from molasses free science. Spores were collected with 50 ml of sterile distilled water containing 0.
Therefore, a vegetative mycelial inoculum was developed in a liquid medium and used as the inoculum for ssf krishna and nokes, 2000. Preparation of inoculum for inoculum preparation, aspergillus niger ipbcc. Polygalacturonase by aspergillus niger using seaweed waste. A comparative study of the preparation and properties of ribonucleic acids of yeast. Identification of fungi of the genus aspergillus hydrolytic enzymes like lipases and amylases 1, 26. Effect of different cultural conditions for phytase. Both strains were maintained on potatodextrose agar medium. Aspergillus niger and aspergillus terreus selected from the ciatej collection, were identified by molecular techniques.
For inoculum preparation, 100 ml of vogels medium containing 0. Thirteen fungal isolates were obtained from soil samples. Enzymatic saccharification of pretreated rice straw by. Purification and characterization of the glucoamylase from. The fungus used for this study was isolated from onion left at room temperature to undergo spoilage.
Inoculum preparation and verification 110 111 spore concentration was adjusted to around 106 conidiaml. Decolorization study on synthetic colorants by using spore inoculum of aspergillus oryzae jsa1. Krishman ps, bajaj the polyphosphatases of aspergillus niger. Standard operating procedure for preparation of aspergillus. The reliability of both methods was assessed by colony counting. Bioprocess optimization for pectinase production using. Inoculum suspensions were prepared from fresh, mature 3 to 5dayold cultures grown on sabouraud agar or potato dextrose. Pdf inoculum standardization for antifungal susceptibility testing. A total of 156 isolates belonging to 8 different species were tested. Validation protocol for hold time study of prepared inoculum.
This wide range of sizes of inoculum suspensions could have a significant influence on mics. Once the culture was prepared in growing culture form, it was subcultured on potato. A fast, practical and reproducible procedure for the. A sterile wireloop was used to dislodge the spore clusters under sterilize conditions and then shaken thoroughly to prepare a homogenized spore suspen. Antifungal susceptibility testing in aspergillus spp. Colour and size of spores which are species dependent influence the od values. Study on the use of agricultural wastes for cellulase. 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. Research article citric acid production by aspergillus. Effect of increasing inoculum sizes of aspergillus hyphae. Pdf morphological development of aspergillus niger in. Spore suspension inoculum was prepared by washing the fungal culture grown on pda plate. Viability increased with time of incubation, but higher production.
The aspergillus niger strain selected for this study needs a vegetative inoculum for ssf, because spore production is minimal on the plate culture. 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. Two methods of inoculum preparation for filamentous fungi were compared. The isolated cultures of aspergillus niger were maintained as stock culture on potato dextrose agar slants. By the spectrophotometric method for inoculum preparation, the best scenario is retrieval of an inoculum that varies between 4. Various agrowastes 1, 2 have been used for of citric acid, aspergillus niger being most commonly used as microbial source 3, 4. Pdf comparison of aspergillus niger spore production on. Nccls document m38p suggests inoculum preparation by spectrophotometric procedure. One hundred eightytwo filamentous fungi pathogenic for humans were used. Pellet growth and citric acid yield of aspergillus niger 110. Mechanical cultivation of aspergillus niger spores in industrial large scale as the fermentation inoculum in citric acid plants was a difficult problem. After that, the spores were dissolved in a solution of tween 80 0. Briefly, the inoculum 112 suspensions were prepared in sterile saline 0.
Development and temperature gradient online monitoring of a. Maximal phytase activity of aspergillus niger was detected in media with 1. The isolates were subcultured from the stock water suspensions on sabouraud agar and on potato dextrose agar. Inoculum standardization is a crucial step in such procedures. The nonidentity of metaphosphatase with apyrase and pyrophosphatase. Inoculum suspensions were prepared from fresh, mature 3 to 5dayold cultures grown on. Influence of inoculum size on phytase production and growth. Sep 01, 2006 aspergillus fumigatus is most frequently isolated from clinical specimens, but other important species include aspergillus flavus, aspergillus niger and aspergillus terreus1. Simultaneous saccharification and fermentation of corn. Some observations on the growth of aspergillus niger asm.
The studies revealed that production parameters ph, inoculum size, substrate. The first method was adjustment of inoculum size by haemocyto. A spore inoculum at a level of 10% was found best for protease production 8. The agreement between the hematocytometer counts and the colony counts cfu per milliliter was 97. The invention discloses a kind of inoculum and preparation method thereof. The study was conducted to investigate the potential of parkia biglobosa fruit pulp as substrate for citric acid production by aspergillus niger.
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. Validation protocol for hold time study of prepared. Identification of the fungal isolate was based on cultural and microscopy characterization following procedures from barnet and hunter 1972. Morphological development of aspergillus niger in submerged. A fast, practical and reproducible procedure for the standardization. Development and temperature gradient online monitoring of. 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.
Alternatives as spore enumeration could be more appropriate. Production of acid protease by aspergillus niger using. Aspergillus sp ufla dca 01, despite not having been totally effective in elucidating species related to a. The culture was maintained on potato dextrose agar pda medium slants and preserved under refrigeration at 4 c. Extraction and purification of protease from aspergillus. The agreement between the hematocytometer counts and the colony counts.
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. Production and characterization of lipases by two new. Production of chitosan by submerged fermentation from. The second choice, specific to the madison chamber will be a final inoculum consisting of 20 ml of 1 x 10. Isolates of all species except members of the genus fusarium were incubated at 35c. In some cases an extended incubation is required for proper sporulation of the. This strain was isolated from citrus fruit and exhibits high extracellular pectinase production 19. Some observations on the growth of aspergillus niger from. Comparative evaluation of two different methods of inoculum. This paper discusses the effects of three aspergillus niger inoculum concentrations 0. Phytase production by aspergillus niger and aspergillus.
Among the inorganic and organic nitrogen sources, ammonium nitrate in concentration of 0. The medium used to prepare the plates is double strength to allow for a 50% dilution once the inoculum is added. 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. Prepare a final inoculum consisting of at least ml of 1 x 10. Antifungal susceptibility testing remains less well developed and utilized than antibacterial testing, the scientific support for its validity has benefited greatly by. When the culture was grown for 12 days in sterile base media containing 50 mg%, 75 mg%, 100 mg% and 200 mg% melanoidin. Aspergillus fumigatus is most frequently isolated from clinical specimens, but other important species include aspergillus flavus, aspergillus niger and aspergillus terreus1. Viability increased with time of incubation, but higher production of. Pda slants preparation potato dextrose agar pda slants were prepared according to the manufacturers instructions in order to cultivate the selected a.
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