ISSN: 1815-1140

Current Issue
Volume 12 Issue 1

Volume 12, Issue 1, Autumn 2018, Page 7-136

Chromosomal abnormalities associated withmale infertility in Baghdad Iraq

Saad Mohamed Nada; Hazim I. AL-Ahmed; HussienJassimObade

Jornal of Biotechnology Research Center, Volume 12, Issue 1, Pages 7-13

In the current study, 142 patients (109 azoospermia and 33 severoligospermia) were studied in order to explore the cytogenetic cause’s background and the hormonal study of male infertility in Baghdad Strip of Iraq. Of the 142 infertile males,14 patients showed abnormal chromosomal karyotypes corresponding to a frequency of 9.85% (14/142), the occurrence of chromosomal abnormalities was only confined to the azoospermia patient group. Patients with abnormal karyotypes represented 12.84% (14/109) of the azoospermia patients. Nine had a 47, XXY karyotype ofKlinefelter's syndrome [47XY;64.28% (9/14)], [47XY+mar;14.28% (2/14)], [46XY,del13(p12) and (del21(p12);7.14% (1/14)], [46XY,tra(5;12, -6); 7.14% (1/14)] and one with chromosomal instability that showed multiple mosaic karyotypes 7.14% (1/14). The hormonal levels of nine patients with Klinefelter's syndrome (KFS) were diagnosed in the present study, and the results showed elevated FSH and LH concentrations compared with control. Whileserum testosterone values were below the control.

Molecular Identification of StaphylococcussppIsolatedfrom Clinical samples

TaghreedKhudhur Mohammad

Jornal of Biotechnology Research Center, Volume 12, Issue 1, Pages 14-24

The analysis of 16S rRNA gene sequences has been the technique generally used to study and confirm the identification andtaxonomy of staphylococci. However, bacterial species cannot always be distinguished from each other using cultural methods. Thus,clinicalsamples were collected from 190 cases only 31 positive for staphylococcal infections withUrinary Tract Infection, Wounds, Burns, Otitis media, diarrhea infections, were applied for microbiological analysis which include: cultures onManitol salt agar and HiCrome UTI Agar medium all the isolates gave positive golden yellowand identify as Staphylococcusspp. DNA was extracted fromStaphylococcussppand the 16srRNA gene were amplified by using specific primer, then sequencing of nucleic acid of genes was performed by machine is AB13730XL, Applied Biosystem, Macro gen company,the DNA sequencing results of flank sense of 16srRNA gene from 31strains of Staphylococcuswas confirm the identification into species level: Staphylococcus haemolyticus, Staphylococcus aureus, Staphylococcus epidermidisAnd Staphylococcus sciuri.Analysis of the sequences appeared that there two substitution(Transversion, Transition) in the Staphylococcusaureusstrains with Sequence IDLC090540.1 location at Range of nucleotide from 4 to 636, 100% compatibility with NCBI while no substitution appeared in the Staphylococcus haemolyticusstrains which have thesequence IDLN998078.1, 99% compatibility with NCBIalso the sequence ID KR812401.1 which related to the strain Staphylococcussciuri not appeared any substitution after sequencing analysis.Types of substitution detected in partial16srRNA gene inStaphylococcus epidermidisstrains 13 Transversion and 5 transition substitutionlocation at range of nucleotide from 6 to 1026 have the Sequence IDKF575160.1 compared with data obtained from Gene Bank, these finding lead to conclusion,our assay allows rapid and confirm the detection to avoid possibility of misidentification of Staphylococcus species based on cultural analysis, the study aimed to propose the partial sequencing of the gene as an alternative molecular tool for the analysis of Staphylococcus species and for decreasing thepossibility of misidentification. New submission of local Iraqi Staphylococcus clinical isolated during the current study show successfully record of four isolate Staphylococcus sciuri,Staphylococcus epidermidis,Staphylococcus aureus andStaphylococcus haemolyticuswith GenBank accession number: KY938530.1,KY938529.1,,KY938528.1, and GenBank: KY938527.1respectivelly.

Effect ofDifferent CellSuspension Culture Densitiesof Physalisangulata L. by Using Plating or Embedding Methods on the Callus Formation

MuthanaMuhamad Al-Mahdawe; TalfanAnad Ahmad; Dhuha Sabah Nadir

Jornal of Biotechnology Research Center, Volume 12, Issue 1, Pages 25-32

The study was successful in establishing cell suspensions cultures derived from the hypocotylcallus of the plant Physalisangulata L. induced in the solid MS medium supplemented with the concentration of 3.0 mg. L-1 2,4-Dichlorophenoxy acetic acid (2,4-D)plus 0.5 mg. L-1 Kinetin(Kin). The culture of the suspensions by different densities (12.36, 13.70, 14.75, 15.98)× 105 cell.cm3 using the platingand embeddingmethods by agar layerin the solidMurashige and Skoogmedium (MS) enriched with the same callus induction concentrations, showed that the plating method exceed theembedding method in the number of cellular colonies which reached the average of 79.4 and 15.0 at the culturing density of 15.98 × 105 cell.cm3, these values were declined with the primary density to reach the average of 14.0 and 11.4 both the plating and the embedding methods respectively, the high density significantly exceed all the rest densities and gave rate of callusprimordia of 56.6 after 28 days from the day of culturing the suspended cells by plating method, and reached 9.8 after 33 days from the day of culturing of the suspended cells by using the embeddingmethod, the transfer of the callus primordia to the MS medium supplemented with 3.0 mg. L-1 2,4-D plus 0.5 mg. L-1 Kin, led to the growth of callus segments and their differentiation to the somatic embryos and their development through all their stages until reaching the shoot formation.

Sequencing of Gliotoxin Genes in Clinical and Environmental Aspergillus fumigatus isolates in Iraq

Russel S. abdulhadi; Nemat J. Abdulbaqi

Jornal of Biotechnology Research Center, Volume 12, Issue 1, Pages 33-39

Gliotoxin is an important virulence factors in Aspergillus fumigatus. The biosynthesis of this mycotoxin is regulated and expressed by the presence of gliP genes. This study aimed to identify Aspergillus fumigatus isolates in clinical and environmental sources with glip genes using conventional PCR and sequence. To achieve this, DNA was isolated from twenty A. fumigatus isolates using commercial kit. The range of the DNA extracted was 65-210 ng/μl with a purity of 1.5-1.9. Species identification of the A. fumigatus isolates was achieved to a high specificity by using tailored primer. The results showed that all isolates had positive results to the primer and all isolates were able to produce gliotoxin. PCR detected the gliotoxin genes, glip in five isolates. The five PCR product samples were sent for sequence analysis and 25 µl (10 pmol) from the forward primer. The results of all the samples indicated have a single band of the desired product of gliP gene of A.fumigatus and the samples sent for sequencing related to molecular weight 190 bp.

Detection of Aflatoxigenic and Non-Aflatoxigenic Isolates of Aspergillus flavus Isolated From Some Clinical and Environmental Sources by HPLC and PCR Techniques

Mushtak T.S. Al-Ouqaili; Mohammed H. Muslih; Salah M. A. Al-Kubaisi

Jornal of Biotechnology Research Center, Volume 12, Issue 1, Pages 40-48

This study aimed to determine the role of polymerase chain reaction (PCR) and High-performance liquid chromatography (HPLC) technique in the discrimination between aflatoxigenic and non-aflatoxigenic isolates of Aspergillus flavus. The isolates were identified based on macroscopical and microscopical characteristics, and extracted aflatoxin was detected by HPLC technique. Furthermore, DNA was extracted from the all isolates and carried out by PCR to amplify target genes encoding for toxin production (nor-1, ver-1 and aflR). The results showed that the genes (aflR, nor-1) were found in 11 (73%) of isolates, while the (ver-1) gene appeared in 10 (67%) of isolates. Both aflatoxigenic and non-aflatoxigenic isolates were also determined depending on the amplification of gene sites in the targeted DNA. HPLC technique has also used with high efficiency to ensure the aflatoxin-producing isolates and to evaluate the level of aflatoxin B1 production for 15 isolates of A. flavus. Ten isolates were able to produce aflatoxin with rates ranged from 0.78 to 45.03 ppm. PCR technique has proved high efficiency in the differentiation between aflatoxigenic and non-aflatoxigenic isolates of A. flavus. Moreover, aflatoxin production was directly associated with gene appearance and gene detection. Also, HPLC technique is a standard and superb technique in identifying and analyzing aflatoxin with high sensitivity and accuracy.

Qualitative and Quantitative Evaluation of Total flavonoids Extracted from Mentha arvnesis Aerial Parts

Muna Abd Mutar; Zainab Yaseen Mohammed Hasan; Najy swady Naser

Jornal of Biotechnology Research Center, Volume 12, Issue 1, Pages 49-55

Mentha arvnesis is an important medicinal plant that contains many groups of secondary metabolites that have extensive biological activity. The flavonoids were extracted from the aerial parts of the mature plant by using reflex extraction, and the aglycan fraction was isolated with ethyl acetate. The flavonoids of the extract were detected qualitatively with TLC technique. The total amount of flavonoids derived from the plant was estimated using a colorimetric method and the total value was calculated using the straight line equation of the standard flavonoid curve (Rutin). The plant was found to be rich in flavonoids, especially rutin, quarrecetin, kaempferol, luteolin and low concentration of coumarin. Total flavonoids were about 0.78 mg for each gm. powdered aerial parts.

Polymorphism of P-selectin (- 2123C/G, rs1800807 C/G) and IL-4 -1098 in a sample of Iraqi Diabetes Mellitus Type 2

Asmaa M. Salih Almohaidi

Jornal of Biotechnology Research Center, Volume 12, Issue 1, Pages 56-63

Diabetes is a major public health problem because it is world’s sixth cause of death. An inflammatory disease that associated with many inflammatory markers, which is a key for complications of vascular endothelial system, literally for high risk populations. Some adhesion molecules were studied in the sera of Iraqi Arab female as P-selectin associated with inflammatory disease and Cytokines. This work tend to evaluate the association between soluble P-selectin level, polymorphism for mutation site 2123C/G (rs1800807) and the role of P-selectin and IL-4 polymorphism in the developing of Diabetes Mellitus type 2. Genetic polymorphism of P-selectin gene was investigated at the position -2123C/G, Present study illustrated three genotypes (GG, GC, CC), with the significant protective effect for G allele and Etiological effect for C allele. The Polymorphism of IL-4- gene at the position IL-4 -1098 was presented with three genotypes (TT, TC and CC), Furthermore the result of C allele frequency (RR=1.45 with EF=0.312) reflect an Etiological effect for C allele with positive association with disease. All present factors in the first view may occur as independent risk factors but after a deeply focusing on results, it seems as if there’s a sequence functions among them, the result in a synergist effect interfere with Diabetes Mellitus type 2

Role of the Anesthetic Determine the Clonidine Effects Mediated by RVLM/-α2-Adrenoceptors on Blood Pressure and Heart Rate in Rats

Imad Hatim Rashid

Jornal of Biotechnology Research Center, Volume 12, Issue 1, Pages 64-78

To examine the hypothesis of a role for α2-adrenoceptors in mediating the mechanism of urethane hypotensive effect whether it's peripheral or central, Wistar rats were anesthetized with urethane or (for comparison) with halothane, to study the influence of urethane that govern the mechanism of central and peripheral α2-adrenoceptors action, on basal BP & HR, and the rise in blood pressure (BP) to the stimulation of caudal pressor area (CPA), when these receptors were either centrally activated by bilateral rostral ventrolateral medulla (RVLM) microinjection of clonidine (30nM), and blockade with any of the clonidine antagonists, yohimbine (500pmol/50nl), and idazoxane (270nM) or yohimbine+idazoxane, or when peripherally activated (of urethane anesthetized rats) by i.v. clonidine (100nmol/kg), which also blockade with idazoxane or yohimbine+idazoxane. The results indicated presence of no anesthetic differences in a partial involvement of α2-receptors-RVLM, vs. a complete involvement of I(1)-imidazole receptors in mediating the hypotensive effects of clonidine. It also indicates α2-/I(1)-receptors synergism in raising the urethane lowering of baseline of SBP to the levels of control or halothane group. In conclusion, the result suggests involvement both of the central and the peripheral α2-adrenoceptors in mediating urethane hypotensive effects.

Morphological and Physiological Response of Potato for Radiation and Salt Stress in vitro

Zainab Abduljabbar Hussain AL-Hussaini; Lameea Azrag Mutlag; Shatha Ayd Yousif; Taghrid Abdul-Jabbar Said

Jornal of Biotechnology Research Center, Volume 12, Issue 1, Pages 79-85

The experiments were undertaken to study the response and behavior of two mutant clones of Potato genotype for salt stress by exposing it to different salt levels of sodium chloride (with electrical conductivity of 8, 10, 12 dS m-1) and compare with those in the control treatment 6 dS m-1. The morphological response was determined by measuring the morphological characteristics of vegetation, root growth, tuber formation. While the physiological response was determined by estimating some ions in the vegetative and root growth. The results showed a significant decrease in the morphological characteristics of the vegetative growth (number of shoots, plant height and dry weight) and tuber formation by increasing salt levels, while the characteristics of the root growth (number of roots, lengths and dry weight) were not affected. There is no significant difference in the behavior of the two clones under saline levels, except for the superiority of vegetative clone (C2) at comparison treatment in the number of shoots (2.00 shoot / plant), and vegetative clone (C1) at comparison treatment and 12 ds m-1 in the shoot length and the percentage of tuber formation (13.40 cm and 100% Respectively). The results also recorded that the root growth of vegetative clone (C2) was a significant accumulation of Na+ and Ca++ at 12 dS m-1 which reached 8.33 and 23.38 mg-1 gm dry weight, respectively, while the accumulation of K+ in vegetative clone (C1) was increased by the root growth in the control treatment which reached 45.03 mg -1 gm dry weight.

Introducing Soybean Oil in Semen Extenders of Awassi Rams

Nameer M. Albiaty; Safaa A. Abed; Hazem J. Alobaidi; Safaa G. Almaliki; Abbas F. Kareem; Mohammad I. Mukhlif

Jornal of Biotechnology Research Center, Volume 12, Issue 1, Pages 86-91

The aims of the study was to investigate the possibility of using an plant lecithin; commercial soybean oil (SO) directly in the components of semen extenders of Awassi rams, storage of semen in chilled technique and the effects of dilution, cooling and storage periods on semen quality. Semen was collected weekly from four Awassi rams by electro- ejaculator. Pooled samples were divided to six equal aliquots and diluted by Citrate egg yolk extender at 37˚C. Treatments were designed on the base of control extender containing 25% egg yolk and four treatments containing different addition of SO (12.5, 25, 37.5, and 50%) and combination treatment of 12.5% egg yolk + 12.5% SO. Treatment tubes were cooled to 5˚C and stored for three days. Semen was evaluated as raw, diluted, cooled and after storage in refrigerator (5˚C) for 1, 2 and 3 consecutive days. Results showed that there were no significant differences among all treatments in progressive motility, dead sperm %, abnormal sperm % and acrosome defect %, while pH were found to be significantly declined (p≤ 0.05) in control group. Significant effects of dilution, cooling, and storage period have been demonstrated with steadily significant deterioration (p≤ 0.05) for all studied characteristics when motility and pH declined while sperm abnormality, dead, and acrosome defect percentages increased. The results clearly indicated successful of using different levels of SO (plant source) as lecithin source instead of egg yolk (animal source of lecithin) without any impacts on the biological characteristics of Awassi ram semen and the process of dilution, cooling and storage periods have deterioration effects on semen quality.

Evaluating the Genotoxicity Enhancement of the Antileukemic Drug 6-Mercaptopurin When Combined With Iraqi Nerium oleander and Narcissus tazetta Extracts in vivo

Bushra Mohammed Amin Mohammed; Mahmood Othman Ahmed

Jornal of Biotechnology Research Center, Volume 12, Issue 1, Pages 92-102

The present study was aimed to evaluate the genotoxicity of the aqueous extracts of Nerium oleander leaves and Narcissus tazetta bulbs each alone and together with the antileukemic drug 6-Mercaptopurin (6MP) in order to investigate the extracts ability to elevate the chemotherapeutic drug genotoxicity which may influence its treatment of cancer. The cytotoxicity test shows that the LD50 of the aqueous extract of Narcissus tazetta was 752.083 mg/kg and for Nerium oleander was 922.023 mg/kg. On the basis of the achieved LD50 values, the doses 92, 46 ,23 mg/kg of Nerium oleander and the doses 75, 37,18 mg/kg of Narcissus tazetta extracts were chosen, depending on chromosome aberrations, Micronuclei and Mitotic index as a powerful cytogenetic assays in bone marrow cells of Swiss albino male mice. The result indicated that Nerium oleander extract alone at the dose 92mg/kg induced significant effect on centromere break and ring chromosome comparing with the negative control (untreated mice) and significantly increased the mean values of chromatid gap and ring chromosome when compared with the positive control ( 6MP ). While only the dose 46 mg/kg and 23 mg/kg of N. oleander aqueous extracts significantly decreased mitotic index and when combined with 6MP it can enhance its antimitotic activity but not significantly. Moreover the extracts alone and when combined with 6MP did not significantly change the total number of red blood micro nucleated cells. For Narcissus tazetta extract, the three experimental doses alone lead to significant increase in chromosomal aberrations like: chromatid break with fragment, chromatid break without fragment, chromatid gap, centromeric break, ring chromosome and dicentric chromosome. While only the dose 75mg/kg had induced significant structural chromosomal abnormalities such as chromatid break with fragment, chromatid break without fragment centromeric break and ring chromosome, when combined with 6MP. The three doses of N. tazetta extract alone, had led to significant reduction in mitotic index compared with untreated control and also its combination with 6-MP significantly decreased the percentage of mitotic index. Moreover, only the doses 75 mg/kg and 37 mg/kg of N. tazetta extracts, when had given alone caused significant increase of the total micronucleated cells. While only the dose 75mg/kg of N. tazetta had induced significant frequency of the total micronucleated cell when combined with 6MP. In the present report, we attempted to establish that N.tazetta and Nerium oleander aquatic extracts enhance the genotoxicity and bioactivity induced by the antileukemic drug 6MP , thus preventing the development of cellular drug resistance which is a major problem that can face cancer patients using this drug .The current study serve the purpose of which is to search for local plants that may contribute to the establishment of novel supportive complementary and alternative medicine (CAM) during the chemotherapy of cancer in Iraq , Further studies are merited to explore this possibility.

Molecular Analysis of Touched DNA Samples Collected From Pistol Surfaces

Asia Abdul Lateef Mahdi; Majeed Arsheed Sabbah; ad Ali Hussain; Sura Nabeel Hameed; Aula Ali Hussein

Jornal of Biotechnology Research Center, Volume 12, Issue 1, Pages 103-107

Molecular analysis of touch DNA samples from pistol surfaces is important for crime scene investigation. In this study touched samples were collected from different parts of pistol. DNA extracted and quantified using different methods, then analyzed by mtDNA sequencing and STR profiling. The results showed that pistol surfaces can be analyzed by mtDNA sequencing but not full STR profiled. These results indicated the need for improving DNA extraction and STR profile kits for analysis low DNA amount.

Treatment of Waste Paper Using Ultrasound and Sodium Hydroxide for Bioethanol Production

Osama F. Saeed; Rafid A. Abdulkareem; Shatha K. Muallah

Jornal of Biotechnology Research Center, Volume 12, Issue 1, Pages 108-114

Bioethanol produced from lignocellulose feedstock is a renewable substitute to declining fossil fuels. Pretreatment using ultrasound assisted alkaline was investigated to enhance the enzyme digestibility of waste paper. The pretreatment was conducted over a wide range of conditions including waste paper concentrations of 1-5%, reaction time of 10-30 min and temperatures of 30-70°C. The optimum conditions were 4 % substrate loading with 25 min treatment time at 60°C where maximum reducing sugar obtained was 1.89 g/L. Hydrolysis process was conducted with a crude cellulolytic enzymes produced by Cellulomonas uda (PTCC 1259).The maximum amount of sugar released and hydrolysis efficiency were 20.92 g/L and 78.4 %, respectively. Sugars released from waste paper were fermented into bioethanol with Saccharomyces cerevisiae. The maximum concentration of bioethanol estimated was 9.5 g/L after 48h of cultivation, the yield and volumetric productivity were 0.454 g/g glucose and 0.2g bioethanol/ L h. respectively. This study of ultrasound and sodium hydroxide treatment may be (we think) it will be a promising technique to develop bioethanol production from waste paper.

Biological Treatment of Used Engine Oil by Single and Mixed Bacterial Cultures Isolated from Soil of Mechanic Workshops

Amal A. Hussein; Saad H. Khudhair

Jornal of Biotechnology Research Center, Volume 12, Issue 1, Pages 115-123

The accumulation of hydrocarbon waste, such as used engine oils in environments, has many impacts on humans and other organisms, therefore many researches were achieved to degrade or remove or consume these pollutants. The aim of the current study is to get a local bacterial isolates has high ability to degrade the spent engine oil as a single or mixed culture. Five soil samples contaminated with spent engine oil were collected from mechanic workshops in Baghdad city to isolate degrading bacteria using Bushnell Hans medium (BHM), pH 7 with 5% of used engine oil. While the growth patterns and gravimetric analysis was used to reveal the ability of these isolates to degrade spent engine oil in liquid BHM medium. The best three isolates A4, B6 and D5 were identified and the optimal temperature and pH for biodegradation of spending engine oil were studied. Also, the consortium culture of three isolates was tested their ability to utilize spent engine oil under the same conditions for single isolate. Twenty five bacterial isolates were obtained from contaminated soil samples and three isolates appeared a maximum degradation rate 74.6, 70.2 and 78.5% respectively. The results from identification tests were showing these isolates belong to Bacillus sp., Acinetobacter sp. and Pseudomonas sp., respectively. The studied three isolates gave the best degradation when incubated at 30°C in BHM medium pH 7. While other results were indicated that consortium cultures are more effective 90.2% than all experiments that used single isolate.

Effect of aqueous extract of Rosemary officinalis on cytotoxicity of CCL4 induced albino male mice

Ruqaya Mohammed Ibrahim

Jornal of Biotechnology Research Center, Volume 12, Issue 1, Pages 124-131

This study focused the line on the effect of aqueous extract of Rosemary officinalis, as well as, effect of toxic compound CCL4, on micronucleus formation and mitotic index assay in albino male mice. This work started at September 2017 at Biotechnology Research center Al-Nahrain University, by using 20 albino male mice. The result indicated that aqueous extract of rosemary caused significant increased in mitotic index and decrease micronucleus formation for two doses tested 50,100 mg/kg in comparison with negative and positive controls, also the results revealed that CCL4 showed significant mutagenic action on biological system of treated mice by increased frequency of micronucleus formation and decreased the percentage of mitotic index in bone marrow cells. Pre-and post –treatment between aqueous extract and CCL4 were also made. The results of pre and post treatment with rosemary extract were also caused a significant decreased in micronucleus formation and increase the percentage of mitotic index for two doses 50,100 mg/kg in comparison with its corresponding controls which caused increased in the frequencies of micronucleus formation and decrease the percentage of mitotic index in bone marrow cells. Conclusions: Rosemary officinalis enhanced immunity, reduced mutagenic effects against cytotoxicity of CCL4.

Effect of Fertility Blend® Administration on the Epididymal Sperm Function Parameters of Vasectomized Mice: Physiological and Genetical Study as Model for Obstructive Azoospermic Men

Haidar A. Al-Ebrahimi; Jasem A. Al-Ethawi; Hazim E. Al-Ahmed; Saad S. Al-Dujaily

Jornal of Biotechnology Research Center, Volume 12, Issue 1, Pages 132-136

Male Fertility Blend® is a new nutrient supplements containing many constituents especially a plant called Dong quai(Angelia Sinensis). The plant extract has been used to facilitate the sperm function parameters. However, the studies concern on its effects on epididymal sperms of vasectomized males and obstructive azoospermia are very rare. Thus, this investigation was designed to elucidate the role of Male Fertility Blend® (MFB) formula on the in vivo epididymal sperm characters and DNA fragmentations of vasectomized male mice as a model for man. In this study, the orally administration of 3.4 µg/ml MFB was used for vasectomized and non-vasectomized mice along 35 days. The results revealed a significant (p≤0.05) increment in certain sperm function parameters of vasectomized mice by using fertility Blend® than that of not treated by this supplement. The percentage of progressive and unprogressive active sperm motility using MFB was significantly (P≤0.05) increased compared with non-treated group. It was concluded that the MFB formula containing different sources of energy and variety of factors that sustain the epididymal sperm of healthy and vasectomized mice. Therefore this supplement can be utilized for males complaining from obstructive azoospermia and other factors of infertility.