Food Biotechnology
Kobra Tajik Toughan; Mohammad Reza Edalatian Dovom; Seyed Ali Mortazavi; Ali Javadmanesh
Abstract
[1]Introduction: Poultry and meat products are the largest sources of non-typhoid salmonella infections in most countries. Studies have shown that raw foods of animal origin, especially poultry and its products, are the main source of contamination of kitchens and restaurants. In terms of growth conditions, ...
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[1]Introduction: Poultry and meat products are the largest sources of non-typhoid salmonella infections in most countries. Studies have shown that raw foods of animal origin, especially poultry and its products, are the main source of contamination of kitchens and restaurants. In terms of growth conditions, these microorganisms are resilient bacteria and easily adapt to their environmental conditions. Salmonella has been known to cause intestinal disease for many years and has been reported as the most important cause of food poisoning. According to Iranian and international standards, there should be no S. enteritidis or S. typhimurium in 25 grams of food. DNA-based methods for the identification and differentiation of Salmonella serovars have been designed and applied using specific primers at the genus and serovar levels. Therefore, they can be used as useful and rapid screening tests, as well as to supplement or replace conventional biochemical and serological tests. Real-time PCR, with the most accurate and reliable results using a fluorescence probe, which of course has a high cost. In this method, sequence specific fluorescence probes are used, and as a result, in the target molecule, screening and determination the presence or even the concentration of specific sequences is possible. Therefore, even in the presence of other types of nucleic acid molecules, the results are obtained quickly and have a high level of specificity. Under these conditions, if specific probes with different florescence dyes are used, even multiple targets can be detected in a single PCR reaction. The aim of this study was to identify S. enteritidis or S. typhimurium by PCR and Salmonella spp. by real time PCR method in poultry products. Material and Method: In total, 45 samples of poultry products, including chicken breast, liver and gizzard (15 samples each) were purchased from different regions of Mashhad and from various companies and transferred to the laboratory in accordance with hygienic standards. For each sample, 25 g of tissue was isolated and homogenized under sterile conditions and DNA extraction was then performed using a DNA extraction kit. The extracted DNA was evaluated by agarose gel electrophoresis. The purity and quantity of DNA extracted from each sample was examined by spectrophotometry method. In the next step, in order to identify the genus Salmonella, the samples were examined by real time PCR. In this method we used an internal control to ensure that negative results are not false negative due to inhibitors. The results of real time PCR showed that out of 45 samples, nine samples were infected with Salmonella. Then, these nine samples were evaluated for Salmonella typhimurium and Salmonella enteritidis infection by conventional PCR method. Result and Discussion: The results showed that out of nine samples that were positive in real time PCR test, seven samples were contaminated with Salmonella typhimurium, of which five samples were related to chicken breast and two to liver. Regarding Salmonella enteritidis infection, out of nine samples, only one sample was contaminated, which was related to chicken breast. Conventional methods have been traditionally used to enumerate target bacteria in food. However, these methods have some limitations and require considerable time and labor. Previous studies have already shown that real time PCR is more effective than conventional bacteriological methods for the detection of Salmonella spp. In a study by Whyte et al. (2002) The presence of Salmonella was assessed by traditional culture methods and by a Salmonella-specific polymerase chain reaction (PCR) test. Salmonella was recovered from 16% of samples using traditional culture methods. In contrast, the PCR assay proved to be more sensitive and detected Salmonella DNA in 19% of the examined samples (Whyte et al. 2002). Results of PCR with specific primers showed that reactions in real time PCR with general primers of Salmonella spp. were done correctly. Despite of accuracy and speed of real time PCR to detect DNA of microorganisms, further studies are developed to have more advantages. Loop-mediated isothermal amplification (LAMP) showed a higher sensitivity of Salmonella detection in compare to qPCR (Vichaibun & Kanchanaphum, 2020). Although LAMP could detect trace amount of Salmonella DNA but primer design for this reaction is very difficult. However, it is important to highlight that non-viable cells can be detected by real time PCR or other DNA-based methods, which does not occur in traditional methods of culture and isolation that require viable cells for quantification (Zeng et al., 2016).
Food Biotechnology
Fatemeh Barmak; Mohammad Bagher Habibi Najafi; Reza Hajimohammadi Farimani; Mohammad Reza Edalatian Dovom
Abstract
Introduction: One of the most important factors in producing yogurt is choosing the right starter. Native isolates of any country are among the genetic resources of that country, which play a major role in the production and development of the organoleptic properties of fermented products. Therefore, ...
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Introduction: One of the most important factors in producing yogurt is choosing the right starter. Native isolates of any country are among the genetic resources of that country, which play a major role in the production and development of the organoleptic properties of fermented products. Therefore, it seems necessary to study the industrial applications of native isolates. In this study, the production of yogurt was investigated by using native starter isolates from traditional Khorasan yogurts and its comparison with yogurts produced with two types of commercial starters. Materials and methods: Six strains of Streptococcus thermophilus and three strains of Lactobacillus delbrueckii subsp. Bulgaricus isolated from traditional Khorasan yogurts were selected. The proteolytic activity of Streptococcus thermophilus strains was determined according to the method of Erkus et al(2012) and the proteolytic activity of Lactobacillus delbrueckii subsp. Bulgaricus was determined according to the method of Nespolo et al.(2010). Milk acidification activity byall examined strains was evaluated according to the method of Erkus et al.(2012). The production property of gamma-aminobutyric acid of native isolates was also evaluated using the method of Lacroix et al. (2013). Yogurt was produced using commercial starters and native isolates. Acidity and pH were measured according to Iranian National Standard No. 2852. Synersis of yogurt samples was measured using centrifugation according to Çelik (2007) method. In order to measure textural properties, a combination of backward extrusion techniques and Texture Profile Analysis (TPA) was used (Yang et al, 2010). Sensory characteristics of yogurt samples were evaluated by 20 panelists using five-point hedonic scale. This study was conducted based on factorial experiment using a completely randomized design. Statistical analysis of data was performed using Minitab Statistical Software (version 17) and ANOVA. Comparison of means was performed using Tukey’s test at the significant level (p < 0.05). Results and discussion: The results of proteolytic activity of Lactobacillus delbrueckii subsp. Bulgaricus showed that the L3 code was weaker than the other two strains. All Streptococcus thermophilus strains were positive protease. All Streptococcus thermophilus strains of the study decreased the pH of the milk to 4.6 in less than five hours, and Streptococcus thermophilus strain S6 code, as the fastest acid producer, decreased the pH of the milk to 4.6within 3 and half hours (Figure 2). The results of GABA production showed that among the streptococcus thermophilus strains, S1, S5 and S6 codes had distinct blue discoloration. Among the Lactobacillus strains, except the L3 strain, two other strains produced green color. Of the 18 samples of the yoghurt, S5L2 and S2L2 samples showed the highest decrease in pH and increased in acidity compared to other samples after being stored refrigerated overnight (P <0.05). The other samples had the same pH and acidity changes as the control samples after overnight refrigeration. The results show the ability of yogurt producion by native starters to compete with those produced by commercial starters in terms of technological characteristics. Thus, samples with codes S2L2, S3L1, S3L2, and S1L1 showed less Synersis of yogurt than commercial starters. This decrease in the Synersis of yogurt might due to the possibility of exopolysaccharide production by these strains. Textural characteristics of yogurt samples were the hardness of the samples in the range of 3 to 4 N, which was significantly different (P <0.05). The lowest hardness was observed for the S3L3, which can be attributed to the poor proteolytic activity of the strain. L3, S6L2, S3L2 and S2L2 had the highest hardness compared to other compounds and control samples (P <0.05). This differences in the hardness of the samples can be attributed to the possible production of some metabolites by the strains. There was no statistically significant difference in the adhesion properties of the samples (p <0.05). Considering the sensory evaluation scores, it was found that in all respects, S3L3 had the lowest score among the other samples (P <0.05). This was in good correlation with the results of the textural properties of the test. Therefore, this compound was eliminated compared to the control samples. Finally, considering the proteolytic behavior of the strains and the ability to produce acid and GABA, as well as the sensory, rheological and physicochemical properties of 18 samples in comparison to the two control samples, Streptococcus thermophilus strains with codes S4, S2, S5, S1, S6, and Lactobacillus delbrueckii subsp. Bulgaricus strains with codes L1 and L2 have the potential to be used as commercial starters in the form of compounds S1L1, S2L2, S5L1, S6L2, S4L2.
Mohammad Ebrahim Goharjoo; Mohammad Reza Edalatian Dovom; Fakhri Shahidi; Farideh Tabatabaei Yazdi; Mohammad Javad Varidi
Abstract
Introduction: Carrot products such as carrot juice and fermented carrot products possess high nutritional value and they are considered as a major source of β-carotene. Carotenoids because of containing conjugated double bonds, have antioxidant properties and provide the natural yellow, orange and red ...
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Introduction: Carrot products such as carrot juice and fermented carrot products possess high nutritional value and they are considered as a major source of β-carotene. Carotenoids because of containing conjugated double bonds, have antioxidant properties and provide the natural yellow, orange and red colors in fruits and vegetables. Due to the outbreak of some problems such as lactose-intolerance and high blood cholesterol especially in dairy products’ consumption, great attention has been drawn toward fermented vegetable products. Lactic acid bacteria (LAB) including important genera: leuconostocs, lactobacilli, streptococci and pediococci are wide-spread and have been divided according to morphological features and fermentation pathway, which utilize glucose. Current knowledge regarding involved microorganisms in vegetable fermentation is still dependent on biochemical and classical data. Nowadays, application of molecular methods in the field of microbial identification has been provided better understanding from fermented foods ecology. Since local starter cultures are considered as precious genetic resources in each country and also they play an important role in production and creation of organoleptic characteristics in fermented products, therefore, the objective of present study was the isolation and identification of lactic flora from fermented carrot with the help of conventional (biochemical) and molecular methods and determination of phylogenetic relationships.
Materials and methods: Following the production of fermented carrot samples, they were packed in plastic container and stored at ambient temperatures (25-27°C). In the next step, total LAB count was performed according to Iranian standard of 5484. Isolation and selection of LAB was done during 32 days with the intervals of 0, 4, 8, 16, 24 and 32. For initial identification of LAB, isolated were subjected to gram staining and catalase tests. Also biochemical tests including growth at 15 and 45C, at NaCl 6.5% and 18%, pH=4.4 and 9.6, were done in order to identify and classify at genus level. Carbohydrate fermentation profiles were obtained for isolates with the aid of 10 sugars. Molecular identification was done with DNA extraction followed by amplification of 16S gene with universal primers (27 F and 1492 R). For sequencing of resulted PCR-products, they were sent to Macrogen Company, South Korea. Phylogenetic tree was plotted with Clustal Omega and Fig. Tree soft wares.
Results and discussion: In the first step, 144 gram positive, catalase negative isolates were screened and selected as presumptive LAB according to gram staining and catalase test and morphological characteristics. Among them, 48 representative isolates were chosen and identified up to genus level according to biochemical tests. Five distinct genera were identified as Pediococci (4.08%), homofermentative lactobacilli (34.69%), hetero fermentative lactobacilli (36.74%), Leuoconostocs (20.41%) and enterococci (4.08%). Carbohydrate fermentation profiles revealed Lactobacilli constitute the highest percent among other genera and also some species like Lb. kimchi and Lb. parakefiri were detected. Growth of lactic acid bacteria experienced increasing trend up to day-16 but thereafter showed decline trend until the end of storage time (day-32). 26 out of 48 isolates were subjected to molecular analysis. Results of sequencing revealed following species: Lb.plantarum (9), Lb. brevis (8), Leu. mesenteroides (4), Lb. casei (1), Lb. paracasei (1), and Lb, pantheris (1). Changes and variation of lactic flora during fermentation stages revealed that at initial stages of fermentation (0- day-8) Leuconostocs sp. were predominant species but disappeared then. In the next stages of fermentation Leuconostocs sp. were replaced by homo-fermentative strains such as Lb. plantarum which was present from the first day up to day-24 but constituted the majority of species on day-16. In the final stage, Lb. brevis dominated the others due to better survival and resistance of this bacterium at the increased acidity level. Phylogenetic tree results revealed three clusters including cluster I (composed of three sub-clusters), cluster II (three sub-clusters) and cluster III (two sub-clusters). Cluster I included two genera: Leuconostocs sp. (mesenteroides) and Lactobacillus (pantheris, casei and paracasei). Cluster II included Lb. brevis and finally cluster III composed of Lb. plantarum.
Saeid Afzali; Mohammad Reza Edalatian Dovom; Mohammad Bagher Habibi Najafi; Mostafa Mazaheri Tehrani
Abstract
Introduction: Yoghurt drink or Doogh has gained great attention in recent years in different countries. On the other hand, this desirable commodity has showed some drawbacks especially from microbial point of view. One of the main problems in Doogh production and storage is considered to be the presence ...
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Introduction: Yoghurt drink or Doogh has gained great attention in recent years in different countries. On the other hand, this desirable commodity has showed some drawbacks especially from microbial point of view. One of the main problems in Doogh production and storage is considered to be the presence and activity of gas producing microorganisms especially in warm conditions and environments. From the consumer point of view, blowing of container is not acceptable and is regarded as a defect. Since lactic acid bacteria (LAB) possess the potential use as adjunct or co-culture in fermented dairy products for inhibition of gas – producing microorganisms’ especially gas-producing yeasts in doogh, inclusion of these bacteria (LAB) can be a solution for this problem in aforementioned product. In this study, antimicrobial effects of two selected strains of Lactobacillus brevis assigned as M2 and M4 isolated from Motal cheese were investigated on the gas-producing and food spoilage microorganisms in Doogh.
Materials and methods: Two strains of lactobacillus namely M2 (Lactobacillus brevis KX572376) and M4 (Lactobacillus brevis KX572378) were selected from traditional Motal cheese isolates. Doogh production was carried out according to National Iranian Standard No. 2453. Each of these two strains was inoculated in Doogh at two levels of 106 and 108 cfu / ml and control sample was taken without inoculation followed by subjecting to microbial and sensory analysis at three temperatures of 4, 25 and 37°C, at intervals of 10, 7 and 7 days, respectively. Microbial profiles including coliforms, E. coli, Mold & yeast and Staphylococcus aureus were determined according to National Iranian Standard Number (2 and 1) –5486, 5234 and 5486, 997 and 6806, respectively. Sensory evaluation was carried out according to Iranian National Standard No. 4691. Sensory evaluation parameters such as taste, texture, color and total acceptance were evaluated by 10 senior food industry students.
Results and discussion: The results showed that sample inoculated with M4106 strain, received the highest score for antimicrobial activity at all three storage temperatures. Results demonstrated that, at 4 ° C, the control sample was contaminated (spoiled) on 50th day and was positive for mold and yeast count, but sample inoculated with M4106 was acceptable (negative) for mold and yeast count. At 25 C, mold growth was detected in control sample on the 14th day of storage, but the sample inoculated with M4106 remained completely un-spoiled until the 21st day. At 37°C, the control sample on day 7 was positive for mold and yeast count, but the sample inoculated with M4106, mold and yeast was not detected until the 14th day. Coliform, Staphylococcus and Escherichia coli counts in M4106 sample were negative. Sensory evaluation was carried out according to Iranian National Standard No. 4691.Sensory evaluation data showed that samples inoculated with M4106 were superior to the control sample. Doogh samples inoculated with Lactobacillus brevis strains experienced acceptable sensory evaluation so that they gained better score than control sample. The highest score was obtained for the M4106 sample. The texture of the Doogh samples showed a decreasing trend at all three temperatures during the storage period. Moisture and pH are factors influencing texture changes during the initial stages of storage. Based on the taste evaluation results, at 4°C, the highest score was related to M4108 until day-20, but from the 20th to the 60th day of storage, the M4106 has gained the highest one. At 25°C until day 7, the highest score belonged to sample M4108 but from this day on, M4106 has been more favorable. At 37°C on production day, highest score for the M4108 sample but on day- 7 and 14, the M4106 took the highest score. Regarding to color index, M2108 has gained the highest score at all 3 temperatures on the production day and for the rest of storage time, the highest scores were obtained for the M4108 and M4106 samples. Maximum total acceptance, was obtained for M2108 on the production day but this was replaced by M4106 for the rest of the time .The results showed that the Lactobacillus strain M4106 strain had the highest antimicrobial activity and the optimum score for sensory evaluation, as well as a significant increase in Doogh shelf life and reduced gas production in the bottle.
Samira Abbaspour Monjezi; Mohammad Reza Edalatian Dovom; Mohammad Bagher Habibi Najafi; Arash Koocheki
Abstract
Introduction: Nowadays, consumers prefer foods produced without synthetic preservatives. These chemical preservatives have been gradually replaced by natural preservatives in formulation of edible films and coating. Since, edible films can be applied as carriers of antimicrobial agents, so, these aforementioned ...
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Introduction: Nowadays, consumers prefer foods produced without synthetic preservatives. These chemical preservatives have been gradually replaced by natural preservatives in formulation of edible films and coating. Since, edible films can be applied as carriers of antimicrobial agents, so, these aforementioned ingredients can be incorporated in such films. Among edible films, protein-based films such as whey protein concentrate (WPC)-based films are more attractive because they also supply valuable nutrients and introduce acceptable mechanical resistance. On the other hand, these films present moderate barriers to moisture due to the hydrophilic nature of whey proteins. Essential oils (Eos) can be incorporated in to edible films in order to compensate (overcome) this defect. Since no published research has been found on integrating mastic tree sap (Pistacia atlantica sub sp. kurdica) essential oil into whey protein edible films, this essential oil was applied for WPC-based film in this research. Some species belong to Penicillium have been known as contaminants of dairy and fruit products. Among Penicillium sp., P. expansum is more popular for causing post-harvest damage of apples. In this study, our objective was focused on mechanical and anti-fungal properties of WPC-based films incorporated with mastic gum essential oil.
Materials and methods: WPC, mastic tree sap and P. expansum were obtained from Multi Milk Company, Kurdistan mastic Gum Company and Persian Type Collection Culture, respectively. Extraction of EO from mastic gum was accomplished using water distillation or hydro distillation with the help of Clevenger-type apparatus for 5 hours to obtain a pale yellow oil. Solution (10%w/w) of WPC in distilled water was prepared. Glycerol (as plasticizer) was added to WPC solution at a ratio of 1:1 WPC: Glycerol. Then concentrations of EO (1000, 2000, 3000 and 4000 ppm) was added to solution and mixed for 2 min. In the next step, some characteristics of film were measured including: thickness and density, water solubility, stability in acidic and alkaline solutions, water vapor permeability and light transmission / film transparency. Some mechanical properties of films such as tensile strength (TS) and elongation at break (%E) of films were also determined.
Regarding microbial assays, following the activation and preparation of fungi spore, MIC was determined using Agar Dilution Method. Determination of antimicrobial activity of film was performed according to film disk agar diffusion assay
Results & Discussion: With increasing essential oil concentration, film thickness exhibited increasing trend which was due to entrapment of micro-droplets of essential oil in film. Along with increasing EO concentration in film samples, WVP declined significantly (P-value
Elham Eshaghabadi; Farajollah Shahriari; Mohammadreza Nassiry; Mohammad Reza Edalatian Dovom; Amin Mirshamsi
Abstract
Introduction: Bread is considered as one of the most important commodity which has provided human nutritional needs for several centuries. In recent decades, bread consumption has witnessed tremendous increase due to rising of other foods expenses. Cereal and bread industries has been always concerned ...
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Introduction: Bread is considered as one of the most important commodity which has provided human nutritional needs for several centuries. In recent decades, bread consumption has witnessed tremendous increase due to rising of other foods expenses. Cereal and bread industries has been always concerned about bread production in an industrial scale along with suitable shelf life, acceptable organoleptic properties and free from any chemical additives and preservatives. Among different methods such as addition of synthetic enzymes, gums, emulsifiers and improvers, sour dough has gained special attention besides shelf life enhancement and improvement of sensorial and nutritional features. The main role of sour dough has been prominent for fermentation and gas production in dough as a result of inclusion of Lactic Acid Bacteria (LAB) as a technology. Fluctuation of process parameters including temperature, dough yield and starter culture composition determine the sour dough quality. The main reason of sour dough application in wheat bread is flavor enhancement. Culture-based microbiological examinations have accompanied with some difficulties in accurate and precise identification of Lactobacilli. So, in recent years, in order to identify precisely, different molecular and PCR assays and gene sequencing have been applied. Thus, in present study, fragment of 253bp from 16 S rRNA gene in Lactobacillus plantarum was amplified with the help of PCR and sequenced in the next step. Also, comparison of nucleotide sequence of this region of traditional Lb. plantarum with nucleotide sequence of industrial starter (Germany) and sequences deposited in Gene Bank was evaluated.
Material and Methods: Sampling: According to sensory properties, sour dough samples were collected from Torbat e jam city and industrial starter sample (sour dough) was prepared from Buker Company, Germany.
Isolation and bio chemical analysis: Ten- fold serial dilutions of samples were prepared and cultured on MRS agar. Gram positive, catalase negative isolates were considered as presumptive LAB.
Molecular Identification: DNA of isolates were extracted according to the procedure of DNA extraction kit (Thermo, K721, USA). In order to amplify the 253 bp fragment of 16S rRNA in Lb. plantarum, specific primers were designed using Primer Premier 5. PCR was performed in Biometra Thermocycler (T- personal, Germany) in 36 cycles. In the following step, PCR products were sent to Macrogen Company for sequencing.
Bioinformatics analysis: Analysis of sequencing results was conducted with the aid of bioinformatics soft wares including CLC Main work bench, Chromas Lite 2.01, MEGA 5 and BioEdit. Phylogenetic tree was designed among different isolates using Neighbor-Joining method (bootstrap=1000) in MEGA 5 software. For determination of genetic distance, Create Pairwise comparison method was used in CLC Main workbench.
Results and Discussion: Quality and quantity of extracted DNA samples were confirmed by Gel electrophoresis. PCR and electrophoresis of PCR products confirmed and proved the accuracy and validity of amplified 253 bp fragment with intended primers. Comparison of Torbat and Germany nucleotide sequences demonstrated that two replacement mutations have been occurred including (A/C) and (T/C) in positions 182 and 205, respectively. These changes resulted in replacement of Histidine by Proline, and Glutamine by stop codon. Nucleotide content of 16S rRNA gene sequence in Lb. plantarum isolated from Torbat e Jam and Germany sour dough samples displayed both sequences contained 49.4% (G+C) and 50.6% (A+T). Phylogenetic results demonstrated that Lb. plantarum isolated from Torbat e Jam and Germany samples were classified in the same sub group. This implies the close phylogenetic relationship between these two lactobacilli. In order to identify Lb. plantarum, 16S rRNA gene was sequenced in sour dough samples of Germany and Belgium and phylogenetic tree showed that Lb. plantarum had the lowest genetic distance with Lb. pontis and highest one with Lb. sanfranciscensis and Lb. salivarius
Neda Nayyeri; Mohammad Reza Edalatian Dovom; Mohammad Bagher Habibi Najafi; Masoumeh Bahreini
Abstract
Introduction : Nowadays, attention to foods free from chemical preservatives is on the rise. Recently, consumers have concerned about foods containing these preservatives in their formulation .Therefore,the use of antimicrobial peptides produced by Lactic acid bacteria (LAB) are strongly highlighted. ...
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Introduction : Nowadays, attention to foods free from chemical preservatives is on the rise. Recently, consumers have concerned about foods containing these preservatives in their formulation .Therefore,the use of antimicrobial peptides produced by Lactic acid bacteria (LAB) are strongly highlighted. Ribosomally-synthesized peptides, which possess antimicrobial properties, are produced by a vast range of organisms including prokaryotes and Eukaryotes.Yeasts are the most important microorganisms which are responsible for fruit juice and soft drink spoilage. In case of growth and production of by-products like CO2, acid and other contaminants, the spoilage will be visible.Lactic acid bacteria isolated from natural, local sources such as dairy products have presented potentially antifungal features against food spoilage fungi. Among these, Rodotorula and Penicillium are regarded as the most critical genera in fruit juice spoilage.The main objective ofthis study was the evaluation of anti-yeast activity of Lactobacillus plantarum isolates from different stages of Lighvan production against Rodotorulamucilaginosa as fruit juice spoilage indicator. In the next step, technological parameters effects were analyzed on antimicrobial potential of cell-free supernatant of these isolates. Finally, we aimed at finding the most effective isolate on aforementioned yeast.Materials and methods: NineteenLb.plantarumisolates, which were identified previously, were subjected to antifungal assay. For this purpose, RodotorulamucilaginosaPTCC 5257 was selected. Agar spot and Well Diffusion Assay (WDA) were applied for antifungal assay in solid and liquid media, respectively.Determination of yeast colony: Following the cultivation of yeasts in Potato Dextrose Broth, it was determined using Spectrophotometer.Preparation of Lb.plantarum cell-free supernatant (CFS) was carried out. In agar spot method, clear zones of inhibition around the spotted colonies were evaluated after 24-48h incubation. In WDA, CFS of Lb.plantarum isolates were poured in wells and clear zones were evaluated around each well after 24-48 h incubation.Technological properties: The influence of different levels of pH (2, 3, 4, 5, 6, and 7) was analyzed on CFS of Lb.plantarum isolates. This assay was done according to Wang et al., 2011. Finally, results were reported using the measurement of clear zone diameter in mm. All experiments were performed in duplicates.The effects of various temperatures were applied on CFS and remaining the antifungal activity was evaluated according to Rouse et al., 2008. Finally, all CFS of Lb.plantarum isolates were subjected to Proteniase K and antifungal properties of CFS were assayed by WDA according to Ghrairi et al., 2005.Results and discussion:Agar spot results showed the highest and lowest clear zone of inhibition related to C28 and LF49 with 16 and 6 mm diameter, respectively. In this method, 11 out of 19 isolates (60%) presented anti-yeast activity with clear zone formation. In comparison between two incubation temperatures (25 and 30◦C), all isolates stronger clear zone in 30◦C than in 25◦C. This was due to enhancement of yeast growth in 25C rather than in 30C. Also, with respect to the mesophilic nature of Lb.plantarum isolates, the possibility of metabolite production are more likely in 30C. It was reported that antifungal activity of LAB is mostly due to synergistic effect of lactic acid and acetic acid. In agar spot, some colony-associated antimicrobial compounds are responsible for antifungal activity. In WDA, 8 out of 19 isolates (42%) were positive for their inhibitory effects. The highest anti-yeast activity was seen at pH=2. It seems that antimicrobial compounds are likely more stable at acidic conditions than at alkaline ones. Among isolates, C28 presented the highest stability at alkaline conditions. With pH increase, the antifungal activity decreased so that no anti-yeast activity was seen at pH=7. Regarding the different temperatures, we should mention that thermal resistance of isolates' CFSwitnessed declining trend with increasing of temperature. This fact implies the presence of low- molecular weight compounds in CFS. Finally, all isolates' CFS was subjected to proteinase K. All isolates have lost their anti-yeast activity after enzyme treatment showing their proteinaceous nature.Conclusion: In WDA, the number of positive isolates showing anti-yeast activity declined in comparison with agar spot. Since some isolates retain their inhibitory activity toward food spoilage yeast at low pH, their CFS can be applied in acidic foods like fruit juice. Also, some isolates showed their antifungal activity at high temperatures (80C and 100C) which are applied for fruit juice pasteurization, so they can be applied in fruit juices as a bio-preservative.
Mohammad Reza Edalatian Dovom; Masoud Yavarmanesh; Fariba Ghiamati Yazdi; Morteza Khomeiri; Neda Nayyeri
Abstract
Introduction : Lactic acid bacteria (LAB) are a group of gram positive, catalase negative bacteria which possess unique metabolic, physiological and morphological characteristics. In addition, lactic acid bacteria are considered as a valuable source of antimicrobial agents including bacteriocins. Bacteriocins ...
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Introduction : Lactic acid bacteria (LAB) are a group of gram positive, catalase negative bacteria which possess unique metabolic, physiological and morphological characteristics. In addition, lactic acid bacteria are considered as a valuable source of antimicrobial agents including bacteriocins. Bacteriocins are defined as non-toxic peptides produced by LAB. "Maskeh" as an Iranian traditional butter carries indigenous LAB which secrete and harbor bacteriocins. Our objective in this study was to evaluate the influence of antimicrobial compoundsproduced by isolated LAB from Maskeh, a local traditional butter, against some pathogenic bacteria suchas: staphylococcus aureus, Listeria innocoa and E. coli using culture-based methods. In the following step, influence of some technological parameters including different temperatures, pHs and proteinase K were determined on stability of antimicrobial compounds in bacteriocins.Materials and methods: Three samples of milk, yoghurt and local butter known as Maskeh were collected from different regions in the south ofKhorasanRazavi, Gonabadcity.Indicator microorganisms and culture condition: In order to activate the indicators, following the cultivation in corresponding and suitable media, they were incubated for 24 hours in suitable temperatures.Survey of antimicrobial activities: In Agar Spot, Antagonistic activities of isolates were evaluated in solid media. Finally clear zone of inhibition was measured in mm. In Well- Diffusion Assay (WDA), positive isolates from previous step, were selected for this assay. In this method, cell-free supernatant (CFS) of isolates were examined for their antagonistic activities. Finally clear zone of inhibition were evaluated around the wells.Determination of antimicrobial compound nature: Effect of proteinase K on CFS: In order to evaluate the enzyme sensitivity of bacteriocin like compounds produced by LAB, CFS of isolates were subjected to proteinase K (final concentration 0.5 mg/ml). Mixture of enzyme- CFS was incubated in 37C for four hours. Finally, the remaining inhibitory activity was determined using WDA.Influence of different temperatures on CFS: Heat sensitivity of bacteriocin like compounds was evaluated with subjecting the CFS of isolates to various temperatures. Again remaining activity of isolates was evaluated with the help of WDA.Influence of different levels of pH on CFS: CFS of isolates was adjusted at different pHs and their inhibitory effects were determined using WDA.Discussion & Results: Following the sequencing, 51 isolates were identified from different steps of Maskeh production. Results showed that 44 out of 51 isolates were effective at least against one indicator. It was clear that E.coli, Staph.aureus, Listeria. innocoa, Lb.plantarum, Lb.sake, Lac.lactis ssp. lactis,Lac.lactis ssp. cremoris were inhibited by 33, 7,11,12,7,35 and 7 isolates, respectively. Among pathogenic indicator bacteria, E.coli was inhibited maximally and regarding the non-pathogenic indicators, Lac.lactis ssp. Lactis was inhibited by the most of the isolates. Among the isolates, Ent.faecium B161 and Str.thermophilus B258 presented the highest inhibitory effect against Listeria.innocoa.Interestingly both these strains had been isolated from Maskeh, implying that they originated the same source. Formation of clear inhibition zone in agar spot method is related to colony-associated antimicrobial compounds like H2O2, lactic acid and other organic acids. Also, the strongest clear zone of inhibition was against Listeria.innocoa. Lb.plantarum B120 showed inhibitory effect against 6 out of 7 indicator bacteria. This strain has been isolated from Maskeh, and only did not affect on Lac.lactis ssp. lactis.In the next step, in WDA, CFS obtained from isolates were subjected to inhibition activity evaluation. In this assay, 39 isolates showed inhibitory activity against at least one indicator and 12 isolates had no inhibition against indicators. E.coli was inhibited by the most of the isolates (11), followed by Listeria.innocoa by 8 and Staph.aureus by 2 isolates. But noticeable point is that the strongest influence was seen against Listeria. In the last step, influence of technological parameters such as : temperature, pH and enzyme were determined on antimicrobial and inhibitory activity of CFS. In this survey, only those isolates were chosen which showed inhibitory effect against Listeria innocoa. Regarding the temperature, with increasing of this factor, the diameter of clear zone had decreasing trend. This means that bacteriocin- like compounds are sensitive to heat. Among analyzed isolates, CFS of two isolates namely Aerococcusviridans M156 and M141 possess the highest sensitivity. In contrast, the highest resistance was related to Aerococcusviridans M165. Evaluation of antagonistic activity of CFS of isolates at different pHsexperienced, in pH between 3-5, growing trend of inhibitory activity which is due to produced lactic acid. Maximum of antagonistic activity was seen at pH=3 and along with pH increase toward the alkaline condition, it decreased.Aerococcusviridans M165 and M141 showed clear zone of inhibition equal to 6.5 and 6, respectively at pH=7.Proteniase K as a proteolytic enzyme, exhibited decomposing influence on antimicrobial effects of all isolates, except two of them. This phenomenon implies the proteinaceous nature of antimicrobial compound in CFS, because of decomposition as a result of proteolytic enzyme. But regarding two isolates, the clear zone did mot destroyed even after enzyme treatment.Conclusion: Some CFS or their producing isolates can be exploited as bio-preservatives; CFS of Aerococcusviridans M 165 can be applied in foods subjected to high heat treatment. In acidic and low pH food, we can use those isolates which theirCFS remain their activity at low pH.
Mohammad Reza Edalatian Dovom; Mohammad Bagher Habibi Najafi; Seyed Ali Mortazavi; Majid Hashemi; Masoud Yavarmanesh
Abstract
The objective of this study was to evaluate lactic flora identification in fresh (one-day) and ripened (90-day) Koozeh cheese samples as one of the raw milk cheeses. In first step, in order to isolate the different genera of Lactic acid bacteria (LAB), selective and differential (corresponding) media ...
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The objective of this study was to evaluate lactic flora identification in fresh (one-day) and ripened (90-day) Koozeh cheese samples as one of the raw milk cheeses. In first step, in order to isolate the different genera of Lactic acid bacteria (LAB), selective and differential (corresponding) media were used as follow: MRS for lactobacilli and pediococci, MRS+ vancomycin for leuconostocs, M17 for lactococci and KAA for enterococci. Totally, 48 single, purified colonies were selected and identified using confirmatory, biochemical and phenotypic tests down to the genus level. Lactobacilli (33.33%), lactococci (12.5%), pediococci (2.08%), enterococci (47.91%) and aerococci (4.16%) were determined as the most abundant genera. At last, carbohydrate fermentation profile using API 50 CH and API 20 STREP kits was used for identification down to species level. Finally, following species were identified: Lactobacillus. plantarum, Lb. brevis, Lb. lindneri, Lb. helveticus, Lb. delbrueckii ssp. delbrueckii, Lb. pentosus, Lb. fermentum, Lactococcus. lactis ssp. lactis, Enterococcus. faecium, Ent. faecalis, Ent. avium, Ent. durans and Aerococcus. viridans. In fresh cheese, the Lactococci and lactobacilli genera were predominant but in ripened cheese the enterococci replaced them. Predominant species in fresh cheese were as follow: Lac. lactis ssp. lactis (44.44%) and Lb. plantarum (22.22%) but Ent. faecium (43.58%), Lb. brevis (12.82%) and Ent. faecalis (7.69%) constituted the major part in ripened cheese. We can mention that these predominant lactic acid bacteria play an important role during ripening and also flavor production in raw milk cheeses like Koozeh. So, more precise identification is very necessary using culture- based molecular and culture- independent methods down to strain level.
Mohammad Reza Edalatian Dovom; Sara Khoshnoudi; Ali Sharif
Abstract
Pistachio nut is one of the most important export crops in Iran. For prevention of rancidity and to ensure good quality, pistachio must be harvested in a short time and be prepared for drying in 24h.The Ohadi variety of Iranian raw dry pistachio nuts was selected for the experiments. Instrumental hardness ...
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Pistachio nut is one of the most important export crops in Iran. For prevention of rancidity and to ensure good quality, pistachio must be harvested in a short time and be prepared for drying in 24h.The Ohadi variety of Iranian raw dry pistachio nuts was selected for the experiments. Instrumental hardness and Sensory characteristics of pistachio nuts such as: flavor , texture and total acceptance were measured in 3 types of packaging (normal air or %21 02 , Vacuum and Oxygen Scavenger), 3 different temperatures (25,35 and 55) during 3 month storage.
Results showed that the effect of temperature, time storage and interaction effects of packaging and temperature , packaging and time storage on instrumental hardness were significant (P
Mohammad Reza Edalatian Dovom; Manuchehr Hamedi; Seyed Ali Mortazavi; Mostafa Mazaheri Tehrani
Abstract
Tomato is one of the most important crops with a yeild of 98. 8 million tons in more than 128,000 hectare of area under cultivation and 3.7 million tons production in Iran. With regard to technological advancement ( improvement, development ) and increasing demand of tomato and their products in Iran ...
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Tomato is one of the most important crops with a yeild of 98. 8 million tons in more than 128,000 hectare of area under cultivation and 3.7 million tons production in Iran. With regard to technological advancement ( improvement, development ) and increasing demand of tomato and their products in Iran and all over the world, it is necessary to know about affecting factors on production, process and storage of this product more precisely. In recent years, many researches have been performed to study physico – chemical and sensory characteristics of tomato in order to forecast necessary parameters in cultivating and selecting of the tomato variety corresponding to their own application ( fresh eating, processing and both of them ). In this research, the effect of variety and storage on physical properties of tomato paste produced from four tomato varieties (Cal .J.n.3 , Early Urbana Y , Early Urbana111 and Peto early C.H) has been determined in one year storage under ambient temperature (average temperature 25 ºC ). The analysis of data on physical properties including :%PWR,RF Index, consistency and color showed that ,Early UrbanaY had the highest consistency , PWR and total scoring for all of the properties Early Urbana Y totally was the best. Cal.j.n.3 variety in the most physical properties had the least changes during storage and from this point of view it was the most stable variety during storage of tomato paste.
Key words:Tomato paste,variety,storage time,physical properties
