Study of genetic diversity in local rose varieties (Rosa spp.) using molecular markers

DOI: 10.7904/2068–4738–VIII(16)–148

Abbas SAIDI1*, Yazdan EGHBALNEGAD1 and Zahra HAJIBARAT1

1Department of Plant Sciences and Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, G.C, Tehran, IRAN

Corresponding author: abbas.saidi@gmail.com, Phone: +982129901964

Abstract. This study was undertaken to evaluate genetic diversity in a germplasm consisting of rose varieties. Genetic distances were estimated using three different molecular marker techniques including: start codon targeted (SCoT), conserved DNA–Derived Polymorphism (CDDP) and directly amplified minisatellite DNA (DAMD). According to the results, the average polymorphism information content was 0.37, 0.36, and 0.36 for SCoT, CDDP and DAMD markers, respectively indicating that the studied marker types were equal in terms of assessing diversity. Cluster analysis using SCoT and CDDP divided the varieties to four distinct clusters whereas DAMD markers data, grouped the varieties into three clusters. There was a positive significant correlation (r=0.92, p<0.01) between similarity matrix obtained by SCoT and CDDP. Results suggested that the efficiency of SCOT, CDDP and DAMD markers had a relatively same efficiency in fingerprinting of varieties. This is the first time that the efficiency of the three molecular markers have been compared with each other in a set of rose samples. The results showed that the studied markers had an appropriate polymorphism and thus were suitable for the study of genetic diversity in rose.

Keyword: Fingerprinting, PIC, molecular marker, genetic distance, correlation.

Alkaline protease producing Bacillus isolation and identification from Iran

DOI: 10.7904/2068–4738–VIII(16)–140

Maryam ARAMESH, Hatef AJOUDANIFAR*

Department of Microbiology, Damghan Branch, Islamic Azad University, Damghan. IRAN

*Corresponding Author: Email:ajoudanifar@gmail.com, Tel: +989124023012

Abstract. Although several microorganisms are known to produce proteases, Bacillus strains are often preferred as major sources for commercial alkaline proteases due to their exceptional ability to secrete large amounts of highly active enzymes. The present study aimed at isolation and molecular identification of alkaline protease producing Bacillus spp. from soil samples of Damghan County in Iran. This research was conducted in the laboratory complexes in the school of pharmacy of Islamic Azad University Damghan Branch. Soil samples were collected from 3 different regions. Streaking plate was done and 9 colonies were selected on the basis of appearance. To identify isolates, biochemical tests were performed. 16SrRNA gene duplicated with PCR and sent to Sinagene Company for sequencing. All isolates were oxidase–negative and catalase–positive. The ONPG tests were negative. 16SrRNA analysis showed that the isolated bacteria belonged to genus Bacillus. The results confirmed that four bacterial isolates had alkaline protease production capacity. It appears that after complementary examinations, the isolated Bacillus strains can be used for production of alkaline protease.

Keyword: alkaline protease, Bacillus, enzyme, biochemical tests.

Effect of Intravenous Infusion of Hypertonic Glucose on the Treatment of Cows with Postpartum Anestrus

DOI: 10.7904/2068–4738–VIII(16)–133

Seyed Fayaz MIRASI OSKOEE1, Hossein HAMALI*2, Reza ASADPOUR2

1Graduated student, Faculty of Veterinary Medicine, The University of Tabriz, IRAN

2Department of Clinical sciences, Faculty of Veterinary Medicine, The University of Tabriz, IRAN

*Corresponding Author: E–mail: hhamali@yahoo.com; Phone: +98 914 114 5575

Abstract. The objective of this study was to evaluate the efficacy of intravenous infusion of hypertonic glucose for recycling of the cows with postpartum anestrus. One hundred true anestrus Holstein–Frisian cows with ≥ 60 days in milk (DIM) according to the ultrasound examinations were selected from a dairy herd located on the suburb of Tabriz. Anestrus cows randomly were allocated into two groups: A&B (n=50). In group A (Gluco–Select synch), cows received 500 mL hypertonic glucose for 3 consecutive days (–3 to 0) IV, 0.021 mg bucerelin acetate on the day 0, IM followed by 25 mg dinoprost tromethamine IM on the day 7. In group B (Select synch), the same protocol was carried out on the cows except that the glucose hypertonic solution replaced by normal saline as placebo. Thereafter all cows were detected for estrus signs for the next 5 days and inseminated based on visual signs. Twenty seven out of 50 cows (54 %) in group A and 8 out of 50 cows (16 %) in group B showed the estrus signs respectively. Conception rates were recorded as A=44.5 %, B=37.5 % for the first insemination and A=B=100 % for the second insemination of cows. The number of insemination per pregnancy was calculated as 1.55 and 1.62 for groups A and B respectively. In the case of estrus rate, the difference between two groups was statistically significant (P≤0/01), but in the cases of conception rate or the number of insemination per pregnancy rate the differences were not statistically significant.

Keyword: Anestrus, Cow, Glucose, Hypertonic, Postpartum.

Protoscolicidal Effects of Bunium Persicum (Boiss) Against Hydatid Cyst Protoscoleces

DOI: 10.7904/2068–4738–VIII(16)–127

Leila DERAKHSHAN1, Sahar EMAMI2*

1Bioconversion Laboratory, Microbiology Engineering and Health Safety, Faculty of Science the Nature and Life, University of Mascara, (UN 2901), BP. 763, Sidi Said, 29000, MascaraALGERIA

2Laboratory of Physical Chemistry of Macromolecules and Biological Interfaces, Faculty of Science the Nature and Life, University of Mascara, (UN 2901), BP. 763, Sidi Said,  29000, MascaraALGERIA

* Corresponding author: haririahmed@yahoo.fr. Tel. 00 213 6 61 72 11 90

Abstract. Medicinal plants have been used for treatment of various diseases. The potent scolicidal activity of medicinal plants has been also known. Thus, the current study was conducted to investigate the protoscolicidal effects of Bunium Persicum against hydatid cyst protoscoleces. Protoscoleces were collected from sheep’s kidney having hydatid cysts. Five concentrations of B. Persicum essential oil (1, 3, 5, 10 and 15 mg/mL) were investigated for 10, 20, 30 and 60 min. Viability of protoscoleces was confirmed using eosin exclusive test. Regardless of the time, the rate of dead protoscoleces was increased with increasing the essential oil concentrations. The most rates of dead protoscoleces (100 %) were observed in 15 mg/mL from 10 to 60 minutes. On the basis findings this in vitro study, it can be advised B. Persicum essential oil as an effective natural scolicidal agent.

Keyword: Bunium Persicum, Hydatid cyst, Protoscoleces, Scolicidal activity.

Effect of pumpkin powder as a fat replacer on rheological properties, specific volume and moisture content of cake

DOI: 10.7904/2068–4738–VIII(16)–116

Aida DADKHAH1, Amir Hossein ELHAMI RAD1*, Reza AZIZINEZHAD2

1Department of Food Science & Technology, Sabzevar Branch, Islamic Azad University, Sabzevar, IRAN

2Department of Plant Breeding, Faculty of Agriculture and Natural Resources, Science and Research Branch, Islamic Azad University, Tehran, IRAN

*Corresponding author: Email: ah.elhami@gmail.com. Mobile: +989151710169, Fax: +985144660889

Abstract. Reducing fat and calorie is an important priority in optimization of bakery products including high–fat cakes. The aim of this research was to evaluate the effects of shortening replacement with pumpkin powder at 0 %, 20 % and 40 % with or without water level increment on rheological properties of cake batter and two properties of produced cake. Pumpkin powder was obtained from pumpkin flesh through drying, grinding and sieving processes. The rheological attributes of the batter and the specific volume and moisture content of cakes were examined using a rheometer, rapeseed displacement method and air–oven method, respectively. Increasing shortening replacement, enhanced viscosity (87.00 Pa.s), while increasing water in shortening–replaced treatments reduced it (11.30 Pa.s). Samples showed shear thinning behavior. In linear viscoelastic range, samples indicated solid viscoelastic behavior. Loss tangent, storage and loss moduli obtained from frequency sweep test, were investigated in three frequencies. In each frequency, with increasing shortening replacement, their viscoelastic properties enhanced. Contrary, increasing water amount in shortening reduced–batter samples, weakened system structuring. With increasing frequency, both moduli of treatments increased and the loss tangent of 20 % reduced–shortening cake without water level increment showed a more similar trend to the control (0 % replacement without increasing water amount). The moisture content of cakes containing pumpkin was greater than control. The specific volume of cake with 20 % replacement without additional water (2.68 cm3/g) was similar to the control and higher than other shortening–replaced treatments. Therefore, this treatment was chosen as the best reduced–fat cake.

Keyword: Cake batter, Pumpkin, Rheological properties, Shortening replacement, Specific volume.

Effect of Spermine, Epibrassinolid and their interaction on inflorescence buds and fruits abscission of pistachio tree (Pistacia vera L.), “Ahmad–Aghai” cultivar

DOI: 10.7904/2068–4738–VIII(16)–105

Fereshte BARAZESH1, Hakimeh OLOUMI2*, Fatemeh NASIBI1, Khosrow M. KALANTARI1

1Biology Department, Shahid Bahonar University of Kerman, IRAN

2Department of Ecology, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, IRAN

*Corresponding author: E–mail: oloumi.ha@gmail.com, Tel: +98343776612

Abstract. Several physiological problems such as abscission of inflorescence buds and fruit, incidence of blank, non–split and deformed nuts leads to diminish yield of pistachio trees. This research designed study the effects of epibrassinolid and spermine application on fruit disorders and abscission on Pistacia vera “Ahmad–Aghai” cultivar. Shoots, of pistachio trees sprayed with Epi–epibrassinolid and spermine, with three replicates. The hormonal treatments consisted of, 0.5 and 1 ppm Epi–Br and 100 and 200 ppm Spm and their combination. Some yield traits including inflorescence and fruit abscission were studied. Fruit abscission diminished significantly in all treatments in comparison with untreated hormonal trees. Epibrassinolid and spermine treatments decreased fruit blankness percentage and ethylene emission in compared with control. Simlutanouse application of epibrassinolid and spermine effectively raised split percentage in compared to each hormonal treatment alone. Chlorophyll content and carbohydrate content significantly increased by spermine 100 and 200 ppm, epibrassinolid 0.5 ppm and combination of spermine and epibrassinolid 0.5 in compared with the control. According to the results, it seems that spermine, epibrassinolid and combination of these hormones could significantly reduce fruit abscission and improve some of the physiological parameters in pistachio trees, probably through the antagonistic effects on ethylene production.

Keyword: Ethylene emission, Fruit and bud abscission, fruit disorder, pistachio yield.

Phytochemical analysis and antioxidant activity of the flavonoids extracts from pods of Ceratonia siliqua L

DOI: 10.7904/2068–4738–VIII(16)–93

Nawel OUIS1*, Ahmed HARIRI2

 1Laboratory of Physical Chemistry of Macromolecules and Biological Interfaces, Faculty of Science the Nature and Life, University of Mascara (UN 2901), BP.763, Sidi Said, 29000, Mascara, ALGERIA

2Bioconversion Laboratory, Microbiology Engineering and Health Safety, Faculty of Science the Nature and Life, University of Mascara (UN 2901), BP.763, Sidi Said, 29000, Mascara, ALGERIA

Corresponding Author: nawel_chim@yahoo.fr, Tel. 00213 6 61 72 11 90

Abstract. This study was carried out to determine the phytochemical profile and antioxidant activity of the carob. The results of preliminary phytochemical assessed showed that the alkaloid bases and salts, flavones aglycones, reducing compounds, fatty acids, polyterpenes, catechin tannins, volatile oils, cardiac glycosides, flavonoids, sterol glycosides and terpenes, amino acids and saponins are classes of chemical groups present in the pods of Ceratonia siliqua. The pods are characterized by 87 ± 1.0 % dry matter, 55 ± 0.2 % total sugars, 4 ± 0.1 % proteins, 7 ± 0.4 % lipids, 4 ± 0.3 ash. We have found that the pod has a 10.56 ± 0.2 Na, 0.75 ± 0.1 Zn, 0.55 ± 0.3 Cu, 41.7 ± 0.2 Mg, 210 ± 0.3 Ca and 1150 ± 0.2 mg/100g K. Pods of carob present 71.6 ± 2.10 mg/100g of flavonoids, 56.51 ± 1.02 mg/100g of alkaloids, 41.25 ± 1.36 mg/100g of saponins and 1.18 ± 0.2 mg/100g of tannins. Flavonoids fractions were extracted using organic solvents with different polarity. The ethyl acetate extract has the highest content of polyphenols and flavonoids 259.4 ± 4.2, 71.34 ± 1.08 mg EGA/g E, followed by the n–butanol extract 62.19 ± 0.13, 53.01 ± 0.11, then the crude extract 15.5 ± 0.21, 12.9 ± 0.17 and finally aqueous extract 13.39 ± 0.27, 11.13 ± 0.62. The flavonoids fractions extracted by ethyl acetate and n–butanol showed the higher antioxidant capacity determined by three methods: free radical scavenging activity, reducing power and liver lipid peroxidation compared to the crude exact and aqueous extract.

Keyword: Carob pod; phytochemical analysis; Flavonoids; Antioxidant activity.

Assessment of immunogenicity of alginate microparticle containing Brucella melitensis 16M oligo polysaccharide tetanus toxoid conjugate in mouse

DOI: 10.7904/2068–4738–VIII(16)–83

Parivash GHADERINIA1*, Reza SHAPOURI2

1Department of Microbiology, College of Science agriculture and modern technology, Shiraz Branch, Islamic Azad University, Shiraz, IRAN

2Department of Microbiology, Faculty of Basic and Medical Science, Zanjan Branch, Islamic Azad University, Zanjan, IRAN

*Corresponding author: Parivash_gh1356@yahoo.com

Abstract. Brucellosis is an infectious disease caused by Brucella bacteria, affecting animals and humans. It is found as an endemic disease in many parts of the world, especially in the Middle East, Central Asian, and South American countries. The objective of this study was to prepare alginate microparticle containing conjugate of Brucella melitensis 16 M oligopolysaccharide (OPS) with tetanus toxoid, and to assess its immunogenicity in mouse, in order to find a new method of confronting brucellosis. In this study, we extracted OPS from lipopolysaccharide (LPS) and conjugated it with tetanus toxoid by amidation method. Then, the obtained conjugate was condensed by gel filtration, followed by emulcification method in iso–octane organic phase to prepare alginate microparticles containing the conjugate. In our study, sera titr of antibodies IgM, IgA, IgG2a, IgG1, IgG2b, IgG3, and total IgG generated against oligopolysaccharide was performed by ELISA test. The final results, considering the obtained immunogenicity and the increased titer of antibodies IgG1 (19 times), IgG3 (19 times), IgG (15 times), and IgM (9 times) after injection of produced microparticle in mouse indicates appropriateness of the produced microparticle as a candidate vaccine. Thus, it could be concluded that alginate microparticles containing conjugates of B.melitensis 16 M oligopolysaccharide with tetanus toxoid can be proposed as a candidate anti–brucellosis vaccine.

Keyword: Brucella melitensis, Microparticle, Tetanus, Alginate, Vaccine.

Evaluation of the quality of the date syrups enriched by cheese whey during the period of storage

DOI: 10.7904/2068–4738–VIII(16)–75

Ahmed HARIRI1*, Nawel OUIS2, Djilali BOUHADI1, Karima OULD YEROU1

 1Bioconversion Laboratory, Microbiology Engineering and Health Safety, Faculty of Science the Nature and Life, University of Mascara, (UN 2901), BP.763, 29000, Sidi Said, Mascara, ALGERIA

2Laboratory of Physical Chemistry of Macromolecules and Biological Interfaces, Faculty of Science the Nature and Life, University of Mascara, (UN 2901), BP.763, 29000, Sidi Said, Mascara, ALGERIA

* Corresponding author: haririahmed@yahoo.fr. Tel. 00 213 6 61 72 11 90

Abstract. This study was carried out to the valuing of the date variety H’mira (very rich’s in sugars and in nourishing elements) and the cheese whey for production of novel syrups. Four samples syrups were manufactured: control represent date syrup obtained after hot extraction at a concentration 20 %, syrups (A) represent control with 10 % of cheese whey, syrup (B) with 20 % and syrup (C) with 30 % of cheese whey. These syrups were stored during 3 weeks at 4 °C. The biochemical, microbiological and sensory evaluations for all syrups were evaluated every week of storage. The biochemical characterization of syrups showed that it’s richness in total sugars, proteins contents, ash and dry mater. The syrup of date enriched with 10 % of liquid acid cheese whey present a good biochemical quality (higher level of sugars, proteins and ash) and important sensorial properties (color, odor and flavor). This syrup can be used for beverage or as medium to fermentation for production a new products.

Keyword: Date, Syrup, H’mira, Cheese whey, Quality, Storage.

Physical-chemical parameters and the level of heavy metals in cow milk in the Baia Mare area

DOI: 10.7904/2068–4738–VIII(16)–69

Aurelia COROIAN1, Vioara MIRESAN1, Daniel COCAN1, Camelia RADUCU1, Adina Lia LONGODOR1, Alexandra POP1, Gratian FEHER1, Luisa ANDRONIE1, Zamfir MARCHIS1*

 1University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Faculty of Animal Science and Biotehnology, Manastur Str. 3-5, 400272, Cluj-Napoca, ROMANIA

Corresponding author: zamfir.marchis@usamvcluj.ro

Abstract. Heavy metals are important pollutants in the environment and can cause problems for the organisms, and the bioaccumulation of heavy metals in the food chain can have adverse effects on human health. Regulations on the legally accepted amounts of heavy metals in food and in the body are very precise. The purpose of this paper was to evaluate the level of heavy metals in cow milk from different localities in the Baia Mare area, considered a very polluted area. The physical–chemical parameters and heavy metals in milk were determined. The determination of heavy metals in milk was performed by inductively coupled plasma mass spectrometry or ICP–MS used to identify and quantify the Pb, Cd, Cu, and Zn elements. The highest concentrations of Pb were observed in samples collected in Firiza locality with an average value of (43.22±0.62) (μg/L) and the lowest concentrations are for the samples analyzed in Săcălăşeni (11.53±0.33) (mg/L) (μg/L). Cd in milk did not show very large variations between localities, so the lowest concentrations were recorded in Săcălăşeni (4.32±0.13) and the highest ones were observed in the locality Firiza (10.93±0.39) (μg/L). The highest concentrations were observed for the Zn in milk, with an interval between (2255.20±7.14) (μg/L) in Săcălăşeni and (3855.80±5.11) (μg/L) for the analyzed samples from Firiza.

Keyword: milk, fat, proteins, Zn, Cd, Cu, Pb.