An Overview And Study Of Koozh Biology Essay
“ Millet ” is a generic term for a heterogenous group of eatage grasses known for their little “ coarse ” grains [ 1 ] . It is an of import minor cereal in tropical and semitropical parts [ 2 ] , India and Africa are considered to be place lands for millets [ 3 ] . India is presently the largest manufacturer of millet [ 4 ] . Traditional fermented nutrient contains high alimentary value and would develop a diverseness of spirit, olfactory property and texture in nutrient substrate [ 5 ] and supply protection against bacterial pathogens [ 6 ] . Agitation of a nutrient stuff in a controlled environment leads to a desirable nutrient merchandise but in uncontrolled conditions consequences in spoilage. Changes that take topographic point during agitation include addition in aminic N, the dislocation of proteins and devastation of any inhibitors that may be present [ 7 ] .
Many traditional fermented merchandises are made from millets both in African and Asiatic Countries. Koozh a Tamil name for a porridge are ready to eat, popular repast or energy / refreshment drink sold in the streets during the forenoon to noon in TamilNadu, India. It ‘s either made from finger millet – Eleusine coracana ( Kezhvaragu in Tamil ) or pearl millet-Pennisetum glaucum ( Cumbu ) flour and broken rice ( noyee ) in a clay pot [ 8 ] .
Koozh readying is a two twenty-four hours uninterrupted procedure shown in flow chart ( Fig.1 ) served from a 12L to 15L plastic or a unstained steel container. The container are moved on a push streetcar or a bike but in some rural markets it is kept in a clay pot laid on moisture sand over a table under thatched roofing. It is served in a 500 to 600 milliliters broad mouthed unstained steel container ( Sembu ) along with concomitants like newly cut onion, pickle ( Oorgai ) , coriander foliage Indian relish ( Kothamalli Thokku ) and dried fried merchandises like- chilis ( Moar milagai ) , turkey berry ( Sundakkai Vathal ) , cluster beans ( Kothavarangai Vathal ) consumed by day-to-day pay earners, laborers and drivers. The managing during readying varies depending on parts later changing their organoleptic belongingss and microbic vegetation of koozh.
Homemade readyings of millets are fed to turning babies as malted flour porridge which is considered “ nourishment for babies ” [ 9 ] .Finger millet after agitation contains important sums of B-complex vitamins, energy, bioavailable minerals, indispensable amino acids, Fe, Ca, fiber, micro nutrientsA and minerals [ 10 ] .
The aim of the survey is to measure the microbic quality of Koozh that is sold in the street.
Materials and Methods
Finger and pearl millet koozh were collected utilizing a 250 milliliter autoclaved broad mouthed prison guard capped fictile containers from market topographic points in Salem, Tamilnadu, India and instantly kept in repast mate ( Milton, Mumbai ) incorporating ice packages. It was transported to the Department of Food Technology, SPIC Bioprocess Laboratory, Anna University, Chennai-25 and subjected to enrichment technique for microbic analysis [ 11 ] . Finger millet koozh samples sold in route side streetcar sellers were bought from four different topographic points located in suburbs of Chennai with autoclaved 500 milliliter containers and microbic analysis were conducted on the same twenty-four hours.
Media and Addendums
The undermentioned media and addendums were used for the microbic analysis: Arginine dihydrolase stock ( M619, HiMedia, Mumbai ) , Bacillus Cereus Agar base ( M833, HiMedia, Mumbai ) , Baird Parker Agar Base ( M043, HiMedia, Mumbai ) , Clostridial Agar ( M497, HiMedia, Mumbai ) , De Man Rogosa Sharpe broth- MRS ( M641, HiMedia, Mumbai ) , E. coli Direct Agar- ECD ( M1357, HiMedia, Mumbai ) , Egg yolk Emulsion ( FD045, HiMedia, Mumbai ) , Egg yolk Tellurite Emulsion ( FD046, HiMedia, Mumbai ) , Listeria Identification agar ( M1064, HiMedia, Mumbai ) , Listeria selective supplement-PALCAM ( FD061, HiMedia, Mumbai ) , M17 Broth ( M1029, HiMedia, Mumbai ) , Plate count agar ( M091, HiMedia, Mumbai ) , Polymxin B selective addendum ( FD003, HiMedia, Mumbai ) , Potato dextroglucose agar ( M096, HiMedia, Mumbai ) , SS agar ( M108, HiMedia, Mumbai ) , Standard home base count agar ( M096, HiMedia, Mumbai ) , Violet red gall glucose agar ( M581, HiMedia, Mumbai ) , Yeast Malt Agar ( M424, HiMedia, Mumbai ) , Peptone ( RM667, HiMedia, Mumbai ) and Agar ( RM026, HiMedia, Mumbai )
Chemicals and Reagents
Benzyl penicillin K salt ( 44197 2W, BDH, UAE ) , Calcium carbonate precipitated ( 17630, Merck, Germany ) , Chloramphenicol ( RM218, HiMedia, Mumbai ) , Gram Stain-Kit ( K001-1KT, HiMedia, Mumbai ) , Sodium chloride ( RM031, HiMedia, Mumbai ) , Streptomycin sulfate ( RM220, HiMedia, Mumbai ) and Hydrogen peroxide solution 50 % Purified ( Merck, Germany ) were used for the microbic analysis.
Determination of pH
Koozh samples pH were analysed instantly after purchase utilizing AP-1 plus, pH Meter ( Susima Technologies, Chennai ) incorporating epoxy organic structure detector pH investigation ( Van London Phoenix Company, Texas, USA ) .
Enumeration of Bacteria, Pathogens and Yeast
All samples were serially diluted prior to microbic analysis. Peptone buffer saline [ 12 ] PBS and CM101 Vortex Mixer ( REMI, Mumbai ) were used for consecutive dilution. The undermentioned selective media were used for numbering by spread home base method – Violet ruddy gall glucose agar for Enterobacteria [ 13 ] , Clostridial Agar for infective Clostridia, ECD Agar for colifroms particularly E.coli, Listeria designation agar supplemented ( supp. ) with PALCAM for Listeria monocytogenes, Bacillus cereus agar supp. with egg yolk, Baird Parker supplemented supp. with egg yolk tellurite for Staphylococcus, Standard count agar ( SPC ) for entire bacterial count, SS agar for Salmonella and Shigella. After hardening of the poured selective media and adding the serially diluted PBS,
it was kept over a PlateMasterTM for SpeedPlatingTM ( LA622, HiMedia, Mumbai ) . Yeast and molds was enumerated on pour home bases of Potato dextroglucose agar ( PDA ) added with filter sterilized Chloramphenicol [ 14 ] .
Bacterial home bases were incubated aerobically at 37oC for 24 Hs but for numbering of infective Clostridia sp, inoculated Clostridial agar kept under Anaerobic Gas Pack Systems ( LE002, HiMedia, Mumbai ) . Yeast and mold home bases were incubated at 25oC for 72 h. Colonies were enumerated manually utilizing a Lapiz Digital Colony Counter ( Medica instruments Mfg. Co. , Mumbai ) .All experiments were carried out in triplicates.
Word picture and Isolation of LAB
MRS and M17 media, were pour plated individually and incubated at 37oC for 24 hours. Colonies on single civilization home bases were examined for difference in morphology like form, size, boundary line, coloring material and lift from the surface of the media. Catalase activity, Gram reaction utilizing the HiMedia ‘s Gram ‘s staining kit and cell motility trial were conducted [ 15 ] . The positive presumptive LAB isolates were named, numbered harmonizing to their difference and quadrant streaked on a fresh home bases. These isolates were grown in selective stock and cryo-preserved at – 80oC for farther trials. MRS, M17 agar media supplemented with 1 % CaCO3 was pour plated with presumptive LAB and incubated at 37oC for 24 hours utilizing the Anaerobic Gas Pack Systems [ 16 ] to place if the isolates could turn in anaerobiotic status and bring forth acid.
Growth at two temperatures -15oC, 45oC and pH – 3.6 and 9.6 [ 17 ] were conducted. The turbidness of the inoculated stock, after incubation at 37oC for 72 hours was considered positive. Gas production from glucose [ 12 ] utilizing selective stock as basal medium and ammonia use from arginine was conducted utilizing arginine dihydrolase broth [ 16 ] .
Result and Discussion:
Koozh is a portion of day-to-day diet and during the specific times of spiritual festivals. Its readying was learnt from their sires and had been practised of all time since. As apparent from figure one, Koozh is a merchandise of millet and rice, from dual agitation caused by a mixture of vegetation affecting bacteriums and barm. The nutrient utilizations of finger millet are confined to flour based merchandises because it was non possible to decorticate the millet similar to cereals. This is chiefly due to the extremely floury endosperm [ 18 ] . “ Pearl millet malting reduces the anti-nutritient phytic acid accordingly increasing the bioavalability of proteins and minerals.Though the mechanism of mousey smell is non known but due to the lessening in pH from the growing of lactic acid bacteriums it is about eliminated ” [ 19 ] .
The agitation mediated by the microbic vegetation would be initiated when the millet is ground to flour, in the pre used milling machine. After adding H2O to the millet flour, manus is used for blending were the human tegument serves as the starting motor vegetation for the first agitation. This nightlong fermented millet slurry kept for 12-16 hours is cooked along with broken rice ( Figure 1 ) . Rice sold commercially in Bangladesh, heat immune spores are a major job because it survives blanching and milling phases [ 19 ] .Cooking inactivates most dirty micro-organisms ; heat immune bacterial endospores may last or even be stimulated to shoot [ 20 ] and cookery non merely kills vegetive cell of viing bacteriums and Fungis but besides activates spores of B to shoot [ 21 ] .
Leting this slurry to ferment overnight, the ensuing merchandise is called Kali. On the 3rd twenty-four hours little portion of the merchandise is transferred into a broad mouthed vas and diluted with drinkable H2O and manus assorted to fix Koozh. A similar merchandise in Inner Mongolia a traditional nutrient, of course fermented by residuary rancid stock, made from millet or rice called Congee ( acidic-gruel ) [ 22 ] .Butter milk a merchandise of diluted curd is added straight to the whole batch at the ratio of 1:12 ( v/v ) or added individually during the clip of functioning.
The pH of the gathered sample was in the acidic, scope from 4.3 – 4.9 ( Table: 1 ) . Kodo ko jaanr a fermented millet drink of the Himalayas, the pH ranged from 3.7- 4.5 [ 23 ] . Koko rancid H2O of Northern Ghana, made from nightlong steeping of pearl millet, the concluding merchandise pH was 3.9A± 0.1 [ 24 ] .In Uganda ‘s western Highlandss, from a mixture of millet and sorghum flour the merchandise called bushera with 3.7A± 0.1pH and sorghum bushera had 4.0-4.5 pH. There was a lessening in figure of coliforms, which may be due to high sourness ( low pH ) as a consequence of acid production by lactic acid bacteriums [ 25 ] .
Without sing the first two samples which was subjected to enrichment technique SPC, LAB and yeast all had a similar norm of 8 log cfu/g value.A co-metabolism between barms and lactic acid bacteriums may be, were the bacteriums supplying the acerb environment, which selects the growing of barms that in bend provide vitamins and other growing factors to the bacteriums [ 26, 27 ] . There was absence of Mould in all the samples.
Gram positive, catalase negative and non-motile isolates were considered as presumptive LAB. Isolates was subjected to assorted conditions for grouping ( Table-3 ) . MRS and M17 supplemented with CaCl2 in both aerophilic and anaerobiotic status produced clearance zone corroborating organic acid production in both Homo and hetero fermentative presumptive LABs.
Out of 60 nine LAB isolates twelve per centum ( 8/69 ) were identified as B remainder were cocci. All isolates were found to be gram positive, catalase negative and non-motile. All grew at 45 oC and produced ammonium hydroxide from arginine. Forty five per centum ( 31/69 ) possessed the ability to bring forth gas from glucose ( heterofermentatives ) , ninety three per centum ( 57/69 ) were able to turn in 15 oC, and forty three per centum ( 39/69 ) in pH 3.6.
The Mills are non cleaned prior to anchoring of the millet which might function as the first beginning of taint. Adhesion of micro-organisms to equipment surfaces has the possible to convey pathogens to nutrient, and this is evident in the nutrient processing industry [ 28, 29 ] . Exposure of pathogens to surfaces may take topographic point either by direct contact with contaminated objects or indirectly through airborne atoms [ 30 ] . Recontamination of the cooked porridge may happen through handling or implements / utensils [ 31 ] . The surfaces of equipment used for nutrient handling, storage or processing are recognized as a major beginning of microbic taint [ 32 ] as bacteriums have the ability to attach to surfaces normally found in the nutrient processing environment, such as polystyrene, hydroxyapatite, glass, gum elastic, and unstained steel [ 33,34,35,36 ] . Earlier surveies indicate that cross-contamination from natural merchandises via custodies, cleaning fabrics or sponges, and utensils used with nutrients that were non subjected to further cookery contributed to the happening of eruptions of nutrient borne unwellness [ 37, 38, 39 ] .
Pathogens presence were confirmed utilizing biochemical trial [ 40 ] . Microbial taint has two constituents: foremost, the saprophytic vegetation required in big population responsible for nutrient spoilage and 2nd, the infective vegetation, a few cells which can do infections and poisoning to worlds and animate beings [ 41 ] .
Listeria and Staphylococcus areus were non detected in any samples. Harmonizing to hypertext transfer protocol: //www.hpa.org.uk/webc/HPAwebFile/HPAweb_C/1259151921557 the infective vegetations were categorised. E.coli was found in all the samples doing it a high hazard class. The 3rd sample did non hold any other pathogen except E. coli, which is a enteral pathogen [ 42 ] , of import member of the coliform group, a portion of the normal vegetation of the bowel of human and craniates, some strains can do stomach flu, diarrhea and urinary piece of land infections [ 43 ] . The major ground may be due to the add-on of non parboiled drinkable metro H2O and handling patterns.
The first and the 2nd sample had Clostridium sp. , Salmonella sp. , Shigella sp at risky count degrees and Enterobacteria, B.cereus in moderate degrees. This is due to the nightlong transit of koozh products.Studies have shown that pathogens such as enterotoxigenicA Escherichia coli, Shigella flexneri, A Salmonella typhimurium, A Bacillus Cereus andA Campylobacter jejuniA are adversely affected when present in traditionalA fermented nutrient [ 44, 45, 46, 47, 48 ] .
The 4th sample had Clostridum sp. at risky and Enterobacteria in moderate hazard class. Food toxic condition by C. perfringens particularly kitchen strains [ 49 ] produces a big sum of heat labile enterotoxins that are cytotoxic and disrupts the membrane of epithelial cells, ensuing in diarrhea [ 50 ] . .Its presence in heat treated nutrients would be due to inadequate cookery or post-processing taint, but the usage of sanitizing rinses may cut down but non wholly take these beings with conformity to HPA, UK.
Fifth had Closridum at moderate hazard and had B.cereus in low degree. Bacillus Cereus is found more efficient in lasting on treating equipments by implementing itself to shoot a measure prior to sanitization [ 51, 52 ] . The 6th sample had merely B.cereus in low degree. B. cereus a nutrient pathogen produces assorted toxins causes enteral diarrhoeal disease to consumer ” [ 18 ] .
During functioning of onion, knife and the cutting board were non cleaned prior to utilize. Hazard analyses carried out in families at Dominican Republic reported that kitchen knives and liquidizers were contaminated with Salmonella spp [ 53 ] . A separate sembu would be dipped with manus and poured in to another cleaned sembu and served. After usage the sembu were washed with prewashed H2O, without detergent and kept inverted for the following helping. Plastic or metal home bases are offered to administer for self service. But in some topographic points merely a spoon would be kept for ego service where the consumer needs to maintain the concomitants in custodies and consume. The spoon would be once more kept back where it was taken without rinsing.
Accompaniments kept unfastened during the clip of functioning on busy location like main roads or route corner, leads to heavy exposure to smoke and dust. Oil fried merchandises that are kept unfastened increases the possibility of taint because oil attracts direct dust and air borne pathogens. This street nutrient is prepared by lower income group with small formal instruction in less hygienic which gives a good chance for pathogens to split, retroflex and multiply.
But the Enteric bacterial pathogens inA foodA and H2O must last the sourness of the tummy before they reach the enteric tractA [ 54 ] and do disease to the host. In decision, koozh contains risky count of pathogens but due to the presence of high LAB count, might posses probiotic belongingss that would stamp down the growing of pathogen that is farther to be studied.
I truly want to thank, University grants committee, India for fiscal and contingent support.