Vol. 22, January, 2009.

1/22 AMELIORATIVE EFFECT OF GAMMA RADIATION ON NACL STRESSED LYCOPERSICON ESCULENTUM L. PLANT GROWTH IN SOIL INFESTED WITH FUSARIUM OXYSPORUM F.SP. LYCOPERSICI

M.A. Rizk and H.W. Botros*

Botany Department, Faculty of Science, Cairo University, Giza, Egypt.

* Plant Research Department, Nuclear Research Center, Atomic Energy Authority.
The present study was carried out to investigate the influence of saline stress and/or gamma radiation on the tomato seedlings development, mycelial growth and sporulation of Fusarium oxysporum. Irradiation of the fungus ameliorated the detrimental effect of salinity and improved the percentage of seedlings emergence and increased the root, shoot lengths and dry weight of tomato seedlings. Also, coupling salinity with irradiation significantly increased the mycelial growth in soil and biomass gain of Fusarium oxysporum up to 2 kGy, above which the growth and sporulation were hardly affected and completely suppressed at 5 kGy. On the other hand, exposure of the tomato seeds up to 4 Gy counteracted the suppressive effect of salinity and increased the growth parameters in presence or absence of the fungus. Fusarium oxysporum f.sp. lycopersici appeared to be tolerant to salinity up to 4.8 EC (millimohse) and highly sensitive to irradiation dose 5 kGy.

 

 

2/22 ADVERSE EFFECTS OF GENITOURINARY INFECTIONS

ON SEMEN QUALITY.

Fathi T. Al-Sabie, Vivek S. Agwan, Abdelhadi Al Najar and Jamal H. Al Amari

Department of Microbiology, Faculty of Pharmacy, Al-Fateh University

for Medical Sciences, Tripoli - Libya.

A total of 1321 semen samples were collected from patient attended a private clinic in the Tripoli city during period from August 2004 to August 2008,. All the samples were studied bacteriologically for presence of pathogenic micro-organisms, by standard microbiological techniques described by world health organization manual. In addition blood samples of all the patients were studied serologically, by venereal disease research laboratory test, to detect or rule out syphilis. Out of all samples 449(34%) contained pathogenic bacteria, 291 samples (22%) contained candidas, 238 samples (18%) had both bacteria and Candida, 185 samples (14%) contained trichomonas, and 158 samples (12%) had bacteria, candidas and trichomonas. Commonest bacterial pathogen isolated was Neisseria gonorrhoeae 162(36%). Serological test was positive indicating infection by Treponema pallidum in 54 (12%) samples. Bacterial infection was seen to cause maximum change in volume of semen 0.1 milliliters to more than 12 milliliters. Infections also caused increase in pH and change in color. Other parameters like increase in pus cells, red blood cells, viscosity and decrease in sperm count, sperm motility; were seen to change more with Neisseria gonorrhoea infections. Candida and trichomonas infections had little effect on semen analysis parameters.

 


3/22 THE MICROBIOLOGICAL AND HEAVY METALS OF PROCESSED CHICKEN MEAT PRODUCED IN EGYPT.

Azza A. Mostafa

Department of Biological and Environmental Science,Faculty of Home Economics, Al-Azhar University.

Poultry products are universally popular and in recent years the consumption of poultry meat has risen dramatically. The present study report on the contamination of processed poultry meat by microorganisms and heavy metals. For assessing contamination, twelve processed poultry meat samples were purchased from supermarket (Cairo, Tanta and Alexandria) The samples were analyzed for the estimation of heavy metals such as Pb, Cr, Co, Zn, Cu and Cd and assessed the microorganisms such as total counts of fungi, yeasts mesophlic aerobic bacteria, coli form, Salmonella spp. and total count of Bacillus cereus. The highest total count of fungi isolated from oriental chicken kofta (330 cfu/g), yeasts (2060 cfu/g) from chicken shawerma, mesophilic aerobic bacteria (706000 cfu/g), from chicken fillet, coliform group (345 cfu/g) from chicken shawerma, Salmonella spp(60 cfu/g) and Bacillus cereus (33 cfu/g) from oriental chicken meat. The concentration of Pb ranged from (0.96 – 87.4 ppm), Cr (3.4 – 46.6), Co (0.0 – 32.6 ppm), Zn (130 -1173 ppm), Cu (12.8 -93.6) and Cd (0.0 – 2.0 ppm).

 

 

4/22 CHARACTERIZATION OF AN ANTIMICROBIAL COMPOUND PRODUCED BY BACILLUS CIRCULANS

Emad A. Abada; Hoda H. EL-Hendawy; Mohamed E. Osman and Mohamed A. EL-Badry

Botany and Microbiology Department, Faculty of Science, Helwan Univeristy, Ain Helwan, Egypt

A Bacillus circulans isolate was obtained from the rhizosphere of Medicago sativa grown in an open field at Helwan governorate, Egypt. This strain and its culture supernatant restricted the growth of a number of Gram positive and Gram negative bacteria as well as fungal species. Of the tested bacteria, Staphylococcus aureus was the most sensitive one, while Serratia marcescens was the most resistant strain. The inhibitory effect of Bacillus circulans towards S. aureus was achieved in 24 h when the former was grown in nutrient broth containing 0.3 % starch and 0.35 % DL- methionine at pH 8 and 30oC. The antimicrobial compound was purified and subjected to spectroscopic characterization by ultraviolet absorption spectrum, the infra red spectrum and analysis by gas chromatography – mass spectrum. The obtained results revealed that the antimicrobial compound is suggested to be 4-(Diphenylmethyl)-6 ethoxycarbonyl-1-phenyl-1H-pyranolo [4, 3-c] pyridine. This study reported that B. circulans strain synthesizes a potent antimicrobial compound. Further work on the cytotoxicity of this compound is in progress to investigate the possibility of using it for further formulation of better drugs.

 


5/22 IDENTIFICATION AND CONFIRMATION OF A BACTERIAL DISEASE, CAUSING NATURAL OUTBREAKS IN FARMED TILAPIA OREOCHROMIS NILOTICUS L.

Amer M. Diab

Botany Department, Faculty of Science, Suez Canal University, Egypt

Bacteria of Vibrio alginolyticus spp. were identified from archive tissues and bacterial cultures from an outbreak of mortalities in tilapia of four big farms in Ecuador. Samples were supplied by the diagnostic facility of Institute of Aquaculture (IOA), University of Stiriling, UK. The isolated bacteria had a biochemical profile similar to V. alginolyticus as well as conventional identification test compatible with this pathogen and the diagnosis was confirmed using Polymerase Chain Reaction (PCR). A sequence procedure was performed for some of the isolated bacteria, and the 16S rDNA sequence (GenBank accession number AY 373027) gave 99% sequence identity to three Vibrio species (V. alginolyticus, V. harveyi and V.parahaemolyticus).

 

6/22 THE MODULATORY POTENTIAL OF IRON CHELATION THERAPY ON IMMUNITY

Nebal M. Darwish and Gihan M. Tawfeek*

Departments of Microbiology & Immunology and Parasitology*,

Faculty of Medicine, Ain Shams University, Cairo.

Iron-overload both aggravates the outcome of infections caused by a variety of microorganisms and exerts subtle effects on immune status by altering the proliferation of T and B cells. Aim: The aim was to investigate the effect of iron-overload on the type of T helper (Th) immunity elicited and the subsequent effect on the susceptibility, course and outcome of Cryptosporidium parvum infection were investigated in the present study. Methods: Separate groups of iron-overloaded (40 mg of iron Dextran / Kg intraperitoneally every other day from the day of infection). Spleens were harvested and an enzyme linked immunosorbant assay (ELIZA) was performed on spleen culture supernatants for in vitro analysis of interferon-gamma (IFN-γ) and interleukin (IL)-4. Iron-overload induced a Th2 cytokine response with significantly higher IL-4 and lower IFN-γ levels compared to infected non-treated control. The infection rate was 100% and the infection was severe and persistent in both immunocompetent and immunocompromised iron-overloaded mice with death of all immunocompromised mice.Iron chelation by desferrioxamine (DFO) enhanced the production of Thl anticryptosporidial immunity (significantly higher IFN- γ compared to infected non-treated control) limiting the severity and clearing the infection in iron-overloaded immuncompetent mice (cure rate 100%) and controlling the infection in immunocompromised mice with cure rate 30%, percentage reduction in oocyst shedding 80.8% and mortality rate 10% at the end of the experiment (30 days post infection) .Conclusion : These data indicated that iron-overload negatively affected Thl-mediated immunity in mice with cryptosporidiosis thus altering the susceptibility course and outcome of infection. Iron-overload represents a risk factor for flaring up of Cryptosporidium parvum infection in absence of any obvious immunosuppressive conditions; hence successful therapy in iron-overloaded hosts depends mainly on iron chelation. The immunomodulatory properties of DFO were elucidated in terms of restoring Thl immunity in iron-overloaded mice.

 

7/22 SEROPREVALENCE OF SYPHILIS IN TRIPOL- LIBYA

Fathi T. Al-Sabie, Vivek S. Agwan, Nebras M. Banana, Neveen F. Abd Al Masih and Asma Al Jilany Al Mallhouf

Department of Microbiology, Faculty of Pharmacy, Al-Fateh University

for Medical Sciences, Tripoli- Libya.

Syphilis is an infectious but curable sexually transmitted disease (STD). This study includes 281 cases showing clinical signs and symptoms suggestive of syphilis, who attended Asma clinic, Tripoli, Libya from 2004 to 2008. These cases included 160 (57 %) male and 121 (43 %) female. All the patients were from 19 to 58 years of age. Blood samples were collected from the patients and subjected to Rapid Plasma Reagin (RPR) test. Out of 160 male patients 34 (21.25%) and out of 121 female patients 9 (7.4%) were found to be RPR test positive. Highest percentage of incidence of syphilis was found in males (36%) and females (41 %); in the age group 19 - 28 years. Out of 34 positive male patients 22 (66%) were married and 12 (34%) were unmarried. All the 9 (100%) positive female patients were married; 8 (89%) non-pregnant and 1 (11%) pregnant.

 

 

8/22 EARLY DIAGNOSIS OF NEONATAL SEPSIS BY AMPLIFICATION OF 16 S rRNA GENE

Nebal M. Darwish, Mohammad A. Mohammad* and Ghada I. Gad**

Microbiology and Immunology Department, Pediatric Department*,

Faculty of Medicine Ain Shams University

Background: Bacterial infection continues to be the major cause of morbidity and mortality in the newborn. Aim: The present study aimed at evaluating the usefulness and practicability of molecular methods in the early diagnosis of neonatal sepsis as compared to the conventional blood culture method. Methods: Forty five newborns with clinical diagnosis of neonatal sepsis, were investigated for objective evidence of sepsis using conventional blood culture method, PCR assay using universal primers for the amplification of the conserved region of bacterial 16S rRNA gene. Results: Blood culture was positive in 35 / 45 (77.8 %) of cases. According to the results of blood culture, neonates were classified into two groups. Group (A): Included 35 neonates with clinically suspected sepsis and blood culture positive results (proven sepsis). Group (B): Included the remaining 10 neonates with clinically suspected infection (CSI) yet the blood culture yielded negative results. Regarding the PCR amplification of 16S rRNA gene, the present study showed agreement between the results of the molecular methods and those of conventional microbiological methods. Both blood culture and PCR yielded 35/45 (77.8%) positive cases .Results were identical for 41 samples (91.1 %), including 33 culture-positive and PCR-positive samples and 8 culture-negative and PCR-negative samples .Only two samples (4.4%), which were culture-positive, gave negative results by PCR whereas two other samples (4.4 %) were identified only by PCR and yielded negative results in culture .Considering the blood culture as the gold standard for diagnosing neonatal sepsis, the sensitivity, specificity, positive predictive value and negative predictive value for PCR were 94.3%, 80%, 94.3 % and 80 % respectively. Conclusion: The ability to rapidly and accurately rule out bacterial sepsis in a newborn infant might have a significant impact on the infant, the family, and the healthcare system. Although culture will not be superseded by PCR-based testing due to the requirement for purified culture isolates in antimicrobial susceptibility testing, molecular techniques could reliably rule out neonatal sepsis in less time than bacterial culture. This, in turn, would allow for the exclusive treatment of neonates with true infections, thus reducing the use of broad-spectrum antibiotics and the potential for infants who are not septic of acquiring drug-resistant bacteria. This approach will also permit shorter hospital stays within the NICU and reduce significantly the overall medical costs to the healthcare system as well as the emotional burdens of the families of these infants.

 

9/22 PRELIMINARY STUDIES ON POSTHARVEST FUSARIUM DRY ROT DISEASE OF POTATO TUBERS

Amira M. Abu-Taleb, Mohamed S. El-Abyad, Nagy G.H. Awad*

and Mohey El-Deen M.A. El-Toamy*

Botany Department, Faculty of Science, Cairo University

* Agricultural Research Center, Giza, Egypt.

Fusarium dry rot is an important postharvest disease of potato tubers, that causes significant yield loss. Variations of Fusarium dry rot incidence were observed among 6 naturally and artificially infected cultivars of potato. The cultivars Oblix and Vangogh were the most susceptible, cv. Frisia was the most resistant, whereas cvs. Baraka, Jaerla and Monaliza showed intermediate reaction with various degrees of response. Fusarium solani was more virulent to potato tubers than Fusarium oxysporum. F. solani has the ability to attack many hosts causing sever infection. Disease severity depended upon potato varieties, pathogen, wound depth and incubation period. There was a direct correlation between lesion diameter and wounding depth as well as incubation period. However, infection site had no significant effect on disease severity. Growth parameters of F. solani were maximum at 25 oC and at 64 % relative humidity. Tissues of potato tubers stimulated growth and spore germination more than sucrose. However, sporulation was abundant on sucrose more than potato tissues. The selected fungicides were efficient against Fusarium dry rot particularly thiabendazole.

 

 

10/22 THE OCCURRENCE OF DERMATOPHYTES AND OTHER KERATINOPHILIC FUNGI IN THE STUDENTS HOUSES OF AIN SHAMES UNIVERSITY, EGYPT

Amal, A.I. Mekawey,

Regional Center of Mycology and Biotechnology

Al-Azhar University, Egypt

A survey of keratinophilic fungi including dermatophytes and other pathogenic forms were carried out in 100 air – dust samples from bedrooms, dinning halls of female student resident houses. Various dermatophytes isolates were obtained the most common namely Microsporum canis, M. gypseum, M. gallinae M. ferrugineum and Trichophyton mentagrophytes. Also, several yeast and/or yeast like species were isolated including C.albicans, C. norvegesis, Geotrichum candidum, Cryptococcus humicola and C. ater.Using dilution plate methods, thirty seven species representing fourteen genera of which Aspergillus niger, A. flavus, A. fumigatus, Rhizopus nigricans, Penicillium chrysogenum, Syncephalastrum racemosum, Fusarium oxysporum, and Alternaria alternata were most frequently isolated.

 

 

11/22 EFFECT OF AFLATOXINS ON ULTRASTRUCTURES OF GEOTRICHUM CANDIDUM

Abdel Razak A. Abou Seadda, Mohamed E. Zain, Ayman F. Ahmed

and Ahmed Abdel-Fattah*

Botany and Microbiology Dept., Faculty of Science, Al-Azhar University, Cairo, Egypt.

*Regional Research Center for Mycology and Biotechnology, Al-Azhar University, Egypt.

Two aflatoxins, B1 and G1, were isolated from Aspergillus parasiticus strain. Geotrichum candidum was grown on mineral medium separately supplemented with 3 and 5 mg/ml of aflatoxins B1 and G1, respectively. The effect of aflatoxins on the ultrastructures of Geotrichum candidum were investigated. The scanning and transmission electron microscopy photographs revealed that the morphological structures and mitochondria of Geotrichum candidum were affected by aflatoxins B1 and G1.

 

 

12/22 PURIFICATION AND CHARACTERIZATION OF NEUTRAL PROTEASE FROM SOIL STRAIN OF BACILLUS SUBTILIS

Mohsen A. Mahmoud, Mohamed H.M. Al-Agamy, Mohamed S. Ashour

and Sameir S. El-Loboudy*

1Microbiology and Immunology Department, Faculty of Pharmacy, Al-Azhar University, Nasr City, Cairo, Egypt

*Botany and Microbiology Department, Faculty of Science, Al-Azhar University, Nasr City, Cairo, Egypt

Protease enzymes of most potent strain, B. subtilis B-6, were purified in 2-step procedures involving ammonium sulfate precipitation, sephadex G-200 and sephadex G-100 gel permeation chromatography. The purification steps of protease(s) resulted in having two proteases namely protease A and B with specific activities of 727.4 and 460.0 u/mg protein/ml respectively.The maximum protease activities for both enzymes were attained at 30 0C, pH 7.2, 1% of gelatine concentration and 0.4 enzyme concentrations. The obtained data showed that the two purified proteases were different enzymes not identical ones. It was suggested that applications of protease(s) of the present purified enzymes in the fields of pharmacy, chemotherapeutic agents and/or industrial purposes and this suggestion, however, needs further investigation.

 

 

13/22 GROWTH AND SURVIVAL OF SOME LACTIC ACID BACTERIAL STRAINS IN CAMEL’S MILK

Elsayed M. Abd El-Wahed

food science Department, Faculty of agriculture, Zagazig University, Egypt.

Growth characteristics of Streptococcus thermophilus, Lc. lactis subsp. lactis, Lc. lactis subsp. cremoris, Lb. delbrueckii subsp. bulgaricus, Lb. acidophilus and Bifidobacterium bifidum in pasteurized camel and cow milk were evaluated. results indicated the growth rate of Streptococcus thermophilus, Lc. lactis subsp. lactis, Lc. lactis subsp. cremoris, Lb. delbrueckii subsp. bulgaricus, Lb. acidophilus were higher in cow milk than in camel’s milk after 12 h incubation and remained high until the end of incubation period. But in case of B. bifidum was higher in camel’s milk than in cow milk, and the death phase did not occur in camel’s milk after 12 h incubation and remained high until the end of incubation period with gradual decrease in pH with increase time of incubation.

 

 

14/22 COMMUNITY ACQUIRED PNEUMONIA IN PEDIATRIC AGE GROUP: ROLE OF ATYPICAL BACTERIA

Amira A.M. Abo Serieh, *Shereen Fawzy, Malak A. Shaheen, Sally A.F. El Sahriggy, and Tharwat E. Deraz

Departments of Paediatrics and *Medical Microbiology and Immunology, Faculty of Medicine, Ain Shams University, Cairo, Egypt

Community-acquired pneumonia (CAP) in infants and children remains a significant problem worldwide. The currently available methods in everyday clinical practice do not allow definitive microbe-specific diagnosis in the majority of patients and initial antibiotic treatment of CAP is usually empirical, customarily covering both typical and atypical pathogens. To date, no local sufficient evidence exists to support this broad coverage. Our aim was to detect the frequency of atypical bacteria (Chlamydophila pneumoniae, Mycoplasma pneumoniae and Legionella pneumophila) as causative agents of CAP in pediatric age group in Egypt for rational implementation of empirical antibiotic therapy. Fifty patients (mean age 3.7±3.5 years) with clinically diagnosed and radiologically proven CAP were included in the study. Sputum specimens were collected from all patients and subjected to standard bacteriological culture and identification for detection of typical bacterial pathogens and PCR technique for detection and identification of atypical bacterial pathogens as causative agents of CAP. Definite bacterial pathogens were identified in 70% of the studied patients; 30% typical bacteria (14% S. pneumoniae, 10% H. influenzae, 4% S. aureus and 2% K. pneumoniae) and 40% atypical bacteria (26% C.pneumoniae, 14% M.pneumoniae). L. pneumophila has not been detected in any of our studied cases. Atypical bacterial pneumonia was found to be significantly higher in children more than 5 years old.   Pediatric patients below 5 years with typical bacterial pneumonia had a severe disease course than those with atypical bacterial pneumonia. Radiological findings showed a limited capacity to differentiate between typical and atypical bacterial pneumonia and between chlamydial and mycoplasmal pneumonia. Our results signify that high percent of children above 5 years have CAP due to macrolide sensitive organisms as C. pneumoniae and M. pneumoniae. Thus it is worthy to include macrolide antibiotics for empirical treatment of CAP in school-age children. 

 

 

15/22 HIGH-RESOLUTION MELTING ANALYSIS REAL-TIME PCR TECHNIQUE FOR DIAGNOSIS OF MYCOTIC KERATITIS

Salwa S. Afifi; *Magda H. Mahran; *Dalia G. Said and *Olfat. A. Hassanin

Department of Microbiology, Faculty of Pharmacy for girls, Al Azhar

University and *Research Institute of Ophthalmology

The goal of this study is to determine a rapid sensitive molecular method for detection of fungal etiologyof clinically important microbial Keratitis, as early diagnosis is crucial for prompt antifungal therapy.Twenty six patients with resistant corneal ulcers were included in this study. After stopping all eye drops for 48 hours prior to sampling, corneal scrapping was done using 23G needle. Microscopic examination of smears, cultures and qualitative Real-Time PCR analysis were performed for all samples. PCR was performed using   universal fungal primer pairs targeted to the 18s rRNA gene. The PCR positive samples were then identified by using genus specific primers for each of Fusarium; Candida; and Aspergillus which detect the presence of sequence variations in target-gene derived PCR amplicons of the internal transcribed spacer (ITS) regions of the r RNA gene. Combination of microscopy and culture gave a positive result for fungus in 7 (26.2%) of 26 samples. By using Real-Time PCR method, fungal infections were detected and identified in 9 (34.6%) of 26 samples of microbial Keratitis at melting temperature (Tm 83.66±2.15). The most common fungal genera were Fusarium followed by Candida and the least one was Aspergillus. In conclusion, Real-time PCR using SYBR Green dye and melting curve analysis can differentiate multiple amplification products and identify the corresponding genus based on the melting temperature of the fragment generated by PCR; this method can be an alternative to culture-based techniques for the rapid diagnosis and treatment of fungal Keratitis.

 

 

16/22 ASSESSMENT OF SPUTUM AND BROCHOALVEOLAR LAVAGE MICROBIAL PATTERN, CELLULAR PROFILE AND INFLAMMATORY MARKERS IN CHILDREN WITH CHRONIC LUNG DISEASES

Asmaa El-Hussieny, *Shereen Fawzy, Malak A. Shaheen, Tarek A. Abdel Gawad, Magda Y. El-Seify and Karima A. Abdel Khalik.

Departments of Pediatrics and *Medical Microbiology and Immunology, Faculty of Medicine, Ain Shams University, Cairo, Egypt

Early detection of lower airways bacterial colonization (LABC) in patients with chronic lung diseases (CLDs) and administration of proper antimicrobials help in delaying or preventing chronic progressive lung damage. Our aims were to identify potentially pathogenic microorganisms (PPMs) colonizing lower airways (LAs) of clinically stable children with CLDs, determine their antimicrobial susceptibility so that protocols for suppressive antibiotic therapy could be formulated, and assess local and systemic inflammatory markers.Bronchoalveolar lavage fluid (BALF) and sputum specimens of 30 clinically stable children with CLD were subjected to semiquantitative bacterial cultures and assessment of local cellular profile. Both BALF and serum Lactate Dehydrogenase (LDH) and Alkaline Phosphatase (ALP) enzymatic levels were determined.Cultures of BALF samples yielded 32 PPMs in 22/30 (73.3 %) patients while culture of sputum samples yielded 13 PPMs corresponding only to 10/22 (45.5%) of these patients. The agreement between BALF and sputum rate was poor (kappa = 0.07, p >0.05) pointing to the more reliability of BALF sampling method over sputum. Gram negative Enterobacteriaceae spp. and Pseudomonas aeruginosa were the dominating PPMs followed by Staphylococcus aureus. Most Gram negative organisms were sensitive to ceftriaxone, imepenem, tobramycin, gentamicin, and amikacin. Most of Gram positive organisms were sensitive to oxacillin, clindamycin and vancomycin. BALF-LDH & BALF-ALP levels were significantly higher than their corresponding serum levels with lack of significant correlation between them. In addition, normal blood total leucocytic count, ESR and CRP were found even in presence of colonization with serious PPM. Our results show that LAs of nearly three quarters of clinically stable patients with CLDs are colonized by PPMs. There is an intense ongoing local inflammation with bacterial colonization with no serum reflection of the ongoing inflammatory process. Clindamycin and tobramycin or gentamicin is the proposed suppressive antibiotic therapy that has to be assessed. 

 

 

17/22 EFFECT OF CHEMICAL, ORGANIC AND BIO-FERTILIZATION ON INULIN PRODUCTION IN HELIANTHUS TUBEROSUS L.

Hussein A. Bosil*, Khamis A. Refa*, Salwa H. El Kashlan*

and Gamal M. Gazal*

Dept. of Horticulture, Faculty of Agriculture , Al–Azahar University, Cairo, Egypt

*National organization for Drugs and Researches

This study was carried out during the two successive growing summer seasons 2004 /2005 - and 2005/2006, under sandy soil condition, to study the effect of chemical, organic and biofertilization on inulin production. Chemical fertilization treatments were 25%, 50% and 100% NPK. Organic fertilization treatments were 15, 20, 25M3/fed. Biofertilization treatments were control, N. fixing, P.dissolving and biomixture. The obtained results from this study indicated that the highest value of inulin content (13.95g/100g) dry weight in tubers was recorded with 100% NPK only, while the highest content of inulin (3.937g/100g) in leaves was recorded with 100% NPK combined with biomixture. However, the maximization of inulin production in plant tubers (14.1 g/100g) was recorded with organic fertilization (25m3/fed) combined with biofertilization (N.fixing). However, organic fertilization (15m3/fed) combined with   biofertilization (N.fixing) resulted the highest inulin content (3.995 g/100g) from plant leaves.  

 

 

18/22 PURIFICATION AND CHARACTERIZATION OF NEMATICIDAL COLLAGENASE PRODUCED BY STREPTOMYCES WARTII

Abdel-Rahman A. Alhumiany

Faculty of Medical Science, Taraba, Taif University, Taif, Saudi Arabia

Purification of the nematicidal extracellular collagenase was carried out on the culture filtrate of S. wartii after 3 days growth on MM1 medium using complete purification procedure. It was possible to bind 76.1 % of the collagenolytic activity by (NH4)2SO4 precipitation at 75 % saturation, which was signaled as partially purified collagenase. The DEAE-Sephadex column chromatography separated the collagenolytic activity into two peaks (A) and (B). The Sephadex, G-100 column of the pooled fractions (A) and (B) resulted in one peak of activity having 11.4 CU mg-1 specific activity, 27.1 fold of purification and 45 % yield. The apparent molecular mass of the homogenous enzyme was approximately 35 kDa using SDS-PAGE and electrophoresis. Characterization of the purified nematicidal collagenase of S. wartii revealed that the optimum enzyme concentration was 20 mg protein ml-1 and the Km was 0.5 mg ml-1, the optimum pH ranged from 7-9 and was thermostable between 10-50 oC. The purified collagenase was completely inhibited by both Hg2+ and PMSF. The inhibition of enzyme by PMSF indicated that it was serine protease with collagenolytic activity. The purified enzyme could hydrolyze collagen and gelatin but was not able to hydrolyze casein, elastin and egg albumin, so it is considered as a true collagenase. The nematicidal activity was evaluated at each purification step, the number of immobilized nematodes increased with increase in purification folds.

 

19/22 EMERGENCE OF CEPHALOSPORIN RESISTANCE AMONG STREPTOCOCCUS PNEUMONIAE ISOLATED FROM HOSPITALIZED EGYPTIAN PATIENTS

Azza A.M. AbouZeid, Yehia A. El-Zawahry, Magdy A. Ghoniem*

and Hayam M. Hamouda**

Botany Department, Faculty of Science, Zagazig University

*Biochemistry Department, Faculty of Veterinary Medicine, Cairo University

**Microbiology Department, National Organization for Drug Control and Research (NODCAR)

Resistance of clinical isolates of Streptococcus pneumonia to β-lactam antibiotics including penicillin and cephalosporins has been reported through out the world with increasing frequency. Monitoring of antibiotic resistance in 52 S. pneumonia strains isolated from in-patients in Fever and El-Sadr hospitals in Cairo, Egypt has been performed. The patients were previously treated with cefotaxime, amoxicillin and penicillin antibiotics. The antibiotic resistance profile of examined strains against seven β-lactam antibiotics (cephatrxyl, cefotaxime, cefizidime, cephalexin, cephazolin, cefepime and penicillin) revealed that 50% of total isolates were resistant to cefotaxime and one strain out of 52 was considered multi-resistant and encoded as (SPR). MICs of the seven tested antibiotics varied greatly between the isolated S. pneumonia resistant strains (SPR) and the sensitive reference strain ATCC 49619 encoded as (SPS). The RAPD-PCR analysis of both tested strains using five arbitrary primers confirmed the close relation since seven similar DNA fragments occurred in case of primer no. 5. Meanwhile, genetic heterogeneity between the two tested strains was obviously detected upon using primer no. 3 where no fragments were produced in case of (SPS) strain. The cellular proteins of both tested strains were verified using SDS- PAGE technique in presence of MICs and MICs/2 of both tested strains. MICs of CTX, CL, KZ and FEP induced the biosynthesis of high molecular weight (81 kDa) cellular protein in (SPR) strain. Similarly, 81kDa protein was induced by MICs/2 of CF and CL antibiotics. Meanwhile, MIC of CTX and MIC/2 of CAZ induced production of another high molecular weight protein (88 kDa). The plasmid profile of the clinical resistant S. pneumoniae strain (SPS) and sensitive reference strain (SPS) revealed the presence of plasmid in both strains with molecular weight 5.125 kbp predicting that cephalosporin resistance gene is harbored on chromosomal DNA. Resolution of chromosomal DNA amplicons amplified by PCR using specific primers for pbp2x gene revealed the occurrence of a band with length 918 bp diverging from the expected length reported in previous reviews suggesting mutation contributing to the emerged antibiotic resistance.

 

 

20/22 FERTILIZERS MANAGEMENT AND N2-FIXERS COMBINED WITH PHOSPHATE SOLUBILIZING MICROORGANISMS AFFECT PEANUT (ARACHIS HYPOGAEA) GROWTH AND PRODUCTIVITY.

Osama N. Massoud and Nadia H. El-Batanony*

Soil, Water and Environment Research Institute, Agricultural Research Center,

Giza, Egypt.

*Environmental Studies and Research Institute (ESRI), Sadat Branch, Menoufiya University, Egypt.

A field experiment was carried out at two seasons 2006/2007 and 2007/2008 to investigate the effect of phosphate dissolving bacteria (Bacillus megaterium var phosphaticum) or AM- mycorrhiza on the growth and yield of peanut (Arachis hypogaea L.) at different fertilization patterns (organic and mineral) in the presence of some nitrogen fixers (Bradyrhizobium sp. and Azospirillum lipoferum). All the growth parameters were significantly increased with organic matter fertilization treatment, compared to mineral fertilization as well as control in the presence of phosphate dissolving bacteria (Bacillus megaterium) and nitrogen fixers than that in the presence of AM mycrrhiza at 45 day of planting. At 80 day of planting, all used biopreparations either in single or paired inoculants enhanced the growth and dry weight of treated peanut plants, the AM infection %, the nitrogenase activity and the NPK of the shoot dry matter especially with the treatment organic matter + AM mycorrhiza + Bradyrhizobium sp. + Azospirillum lipoferum. Moreover, AM mycorrhiza was more effective than B.megaterium. Phosphorous became more available with AM mycorrhizal inoculation (0.096 % and 0.418) than Bacillus megaterium inoculation (0.056 % and 0.332) at 45 and 80 day of planting, respectively during 1st season. The significant highest increase in N and P % was in organic matter + AM mycorrhiza + Bradyrhizobium sp. + Azospirillum lipoferum (2.53 N %, and 0.219 P %) and (2.507 N %, and 0.709 P %) at 45 and 80 day of planting. Furthermore, AM mycorrhizal inoculation, recoded significant nitrogenase activity (1.291 and 5.227 mmol/C2 H4 /h/ g Nod.D.Wt at 45 and 80 day, respectively) higher than Bacillus megaterium inoculation (1.275 and 4.630 mmol/C2 H4 /h/ g Nod.D.Wt at 45 and 80 day, respectively). In addition, the results obtained revealed that, the organic matter application significantly increased pods number per plant, weight of 100 seeds (g) and yield (ton/fed) more than the mineral fertilization as well as control. It gave highest mean values 42.833, 69.882 and 1.258, respectively. productivity of peanut plants became enhanced with AM mycorrhizal inoculation than B. megaterium inoculation as indicated by the significant highest mean values of pods number per plant, weight of 100 seeds (g) and yield (41.111, 65.113 g and 1.191 ton/fed, respectively). So, it could concluded that the combinedinoculation of AM mycorrhiza and N2-fixers was more efficient in phosphorus solubilization than the inoculation with B. megaterium in the presence of organic matter (compost) fertilization than mineral fertilization.

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