Current Research in Microbiology and Biotechnology

ABSTRACT: Present study focused in the difference expression of Aflatoxin B1 Regulatory Gene in clinical and environmental Aspergillus parasiticus isolates was subjected to RNA isolation, cDNA synthesis and PCR amplification, result appeared clean bands upon amplification with aflR1 primers with Product size 798bp. The ability of Aspergillus parasiticus confirmed the presence of aflR2 gene in Product size 400 bp in all the studied isolates with different gene expression The present study showed that there was direct correlation between aflr gene expression and source of the examined isolates. The results showed that the influence of source of isolate in aflr gene expression was significant. It appeared that aflr gene expression increased at the Aspergillus parasiticus isolated from Zea mays with gradual gene expression in all Aspergillus parasiticus isolated from grains of Zea mays with no clear expression in the Aspergillus parasiticus isolated from Rice grain. These results indicate the suitable molecular method for detecting and controlling the spread of Aspergillus parasiticus in the grainer and in hospitals to avoid infection.

KeyWords: Aspergillus parasiticus; Aflatoxin B1 ; gene expression; aflR.

How to cite: Nemat J.Al-jady. Expression of Aflatoxin B1 Regulatory Gene in Environmental Aspergillus parasiticus Isolates Using Real-Time PCR. Curr Res Microbiol Biotechnol. 5(4) 2017: 1106-1111

ABSTRACT: This study aimed to assess the efficiency (hydrogen peroxide) to resist wilt plants tomato caused by pathogenic fungus Fusarium oxysporuim under laboratory protected and field and home conditions. The results of the effect of treatment hydrogen peroxide on the growth of pathogenic fungus F. oxysporum under laboratory conditions that all concentrations tested may decline significantly linear growth and dry weight of the biomass of the fungus pathogen rate and the number of spores have treatment surpassed 200 ppm of hydrogen peroxide sigificantlly on the other to reduce the rate of growth linear and dry weight of the biomass and the number of spores where he recorded 26.0 mm, 0.0 mg, 10 x 0.46 6 / cm 2, compared with the control treatment (0.0 ppm of hydrogen peroxide which recorded 45.67 mm, 0.33 mg, 10x 9.17 6 Spore / cm2, respectively. the results also showed that the treatment of 200 ppm of hydrogen peroxide has achieved the highest percentage inhibition of the growth of the dry weight of biomass which recorded 43.06% and 100%, respectively. results of the effect of three concentrations of hydrogen peroxide (50 ,100 ,200) ppm on some tomato plant growth standards under protected cultivation and field conditions that all treatments have improved significantly, "most studied growth standards compared with two treatment, control and pathogen treatment in addition to the reduced significantly of the percentage of wilting plant.

KeyWords: Fusarium oxysporuim sp., hydrogen peroxide, tomato, pathogenic, biomass.

How to cite: Mena waleed Hatem. Evaluation of the efficiency of hydrogen peroxide against Fusarium oxysporum, which causes fusarium wilt on tomato plant. Curr Res Microbiol Biotechnol. 5(4) 2017: 1112-1116

ABSTRACT: Cutaneous leishmaniasis (CL) is caused by a protozoan parasite of the genus Leishmania that infects macrophages of many mammals including humans. Their infection induces both humoral and cellular immune responses, but the balance of their expression varies with the type of the disease. The aim of the present study is to determine levels of serum chemokines that include granulocytes-macrophages colonies stimulating factor (GM-CSF), intercellular adhesion molecule1 (ICAM-1), interferon-gamma inducible protein-10 (IP-10) and vascular endothelial growth factor (VEGF) in Iraqi patients with CL lesions which can be considered as immunological markers for CL infection. Ninety people were included in the present study; sixty of them were infected with CL lesions while 30 uninfected people were considered as control. Patients were diagnosed on the basis of clinical and parasitological criteria. Sera (GM-CSF, ICAM-1, IP-10 and VEGF) levels were determined by ELISA using a quantitative sandwich enzyme immunoassay technique. The results showed no significant differences between males and females (p>0.05) in the infection with CL, while highly significant difference (p<0.01) for distribution of CL lesion according to site of infection were recorded. The prevalence of multiple lesions was more than single lesions with a highly significant differences (p<0.01) .Sera levels of GM-CSF, ICAM-1, IP-10, and VEGF were significantly higher in patients group than healthy subjects (P< 0.01).

KeyWords: Cutaneous leishmaniasis, Chemokines, GM-CSF, ICAM-1.

How to cite: Ali N. Yaseen. Determination of some Sera Chemokines in Iraqi Patients infected with Cutaneous Leishmaniasis. Curr Res Microbiol Biotechnol. 5(4) 2017: 1117-1121

ABSTRACT: Infections with multi-drug resistant strains become a big problem specially the nosocomial infections, which can cause death especially for the debilitated older adults. The aim of this study is to find a better way to kill multi-drug resistant staphylococcus aureus instead of the orthodox medicine, which become scarcely effective. The multi-drug resistant organisms are undergo progressing due to the continuous evolution of the antimicrobial patterns. This problem had directed the researchers to use alternative therapy. In this work ozonated water and Bacillus lentus have been chosen to study their activity and the merits of each in comparison with the traditional antibiotics.

KeyWords: multi-drug resistant, Bacillus lentus, Ozone solution, alternative therapy, traditional antibiotics.

How to cite: Samarah Jafar Alwash. Comparison between biological and chemical methods to kill multi-drug resistant Staphylococcus aureus. Curr Res Microbiol Biotechnol. 5(4) 2017: 1122-1125

ABSTRACT: Capsule of K. pneumoniae is a polysaccharide in structure, it’s one of bacterial virulent factors. In this study we focused to finding the best capsular antigen from different isolates led to stimulate immune response, throughout measuring several biochemical and immunological parameters. Thus six K. pneumoniae isolates from different specimens (urine, wound and burn swabs, sputum, blood, stool) were used to extract and partially purified the capsules by organic solvents (ethanol, chloroform, butanol) and strong alkaline. The results of chemical analysis showed that the carbohydrates concentration of the partially purified CPS extracts were 115(87.1%), 140(90.7%), 150(89.8%), 163(88.3%), 135(88.8%), 130(90.9%)µg/ml, whereas the concentrations of protein in the same extracts were5(3.7%),3(1.94%),4(2.39%),5(2.7%),2(1.31%),1 (0.69%) µg/ml, while the concentration of lipids 12(9.09%), 11.2(7.2%), 13(7.7%), 16.4 (8.8%), 15(9.8%), 12(8.3%)µg/ml ,with(0.6, 0.8, 0.6,0.7, 0.5, 0.7) ng/µl nucleic acid respectively . In vivo study revealed that, the concentration of anti K. pneumoniae CPSs (IgG) in serum of rabbits were elevated significantly (P≥ 0.05) in all groups (108.64±5.38 , 168.52±3.20, 129.63±4.27, 196.30±7.70, 102.47±10.10 ,and 121.23± 5.58)ng/ml, as well as the level of IL-4, IL-10 and TNF-α ,were (48±9.16, 59.66±8.83, 58±7.50,64± 1.52, 47±5.50 and 56±7), (42.33±4.40, 41.33±1.85, 42±1.73, 35.3±2.02 ,43±5.50and 42.33±0.88) and (307±1.73, 366.3±6.43, 501.66±6, 616.6±2.18, 766.3±13.73 and 835±22.91) pg/ml respectively for that K. pneumoniae CPSs extracted from urine, wound, burn, sputum, blood and stool respectively, in comparison with control. But IL-2 level shown a significant elevation (P ≥0.05) in some groups which were (74± 6.24, 68±17, 54±2.08, 53.66±15.38) pg/ml for that CPSs of sputum, wound, burn and stool respectively, while the level of IL-6 showed a significant elevation (P ≥0.05) in groups of(sputum, burns, blood and stool )were(417±3.71, 294±22.27, 263.3±21.85 and 150±16.07)pg/mlrespectively. The positively increasing of WBC counts, which effect on the phagocytosis by increasing the PI compared with control. Thus the challenge test with virulent K. pneumoniae, showed the ability of CPSs extracts to protect the treated rabbits against K. pneumoniae infection.

KeyWords: K. pneumoniae, Capsular polysaccharide, K. pneumoniae capsule.

How to cite: Rand M. Abd Al-Rhman et al. Assessment of K. pneumoniae capsular antigens isolated from different clinical specimens in stimulation several immune parameters. Curr Res Microbiol Biotechnol. 5(4) 2017: 1126-1133

ABSTRACT: Three types of local algae (Chlorella sp., Oscillatoria sp., Scenedesmus sp.) were isolated from Iraqi aquatic environments and were cultivated and grown to produce biomass. The effect of these algae was studied as a natural vegetarian food source on the growth of fishy brown fish fingerlings and investigated the effect of these algae by mixing them in specific proportions with a sample of pre-prepared forage Stander Media(SM) that used in fish farming and production. Eight pools with ten fingerlings in every pool were prepared and each two were supplied with specific forage and considered to be a treatment, treatment A1 (25% algae+75%SM), treatment A2 (50% algae+50% SM) and treatment A3 (75% algae+ 25% SM). The BOD, pH and temperature were measured daily and the survival rate measured every week for each treatment. The experiment continued for ten weeks, during which changes were observed in the rate of weights, lengths and survival of the fish sample for each treatment. In this study it was found that A2 was the best treatment used for improving the growth of fishes. The aim of the study is to identify the importance of algae and their potential use as alternative natural forage and to invest it as promoters for the growth of important aquatic organisms such as fishes.

KeyWords: Cultivation algae, Fish growth, Fingerlings, Alternative forage.

How to cite: Ibtisam Fareed Ali Karm. Cultivation of Some Local Algae from Iraqi Environment and Study its Impact on the Growth of Fishes. Curr Res Microbiol Biotechnol. 5(4) 2017: 1134-1137

ABSTRACT: This study investigated the potential in vivo antagonistic behavior of human derived probiotics; Bifidobacterium longum Bl and Lactobacillus acidophilus La against infection with invasive food born Listeria monocytogenes Lm in murine model. Mice Pre-feeding with probiotics conferred a significant protection against orally challenged mice with virulent isolate of Lm. A significant reduction in the level of Lm colonization of the gastrointestinal tract in mice was observed in the probiotics pre- colonization in comparison to the control animals, in that the Lm viable count decreased by almost two log cycle, and this reduction was accompanied by a marked elevation in the level of sIgA and serum IgA, particularly in mice pre-colonized with Bl, as the level of sIgA rose about 3.7 folds. Probiotics pre- colonization also stimulated phagocytic activity, the peritoneal macrophages were pronounced a significant (P< 0.05) enhancement in the phagocytic function, the phagocytic index (PI) increased to 65.7 ± 3.95 % and 55.2 ± 3.1 % respectively, when Bl and La were orally administrated to mice. The antilisterial activities appear to be non – lactic acid dependent behavior.

KeyWords: Probiotic, Lactobacillus, Bifidobacterium, L. monocytogenes, Antagonistic effects, immunomodulatory activity.

How to cite: Shadan A. Alwendawi. The Suppressive effects of Human Derived Probiotics Ingestion against Listeria monocytogenes in Murine Model. Curr Res Microbiol Biotechnol. 5(4) 2017: 1138-1145

ABSTRACT: The pathology resulting from Staphylococcus aureus MRSA infections is of great importance as one of the community associated bacterial infections and its ability to produce biofilms. Studies have focused on monospecies biofilm production, therefore, this study was designed to detect multispecies biofilm production. Biofilm production was evaluated by Microtiter plate method using crystal violet stain. Different clinical isolates including Staphylococcus aureus (MRSA), Candida albicans and Pseudomonas aeruginosa. Dual species included S. aureus MRSA with C. albicans and S. aureus MRSA with P. aeruginosa. Strong biofilm production of S. aureus MRSA showed the highest percentage 70% , while in dual species with C. albicans it showed 25% and with P. aeruginosa 50%. These results show the different behavior in biofilm production of bacterial species with other bacteria or yeasts in combinations, which may affect the mixed infections and the ability of such microorganisms to resist antibiotics.

KeyWords: Biofilm; Interactions; Pathogenic bacteria; Yeasts.

How to cite: Ghadah Mohommed Saleh et al. Biofilm Production of Staphylococcus aureus (MRSA) and its interaction with each Candida albicans and Pseudomonas aeruginosa. Curr Res Microbiol Biotechnol. 5(4) 2017: 1146-1150

ABSTRACT: Synthesis of iron bio-nanoparticles was done by using Juglans regia water extract in a simple method using Ferric sulfate as un-oxidizing agent. Characterization of Iron nanoparticles was performed using UV, XRD, and FTIR. The diameter of iron nanoparticles was about 21 nm. In vitro anti-inflammatory activity was detected by using albumin denaturation assay, membrane stabilization, at different concentrations of bio-nanoparticles in comparison with standard drug. The results showed that heat induced haemolysis of erythrocyte was significantly inhibited at the concentration of 400 μg/ml, also the highest albumin denaturation inhibition appeared at 400 μg/ml.

KeyWords: bio-nanoparticles, UV, XRD, FTIR.

How to cite: Hanen Abdulalsalam et al. Evaluation of anti-inflammatory effect by using iron nanoparticles prepared by Juglans regia water extract. Curr Res Microbiol Biotechnol. 5(4) 2017: 1151-1156

ABSTRACT: Laccase enzyme from P. aeruginosa SR3 was produced in lab scale fermenter. It has shown from the results that maximum laccase production was observed after 24 hr of incubation, and the specific activity was reached to 6214 U/mg protein after 24 hrs of incubation, whereas the production was increasing when xylene concentration was nearly depleted from the medium after 24 hr. Immobilization of enzyme was conducted in two methods, entrapment by calcium alginate and agar-agar with maximum immobilization efficiency 49% and 32%, respectively and covalent method by chitosan and gelatin with maximum immobilization efficiency 63% and 50% respectively. The highest rate of enzyme specificity found with oxidation of O-tolidine, while gallic acid, vanillic acid are poor substrates for laccase. The decolorization capability of immobilized laccase on chitosan loaded with black dye at concentration of (30 mg/l) in packed bed-reactor operated as a batch culture system for 3 hrs. Decolorization rate was stabilized approximately after 2 hr with removal efficiency 97.3% of dye. Decolorization of black textile dye was performed also by operation of packed bed bioreactor continuously at different flow rates (15, 30 and 45) ml/hr and HRT (7.9, 3.9 and 2.6) hr. respectively. Decolorization efficiency of black textile dye with immobilized laccase in a bioreactor increased at flow rate 15 ml/ hr., and reached to 71% after 2.5 hr.

KeyWords: Pseudomonas aeruginosa, fermenter, laccase enzyme, immobilization, dye, bioreactor.

How to cite: Sahar I. Hussein et al. Decolorization of Textile Dyes in Packed Bed-Reactor Using Batch and Continuous System by an Immobilized Laccase Produced from Local Isolate of Pseudomonas aeruginosa SR3. Curr Res Microbiol Biotechnol. 5(4) 2017: 1157-1166

ABSTRACT: Multiple sclerosis (MS) is an autoimmune disease characterized by losing the myelin sheath surrounding axons in the central nervous system, although the etiology of the disease still unclear, but many of environmental factors such as infectious diseases might contribute in inducing the disease. One of these infectious diseases is cytomegalovirus. The current study involved examining 45 patients with multiple sclerosis (MS) and 25 apparently healthy persons as control, to detect the anti-Cytomegalovirus antibody in their sera by a rapid test and ELISA technique. Our results showed that three sera sample were positive in patients with MS (6.66%), which suggest that cytomegalovirus may be one of the triggering agent to multiple sclerosis.

KeyWords: autoimmune disease, multiple sclerosis, viruses, Cytomegalovirus.

How to cite: Dunya fareed salloom et al. Detection of Cytomegalovirus in a sample of multiple sclerosis. Curr Res Microbiol Biotechnol. 5(4) 2017: 1167-1169

ABSTRACT: Epidemiology is the study of the distribution and determinants of health-related states or events (denominators), including diseases and subclinical cases. Various methods can be used to carry out epidemiological investigations: surveillance and descriptive studies can be used to study distribution; analytical studies are used to study determinants. It is the cornerstone of public health, and shapes policy decisions and evidence-based practice by identifying risk factors for disease and targets for preventive healthcare. Opportunistic psychrotrophic, biofilm-producing and multidrug-resistant foodborne Carbapenemases producing or Extended Spectrum Beta Lactamase Resistant Klebsiella pneumoniae (ESBL/KPC) strains are pro-growing threat worldwide. This epidemiological surveillance focused on isolation and Identification of K. pneumoniae especially ESBL producers from Cows mastitic milk in Baghdad. Study design including collection and processing of forty five samples of mastitic milk from clinically infected Cows from regions of Abu-Ghraib, Al-Fudhaliyah and Al-Sadrya (fifteen samples from each region) during November (2016) to March (2017), in which they collected and processed according to modified dairy microbiological methodology in veterinary public health laboratory by McConkey agar. Gram stain, catalase, oxidase and negative staining technique with nigrosine and safranin for capsule detection. Electronic RapID™ ONE (4 hours) biochemical panel micro-tubes strep identification system compendium with reference colors chart and online confirmation microcodes data base software. Gold standard double staining technique, Microtiter Plate Assay for biofilm formation with methylene blue and safranin dyes. Antibiotics Susceptibility Pattern by Kirby-Bauer technique or disk diffusion method was proceed according to instructions of clinical laboratory standards institute (CLSI) or national committee for clinical laboratory standards (NCCLS) by using a Muller-Hinton agar and McFarland opacity tubes for checking resistance profile of isolates. Double diffusion inhibition technique or Oxoid Cefpodoxime Combination Kit determined ESBL resistance activity. Data were analyzed for significant differences by statistical package for social sciences software (IBM SPSS) in which a Chi-square was used. The results verify detection of twenty two strains (48.88%) of phenotypically indole negative K. pneumoniae out of forty five samples: twelve strains (26.66%) from Abu-Ghraib: eight strains (17.77%) were ESBL producers; five strains (11.11%) from Al-Fudhaliyah: two strain (4.44%) was ESBL producers and five strains (11.11%) from Al-Sadrya: two strain (4.44%) was ESBL producers. In conclusion, data verified isolation and identification of K. pneumoniae in Baghdad, in which ESBL producers were detected, thus I recommend confirmation of isolates by multiplex-PCR, monitoring of subclinical infected & carrier Cows, milk handlers, environment, utensils, equipment, milk cans, vectors, etc. through application of sanitation practices and hazard analysis critical control points (HACCP) strategies to overcome or reduce these public health problems.

KeyWords: Klebsiella pneumoniae, ESBL, Mastitis.

How to cite: Ali Hassan Ahmed Al-Shammary. Epidemiological Pattern of Extended Spectrum Beta Lactamase Resistant Klebsiella pneumoniae (ESBL) isolated from Mastitic Milk in Baghdad. Curr Res Microbiol Biotechnol. 5(4) 2017: 1170-1177

ABSTRACT: Algae as a natural therapy recently drew attention because its limited side-effects. In the present investigation, after phytochemical analysis of Spirulina platensis; different concentrations of S. platensis aqueous extracts were investigated in vitro and in vivo for antimicrobial effect against pathogenic Pseudomonas aeruginosa as gram negative bacteria; and Staphylococcus aureus as gram positive bacteria, which commonly associated with wound infections. After using Agar well diffusion method, variable inhibition zones were observed at 100mg/ml of extract compared with reference antibiotics (Ceftazidime for P. aeruginosa and Azithromycin for S. aureus). By using open wounds procedure as a clinically in vivo wound model that contaminated with P. aeruginosa and S. aureus, treatment with the extract showed accelerated epithelial regeneration compared with reference antibiotics, and infected wounds without treatment. Through the results of in vitro and in vivo experiments, we observed that Spirulina platensis aqueous extract has an effective antibacterial activity which a proved by accelerating the wound healing process.

KeyWords:Antibacterial; In vitro; In vivo; Spirulina platensis.

How to cite: Thura M.H. Alyasiri et al. In vitro and In vivo Antibacterial Activity of Spirulina platensis. Curr Res Microbiol Biotechnol. 5(4) 2017: 1178-1183

ABSTRACT: To evaluate the expression of the MAPK1 and MAPK4 genes under salinity stress, four wheat genotypes were selected form our previous study based on potassium/sodium ratio include Iba99, Axad9, Uruk and Furat genotypes. Seeds were germinated in growth chamber, and then Plants were grown in hydroponic system until the third leaf was emerged (after 2-3 weeks). Wheat plants were supplied with nutrients solution containing nutrients solution. Four concentrations of salinity including control (distilled water) 100,150 and 200 mM NaCl were used.RNA was extracted from leaf tissues, and converted into complementary DNA (cDNA). Two primers pair (MAPK1 and MAPK4) genes with Tubulin as internal standard for normalization were used.These results indicate that the MAPK1 and MAPK4- mediated plant tolerance to salinity. The expression analysis of the MAPK1 and MAPK4 genes illustrated that the genotypes Uruk and Furat are salt tolerance and Iba99 and Axad9 genotypes are susceptible to salinity. RT-PCR is a powerful method to identify gene expression in different tissues. Therefore, examination the differences in gene expression of bread wheat genotypes under salinity stress could provide more knowledge about the variation between susceptible and tolerant wheat genotypes.

KeyWords:Gene expression, MAPK1 gene, MAPK4 gene, Salinity stress, Hydroponic system.

How to cite: Kamil M. AL-Jobori et al. Expression Pattern of Wheat Mitogen-Activated Protein Kinase (MAPK1 and MAPK4) Genes under Salinity Stress. Curr Res Microbiol Biotechnol. 5(4) 2017: 1184-1195

Curr Res Microbiol Biotechnol

CRMB :::: Vol.5, No.4, July - August 2017