Showing papers in "Mesopotamia Journal of Agriculture in 2009"

Effect of microbial transglutaminase treatment on soft cheese properties

Abstract: Microbial transglutaminase (MTGase) was isolated from the bacteria Streptoverticillium mobaraense and used to improve the yield and properties of soft cheese manufactured from cow milk by enhancing the cross-linking reaction among milk proteins. The enzyme was applied at different concentrations and at various addition times. Results indicated that the addition of the enzyme before rennet prevented milk coagulation while the simultaneous addition of MTGase and the rennet significantly decreased curd strength and cheese hardness and increased the loss of proteins and fat in the whey. It was found that the most suitable way of adding the enzyme was after coagulation and curd cutting. This treatment enhanced the cross-linking of whey proteins with the curd proteins which was proportional to the concentration of the added enzyme up to 60 enzyme units/liter. The crosslinking reaction was verified by SDS-PAG electrophoresis experiment which showed a high molecular weight band accompanied by a decrease in the density of αs-casein, β-casein and whey proteins bands. Transglutaminase affected the produced cheese composition noticeably by increasing protein content, total solids and decreasing the protein content of the whey. Sensory evaluation of the obtained product showed that the enzyme treated cheese was superior to the untreated cheese throughout all storage period (8 days). INTRODUCTION Cheese production is rising sharply and so is whey. Several billion pounds of fluid whey are produced annually as a by-product all around the world. It is not efficiently utilized and more than 80% of this whey ends up as pollutant. In conventional processes for cheese making, a considerable amount of protein is lost with the whey, as protein is a valuable component of milk and milk products. One of the efforts to reduce cost of cheese production has been to keep the whey proteins from being lost in the manufacturing process. Several methods have been proposed to recover whey proteins. They included the concentration or drying the whey proteins and adding it to the cheese curd, heat or pressure treatment of the milk, and concentration of the milk by ultrafiltration, evaporation or reverse osmosis (Ernstrom et al., 1980; Hinrichs, 2001; Kosikowski, 1977; Lo and Bastian, 1998; O’Reilly et al., 2001). Most of these methods are inefficient due to the lost of whey proteins during curd pressing stage. One of the most attractive methods of recovering whey proteins is the enzymatic treatment of milk or curd with transglutaminase which links whey proteins with curd proteins leading to a noticeable increase in protein and yield of the cheese accompanied by a decrease in whey protein content. This treatment has economical, nutritional and environmental importance. Part of Ph.D. Dissertation of the second author. Received 30/12/2008 Accepted 25/3/2009 Mesopotamia J. of Agric. (ISSN 1815-316X) Vol.(37) No.(4) 2009 Transglutaminase (Protein-glutamine : Amine γ-glutamyl transferase, EC: 2.3.2.13) catalyzes the acyl-transfer reaction between the γ-carboxamide group of peptide-bound glutamine residues and various primary amines, including the εamino group of protein lysine residues. The cross-links introduced by this enzyme change the protein structure and improve its functional properties, like texture, viscosity and water-holding capacity without decomposing the nutritional quality of the lysine residue (Seguro et al., 1996). The enzyme is found in almost all eukaryotic and prokaryotic organisms. It has been approved by food industry to improve quality of many foods such as meat, fish, soy products, yogurt, ice cream and cheese (Zhu et al., 1995; Motoki and Seguro, 1998; Kuraishi et al., 2001). Several factors may affect the efficiency of transglutaminase in stimulating the cross-linking of whey proteins with the curd proteins such as temperature, pH, concentration of the enzyme, heat pretreatment of the milk, availability of cofactors and the step in cheese manufacture at which the enzyme is added. The present work was aimed to investigate the effect of MTGase concentration and its addition time on the physical and sensory properties of the produced cheese as a function of storage time. MATERIALS AND METHODS Chemicals: N-carbo-benzoxy-L-glycine, L-glutamic acid γ-monohydroxamate and hydroxylamine were obtained from Sigma Chemical Co. Calf rennet was from Chr. Hansonsۥ Laboratories, Copenhagen. All other chemicals were reagent grade. Bacterial cultures: A lyophilized culture of Streptoverticillium mobaraense (DSM40847) was obtained from Deutsche Sammulung Von-Micro-Organismen Und Zelkulturen GmbH (DSMZ). The culture was maintained on slants of malt extract agar at 30 oC for 6 days and stored at 4 oC. The bacteria were transferred from the slants to 100 ml of the propagation medium used by Ando et al. (1989) and contained 0.5% glucose, 0.2% peptone, 0.2% K2HPO4 and 0.1% MgSO4 at pH 7 in 500 ml Erlenmeyer flasks. Propagation was conducted in a shaker incubator at 30 oC and 100 rpm for 2 days. The culture was then transferred to the enzyme production medium which was consisted of 2% starch, 2% peptone, 0.2% yeast extract, 0.2% K2HPO4, 0.1% MgSO4 and 0.05% glycerol. Incubation conditions were as described above. Enzyme isolation and precipitation: At the end of the incubation period (3 days), the culture was filtered through Whatman no.1 filter paper and the filtrate was cooled to 0 oC. The enzyme was precipitated from the filtrate by adding one volume of cold acetone (-15 oC) with gentle stirring. The precipitate was dissolved in Tris-acetate buffer (0.1 M, pH 6) and lyophilized. Determination of transglutaminase activity: The activity was measured by the colorimetric hydroxamate procedure with N-carbo-benzoxy-L-glycine as the substrate according to the method of Folk (1970). Reaction mixture containing 50μl of the enzyme, 350 μl of Tris-acetate buffer (0.1 M, pH 6), 25 μl of 2 M hydroxylamine and 75 μl of 0.1 M CBZ-L-GLN-GLY, was incubated at 37 oC for 10 minutes and then the reaction was stopped by adding 500 μl of 15% trichloroacetic acid (TCA) containing 5% FeCl3. The absorbance was measured at 525 nm using CECIL 3021 spectrophotometer. One unit of transglutaminase Mesopotamia J. of Agric. (ISSN 1815-316X) Vol.(37) No.(4) 2009 activity was defined as the amount of enzyme which causes the formation of one micromole of hydroxamic acid per min at 37 oC. A calibration curve was prepared using L-glutamic acid γ-monohydroxamate. Soft cheese manufacturing: Whole fresh cow milk (containing 3.7% fat, 3.38% protein and 12.44% total solids) was pasteurized at 72 oC for 15 seconds, cooled down to 35 oC then calcium chloride (0.3 gm/10 kg) and the rennet were added followed by incubation for 40-45 minutes for coagulation then the curd was cut and left for 5 minutes. The separated whey was drained out and the salt was added ( 3% of curd weight) with mixing for five minutes. The curd was then transferred into plastic molds and pressed (2 kg/cm) for two hours. The produced cheese was then stored at 4 oC. Treatment with MTGase: The enzyme with concentrations ranging from 12-72 unit/liter milk was incorporated into the cheese at three different stages of manufacturing. In the first treatment, MTGase was added to the milk and incubated at 35 oC for 30 minutes before adding the rennet. In the second treatment, the enzyme was added with the rennet at the same time. In the third treatment, the enzyme was added after coagulation and curd cutting using the same concentrations. Chemical analysis of milk and cheese: Fat content of milk was estimated using Gerber method according to British Standards Institution (1969). Milk protein was estimated by Micro Kjeldahl method according to AOAC (2000). Total solids were measured by a hydrometer (Quevenne). pH of cheese was measured using a pHmeter (WTW-530). Titratable acidity was estimated according to AOAC (2000). To detect the formation of cross-links, SDS-PAG electrophoresis was conducted for cheese and whey proteins by using vertical electrophoresis apparatus (Jookoh Co. LTD) following the procedure described by Laemmli (1970). Determination of curd tension and cheese hardness: A texture analyzer with TA7 plate-shaped probe was used for curd tension measurement while a TA16 conical-shape probe was used for the measurement of cheese hardness. The force required to penetrate 20 mm of the sample at a speed of 1 mm. sec was estimated. Sensory evaluation: Sensory evaluation of the produced cheese was carried out for flavor, texture, holes, consistency, color and bitterness after 1, 4, and 8 days of storage. Statistical analysis: The data were analyzed by using the Complete Random Design (CRD). Sensory evaluation data were analyzed by using Factorial Complete Random Design (FCRD) according to SAS (2001). RESULTS AND DISCUSSION Soft cheese was manufactured using TGase-treated and untreated milks. Three treatments of the enzyme addition with various concentrations were tested. Addition of MTGase prior or with the rennet addition: The addition of the enzyme to the milk prior to rennet addition caused the prevention of milk coagulation. The second treatment included the addition of MTGase to the milk with the rennet at the same time. This treatment caused a significant drop in the curd and cheese strength accompanied by a noticeable loss of protein and fat in the whey with all concentrations of the added enzyme (Table 1). The curd strength is considered as an important parameter in the cheese manufacture process due to its Mesopotamia J. of Agric. (ISSN 1815-316X) Vol.(37) No.(4) 2009 direct effect on cheese properties. Weak curd always produces cheese with low quality and yield due to the loss of noticeable amounts of protein and fat in the

Extraction and characterization of polyphenol oxidase from apricot, apple, eggplant and potato

Abstract: Polyphenol oxidases (PPOs) from the fruits of apricot, apple, eggplant and potato tubers w ereextracted by homogenization with potassium pho sphate buffer followed by precipitation with 1.5 volume sof cold acetone. Some properties of the isolatedPPOs were studied. The optimum pH for potato PPO activity was found to be 6.4 while it was 7 for the other fruits PPOs. Optimum temperature of activit y was 20 o C for apricot and apple PPOs, while it was 22 o C for eggplant and potato PPOs.

MICROPROPAGATION OF GARDENIA Gardenia jasminoidesBY USING SINGLE NODES

Abstract: The present study was carried out to study the micropropagation of Gardenia (Gardenia jasminoides) by using single nodes excised from soft cuttings using half strength MS salts, 30gl sucrose, 7gl Agar and different concentrations of plant growth regulators in culture medium. Results of the experiment at initiation stage revealed that the culture of single nodes of Gardenia on a medium containing 5mgl BA gave the highest number of shoots (2.2 shoots/explant). Concerning the interaction, a nutrient medium containing (2 mgl BA+ 0.4 mgl IAA) gave the highest values of average number of shoots and leaves and length of new shoots (1.6 shoots/explant, 2 leaves/explant, 1.6cm respectively). At vegetative multiplication stage, the results showed that the medium supplemented by 2 mgl -1 BA gave the highest values of average number of shoots and leaves and length of new shoots (2.20 shoots/explant, 3.20 leaves/explant, 1.80cm respectively), whereas the medium supplemented by 0.3 mgl IAA gave the highest number of shoots and length of new shoots. In order to promote the shoots produced at vegetative multiplication stage to root, it was noticed that the treatment of 8mgl NAA gave the highest percentage of rooting (90%) and the medium supplemented by 12mgl IAA gave (100%) rooting. Plantlets obtained were transferred to pots and acclimatized with 95 % success. INTRODUCTION Gardenia jasminoides or common Gardenia is a member of the family Rubiceae and belongs to the genus Gardenia. There are over than 200 species of Gardenias. Two species are of primary importance, Gardenia jasminoides, containing many cultivars, and Gardenia thunbergia, grown primarily as a rootstock. Gardenias can be used as screens, hedges, borders or ground covers. They may also be used as free-standing specimens or in mass plantings. Cultivars of Gardenia jasminoides can be propagated by cuttings or grafting. Cuttings can be taken in any time during the year. Propagation could be done by grafting scion from a desired cultivar to a seedling rootstock of Gardenia thunbergia. Rootstock seedlings, however, they are difficult to obtain due to problems in seed germination. Plant tissue culture is the growth of plant organs or tissues in aseptic culture where the environment as well as the nutrient and hormone levels for growth are tightly controlled (Razdan, 2003). Pontikis and Sapoutzaki(1984) were able to get shoots grown on terminal buds of troyer citrange seedlings grown on MS medium enriched by 0.5 mglBA, 0.1mgl IBA and 0.1 mgl GA3. Pasqual and Audo (1989) promoted vegetative multiplication of tri-foliate orange shoots by using MS medium supplemented by 2 mgl BA and 1mgl NAA. Kyoichi et al. (1987) found the best response of shoot tip of pear cultivars (Bartlett, Lelectierr, Silver Bell and Lafrance) when cultured in Received 5 / 8 / 2008 accepted 31/ 12 / 2008 Mesopotamia J.of Agric. (ISSN 1815-316X) Vol. (37) No. (3) 2009 MS half strength medium with 8.88-4.4 Mm of BA. Werner and Boe (1980) found that MS medium with half salts strength and 0.5mgl BA was the optimum medium in M7 apple rootstocks multiplication and they could isolate 13 shoots from one shoot. Fiorino and Leva (1986) Berenguer and Gonzales (1990) and Rugini (1990) had emphasized that number of shoots reached to 3 shoots/explant by using half strength MS medium supplemented with zeatin, GA3 and IBA in olive propagation. Mante and Tepper (1983) and Hatamla and Al-Khazaela (1990) found that the growth regulator BA with concentration of 1-4 mgl is the best concentration in multiplication stage, While Hamad (1996) and Mendes et al. (1996) noted that concentration of 4.5,5 mgl of BA is the best in multiplication to propagation of Musa tektiles in MS nutrient media. Moretti et al. (1991) obtained a high percent rooting when pear shoots cultivar (Kaiser) cultured on MS media with half strength supplemented with IAA at 0.49 Mm. Bader et al. (2000) found that perfect rooting (100%) can be obtained at growth shoot culture with pear stock (Pyrus calleryana) in multiplication stage on MS medium half strength with 6.66 Mm NAA which gave 2.7 root/shoot. Abdullah et al. (2003) stated that many plantlets were obtained by culturing shoot cuttings of Gardenia in MS nutrient media with 30gl sucrose, 7 gl agar and different concentrations of BA and IAA .The best combination was 1mgl BA with 0.5mgl IAA. This treatment gave the best shoot growth suitable for rooting in primary and secondary culture by reculturing the rootstock cutting every 6 weeks and for many times. The objectives of this study were to evaluate the affects of explant type, MS salt strength and different type and concentration of CKs and auxins on in vitro propagation. MATERIALS AND METHODS This experiment was carried out at the laboratory of plant tissue culture of Horticulture Department, College of Agriculture, University of Dohuk between June, 2006 and January, 2007. Plant Materials [Source of Explants and Explant Preparation]: Shoots of Gardenia jasminoides 15cm in length were collected from plants grown in the greenhouse of the Department of Horticulture/College of Agriculture at University of Dohuk. Immediately after collection, the shoots were kept in polyethylene bags and taken to the laboratory. After leaves removal, selected shoots were washed under tap water for 1 hour, followed by tap water and liquid soap for 20 minuets, followed by three – five minute rinses in sterile distilled water. Then they were cut into shorter sections (1.5 cm) long including the single nodes with axillary bud. To decrease tissues browning, shoot were placed in cold antioxidant solution containing (150 mgl) citric acid and (100 mgl) ascorbic acid for 20 minutes followed by 5 minute rinses in sterilized distilled water (Mohammed and Omer,1990 and Olivares et al, 1990 ). The shoots were disinfect by immersion in the solutions of Mercuric Chloride (HgCl2), (0.1%) w/v for 10 minutes. The disinfect tissues [explants] were rinsed 5 times with sterilized distilled water, and the ends of explants exposed to sterile solution were trimmed. The nutrient media contained half inorganic and organic constituents according to (Murashige and Skoog, 1962). Then 30 gl of sucrose, 100 mgl of myo-inositol, and different Mesopotamia J.of Agric. (ISSN 1815-316X) Vol. (37) No. (3) 2009 vitamins in addition to several studied growth regulators were added according to the purpose of the experiment and further than containing 7 gl agar. In initiation stage: Different concentrations of BA and IAA were tested to find out their effect on culture initiation both alone and when combined together. BA was used at 0, 2, 3.5 and 5 mgl and IAA at 0, 0.2, 0.4 and 0.6mgl alone or in combination. Ten explants were cultured (an explant in each test tube for each concentration). They were incubated on 24±1°C under light conditions of 16 light hours 1000 lux and 8 darkness hours. The results were recorded after 4-6 weeks from planting. In multiplication stage: BA was tested at concentrations of 0, 2, 3.5 and 5 mgl and IAA at 0, 0.1, 0.2 and 0.3mgl alone or in combination. GA3 was added to MS medium with 3mgl to all the treatments including control treatment. In rooting stage: when the explants reached 2-4cm in length, they were transferred into half strength MS medium containing different concentration of IAA and NAA (one shoot for each test tube and ten test tubes for each treatment). The effect of IAA and NAA added to the culture medium on shoots rooting was studied by carrying out several separate experiments by adding NAA with ( 0,2, 4, 6 and 8) mgl and IAA with (0,3,6,9 and 12) mgl ,all these treatments were examined in half strength salt. Number of rooting shoots, root number /shoot and root length (cm) were recorded and this evaluation was performed on a weekly basis for 4-6 consecutive weeks. At the end of six weeks, the results were compiled, averaged and expressed as a percentage or number for each treatment. In acclimatization stage: After 6-8 weeks from Gardenia shoots rooting, several plantlets were selected from those that formed good vegetative and seedy growth. They were washed under tap water to remove agar from the roots which might be a source of contamination. It is important to avoid cutting any part of the roots during washing. They were then put in Benlate fungicide solution (0.1%) and then planted in plastic pots filled with a sterilized mixture of peat moss and river soil (2:1). In order to maintain high humidity in culture environment, the pots were covered with a light plastic cover which permits light passing and contains many openings to permit air entrance. Plants were watered and given a solution containing MS salts with 0.25 of original power. The plastic cover was removed from time to time after two weeks from planting. After four weeks, the transplants were transplanted after being sprayed with Benlate fungicide (0.1%) as required. Statistical Analysis: Experiments were designed as Randomized Complete Block design (RCBD).Ten test tubes were used for each treatment. Data scored on percentage were subjected to arcsine transformation before analysis and then converted back to percentage for presentation. Significant differences among mean values were separated using Duncan multiple range tests at P≤0.05 (Duncan, 1955). RESULTS AND DISCUSSION Initiation Stage: Figure (1) shows that the effect of different concentrations of IAA, BA and their interaction on response percent of the single nodes excised from the soft cuttings of Gardenia cultured on half strength MS salts. Control treatment gave the lowest response percent (60%), whereas the highest concentrations of BA (3.5, 5 mgl) gave the highest response percent (100%) which was superior upon Mesopotamia J.of Agric. (ISSN 1815-316X) Vol. (37) No. (3) 2009 the other concentrations. Concerning the interaction between BA and IAA concentrations the figure shows that the treatment of 2 mgl of BA + 0.2,0.4 mgl of IAA ,3.5 mg

Effect of some additives on bacteriological properties of local meat product (basturma) during storage

Abstract: Basturma is a traditional meat product manufactured to preserve meat in Iraq. The objective of this study was to determine the influence of adding ascorbic acid, nitrate and nitrite, sodium phosphate, acetic acid and lactic acid on the survival of some microflora during storage period under natural climate conditions. Results demonstrated that the used additives, reduced the growth of some microorganisms studied and eliminated the others during storage depending on the type and concentration of additives. In general, the use of different concentrations of ascorbic acid, nitrate and nitrite and sodium phosphate decreased the total count, coliform, lipolytic and proteolytic bacteria, along the period of storage. When higher concentrations of different additives were used, a complete inhibition of all types of bacteria. The results showed similar patterns at the last months of storage. The treatment of meat trimming before grinding with 5% of both acetic and lactic acids generally led to the decrease of total count and coliform and a very weak growth of proteolytic bacteria and a complete inhibition of the counts of lipolytic bacteria . INTRODUCTION Basturma, an Iraqi meat product , is produced in a large amount in various parts of the country, especially in Mosul city (Nineva province). It is similar to Turkish dry-fermented sausage \"Sucuk\", Spanish dry fermented sausage \"Chorizo\" , which is havily smoked , and Greek traditional sausage . The Greek product has to be kept until consumption under cold storage and is characterized as fresh non cooked (like Iraqi basturma) and manufactured under the addition of salt, phosphate, nitrite, ascorbic acid, sugars and different seasoning . They may be partially dried or smoked and should be consumed only after heat processing (frying or grilling) (Papadima and Blouka,1999 and Ambrosiadis et al.,2004) Basturma is highly regarded among consumers in local markets. However, traditional product does not usually achieve the uniform and consistent quality. This problem seems to be related to the fact that the changes that constituents undergo thoughtout ripening process are not fully understood. These changes contribute to the typical organolebtics of the product. Basturma in Iraq is produced from lamb, tail fat, salt and spices, however no others additives such as ascorbic acid, nitrate, nitrite, phosphate and organic acids.....etc. Other are added. The production is preferably during the coldest months of the year (autumn, winter and spring) and should be consumed only after heat processing (frying). The basturma is fermented during the partially drying process under climatic Part of Msc thesis of the first auther Received 14/1/2009 accepted 18/2/2009 Mesopotamia J. of Agric. (ISSN 1815-316X) Vol.(37) No.(4)2009 conditions. The fermentation was occurred by the inoculation from the raw material or environment during the manufacturing occurred with Turkish sucuk (Bozkurt and Erkmen,2002). Combination of drying and fermentation is known as the ripening period. The main changes take place during this period affects the quality attribute of basturma and continue in the storage period. The ripening of basturma during storage may be represented by a set of reactions involving proteins, lipids and carbohydrates and many different types of end products are formed, this means that there are several possible sources of contaminations of basturma. Research on basturma is very limited in the literature. The studies carried out on this product until nowadays refer only to some chemical characteristics, and there is a lack information about the microbiological changes that occur throughout its storage. Meat and meat products represent an important part of the human diet. Meat is considered to be suitable conditions for the growth of hazardous microorganisms. Microbial contamination can lower the quality of fresh minced meat, shorten its shelf-life. Dorsa, et al. (1998) demonstrated that the most important factor contributing to source and level of microbial contamination of ground beef was the microbial state of raw beef material used for grinding. However, it could be noted that the content of water and nutritional needs in different meat is considered suitable condition for the growth of microorganisms and may affect the meat safety or produce many toxic compounds (Phillips et al., 2001). For this reason several techniques have been used to preserve meat products and to minimize or to inhibit the growth of these microorganisms. Many researchers began to refocus attention on the use of some additives like acidulants and other material in the processing of meat trimming destined for ground meat, as a means to provide additional safety for the ground meat-product. Antimicrobial effect of various additives with different concentrations was added to the meat products (Dorsa et al., 1998 and Ellebracht et al., 1999). An organic acid that has been frequently studied on meat tissus is acetic acid (Hardin et al., 1995). Acetic acid has effective antimicrobial capabilities due to its ability to lower the pH and cause instability of bacterial cell membrane (Luck and Jager , 1998). Total count bacteria is considered a good indicator of spoilage of foods like meat products. It is noticed that the use of additives like preservatives and organic acids decrease significantly the total count in ground meat (Stivarius et al., 2002a). The addition of some preservative materials greatly affects the decreasing coliform bacteria. Pohlman et al. (2002) mentioned that adding sodium phosphate to meat decreased total bacterial count. When 10% sodium phosphate is used, the amount of total bacterial count decreased from 7.18 to 6.57 log cfu/g, E.coli from 6.69 to 5.92 log cfu/g and coliform bacteria from 6.33 to 5.66 log cfu/g. However, Stivarius et al. (2002b) found that the addition of some kinds of organic acid, like acetic acid to meat and meat products decreased E.coli and coliform from5.89 and 5.70 to 4.64 and 4.23 log cfu/g, respectively. They also find that the addition of lactic acid (5%) decreased E.coli and coliform from 6.69 and 6.33,6.03 and 5.63 log cfu/g, respectively. Mesopotamia J. of Agric. (ISSN 1815-316X) Vol.(37) No.(4)2009 Anderson and Marshall (1990) showed that citric acid in combination with acetic acid, lactic acid and ascorbic acid reduced aerobic plate counts, E.coli and Salmonella Typhimurium. Nitrate and nitrite are considered a preservative materials and they affect the growth of some microorganisms (Lucke, 1998). Sodium nitrite is widely used as an additive in cured meat where it has important antimicrobial and antibotulinal properties. Therefore, due to the lack of documentations on the effectiveness of adding some additives, the objective of this study was to evaluate the effects of adding ascorbic acid, nitrate and nitrite, sodium phosphate and some organic acids on some bacteriological properties during storage period (ripening period) of basturma. MATERIALS AND METHODS Preparation of samples: The basturma which was used in this study was prepared according to the traditional procedures and the fermented and drying steps are carried out in rooms subjected to natural climate conditions with sufficient air circulation. The lean meat (lamb) and tail fat (about 20%) was cut in small pieces (about 2×2×2 cm), then minced. Minced meat (about 4 mm), minced fat, salt (NaCl about 1.5%) and different spices, especially used in the production of basturma in local markets are mixed (hand kneeded) and stuffed after adding the additives into natural animal casings (intestine, locally named \"Sundaweel\") of about 4-6 cm in diameter and 30-40 cm length. The casings are obtained by cleaning and stripping off the mocus, and muscular layers and usually preserved by drying. After stuffing the basturma spicked over some parts of the surface to allow the entrapped air to escape and after that stored for some hours under heavy materials, then hanged on ropes in storage room. Treatments and sample processing: The treatments of this study include (1) an untreated control, (2) addition of ascorbic acid, 100, 200 and 300 mg/kg, (3) addition of nitrate and nitrite, 300+100, 400+150, and 500+200 ppm, (4) addition of ascorbic acid in combination with nitrate and nitrite, 200 mg/kg and 400+150ppm, respectively, (5) addition of sodium phosphate, 0.1, 0.2 and 0.3%, (6) 5kg of meat trimmings is dipped for two minutes in 5% solution of lactic acid and acetic acid separately, then minced to prepare the basturma. Microbiological analysis: The microbiological analysis of all samples are evaluated for total bacteria count coliform, proteolytic and lypolytic bacteria, using the methods mentioned by Harrigan et al., (1976). The bacterial growth is determind as colony forming units (cfu). The microbiological analysis was: Total bacterial count which determined using nutrient agar (Himedea lab. Pvt. limited, India) incubated at 30C for 2 days, coliform on MacConkey agar which contained bile salts (Himedea lab. Pvt. Limited, India) incubated at 37C for 2 days, proteolytic bacteria on nutrient agar with 10% of sterilized skim milk incubated at 30C for 3 days and lipolytic bacteria on nutrient agar with 10% olive oil incubated at 30C for 2 days, then the growth colonies were dipped in zinc sulphate solution and enumerated. All different counts were plated onto growth media in duplicate. Mesopotamia J. of Agric. (ISSN 1815-316X) Vol.(37) No.(4)2009 Statistical analysis: The experiment was conducted in duplicate. Data were analyzed by using the complete randomized design (CRD).The Duncan multiple range test, at level of (P<0.05), was used to detect differences among means. The data were analyzed using the SAS program (Statistical Analysis System)(2001). RESULTS AND DISCUSSION Changes in different kinds of bacteria in stored basturma: Some kinds of bacteria were studied in basturma product from the beginning of manufacturing and d