World Journal of Experimental Biosciences

40-44

Authors: Zainab M. Hamid, Silva A. Yakoub Zokian

This study attempts to identify genetic diversity among four genotypes of pomegranates cultivated in Diyala city, Iraq and find the genetic polymorphism among them by using DNA markers generated by polymerase chain reaction (PCR). Total genomic DNA of genotypes studied was extracted from dry young leaves by using cetyltrimethyl ammonium bromide (CTAB) procedure. Molecular analysis was performed by using twelve random markers in random amplified polymorphic DNA (RAPD-PCR) technique. The analyses based on five primers OPH – 08, OPH – 13, OPH – 18, RI – 3 and RI – 16, which used in amplification and polymorphisim for the four cultivated genotypes studied. The genetic polymorphisms value of each primer was determined and ranged between 31 to 100%; primers OPBA–3 and OPBB–4 produced the highest percent of genetic polymorphism compared with primer OPH–13. The genetic diversity and the isolation of the four cultivated genotypes of pomegranates confirmed obviously by RAPD-PCR technique.

50-56

Authors: Marwa. H. Al-Khafaji and May. T. Flayyih

Three hundred milk and cheese samples were collected from Baghdad markets. Two hundred staphylococcal isolates were isolated from milk and cheese samples. Staphylococcus aureus was predominant species (97 isolates (48%)) followed by S. chromogenes (82 isolates (41%)) and S. epidermidis (21 isolates (11%)). The antibiotic susceptibility of coagulase positive staphylococci (COPS) and coagulase negative staphylococci (CONS) isolates to 3 antibiotics (Methicillin, Tetracycline and Vancomycin) was determined using disc diffusion method. The results revealed that 80 isolates of S. aureus (82.47%) were resistant to methicillin (MRSA), while 8 isolates (8.24%) were resistant to vancomycin (VRSA) and 18 isolates of S. aureus (19%) were resistant to tetracycline. Sixty four CONS isolates (62.13%) were resistant to methicillin, 28 CONS isolates (27.18%) resistant to tetracycline, and 5 CONS isolates (4.85%) resistant to vancomycin. Deoxyribo nucleic acid (DNA) was extracted from staphylococcal isolates. The genetic determinants of methicillin resistance genes (femA and mecA) were amplified using monoplex PCR technique to identify methicillin resistant (mecA+) and susceptible (lacking mecA) staphylococci and to identify S. aureus (femA+) and coagulase negative staphylococci (lacking femA). Ninety six S. aureus isolates (98.96%) were attributed as harbour femA gene. The mecA gene was detected in 91 (93.81%) MRSA isolates, while it was detected in 70 (67%) CONS isolates.

61-68

Authors: Najlaa N. Yaseen1*, Harith J. F. Al-Mathkhury1

In present study 74 specimens of urine were collected from patients suffering from urinary tract infections. Fifty (67.56%) isolates were identified as Escherichia coli. 78% of isolates were identified as extended spectrum beta lactamases (ESBL) producer. Antibiotic susceptibility test was done and ceftazidime was selected to complete this study by implying stress at sub-MIC on isolate harbor high number of resistance genes (N11) and compared with sensitive isolate (S). Only four β-lactamase coding genes were detected; blaTEM, blaPER, blaVIM and blaCTX-M-2 and N11 had blaTEM, blaPER, and blaVIM. It was found that the resistant isolate did not form biofilm when compared with the sensitive one, which formed moderate biofilm. In addition, ceftazidime stress reduced the ability to produce slime layer and affected the viable bacterial count in combination with pH and temperature stresses.

74-79

Authors: Ahmed, Dalia Azhar, Hassani, Hayfaa H

identified by bacteriological and biochemical assays and confirmed by vitek 2 identification system. It was found that 20 (66.6%) isolates were identified as P. mirabilis and 10 (33.3%) isolates were P. vulgaris. Susceptibility of P. mirabilis to cefotaxime was done, Notable, 13 (65%) P. mirabilis isolates were resistant to cefotaxime, whereas, 6 (30%) P. mirabilisisolates were extended spectrum beta lactamases producers. MIC value of cefotaxime was estimated to 7 isolates (512 μg/ml), and the MIC was 4096 μg/ml for other isolates, One isolate had MIC equal to 128 μg/ml while, another isolate showed MIC equal to 8192 μg/ml to cefotaxime. DNA was extracted from 8 P. mirabilis isolates that was resistant to cefotaxime and 2 isolates were sensitive to cefotaxime. The blaCTX-M-2 gene was detected by polymerase chain reaction (PCR). The 8 cefotaxime resistant P. mirabilis isolates and one cefotaxime sensitive isolate carried blaCTX-M-2 gene, while the cefotaxime sensitive isolate did not harbored the gene. Furthermore, this study confirmed that blaCTX-M-2 gene was carried on chromosomal DNA, by extraction the DNA from 10 P. mirabilis isolates having blaCTX-M-2 gene. DNA profile showed chromosomal DNA band only. Curing of blaCTX-M-2 gene was done, 4 cefotaxime- resistant P. mirabilis isolates were treated with ethidium bromide as a chemical curing agent in different concentrations. The results confirmed that blaCTX-M-2 gene was carried on chromosomal DNA.

84-88

Authors: Abeer Mohammed Ali Al-garawyi

Ampicillin hydrolyzing class C β-lactamase (AmpC) that confer resistance to extended spectrum cephalosporins and never inhibited by β- lactamase inhibitors. Results of this study confirmed that 37 (41.1%) isolates of Klebsiella pneumonia from 90 isolates tested for cefoxitin susceptibility by disk diffusion method. AmpC β-lactamase producers obtained from clinical isolates; 13 (35.1%) from urine, 10 (27%) from blood, 10 (27%) from wound, and 4 (10.8%) from vagina. AmpC β-lactamase producer isolates were confirmed in for cefoxitin resistance 37 K. pneumoniae isolates by modified three dimensional test and AmpC disk test. From 90 isolates, plasmid encoded AmpC genes were detected by multiplex PCR in 30 (33.3%) of the K. pneumoniae isolates. Of these, plasmid encoded AmpC genes belonging to the MOX family were detected in 14/30 (46.7 %) isolates. Gene of the family ACC type was present in 2/30 (6.7%) isolates. While, the other isolates 14/30 (46.7%): 7(23.3%), 4(13.3%), 2(6.7%) and 1(3.3%), isolates contained another families like DHA, FOX, CIT and EBC, respectively.

94-99

Authors: Zainab Zamel Khalaf and May Talib Flayyih

Escherichia coli is natural and essential part of the bacterial flora in the gut of humans and animals. E. coli produces colicin, proteinaceous toxin that could inhibit the growth of similar or closely related bacterial strains. The production incidence of four colicin types in E. coli were tested in 31 bacterial isolates isolated from different sources (urine, stool, blood, ear swab and sewage water) by using polymerase chain reaction of specific primer for each colicin type. Production of colicin E1 was detected in four bacterial isolates (13.33%), three of them isolated from urine samples, while the last isolated from sewage water. Colicin M was detected in 15 colicinogenic isolates (50%), 12 isolates isolated from urine, 2 from sewage, while last one from stool. However, colicin E3 and E9 were not detected in all colicinogenic isolates. E. coli isolated from human urinary tract infections showed high incidence of colicin M and E1 production, respectively

1-5

Authors: Farah Mohommed Salehand Ghadah Mohommed Saleh

Pseudomonas aeruginosa is emerging opportunistic clinical pathogens. Clinical isolates of P. aeruginosa resist wide spectrum of antibiotics and form biofilm. The comparison study between clinical and environmental of P. aeruginosa in terms of biofilm formation and antibiotic resistance is very scanty. Thus, in current study microtiter plate technique was used to measure the biofilm formation by several clinical and environmental isolates. Moreover, the antibiotic susceptibility of these bacteria was evaluated by VITIK 2 techniques. The relationship between the antibiotic susceptibility and biofilm formation was evaluated for clinical and environmental isolates. Clinical and environmental isolates of P. aeruginosa produced a good amount of biofilm but the clinical isolates produced higher amount of biofilm as compared to environmental isolates. Resistance to antibiotics by clinical isolates was higher than resistance to antibiotics by environmental isolates and the minimum inhibition concentration (MIC) of most antibiotics to clinical isolates were higher than MIC against environmental isolates. Little relationship was observed between the biofilm formation and antibiotic resistance in case of clinical isolates, while no relationship was seen between the antibiotic susceptibility and biofilm formation. It can be concluded that the clinical isolates produced biofilm higher than environmental isolates. The relationship was seen only between the biofilm produced by clinical isolates and antibiotic susceptibility.

14-17

Authors: Hind J. Al-Ubaidy, Khalid A. Rasheed

The current study included an assessment the ability of Ceratophyllum demersum Phytoremediation of cadmium in accumulative concentration by measuring the heavy metal in the tissues of the plant to check the ability of C. demersum to reduce or eliminate the harmful and toxic effect, which resulting from this type of pollutants in the most important ecosystem. In present study the plant was grown in natural conditions for two months and treated to two concentrations of cadmium (3.714, 4.952 mg/l). The samples of water and a certain weight of the plant were taken before and after treating with cadmium chloride in order to estimate the level of cadmium in these samples before and after treatment. The residual amount of cadmium in the water and also in the plant has also been calculated at different time intervals (3, 6 and 9 days). The present study showed that cadmium accumulated rapidly in the plant and increased dramatically with time, thus the highest level of cadmium was observed at day nine. C. demersum showed high ability to remove cadmium and was at the ninth day of the experiment and that was 1.53, 1.089 mg /l) when exposed to 3.714, 4.952 mg /l of cadmium respectively. The current study proved strongly the ability of C. demersum to eradicate the cadmium in ecosystem.

22-25

Authors: Areej Abbas Zabbon

The impact of immunity on spermatogenesis is documented in the literature. This study highlighted the influence of Pseudomonas aeruginosa hyper-immunization on male rabbit's testicular spermatogenesis as the effect of immune response on sex hormones changes. Morphometric criteria were used to evaluate spermatogenesis. The level of hormone was estimated by enzyme-linked immunosorbent assay (ELISA). We found that male rabbits vaccinated with the bacteria showed a highly significant (P<0.001) decreased in spermatogonia cell number (11.46±8.11 in test vs 45.19±10.76 in control). In addition there was a significant increase (P<0.001) in spermatocytes cell number represented by pachytene and zygotene cells in vaccinated animal as compared to control (88.5±8.11 in test vs 54.14±10.13 in control). In concomitance with these changes, there was a significant elevation in level of testosterone in immunized groups (3.55±3.86 in test vs 0.98±0.8 in control). While, no significant change was observed in progesterone (2.00±0.80 in test vs 1.83±0.79 in control) and estradiol (21.42±14.02 in test vs 19.31±3.72 in control) levels. The histopathological changes included detachment of spermatogonia in the test group was observed. The increase in spermatogenesis and the changes in hormones level as well as histopathological changes suggest that the immune mechanisms impact on spermatogenesis in males rabbits.

30-35

Authors: Nada Abdulrahman F. Al-Easawi, Muhammad Nafea Ali Al-Azzawi

The effect of shisha smoking on liver parenchyma tissues were evaluated in (120) male adult mice. The animals were divided into four groups according to kind of exposed smoke (cigarette, shisha and mixture of cigarette and shisha). The mice exposed to fresh air considered as control group. A special inhalation chamber designed locally was used to expose the animals to different kinds of smokes. Exposure to cigarette smoke was done for 5 min/day, while exposure to shisha smoke was done for 15 min /day for 4, 8, 12 weeks. The tissues of control and exposed groups were processed for histological study. The results showed different histological changes such as hepatocytes degeneration, congestion, sinusoid dilation and infiltration of inflammatory cells in liver parenchyma tissues. These changes observed clearly when the period of exposure increased especially in groups exposed to shisha smoke and shisha plus cigarette smoke for 12 weeks. The results showed that shisha smoke causes mild damage in liver parenchyma tissue and this damage increased concomitantly with the increase of duration of exposure.