Journal of Biotechnology Research Center

Abstract
The study was Conducted for stimulation of seeds germination of eggplant (Solanum melongena L.) and measure vegetative growth in vitro under the influence of garlic (Allium sativum) alcoholic extract and the plant growth regulators, 0.5 mg/L Naphthalene acetic (NAA) and 1.0 mg/L Benzyl adenine (BA) and the MS and B5 media. The results showed that the presence of garlic plant extract in the media have a positive effect on the germination of seeds of eggplant, which gave the highest germination 10 seed in concentration 4 mg/L in the all of MS media and B5 media full force without growth regulators, but the presence of growth regulators with the extract gave the same result under study containing extract contained (0,2)mg/L. For the length of branches showed extract 4mg/L significant effect for of the rest of the transaction experience, which gave the highest along the branches in the MS media full force and MS half strength macro and micro elements, (7.66,7.60)cm, respectively, followed by concentration of extract 2mg /L in media used in the experiment and without growth regulators.

Abstract
Callus induction of Nerium oleander young leaves explants was studied using Murashige & Skooge (MS) Medium supplemented with 2,4-dichlorophenoxy acetic acid( 2,4-D 0.0, 0.5, 1.0, 2.0) mg/l with Benzyle adinine (BA0.0, 0.5 or 1.0, mg/l). comparative study was made for qualitative determination of some active compounds between the initiated callus and leaves of fieled plant (intact plant). Results showed that 2,4-D at 1.0 or 2.0 mg/l with BA at 0.5 or 1.0, mg/l were the best for callus induction that gave the highest fresh and dry weight. Qualitative detection showed that (Alkaloids, Flavonoids, Terpenoids, Tanins, Saponins, and Cardiac Glycosides) the main active compounds were found in callus and leaves of field-grown plant.

Abstract
The interest in bio-ethanol increase as biofuel, it seems to be a good alternative for fossil fuels. The growth of yeast Saccharomyces cerevisiae is influence in micronutrient like ions. This research aims to evaluate the influence different amount of magnesium as co-enzyme in growth and fermentation of yeast S. cerevisiae which isolated from different local sources. The batch fermentation process was followed at different magnesium concentrations as magnesium chloride at incubation temperature 30C○ and pH 4.5, best supplement concentration was 1.25 gm/l by measuring the mass of the released carbon dioxide. Supplement media with magnesium prolonged exponential growth, reduced the decline in fermentation activity resulting in reduced the time required for the conversion glucose into ethanol.

Abstract
This study has been conducted to knowing the effect of different concentration of alcoholic extract 2,4,12% and the cold aqueous extract 12,15,25, 23% of the fruits of Melia azedarach on the ability of the adults of Oryzaephilus surinamensis to penetrate the filling sacks (jute and poly propline)and on them the loss percentage in the seeds rice weight of the stored grains. Aqueous have shown the lowest percentage of losing weight of the seeds that were sacked in poly propline sacks that where treated with the alcoholic extract at the concentration of 12% which was 1.88% where by the cold watery extract has given the lowest percentage of losing weight of the sacked seed at the concentration of 35% which was1.02%after six months of storage when treaded jute sacks with insecticide actellic at concentration 0.01% (recommend concentration) has given the lowest percentage of losing weight of sacked seed was 0.36% after 6 months of treatment. Therefore we suggest using the treated with the alcoholic extract at the concentration of 12% weight of the sacked seed at the concentration of 35%.

Abstract
The present study was conducted in the fish diseases laboratory of College of Veterinary Medicine-University of Baghdad to investigate the effect of magnetized treated water on common carp Cyprinus carpio infected with Aeromonas hydrophila.
Total of 80 specimens of common carp Cyprinus carpio were used ranges between 142-156 gm in weight and between 22-27cm in length. Fish were distributed randomly with ten fish to every aquarium upon four. First treatment considered control treatment free of magnetized water while the second treatment contained magnetized treated water with 500 gauss, third treatment contained magnetized treated water with1000 gauss, forth treatment contained magnetized treated water with1500 gauss with two replicates for each treatment through the period of experiment attain four weeks. The experiment represented by induce the infection by intramuscular injection of the bacteria in the site of left behind the front edge of the dorsal fin with dose of 107 cfu/100 gm of fish body weight. Blood picture were examined weekly and description of the clinical and macroscopic signs and survival

rate. Statistical analysis results show presence of significant differences p≤0.01 of blood picture results of magnetized treated water treatment compared with control, experimental results indicate to presence of improvement of clinical and macroscopically signs and increase of survival rate of magnetized treated water compared with control.

Abstract
Identified and studied the mechanism of growth kinetic for some types of algae stuck on different materials surfaces which sank in Tigris River as medium for growth such as awash surface for boats, aquatic plants, and clay . The results shown participation of the algae: Gyrosigma acumintum, Firgallares capucina var -valvaire, Diatoma elongatum and Oscillatoria sancta which were stuck on awash surfaces in the Tigris River. Results shown also the dominion of diatoms algae Bacillariophyce Class (Cymbella cistula, Diatoma elongatum, Firgallares capucina. var -valvaire, Melosira granulata. var.connective, and Nitzschia )which stuck on the awash surface of cane by 76.4% more than remnant of algae. The class of Cyanophycea (Oscillatoria sancta and Merismopedia glauca ) more dominion by 11.7% than remnant of algae. The class of Chlorophycea (Spirogyra borgeana) more dominion by 5.8% than remnant of algae.
الكلمات المفتاحية: الطحالب الملتصقة، المغمورة، آلية النمو، النباتات المائية
The characteristically of Penale's order algae dominion for growth by comparing with Centrals' order algae which consists of diatoms algae (Bacillariophyce class). The characteristic of cane more effect than clay and boats by adhesion of most algae on it during the period of growth on awash surfaces in Tigris River. The concentration value of ammonia and nitrate are 22.4 and 16.8 ppm respectively for test of the clay sample which assist for growth of algae such as Oscillatoria boryana and Oscillatoria sancta. The study identified 25 species for 18 genuses for different algae with dominion ability of Penale's order (diatoms algae class) more than remnant of algae for growth.

Abstract
The present study was carried out to detect the active constitutions extracted from Rosmarinus oficinalis leaves by 70% methanol, the dried weight of aqueous extract (F1) were 12%(w:w) contain polyphenols represented by phenols, flavonoids and coumarins as a major compounds , while the dried weight of organic extract, [chloroform(F2)] was 2.8%(w:w) represented as terpenoids, tannins and coumarins detected by qualitative chemical analysis. Antimicrobial activity of Polyphenols and terpenoids were tested against four pathogenic bacteria isolates including: Escherichia coli; Staphylococcus aureus, Pseudomonas aeruginosa and Bacillus cereus ,which grown on Mueller Hinton agar (MHA) plates and then treated with different concentrations of rosemary extract (F1) and (F2) ranged from 0.15-1.2mg/ml, the results revealed that the higher inhibition zone obtained by F1 were 35mm in S. aureus, 24mm in B. cereus and lowest inhibition were 16mm in E. coli and P. aeruginosa at concentration 1.2mg/ml, and the inhibition were depends on extract concentrations. The MIC value of F1 was 600 µg/ml against S. aureus, 1200 µg/ml against B. cereus and 2400 µg/ml against E. coli and P. aeruginosa, while the MICS values of F2 were 2400 µg/ml against S. aureus and P. aeruginosa, and 4800 µg/ml against E. coli and B. cereus. Furthermore calculating index fractional inhibitory concentration (∑FIC) of rosemary extracts (F1 or/and F2), the ∑FICA ranged from synergistic to additional on E. coli and P. aeruginosa respectively, antagonistic on S. aueras and B. cereus, while the ∑FICB index showed synergistic effects on all tested organisms except the additional effect on P. aeurginosa on the basis of resultant zone of inhibition. The anticancer effects of (F1) were used on three cell lines, human larynx epidermoid carcinoma (Hep-2), mammary adenocarcinoma (AMN-3), rhabdomyosarcoma (RD) and one normal cell line, Rat embryo fibroblast (REF) at different concentrations ranged from 62.5-1000 µg/ml. The highest inhibition rate (IR) was 82.41%, 85% on RD, 52.62%, 75.48% on Hep2, 39.14% and 49.75% on AMN-3 at concentrations 500 and 1000µg/ml after 72h exposure time respectively, while IR of Ref cell line exhibit low inhibition rate ranging from 5.8 to 15.7% after 72h in the same conditions.

Abstract
The cytogenetic effects of taking oral contraceptive pills OCPs that contain steroid hormones (estrogen and progesterone) during pregnancy on some aspects of fetus and newborn have been previously reviewed. The objectives of this study are to detect the effect of using different doses of progestin and estrogen by pregnant female mice on the chromosomal aberration CAs and mitotic index MI in their male fetuses. Thirty female mice were divided equally into three major groups: two experimental G1 and G2 and control group according to the level of the dose. Administration of COCPs (either 0.034 or 0.068 mg/kg b. wt. /d dissolved with 0.1ml distilled water) orally once a day, were started at the first day of gestation and continued till day 14 for the experimental groups G1and G2 respectively. When the new pups in each experimental and control group reach two weeks of age, those male offspring 10animals/ group were used for chromosome preparation, while the control group divided into two groups: ten animals represented a negative control given distilled water only, while the other ten animals injected intra peritoneal with mitomycin–C 2mg/Kg represented positive control (once dose). The statistical analysis of mice bone marrow cells have no significant decrease in the mitotic index of the pups belongs to G1 mothers in comparison with negative control group, but it shows a highly significant differences P<0.01 as compared with that of positive control group . Although the pups belong to G2 mothers showed a slight decrease in the mitotic index in comparison with negative control group. Differences were non- significant with negative control and highly significant P<0.01 with positive control group. In addition, the mitotic index of spleen cells of the pups belongs to G1 mothers noticed no significant decrease in comparison with negative control group, however, it shows a significant differences P< 0.05 as compared with positive control group. In the pups belongs to G2 mothers there were significant decrease P< 0.05 in mitotic index in comparison with negative and positive control groups. The results showed that the COCPs caused an increase in chromosomal aberrations CAs with increasing of steroid hormones concentration. In G1 group the CA equal to 19.91%. This reducing in CA was not significant in comparison with those of the negative control which gave 14.33%; this percentage was increased in the result of CAs of the mice treated with mitomycin –C (MMC) which was 48.58%. Treatment with 0.068 mg/kg B wt. of COCPs gave CA equal to 17.18%. This value was not significant in comparison with those of the negative control which was 19.91%. Higher doses of OCPs may cause cytogenetic effects in chromosomal aberrations CAs and decrease the mitotic index (MI) in the bone marrow cells and spleen cells of offspring belonged to mothers treated with oral contraceptive during their pregnancy for 14 days.

Abstract
This study aimed to investigate the regeneration capability of cell suspension derived from hypocotyl callus of the vegetable plant, muskmelon, Cucumis melo L. Culture of the densities (7.5, 9.8, 13.0, 11.2, 7.8, 4.2 )× 102 cell/ml produced callus primordia which formed typical callus culture successfully. This green – yellowish color and semi-compact callus regenerated shoots on agar solidified MS medium supplemented with 2.0mgL-1BA. These regenerates rooted readily in MS0 medium. They were adapted and routinely transferred to soil. The conclusion of this work that muskmelon plants have a good response to in vitro culture with no need to specific requirements.

Abstract
The present study describes plant regeneration via somatic embryogenesis from cotyledonary node callus. Callus cultures were derived from cotyledon node segment excised from Trigonella foenum-graecum L. axenic seedlings grown in agar solidified Murashing and Skoog's(MS) medium supplemented with Benzyl adenine BA at 2.0 mg l-1 and Naphthalene acetic acid NAA at 1.0 mg l-1. These cultures were stimulated to form somatic embryoids at 32% frequency of in agar-solidified MS medium containing NAA at 0.5mg l-1 in the presence of 15% v/v coconut water. Light microscopy examination of sections of heart embryos stage proved the presence of vascular system. Transfer of other developmental stages of embryos to the same induction medium led to the development of secondary somatic embryos and embryoids into plantlets in agar-solidified MS medium lacking hormones. They were readily rooted in MSO and successfully transferred to soil.

Abstract
Protoplasts were isolated from leaf mesophyll of sugarbeet (Beta vulgaris L.) axenic seedlings. Eight enzyme mixtures were tested for cell wall degrading ability. The efficient enzyme solution was mixture "II" that consist of 1.5% Cellulase RS, 2% Cellulase R10, 1% Macerozym R10 and 0.1% Pectolyase Y23. This mixture was efficient in releasing protoplasts and gave high yield of a density 7.3 × 104 protoplast / ml. These isolated protoplasts were viable 93%, their sizes ranged from 13 up to 52 µm, vacuolated and unvacuolated. This finding enable workers to focus on somatic hybridization through protoplast fusion to overcome some barriers facing gene transfer to improve plant species such as transfer of N2 fixation ability to non-legumes and producing resistant varieties to biotic stress and other traits.

Abstract
The aim of our study was to determine micronucleus MN frequency and nuclear division index in peripheral blood lymphocytes of patients diagnosed with Acute Lymphoblastic Leukemia ALL, who had undergone chemotherapy. Patients were treated with nine of drugs, which included Vincristine , Methotrexate , Cytosar-U, L-asparaginase , Etoposide, , dexamethasone (Decadron) , Indoxan , Steroids. The study was carried out on fifty Iraqi patients (34 Male, 16 Female), aged 2-70 years with Acute Lymphoblastic Leukemia ALL. These samples included 20 pretreatment aged 7-70 years, 15 under treatment aged 2-57years and 15 relapsed aged 9-40 years, compared with a sample consisted of 50 apparently healthy normal individuals collected randomly from population living in Baghdad aged 3-75 years. Results of the of MN in the human lymphocyte observed a significant increase p<0.05 in the males and females of human peripheral blood lymphocytes of patients with Acute Lymphoblastic Leukemia, before and after the chemotherapy as compared with the control. While, a significant decrease P> 0.05 in nucleic division index NDI was observed in the human peripheral blood lymphocytes of Acute Lymphoblastic Leukemia Patients (males and females), before and after the chemotherapy as compared with the control. In addition, the results of MN and NDI were compared in the genders of the groups studied. In conclusion, the results indicated that there is a possibility of using the changes in the mean of MN frequency and NDI as biomarkers for the assessment of DNA damage in the human peripheral blood lymphocytes of patients with Acute Lymphoblastic Leukemia ALL before and after the chemotherapy treatment and the increase frequencies of MN in ALL patients indicate the effect of antileukemic agents in inducing somatic genetic damage

Abstract
The aim of our study was to determine micronucleus MN frequency and nuclear division index in peripheral blood lymphocytes of patients diagnosed with Acute Lymphoblastic Leukemia ALL, who had undergone chemotherapy. Patients were treated with nine of drugs, which included Vincristine , Methotrexate , Cytosar-U, L-asparaginase , Etoposide, , dexamethasone (Decadron) , Indoxan , Steroids. The study was carried out on fifty Iraqi patients (34 Male, 16 Female), aged 2-70 years with Acute Lymphoblastic Leukemia ALL. These samples included 20 pretreatment aged 7-70 years, 15 under treatment aged 2-57years and 15 relapsed aged 9-40 years, compared with a sample consisted of 50 apparently healthy normal individuals collected randomly from population living in Baghdad aged 3-75 years. Results of the of MN in the human lymphocyte observed a significant increase p<0.05 in the males and females of human peripheral blood lymphocytes of patients with Acute Lymphoblastic Leukemia, before and after the chemotherapy as compared with the control. While, a significant decrease P> 0.05 in nucleic division index NDI was observed in the human peripheral blood lymphocytes of Acute Lymphoblastic Leukemia Patients (males and females), before and after the chemotherapy as compared with the control. In addition, the results of MN and NDI were compared in the genders of the groups studied. In conclusion, the results indicated that there is a possibility of using the changes in the mean of MN frequency and NDI as biomarkers for the assessment of DNA damage in the human peripheral blood lymphocytes of patients with Acute Lymphoblastic Leukemia ALL before and after the chemotherapy treatment and the increase frequencies of MN in ALL patients indicate the effect of antileukemic agents in inducing somatic genetic damage

Abstract
Leucaena leucocephala is a tree species used for several agricultural purposes in Mediterranean region. The seeds of these species exhibit dormancy causing delayed germination. A series of experiments evaluated the effects of various treatments on the germination of leucaena leucocephela L. seeds. Germination of fresh seeds was 46% but it was improved (to about 70%) by mechanical scarification using sand paper. A germination of about 60% was obtained when dry seeds were pre-chilled at 4 ̊C for 2,3 and 4 weeks prior to germination that various storage periods of pre-chilling had no influence upon germination enhancement of seeds. Exposure of seeds to -18 ̊C inhibited germination in comparison to the control. There was no significant increase in germination percentage after cold treatment. Maximum improvement being achieved when seeds were kept dry under alternating cold/ warm temperatures for 8days at two day intervals prior to germination, since germination percentage increased to 73%. A significant promotion was recorded when seeds treated with 100 mg/l GA3 prior to germination and that pre-chilled treatment improved germination percentage compared with those un-chilled the maximum (70%) at 100 mg/l of GA3.The effect of nutrient media on seed germination was studied on MS and B5 media after being stored for one week in refrigerator (at 4°C) or in the freezer (-18°C). The germination percentage of Leucaena seeds cultured on MS medium showed a significant increase over the control for those stored one week at 4°C, when the effect of B5 medium on seed germination was found to be similar to the control (50%) for seeds stored one week at 4°C. These results demonstrate that alternating temperatures (cold/warm) provide faster and highest germination percentage and could be secured at low cost which would be adequate to large scale treatment of Leucaena leucocephala seeds.

Abstract
Leucaena leucocephala is a tree species used for several agricultural purposes in Mediterranean region. The seeds of these species exhibit dormancy causing delayed germination. A series of experiments evaluated the effects of various treatments on the germination of leucaena leucocephela L. seeds. Germination of fresh seeds was 46% but it was improved (to about 70%) by mechanical scarification using sand paper. A germination of about 60% was obtained when dry seeds were pre-chilled at 4 ̊C for 2,3 and 4 weeks prior to germination that various storage periods of pre-chilling had no influence upon germination enhancement of seeds. Exposure of seeds to -18 ̊C inhibited germination in comparison to the control. There was no significant increase in germination percentage after cold treatment. Maximum improvement being achieved when seeds were kept dry under alternating cold/ warm temperatures for 8days at two day intervals prior to germination, since germination percentage increased to 73%. A significant promotion was recorded when seeds treated with 100 mg/l GA3 prior to germination and that pre-chilled treatment improved germination percentage compared with those un-chilled the maximum (70%) at 100 mg/l of GA3.The effect of nutrient media on seed germination was studied on MS and B5 media after being stored for one week in refrigerator (at 4°C) or in the freezer (-18°C). The germination percentage of Leucaena seeds cultured on MS medium showed a significant increase over the control for those stored one week at 4°C, when the effect of B5 medium on seed germination was found to be similar to the control (50%) for seeds stored one week at 4°C. These results demonstrate that alternating temperatures (cold/warm) provide faster and highest germination percentage and could be secured at low cost which would be adequate to large scale treatment of Leucaena leucocephala seeds.

Abstract
The present study was aimed to study the optimum conditions of producing yeast biomass and lytic enzymes, and removal of total solids from dairy waste water. Results showed that dairy waste water serve as a good substrate, enabling the growth of Saccharomyces cerevisia S4, which produced a considerable amount of yeast biomass. Maximum production of biomass was 26g/l obtained after 96h fermentation, at pH6, temperature 30°C and dairy waste concentration of 2.5% w/v. The maximum activity of α-amylase and protease were (67.7, 60.2)U/mg, respectively obtained when cultured the yeast in dairy waste water 2.5%w/v at pH6 and incubated for 120h at 30°C. The maximum reduction of total suspended solids and total dissolved solids were (44.1,53.6)% respectively observed after 96h of fermentation period at 30°C.

Abstract
The antimicrobial activity for Thevietia peruviana was evaluated by measuring inhibition zone diameter in agar using well diffusion assay. The aim of the present study was to evaluate the antimicrobial potential of Thevietia peruviana leaf extract as compared with callus extract against some bacterial strains and fungi. The results showed that the addition of 2,4-D at the concentration of 9 mg/l, and 0.1 mg/l of kinetin led to obtain callus weight reached 800 mg. It was noticed that the reduction of 2,4-D concentration up to 6 mg/l resulted in compact and green pieces of callus. The optimal weight and friable callus was obtained at 9 mg/l. Among the susceptible bacteria are the gram negative pseudomonas aeruginosa seemed to be sensitive against all concentration of Thevietia peruviana leaf and callus extracts, While Escherichia coli showed resistance with all concentrations of extracts. It was noted that the extracts were more active against gram positive staphylococcus aureus, as compared with other bacterial species. Results of this study revealed that callus extract of Thevetia peruviana possess higher activity in comparison with leaf extract against gram positive bacteria (Staphylococcus aureus, Bacillus cereus) and gram negative (Pseudomonas aeruginosa). Finally all the bioextracts were well stable at room temperature during the period of the study and did not show any reduction of activity against the bacterial strains used in this study experiments.

Abstract
The ability of five Aspergillus flavus that produce Aflatoxin B1 have been detected using coconut medium as substrate. Chromatographical analysis by TLC and HPLC revealed that, three out of five isolates were a good producer for the Aflatoxin B1. In this study, rapid assessment of five isolates of A. flavus was accomplished using an indigenously designed primer pair for the Aflatoxin regulatory gene aflR in polymerase chain reaction (PCR). Specificity was assayed in pure culture systems using DNA extracted from five different A. flavus isolates as PCR template. Positive amplification was achieved only with DNA from A. flavus that produce Aflatoxin B1.