جستجوی مقالات مرتبط با کلیدواژه « polymerase chain reaction » در نشریات گروه « صنایع غذایی »

Studying the microbiota of food materials is important from sanitary, spoilage and technological perspectives. Due to manual harvesting, grading and packaging of date fruit (Phoenix dactylifera L.), it's not surprising to find bacteria of human origin in date fruit products. On the other hand, microbes present in date palm, especially high moisture content fruits, could produce lactic acid and acetic acid and make the fruit sour, so this kind of microbes potentially could be useful especially in food fermentation and vinegar production. In this project, seven varieties of date fruits were examined chemically and microbiologically. Total solid and acidity of dates were between 72% to 86% and 0.06% to 0.78% respectively. Isolated bacteria were identified by 16S rDNA amplification and restriction analysis (ARDRA), sequencing and 16S rRNA gene comparison. The results show Staphylococcus epidermidis, Aerococcus, Bacillus and Leuconostoc mesenteroides are the present bacteria in local date fruits.

Dairy products can be a rich source of diverse lactic acid bacteria (LAB) with high functional properties, such as exopolysaccharide (EPS) production. EPS are high molecular weight polymers that are composed of sugar units and are secreted by microorganisms into the surrounding environment. Produced EPS can be used as an additive by a health effect and texturing properties. In this study, exopolysaccharide producing LAB (ropy and mucoid colonies) were isolated and identified from raw sheep milk and sheep yogurt that made in rural areas of Urmia. For this purpose, lactic acid bacteria cultured on MRS agar and M17 agar media and then isolated on the basis of the ability to produce exopolysaccharides to study the diversity of species by phenotypic methods (Gram stain, biochemical and physiological tests) and then identified by polymerase chain reaction (PCR). The 4 strains of LAB isolated were Gram-positive as well as catalase negative and were able to produce large amounts of EPS. The amounts of bonded and abandoned exopolysaccharides which measured by phenol/sulfuric acid method, were 40.28±0.2 to 65.26±0.47 mg/L and 105.68±3.2 to 136.35±0.2 mg/L, respectively. Sheep yogurt which manufactured traditionally in West Azerbaijan province, contained exopolysaccharide producing strains that can have the potential to use in the dairy industry.

Bacteriocins are ribosomally synthesized proteins/peptides that have bacteriocidal or bacteriostatic trait against genitically closed species. Bacteriocins have role as biopreservative and increase of the safety of food products. Study on native strains of lactobacillus plantarum (DL2 ¡BL1 ¡L28 ¡L27 ¡ L25 and EL3) that their growth inhibition ability had been analyzed, was performed. after approving inhibition ability of strains by disk diffusion in previous study, to investigate bacteriocin encoding gene, polymerase chain reaction (PCR) was performed by specific primers. The correspondant region was amplified in L25, BL1, EL3 and L28 strains. Using designed primers and sequencing could be shown that EL3 strain contained the structural genes for the bacteriocins plnEF, plnJK and plnN, howover, the Lb. plantarum strain BL1 and L28 contained only the structural genes for plantaricin EF production. sequences showed ٪99 similarity with bacteriocin genes of Lactobacillus plantarum strains recorded in Gene Bank database. The sequences were submitted at NCBI with accession numbers of EL3 (KT028600), L28 (KT028601) and BL1 (KT028602).

Pistachio nut is one of the popular tree nuts. Among the different species of the genus Pistacia, only the fruits of Pistacia vera attain optimal size to be acceptable to consumers as edible nuts. Contamination of pistachio by Aspergillus species and their mycotoxins is the most important problem for consumption and export of this product. Aflatoxins are potent toxic, carcinogenic and mutagenic secondary metabolites primarily produced by two fungal species, Aspergillus flavus and Aspergillus parasiticus. Aspergillus flavus produces AFB1 and AFB2, while Aspergillus parasiticus produces AFB1, AFB2, AFG1 and AFG2. Among four main groups of aflatoxins, AFB1 is the most potent carcinogenic compound. Therefore, identification of toxigenic fungi is necessary for evaluating the foods quality and the presence of mycotoxins. The current methods being used for assessing fungi presence in foods based on cultivation methods and microscopic characteristics are time-consuming and labor-intensive. Recently, molecular techniques such as polymerase chain reaction (PCR) due to high sensitivity, specificity and rapidity has been introduced as powerful tools for detecting toxigenic fungi. Many genes involved in the biosynthesis of these mycotoxins have been identified and their DNA sequences have been published. PCR methods can be used to detect of aflatoxigenic Aspergilli based on structural genes (nor1, ver1 and omtA) encoding key enzymes in aflatoxin biosynthesis pathway and the regulatory gene aflR.
Pistachio samples were collected from different cultivation regions of two towns including Gonabad and Feyzabad. Samples were packed in sterile plastic bags and immediately transferred to the laboratory. The moisture content of samples was determined using thermal method and drying in at 95-100°C. Among fungal isolates 30 Aspergillus genus were detected and purified by cultural-based methods using PDA (potato dextrose agar) medium. Colonies of the fungus were transferred to PDB (potato dextrose broth) medium and incubated for 5 days at 28°C with shaking at 150 rpm. The mycelium was frozen in liquid nitrogen and ground to a powder for later DNA isolation. DNA was extracted with CTAB (cetyl trimethyl ammonium bromide) extraction buffer, then was purified with organic solvents such as chloroform/isoamyl alcohol and finaly was precipitated by isopropanol. Aspergillus genus were detected using polymerase chain reaction by specific primer pair Asp1/Asp2 for amplification of 18S rRNA region. Furthermore, aflatoxigenic genes were detected by three sets of primers (APA-450/APA-1482, ver1/ver2 and OMT-208/OMT-1232). PCR was performed in a volume of 25 µl containing 0.5 µl of each primer, 12.5 µl of Taq DNA polymerase master mix red, 10.5 µl of sterile distilled water and 1 µl of genomic DNA as template. A PCR consisted of an initial denaturing step of 5 min at 94°C followed by 35 cycles (30 s at 94°C, 35 s at 65°C and 40 s at 72°C) finished by a final extension step at 72°C for 10 min. The PCR products were analyzed by electrophoresis on a 1% agarose gel in TBE.
Among fungal isolates 30 Aspergillus genus were detected using microscopic characterstics and colony color. Under the microscope, conidia were one-celled, spherical, hyaline or pigmented and they formed long chains. 12 and 4 out of 30 samples had omtA and ver1 genes respectively. No observation was found for aflR regulatory gene in the fungal isolates. The results showed that although some isolates had one or two structural genes in the aflatoxin biosynthetic pathway, they could not produce aflatoxin due to not having any aflR gene. Coefficient of correlation was calculated to find the relationship between the existence of Aspergillus molds and aflatoxigenic genes in pistachio. The statistical results indicated that there is a significant correlation between the enumeration of Aspergillus molds and the existence of genes (omtA and ver1) in different moisture domains (p> 0.05) while no significant correlation was identified between the enumeration of Aspergillus molds and the existence of genes in different domains of enumeration of mesophilic bacteria, yeasts and molds. Contamination of nut seeds by fungi occurs during growth, harvesting, transport and storage. The production of aflatoxin is affected by different factors, such as genetic properties of the producing fungi, temperature, moisture content, the chemical composition of food and antimicrobial agents produced by other microorganisms. Water stress and temperature are the most relevant environmental factors which influence fungal growth and mycotoxin production. Other studies showed that there was a good correlation between the expression of an early structural gene (aflD) and aflatoxin B1 production in peanut seeds. Also previous studies have shown that there was a significant relationship between A.flavus contamination in the peanuts and pistachio with high humidity (p> 0.05). Since other factors such as temperature, pH and chemical composition of pistachio can affect the existence of Aspergillus molds and expression of aflatoxigenic genes, the influence of these factors on existence of Aspergillus molds and genes involved in aflatoxin biosynthesis pathway need to be investigated.

Human breast milk is known to be the best food for rapidly growing infant scince breastfeeding protects the newborn against some diesease such as infection disease،asthma and allergy. This effect may be due to the useful and natural microflora of breast milk which could be depending on diets of mothers. For the first time، this study accomplished to evaluate microflora of mother’s breast milk along with sepration and identification of local bacteria that could be as a candidat for bacteriotherapy and possessing probiotic propertiesin Iran. In this study،309 different colonies were isolated from 20 breast milk samplesof mothers who were registered in Mshhad. The isolated were selected based on the results of catalase test، gram stain، and bacterial morphology. The results showed that 40 out of 309 selected bacteria were catalase negative and gram positive stain. The whole genomes of 16s ribosomal DNA of isolated colonies were amplified using polymerase chain reaction for identification down to the strain level. According to the results، the breast milk contains various species of enterococci، Streptococci، Staphylococci and Lactobacilli. The most abundant bacteria، enterococcus faeciumwasisolated from 50% of samples representing 47. 5% of the total isolated bacteria. Generally 10 and 27. 5 percentage of isolated bacteria were La. rhamnosusLc 705and Streptococcus salivarius، respectively، that were isolated from 20 and 35 percent of milk samples،respectively.