Journal of Biotechnology Research Center

This study was conducted to investigate the contamination of dairy ice cream (local and imported) located in the city of Baghdad markets with Listeria monocyogenes L. (monocyogenes (bacteria by examining 50 samples of them during the months of July and August of 2015. Initial diagnosis was conducted on the agar media PALCAM (Polymyxin Acriflavin Lethium Chloride Ceftazidime Aesculin Mannitol Agar) a melanocyte selective differential media. Diagnosed isolates have confirmed their specific ownership to L. monocyogenes bacteria based on phenotypic agricultural traits and biochemical conventional methods tests like Catalase, oxidase, Motility and Hemolytic tests in addition to the use of modern diagnostic techniques represented both API Lesteria KIT which consists of 10 biochemical tests and CLEARVIEWTM technique that depend on the bacterial antigen reaction, especially flagella antigen as one of the advanced methods in the disclosure it. The results shown the presence of L. monocytogenes bacteria by 6% in the packaged and non- packaged local ice cream by conventional methods of isolation and diagnosis, while its ownership amounted to 4% in the same products when using Listeria KIT API and CLEARVIEWTM techniques, these bacteria did not detected the imported ice cream using the methods referred to above.
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The aim of this study was to extract basil seed gum and to examine the physicochemical properties. The pH value, relative density of 1% gum solution, bulk density, tap density, Hausner's ratio, compressibility index and loss on drying of the gum are: 6.3, 0.977 g/cm3, 0.30 g/cm3, 0.32g/cm3, 1.06, 6.25% and 7.2% respectively, and for the Arabic gum as a comparison gum were: 4.5, 0.6641 g/cm3, 0.68 g/cm3, 0.90 g/cm3, 1.32, 24.44%, 4.8% respectively. The melting point of the basil seed gum was high compared to the Arabic gum which was 290 and 275 °C respectively. The gum shows high swelling ability with water where the swelling index reached 5.6% and it was less than the swelling index in 0.5% concentration of sodium hydroxide solution reaching 5.0%. Basil seed gum characterized as a good organoleptic properties as it was tasteless and odorless, creamy color shaped like hygroscopic powder. It also shows positive results for reducing sugar and protein. The presence of small amount of protein in gum was very important for enhancing emulsification properties, fat holding capacity and other functional properties.

Police investigators across the world use the genetic tests for incrimination to suspects as well as judges' use these tests for convictions. Genetic tests were accurate and sensitive for low DNA concentrations. Criminals may did not leave any visible evidences behind their crimes but they may leave some invisible cells when they touch surfaces can lead to their identity. Scientists try to improve methods for cells collection and DNA extraction in order to get suitable DNA quantity for genetic analysis. The DNA that isolated from touched surfaces as Touch DNA. Several factors play major roles in getting suitable DNA amount for genetic analysis those were related with the touch person, others related to the nature of the touched surfaces, cells collection and DNA extraction methods.

Caraway (Carum carvi L.) was an important medicinal plant, which contains a lot of secondary metabolites with wide bioactivity. The dried seeds plant was extracted overnight with four solvents separately: 70% methanol, 70% ethanol, 70% n-hexane and water by continuous hot extraction using Soxhlet apparatus for the three solvents and by maceration with the last. Chemical detection for major active constituents in the four extracts was performed. The flavonoids was isolated from the crud plant using reflux, and subjected to thin layer chromatography (TLC). The aglycon moiety was extracted with chloroform.The extracted material was augmented by using high performance liquid chromatography (HPLC).The result of this study was indicated that each solvent extracts different compounds than the other and the plant was rich with flavonoids specially Rutin, Quarecetin kaempferol, Luteolin and less amount of Cumarins. The total flavonoid in each one gram dried caraway seed is 0.78mg.

A field study was conducted during spring season of 2015 at experimental farm of Field Crop Department, College of Agriculture, Baghdad University in order to study the effect of irrigation and concentrations of ascorbic acid (ASA) in some morphological characteristics for two cultivars of Zea mays L. Split Complete Block Design with three replications the main plots were irrigation treatment, the control treatment (irrigation 50%of the water available),75%, 50% of the amount of control water treatment the subplots were a combination of ASA concentrations (0,100,200 ppm) and two cultivars (fajer,5018). Irrigation treatment 75% of the treatment (control) gave the average root dry weight 39.11 g.plant-1 and leaf area 4406 cm2 leaf area index 3.15 and chlorophyll content 44.23 micog-1 fresh weight and dry matter to flowering to 50% 127.60 g.plant-1, no significant differences from irrigation treatment (control) started gave higher average rate of 41.22 g.plant-1 and 4753 cm2 and 3.39 and 135.00 g.plant-1 respectively, while the irrigation treatment gave 50% of the treatment measurement lowest average for studied measurements. Outweigh the concentration of 200 ppm by increase the average height plant 178.65cm and stem diameter 8.44cm and leaf area 4203 cm2 and leaf area index 3.00 and chlorophyll content 45.26 micog-1 fresh weights and dry matter to flowering to 50% 128.70 g.plant-1. while 5018 gave the highest plant height, leaf area, leaf area index, chlorophyll content, dry weight matter to flowering to %50 and root dry weight. The interference between irrigation treatments and two cultivars were significant in stem diameter and leaf area with its index, while the interference between irrigation treatments and ascorbic acid was in leaf area and its index. The interference between irrigation treatments with two cultivars were significant in stem diameter, leaf area with its index, while the interference between irrigation treatments and ascorbic acid ASA was in leaf area and its index. Therefore, we recommend possibility treatment of Zea may L. plant with ASA to improve capacity of water stress and recommend possibility of irrigation water by 75% of a need of the full irrigation (50% depletion of available water) without significant effect on same morphological growth.

The bacteriological study was included isolation and identification aquatic bacteria of dominant species in Tigris River. The period of the study for one year which starting from June 2015 until 2016 May which include 8 stations which distributed as follows : two station in the each site with space 50-meter between them, which have all of the following districts counties run through AL-Azizia AL- Zubaydiah, AL-Numaniyah and AL-Akut distributed as two repeaters within Wasit province. Identifications methods using the biochemical tests and verify the identification by API20E system. Results have pointed to the appearance of the isolation of 22 belonging to the following genera and species in the river water samples involve: Aeromnonas veronii , Aeromonas punctata , Bacillus subtilis , Exiguobacterium profoun , Pseudomonas alcaligenes , Enterobacter cloacae , Aeromonas caviae , Pseudomonas fluroescen , Bordetella spp , Photobacterium damala , Aeromonas hydrophila , Acinetobacter bumannii , Serratia liquefciens , Moraxella spp , Buttiaxella agrestis , Shigella spp , Chryseomonas meningosept , Pasteurella spp , Erwinia spp , Escherichia coli , Enterobacterium hormaechei , and Escherichia vuleneris. Temporal variation of aquatic bacteria in the study area was include: 16 bacterial isolate at June 2015 (27.5 %) 16 isolate at September (27.5%), 12 isolate at December (20.6 %) and 14 isolate in the May 2016 (24.1%). This study discover that station 1 was the most contaminated because domestic sewage and cattle feces within agricultural activities.

Chitosan, the deacetylated form of chitin, is a polysaccharide formed primarily of repeating units of D-glucosamine. It is a versatile biopolymer exhibiting various unique biological properties hence finds wide applications in food, biomedical and chemical industries. This study was conducted to evaluate optimum condition for chitosan production from Aspergillus oryzae SU-B2. The result have showed the best concentration 2% glucose as carbon sources, while 0.5 %(NH4)2SO4 and 0.1% yeast extract were optimum concentration as nitrogen sources. The optimum pH was 5.5 whereas the optimum temperature is 30˚C with incubation time 120h and agitation rate 150 rpm. The high yield of chitosan 10200 mg/ml was obtained with average increase 207.2%.

Study the effect of two nanoparticles compounds namely Silver nitrate nanoparticle (AgNO3-NPs) and Titanium dioxide nanoparticle (TiO2-NPs) was used in MS medium on plantlets growth and microtuberization of three potato cultivars (Arizona, Riviera and Burren) in vitro. The results showed significant differences between potato cultivars in the response of tissue culture and microtubers production. Arizona cultivar gave highest length, fresh and dry weight of plantlets and number of micro tubers production. It done 13.61 cm/plantlet, 1283.48, 145.04 mg/plantlet and 2.83 micro tuber/plantlet respectively. Riviera cultivar gave a highest weight of microtuber/plantlet and Burren cultivar done a highest diameter of micro tuber and they significant compared with Arizona variety. The result also showed that 2 mg/l of AgNO3-NPs caused significant increase in height, fresh and dry weight of plantlet, weight and diameter of micro tuber, 15.05 cm/plantlet, 1021.44, 13897 mg/plantlet, 317.35 mg/ micro tuber and 6.07 mm/micro tuber respectively. The 2mg/l of TiO2-NPs gave a highest number of micro tuber achieved 2.48 micro tuber/plantlet; it was statistically significant compared with the control and 2mg/l of AgNO3-NPs, as well as showed significant interactions between treatments and varieties in the most of characterization studies.

Cytarabine (Ara-C) is widely used as an effective chemotherapeutic agent, essentially in the treatment of acute myeloid leukemia. Albino male mice (Mus musculus) are used in this study to determine in vivo toxicity of the drug on spermatogenesis and sperm abnormality. Mice are divided into two groups; the experimental group received a dose of 0.33 mg/kg/day Ara-C for seven days, while the control group received normal saline for 7 days. Results indicated a significant increase in head abnormalities percentage (p≤ 0.05) in comparison to control group. Testosterone hormone concentration was significantly decreased (1.14 ± 0.12) ng/ml in treatment group in comparison with control group (2.41± 0.22) ng/ml. Histopathological changes revealed detectable spermatogenesis degeneration. Treatment caused spermatocytes disruption, presence of necrotic cells and appearance of spermiophages with slought cells in several seminiferous tubules. Leydig cells morphology are mostly intact; however, interstitial area appeared hypercellular. It is concluded that Ara-c exposure highly destructive to seminiferous tubule epithelium, which may lead in depressed mice fertility.

Phenol oxidases (laccase), copper-containing oxidase enzymes, are found in many plants, fungi and microorganisms. Many factors that influence laccase production from Azotobacter chroococcum are determined in this study, these factors are: carbon source, nitrogen source, pH, incubation temperature and incubation period. The results showed that the best carbon and nitrogen sources for laccase production are sucrose and yeast extract respectively, and the best factors for laccase production conditions are pH 7.0 with an incubation period of 6 days at temperature 30 Cᵒ.

The present study aimed to study the cytotoxic effects of three Iraqi traditional medical plants extracts using tissue culture teqnique on the human hepatocarcinoma HepG2 and mouse cell L20B cell lines. Glycyrrhiza glabra L. root, Morus nigra L. and Urtica urens L. leaves were extracted using 99% ethanol solvent. Seven crude concentrations were prepared by serial dilution, with concentrations of 3.9, 7.81, 15.62, 31.25, 62.5, 125 and 250 mg/ml, respectively. These were added to the microtiter plate containing 1x105 cells/well and 200 µl of the medium. The seven concentrations were used in triplicate to investigate their cytotoxic and anti-proliferative effects. The analysis of chemical composition of these plant extracts were determined by fast liquid chromatography (FLC). The extract of Urtica urens L showed the highest potent cytotoxicity in the HepG2 and L20B cell lines, while both G. glabra L. and Morus nigra L. crude extracts showed the lowest cytotoxicity. All concentrations of crude extracts showed different cytotoxic activity in vitro.

Abstract
Carbapenemases are clinically important because they destroy and may confer a resistance to carbapenems, severe infections caused by carbapenemase producers is associated with increased mortality. To achieve this goal, 180 samples were collected from different clinical sources included 92 urine, 33 smears of wounds, 13 smears of burns and 42 sputum. The samples were taken from patients attended Al-Yarmouk Teaching Hospital and Ibin Baladi Hospital in Baghdad Governorate. Diagnosis of bacterial isolates was done depending upon the microscopic examination, the cultured characteristics and biochemical tests. DNA extracted from 84 samples. Accordingly, detection of blaKPC2 gene was conducted by using specific primers for amplification of blaKPC2 gene. Moreover, the sequencing of 910 bp for blaKPC2 gene was performed by the biotechnology lab. at the National Instrumentation Center for Environmental Management (NICEM). Such test has been implanted by using 3730XL as a DNA sequences. The obtained results were analyzed by blast at the National Center Biotechnology Information (NCBI) and detect polymorphism in blaKPC2 based on the Bio Edit. Consequently, 94 variations between 47 transversions, 43 transitions and 4 deletions nucleotide were noticed. In a sense the test showed 79% under sequence ID gb|CP009872.1| from 3037673 -3038166 number of nucleotide from K. pneumoniae subsp. pneumoniae strain KPNIH30 of Gene Bank, score (329) and expect 5e-86 with the wild type of blaKPC2 gene from Gene Bank. Finally, the results illustrated polymorphism between local strains of
K. pneumoniae isolated from Iraqi patients and strain of K. pneumoniae obtained from Gene Bank.

Abstract
In the current study, synthesis and characterization of Zinc oxide nanoparticles (ZnONPs) and its application as anti-pathogenic bacteria were investigated. ZnO which has been prepared by using aqueous of green tea leaves extract (Camellia sinensis) as a reducing agent. The wavelength range was measured by Ultraviolet–visible spectroscopy (UV-Vis) for monitoring the formation of the nanoparticles, which showed sharp peak at 360 nm. The average size and shape of the nanoparticles were detected by using Atomic Force Microscopy (AFM) which was 88 nm with spherical shape. Fourier transform–infrared (FTIR). FT-IR spectra was documented for the ZnO nanoparticles synthesized by green tea extract to detect the biomolecules involved in the synthesis process. The antibacterial activity of crystal Zinc Oxide (ZnO) nanoparticles was explored against pathogenic bacteria that included Escherichia coli, Staphylococcus aureus, and Acinetobacter baumannii. The antibacterial test was conducted in solid media using different concentrations of ZnO and disk diffusion method, 100 µg/ml presented the best antibacterial activity, and further studies on the damage of bacterial genomic DNA of Escherichia coli and Acinetobacter baumannii were carried out using gel electrophoresis exposed the DNA fragment bands, this activity may be caused by the interactions between the surface charge of cell and nanoparticles. Reactive oxygen species (ROS) properties of the particles might disturb cell wall and great antimicrobial action

Abstract
The present study was planned to evaluate some cytotoxic effect of aspartame intake trough determining the apoptosis cells from peripheral blood and sperm head abnormality from cauda epididymis. After one week of administration of various doses of asparatame (3.5,35,350 mg/kg) to mature albino male mice and distilled water treated group was served as control. The results were statistically analyzed to compare the data of individual dose with that of the control, apoptotic and necrotic cells as well as sperm head are determined. The results showed that induce apoptosis cells in all concentrations (3.5,35,350 mg/kg) and necrotic cells in doses (35, 350 mg/kg) while late apoptosis in doses (35, 3.5 mg/kg) on the other hand, asparatame is increased in sperm head abnormality in (350, 35 mg/kg) dose. According to these results, it was possible to conclude that aspartame have a cytotoxic risk. Therefore, it is necessary to be careful when using such materials in food and beverages as a sweetener.

Abstract
The present study aims to evaluate the effects of methotrexate (MTX) with and without vitamin A (Vit. A) on some biochemical parameters and histological structure in male rabbits liver. Twenty male rabbits weighing 1250-1480 gm were divided into four equal number groups. The first group was given 2 ml distilled water as control group. The second group was given MTX (20 mg/kg), the third group was given Vit. A (5000 IU), while the fourth group was given MTX (20 mg/kg) +Vit. A (5000 IU) in alternative days. Following four weeks of treatment, lipid profile total cholesterol (TC), triglycerides (TG), high-density lipoprotein (HDL), [low-density lipoprotein (LDL) and very low-density lipoprotein (VLDL)]; in addition to thyroid hormones triiodothyronine (T3) and thyroxin (T4)] and liver enzymes [glutamic pyruvic transaminase (GPT) and glutamic oxaloacetic transaminase (GOT)] were determined in the serum. Also, the histological examination of liver of all the experimental groups were carried out. The results were revealed that the treatment with MTX caused a significant P≤0.05 increases in TC, HDL, LDL, T4, and GPT when compared with the control group. The treatment with Vit. A did not cause any significant P≥0.05 differences in all the studied parameters. The MTX+Vit. A treated group showed a significant P≤0.05 increases only in GPT compared with the control group; while a significant P≤0.05 decreases was found in TC, HDL, T3, T4, and GOT when compared with the MTX treated group. The histological examination of the liver sections showed that MTX administration caused major histological changes in comparison with the control such as inflammatory cell infiltrations, vascular congestion, sinusoidal dilatation and granular degeneration of hepatocytes. Treatment with Vit. A showed a typical structure in liver tissue. While in MTX+Vit. A group, the histological changes were less severe than those in the MTX treated group; these changes were granular degeneration of hepatocytes and sinusoidal dilatation at low levels. The overall results of this study confirmed that administration of Vit. A decreased the side effects of MTX; this protective effect of Vit. A may have clinical applications in chemotherapy.

Abstract
There have been numerous genetic causes of obesity specifically leptin, genetic variants of the leptin receptor gene (LEPR Gln223Arg) which appeared to be polymorphismized (A>G; rs1137101) have been implicated in the pathogenesis of obesity in several populations, although no association has been evidenced in other regions in the world. In this study, the association between LEPR Gln223Arg polymorphism with Body Mass Index (BMI) and plasma leptin levels in obese diabetic and obese non-diabetic adults who are randomly selected from Erbil city is evaluated. Blood samples were collected for DNA extraction, and plasma leptin measurements. LEPR (A > G; rs1137101) genotypes were identified by a PCR- RFLP. The results show that plasma leptin concentrations increased with body mass index, and in obese diabetic group was more than two-fold increases p=0.001 when compared to those of obese non-diabetic patients. LEPR (A > G; rs1137101) gene polymorphisms did not found associated with BMI in the whole studied population. Furthermore an increased frequency of the GG genotype in the female control group 32.1 compared to obese group 19.2, but the frequency did not significant (OR= 1.38: 95% CI; 0.74-2.03, P=0.07 ) which indicated that this genotype might be associated with a protective effect against obesity in female only and that this effect was independent of diabetes. Further analysis of a larger population is required to confirm the biological relevance of this polymorphism for obesity in the Kurdish population.

Abstract
Hair can be a valuable source of DNA especially in forensic casework and for noninvasive studies of human, when blood samples not available. This study emphasizes the impact of hair dyes on DNA sequence analysis. Samples collected from forty Iraqi families; each sample was divided in to two parts hair follicles and hair shaft. DNA extracted by using two different techniques, Phenol-chloroform (organic) method and prepFiler forensic DNA extraction kit. After quantification of DNA by real time PCR to confirm the exact DNA yield, Mitochondrial DNA (MtDNA) hypervariable region 1 successfully amplified from (50% of samples include hair follicle and 20% samples include shaft only), which all extracted by organic method. Whereas by using prepFiler kit the ratio of amplification success reach to 95% of samples included hair follicles, but there wais no DNA outcome from hair shaft by using this kit. Our results demonstrate that treated hair by dyed or henna had a significant influence on the sequence analysis results. Organic method was an appropriate method for extraction DNA from hair shaft, since this method used for extracting the old and degraded samples. While prepFiler DNA extraction kit was more convenient for isolation DNA from, hair samples included follicles only with excellent result.

Abstract
Female Fertility Blend ® (FFB) is one of new nutritional supplement that used to enhance the fertility status in women. This supplement containing vitamins, minerals, enzymes, amino acids, all may improve the oocytes quality and ovarian function; at the same time protect oocytes from free radicals damage. The aim of the study is to examine the in vivo effect of FFB on oocyte quality, and in vitro fertilization rate (IVFR), and embryonic development (ED) at early cleavage stages using the mice as a model for human being. Therefore, two groups of mature female mice were involved (20 mouse each). The treated group is daily orally administrated by 3.4mg/kg /body weight from FFB for 10 days and the other groups (the control) were treated with FFB- free distilled water only for the same period. Oocytes were collected and an epididymal sperms from mature fertilized male mice are obtained and in vitro fertilization (IVF) is done. Following 24 and 48hrs from IVF, the FR and ED rate are recorded. This results showed a significant (P<0.05) differences in fertilization rate and embryonic development when treating the female mice with FFB compared to control group. It is concluded that the FFB treatment has a great improvement in oocytes maturation and in vitro fertilization and embryonic development status