Vol. 6, September, 2003

1/6   INCIDENCE OF MICROBIAL INFECTION AMONG DRUG ABUSERS

M.S. Ashour, N.G.M. Abou EL Azayem*, M. El Tahan**, W.R. Hablas*,

F.H. El Tahan***, W. El Tayeb and A.A. El Sharif.

Microbiology Dept., Faculty of Pharmacy, Al-Azhar University.

* Clinical and Chemical Pathology Dept., and **Cardiology Dept., Faculty of Medicine, Al-Azhar   University.

*** Microbiology Section, A.R.C.

In this study one hundred and fifty addicts were examined. All addicts were asked to fill questionnaire sheet to obtain the demographic data and risk factors for exposure to infections. Also they were subjected to clinical and electrocardiogram (ECG) examination, testing for anti HIV-1/-2, anti HCV, and HBsAg. Blood samples, pus from abscesses, sputum, throat swab and corneal swab, were microbiologically examined for bacterial and fungal infections. Urine samples were subjected to detection of the main 5 groups of narcotics; opiates, amphetamines, cannabinoids, barbiturates and benzodiazepines. Microbiological examination of 39 samples of different illicit drugs including brown heroin, white heroin, and hashish (herb and cigarettes) was done. All fungal isolates were examined and identified using image analyzing system. Results revealed that the duration of addiction increased the risk of bacterial and fungal infections. The route of administration was the main risk factor affecting contracting viral infections in addicts. Invasive infections caused by bacteria and fungi are common complications of drug abuse. It was noticed that there is a relation between the fungal species isolated from the used drugs and those isolated from sputa of patients with reproductive cough.

 

2/6 ISOLATION AND CHARACTERIZATION OF SOME STREPTOMYCES SPP. FROM TOUSHKA’S EGYPTIAN SOIL; WITH PRELIMINARY INDICATION OF UNUSUAL N2 – FIXATION IN A LOCAL STRAIN

M.A. Hewedy

Bot. Dept, Women College for Arts, Sciences and Education

Ain Shams University, Heliopolis, Egypt.

Four Streptomyces strains isolated from Toushka's soil-Egypt were investigated. Morphological cultural, physiological and chemotaxonomic characteristics of the strains are described. They were proposed to be: St. ochraceiscleroticus, St. chibaensis, St. canarius and St. aurantiacus. St. chibaensis has high capacity than other strains to dissolve phosphate, degrade chitin, antifungal activity, neutralism relation among N-fixers bacteria and give preliminary indication of unusual N­2-fixing system.

 

 

3/6 PROTEASE PRODUCTION BY STREPTOMYCES PSEUDOECHINOSPOREUS AND BACILLUS MEGATERIUM

K.M.A. Abo Shanab, N.A. Hassouna, M.A.M. Yassien, M.M.M. Abo El-Wafa

Department of Microbiology and Immunology, Faculty of Pharmacy, Ain-Shams University, Cairo, Egypt

A total of 335 isolates of Streptomyces species and 489 of Bacillus species were recovered from 97 soil samples collected from different localities in Egypt. The protease productivity of the collected isolates was determined and the highest protease production isolates, identified as Streptomyces pseudoechinosporeus and Bacillus megaterium, were selected for further studies. The effect of different factors on the protease production by the selected isolates was studied. The results showed that maximum protease production of Streptomyces pseudoechinosporeus and Bacillus megaterium was obtained at incubation temperature 30oC and 37oC, and after 72 hrs and 48 hrs incubation period, respectively. In addition, the use of a modified basal protease production medium A (MBPP-A) for Streptomyces pseudoechinosporeus, and medium B (MBPP-B) for Bacillus megaterium caused 1.8-fold and 2-fold increase in protease production, respectively, as compared with that of the basal protease production medium. UV variants of high protease productivity, isolated from Streptomyces pseudoechinosporeus, and Bacillus megaterium isolates, produced 1.3-1.4 and 1.5-1.8 times more protease enzymes , respectively, as compared to that of the parent strain.

4/6 SELECTION OF A NOVEL OSMOTOLERANT BAKER’S YEAST SACCHAROMYCES CEREVISIAE STRAIN FOR COMERICAL BAKER’S YEAST PRODUCTION

A.N. Ibrahim**, A.M. Shatta**, M. Fadel* and A.S.H. Shafiek***

* Microbial Chemistry Department, National Research Center, Dokki Cairo, Egypt

** Agricultural Botany Department; Agriculture Faculty, El-Azhar University.

*** El-Salam Baker’s Factory. Grand Cairo Bakers, Egypt.

Eighteen baker’s yeast (S. cerevisiae) strains including three strains employed for commercial baker’s yeast production in Egyptian Factories were tested for selection of osmotolerant strain to be used in commercial baker’s yeast production. Item considered for selection were biomass yield, trehalose content and raising powers in high sugar or high sugar and salt concentrations doughs as well as total viable cells. Promising results were achieved with S.cerevisiae A1. Cultural and nutritional conditions including pH, temperature, sugar concentrations, urea level, antifoam, trace elements and growth stimulators were studied. The selected strain is scaled up in 500 L, 12m3 and 100m3 fermenters for commercial baker’s yeast production in Egyptian baker’s yeast factory. The commercial product of S.cerevisiae Al was compared with that of S.cerevisiae DGI 342 under the same amounts of nutrient and aeration in the growth medium. The following data were achieved 115 g fresh yeast cells (32.6% dry matter) /L involved 12.36% (w/w) trehalose and 15 x 109 total viable cells /g fresh yeast, gave raising powers 950, 435 and 415 Cm3 CO2/ 1st h. in lean, high sugar concentration or high and salt sugar concentrations doughs respectively. Compared to 95 g fresh yeast cells (30.2 % dry matter) /L. involved 9.6% (w/w) trehalose and 14.4 x 109 total viable cells/g fresh yeast gave raising powers 850, 360 and 340 Cm3 CO2 / 1sth for the above tested dough respectively for S. cerevisiae DGi 432.

 

 


5/6 CHARACTERIZATION OF CRUDE PROTEASE ENZYMES PRODUCED BY STREPTOMYCES PSEUDOECHINOSPOREUS AND BACILLUS MEGATERIUM

K.M.A. Abo Shanab, N.A. Hassouna, M.A.M. Yassien, M.M.M. Abo El-Wafa

Department of Microbiology and Immunology, Faculty of Pharmacy,

Ain-Shams University, Cairo, Egypt

Streptomyces pseudoechinosporeus and Bacillus megaterium are characterized by their high protease productivity. Characterization of crude protease enzymes produced by these isolates was carried out through studying the influence of different factors on their proteolytic activity. The obtained results showed that the optimum incubation temperatures for maximum proteolytic activity of crude enzymes produced by Streptomyces pseudoechinosporeus and Bacillus megaterium were 40oC and 37-40oC, respectively, while, the optimum pH range was 7.2-7.5 and 6.8-7.2, respectively. It was also observed that the presence of 10 µg/ml of Ca++ or Mg++ increased the proteolytic activity of crude enzyme of Streptomyces pseudoechinosporeus and Bacillus megaterium. While , the presence of Mn++ at concentration 10 µg/ml increased only the proteolytic activity of the crude enzyme produced by Bacillus megaterium. In addition, maximum proteolytic activity of the crude enzymes of both strains was observed in the presence of 0.1% gelatin, however, further increase in the gelatin concentration causing decrease in the proteolytic activity. As regard to thermal stability of the protease enzyme, it was found that proteolytic activities of the crude enzyme produced by Streptomyces pseudoechinosporeus and Bacillus megaterium was not affected when incubated for 2 hours at temperature up to 50oC.

 

6/6 SCALING UP A NEW OSMOTOLERANT BAKER’S YEAST SACCHAROMYCES CEREVISIAE 5X STRAIN FOR COMMERICAL PRODUCTION IN EGYPTIAN YEAST FACTORY

A.N. Ibrahim*, A.M. Shatta *, M. Fadel** and A.Sh. Shafiek ***

*Agricultural Botany Department, Faculty of Agriculture, Al –Azhar Univeristy, Egypt.

** Microbial Chemistry Department, National Research Center, Cairo, Egypt.

*** El- Salam Yeast Factory, Grand Cairo Bakeries, Cairo, Egypt.

            A new baker’s yeast, S. cerevisiae 5 x strain was selected among eighteen baker’s yeast strains to tolerate high sugar or salt concentrations incorporated in the flour. The strain was propagated in 500 L and 12m3 fermenters containing low or high sugar cane molasses media under batch fermentation conditions. The propagated culture was scaled up in 100m3 fermenter for seed yeast production to inoculate 100m3 fermenter for commerical bakers yeast production under increamental feeding fermentation system with high aeration. Comparison was carried with commercial baker’s yeast strain S. cerevisiae DGi 432.The results obtained were 186 g fresh yeast weight/L containing 32.2% dry matter, 11.2% (w/was) trehalose and 15.2 x 19 total viable cells/ g fresh weight, and the raising powers were 900, 420 and 395 Cm3 CO2/ 1st h produced in lean , high sugar and high sugar and salts doughs respectively for S. cerevisiae 5x. These data are compared with 150 g fresh yeast weight /L containing 29.8% dry matter, 9.6% (w/w) trehalose and 13.2 x 109 total viable cells, and the raising powers were 800, 370 and 335Cm3 CO2/ 1st h. obtained for S. cerevisiae DGi 342. The load of wild yeast and bacterial counts were less in S. cerevisiae 5x product than that of S. cerevisiae DGi 342.

 

7/6 EFFECT OF ZOOPLANKTON GRAZERS ON BACTERIAL INDICATORS OF FAECAL POLLUTION AND PATHOGENS IN NILE WATER AT EL-AMIRIA, CAIRO, EGYPT.

R.A. El-Bassat and S.A. Abdallah*

National Institute of Oceanography and Fisheries, Inland Water and Aquaculture Branch, El-Kanater Research Station

* Botany Dep., Faculty of Girls, For Science, Education and Arts,

Ain Shams University

Bacterial consumption by zooplankton grazers was measured in situ in the Nile water during 24 hours grazing experiment. The hypothesis of getting rid of pathogenic bacteria and faecal pollution indicators by using zooplankton grazers was tested during the present study. The main consumers over the study period were Free-Living Protozoa and Rotifera. In Absence of grazers, the bacterial heterotrophic plate counts of total coliform, faecal coliform, faecal Streptococci, Pseudomonas aeruginosa and Yersinia enterocolitica were increased at both 22°C and 37°C, showing their normal growth rates. On the other hand the reduction rates for the tested faecal pollution indicators in presence of zooplankton predators were remarkable (up to 99%), while the pathogenic isolate, Yersinia enterocolitica was not detected after 6 hours of the experiment. Univariate measures ANOVA showed significant relations between bacterial Heterotrophic Plate Count (HPC) and 3 taxonomic groups of grazers (Protozoa, Rotifera and Copepoda). It was concluded that, Nile water suffers from signs of faecal pollution and as a main source of drinking water for Egypt; it needs an appropriate environmental management to control many health problems and diseases.

 

 

8/6 DIFFERENT CYTOKINES PROFILES IN SPLEEN CELLS AND LIVER GRANULOMA OF SCHISTOSOMA MANSONI EXPERIMENTALLY INFECTED MICE DURING DISEASE DEVELOPMENT

N.M. Al-Ajmi*, N.F. Bouzubar*, M.A. Ramadan** and H.A. Al-Mukhazim***

Medical Laboratory Technology*, Parasitology** Department of Medical Laboratory Technology, College of Health Sciences, PAAET, Kuwait and Medical Laboratory Department***, Sabah Hospital, Ministry of Public Health, Kuwait.

To determine the importance of Th1 and Th2 cells in modulating granuloma formation, mRNA transcripts for Th1 (IL-2 and IFN-γ) and Th2 (IL-4 and IL-5) cytokines were assessed by the molecular technique of in situ hybridization in the liver granuloma. The molecular studies showed few number of cells expressing mRNA transcripts for IFN-γ whereas, considerable number of IL-2 cells were present in the liver granuloma at 6 weeks(wks) post-infection (p.i.). Complete disappearance of IFN- γ expressing cells were found when the disease progressed to 13 wks p.i. Conversely, very high number of cells expressing mRNA transcripts for IL-4 and fair number of IL-5 cells were present at 7 wks p.i. with a maximum level of IL-4 cells at 13 wks p.i. These in situ molecular studies of the liver tissues, demonstrated that Th1 cells were present at the very early granuloma development. Moreover, Th2 cells were required for its full development. The main interesting finding was the number of cells expressing mRNA for IL-4, was very high and it might exceed the total number of lymphocytes in the granuloma. Lymphocytes from experimentally infected mice-spleen cells were cultured in vitro with media only or with S. mansoni soluble egg antigen (SEA) and the same cytokines of lymphocytes supernatant were measured by ELISA assay. The levels of IFN-γ and IL-2 were high at 6 wks. p.i., with a slight decline of IFN-γ and increasing amount of IL-2 at 10 wks p.i. Spleen lymphocytes of fully formed granuloma secreted high levels of IL-4 and IL-5. The results suggest that the development of schistosome egg-induced liver granuloma is a complex process and both Th1 and Th2 cell subsets sharing with other inflammatory cells (non lymphocytes), may play an important role in regulating and modulating the immuno-pathology of granuloma formation and the subsequent hepatic fibrosis.

 

 

9/6 BIODEGRADATION OF CUTIN BY PSEUDOMONAS AERUGINOSA

N. E. Yousef, M. Okasha and H. Abd Latief

Department of Microbiology, Faculty of Pharmacy, Zagazig University,

Zagazig, Egypt.

Degradation of cutin and detection of plasmid mediated cutinase production in ten bacterial isolates of Pseudomonas aeruginosa were achieved. The ten isolates were screened for their ability to produce cutinases (cutin-degrading esterases). Initially, esterases activities of the culture filtrates of isolates grown in basal mineral salt media (BMS) supplemented with 0.4% apple cutin, were determined by spectrophotometric assay utilizing the model substrate p-nitrophenyl caproate . Cutin degradation and enzymes production were confirmed by HPLC and GC. Hydrolytic product of cutin degradation were identified as 18-hydroxyoctadeca-9, 12-dienoic acid; 10, 16-dihydroxyhexadecanoic acid; 3,methoxyhydrins derived from 9,10-epoxy-18- hydroxyoctadeca-9, 12-dienoic acid; 4, methoxyhydrins derived from 9,10-epoxy- 18- hydroxyhexadecanoic acid and 5,9,10,18 –trihydroxyoctadecanoic acid in comparison to a standard GC profile. The culture filtrates of 8 tested Pseudomonas aeruginosa isolates exhibited esterase activity while 2 isolates showed no enzyme activity. Time coarse study in basal mineral salt solution supplemented with apple cutin indicated maximum cutinase activity at the begining of stationary phase. Cutin degrading Pseudomonas aeruginosa isolates were found to have plasmids encoded enzyme cutinase production and cutin degradation since plasmid-cured Pseudomonas aeruginosas isolates displayed no enzyme activity. The extracted bacterial plasmid DNA of cutinase-producer showed molecular weight over 12.0 Kb whereas cutinase-deficient cells had plasmids of about 4.0 Kb . Cured cells lack these plasmids.

 

10/6 EFFECT OF LEAD ON GROWTH AND PHOSPHATASE ACTIVITY OF MOSS BRYUM PSEUDOTRIQUETRUM GROWN IN AXENIC CULTURE.

A.M. Al-Shehri

Biological Sciences Department, Faculty of Science, King Khalid University, Abha, P. O. Box 9019, Saudi Arabia

Effect of lead on growth rate and phosphatase activity of Bryum pseudotriquetrum was examined. Dry weight, chlorophyll content and shoot length of protonemal culture decreased gradually with increasing concentration of lead nitrate and lead acetate. At low concentrations of lead salts (50 mg. l-1) the protonema was healthy green with numerous chloroplasts, then it became pale-green to brown with increasing concentrations (75 to 125 mg. l-1). On the other hand, the time of bud initiation was not significantly affected by lead salts. Phosphatase activities response to different lead salts were broadly similar for phosphomonoesterase (PMEase) and phosphodiesterase (PDEase). Both were markedly inhibited at 10.0 mM lead nitrate and lead acetate.

 

11/6 INFLUENCE OF CULTURAL CONDITIONS ON PROTEASE PRODUCTION BYAEROMONAS HYDROPHILA.

S.M.A. Taha and S.A. Meligy

National Center for Radiation Research and Technology, P. O. Box 29,

Naser City, Cairo, Egypt.

Protease production by A. hydrophila was influenced by incubation temperature, aeration, pH and growth media. The highest optical density and protease production are observed at 28°C and pH 7.0 under high aeration (200 rpm). Also, the addition of skim milk to the culture medium increased the growth and protease production by A. hydrophila at both temperatures of incubation (4º and 28ºC). However, the results showed that the production of protease was high when the medium was supplemented with lactose or glucose as carbon sources and with peptone or yeast extract as nitrogen sources. Protease production by A. hydrophila in foods was studied. All tested foods supported the growth and protease production at refrigeration (4ºC) and room temperature (~20°C). So, the present data suggest that prolonged cold storage of foods should not be encouraged.

 

“Review Article”

12/6 OCCULT HEPATITIS B VIRUS IN CHRONIC HEPATITIS C INFECTION

Z.N.A. Said

Department of Microbiology, Faculty of Medicine (for Girls),

Al-Azhar University

Viral hepatitis exists throughout the world and is a major global public health problem (Nakaiet al., 2001). Hepatitis B virus (HBV) and hepatitis C virus (HCV) are responsible for the majority of chronic hepatitis, liver cirrhosis and hepatocellular carcinoma (HCC) cases worldwide (Sagnelli, 2000). Concurrent infections with both viruses are increasingly recognized in patients with both acute and chronic hepatitis (Kaoet al., 2002). The prevalence of patients with HCV and HBV coinfection has been described as high in geographic areas with a high endemic level of both infections such as Western Asia and the Mediterranean Basin (Bukhtiari et al., 2003).

The interaction between HCV and HBV has so far been poorly investigated. Little is known about the clinical presentation, the natural history and the response to antiviral treatment of liver diseases associated with HBV and HCV coinfection (Sagnelli et al., 2000).

Hepatitis B virus (HBV) infection in patients who lack detectable hepatitis B surface antigen (HBsAg) is called occult (hidden) infection (Cacciolaet al.,1999). In recent years, it has become evident that HBV can be transmitted by individuals with occult HBV infections with few or no signs of ongoing viral replications (Jenntetet al., 2002). Chemin et al. (2001), showed that a significant proportion of patients with hepatitis of unknown aetiology was in fact infected with HBV, all were seronegative for HBsAg whereas several had detectable anti-HBc antibodies. Several recent studies have indicated that occult HBV infection can be found in patients with chronic HCV infection at various frequencies (50 to 87%) (Kao et al., 2002 and Giannini et al., 2003). The high prevalence of this occult infection in such patients has been suggested to have clinical implications in the pathogenesis of HCV induced Chronic Liver Disease (CLD).

 

13/6 EXPRESSION OF COLD ADAPTIVE ENZYMES OF PSEUDOMONAS FLUORESCENS

W.H. Mostafa, H.H. Radwan and A.M. Hashem*

Department of Microbiology and Immunology, Faculty of Pharmacy, Helwan University and *Cairo University.

A total of 693 microbial isolates, locally isolated from industrial, domestic refrigerators and freezers and tested for their ability to grow at temperatures lower than 10oC. Only 37 isolates could grow at 7oC, all isolates were psychrotolerant and non-of them were a true psychrophiles. The isolates were screened for their productivity of proteinase, lipase and nitrate reductase enzymes at low temperature. Pseudomonas fluorescens was the only isolate that produces the three enzymes at 7oC. The bacterial strain was able to produce two lipases (Lip1, Lip2), two proteinases (Pro1, Pro2) and two nitrate reductases (NR1, NR2) enzymes. The two isoenzymes of both lipases and proteinases were of different characters. Lip1 and Pro2 were expressed in higher yield at 7oC than at 37oC, while Lip2 and Pro1 were produced in greater amount at temperature 37oC. On the other hand, both NRs were expressed at either temperatures with the same ratio. The enzymes were purified from the culture filtrate in a two-steps purification procedure by ion exchange chromatography. The six enzymes were purified till homogeneity as tested by SDS-electrophoresis, and the molecular weight for the purified enzymes, Lip1, Lip2, Pro1, Pro2, NR1 and NR2 were 80, 95, 24, 45, 130 and 180 kDa respectively. The total amino acid analysis of the four enzymes (Lip1, Lip2, Pro1 and Pro2) revealed the presence of 2 lysine and 3 phenyl alanine residues in Lip1 more than that present in Lip2; indicating that Lip1 was not a degradative product by autolysis of Lip2. Western blot of the four enzymes using a polyclonal antibody preparation directed against Lip1 and Pro1 showed that there were no common epitopes between the two lipases but there was a common epitopes between the two proteinases. The optimum activity of both lipase was around pH 6.5, while for both proteinases was 8.0 and for NRs 7.0. Furthermore, the incubation of pure Pro2 at 37oC leads to autoproteolysis of the enzyme and a new band at 24 kDa appeared and became more obvious by further incubation. The optimum temperature of both Lip1 and Lip2 activities were 28oC and 40oC respectively. Whereas, the optimum temperature for the two proteinases was 40oC, and for the two NRs ranged from 20 -50 oC. Except Lip1 all enzymes showed similar thermal stability up to 45oC, where Lip1 lost about 50% of its activity at 40oC. The cold adaptation phenomenon was further confirmed by comparison of two other Candida lipase enzymes. These lipases were obtained from Candida lipolytica (CLL) and from Candida albicans (CAL). The former remains active at temperature lower than 10oC and the second loses its activity at temperature lower than 20oC.

14/6 NOCARDIA: A NEGLECTED LUNG PATHOGEN

M. Abd El-Alim

Clinical Pathology Department, College of Medicine, Ain Shams University

Nocardia represents a rare opportunistic pathogen that has been isolated in both animals and man. A definitive diagnosis of Nocardiosis requires the isolation and identification of the organism. Delay in establishing the correct diagnosis is common due to the non-specific and diverse clinical presentation and the inherent difficulty in cultivating Nocardia. The primary aim of the present study was to clarify the representative Nocardia isolated from sputum of patients suffering from pulmonary diseases. In this study, we examined 300 sputum samples. These sputum samples were taken from patients with pulmonary diseases at the Ain Shams university hospitals. Nocardia was isolated from7 samples (2.3%) out of the 300. Our strains were provisionally assigned to the genus Nocardia according to morphological criteria. Direct smears were taken from all the samples and were stained with Gram’s and Zeihl-Neelsen stains. Furthermore, these isolates were studied to establish their taxonomic status. We inoculated all the samples on buffered charcoal yeast extract agar and on Lowenstein – Jensen slopes. The isolates were identified by standard biochemical tests for Nocardia. Antibiotic susceptibility tests on Muller-Hinton agar was performed using different antibiotic disks. Antimicrobial agents demonstrated to be active against all species were Amikacin, Amoxacillin–Clavulinic acid and Trimethoprim – Sulfamethoxazole. After revision of morphological features and by comparing the susceptibility pattern with that of Nocardia, four (57%) out of the 7 isolates were primarily identified as N. asteroides and three (43%) as N. farcinica. We recommended that clinicians should consider pulmonary Nocardiosis especially when patients with chronic respiratory infection fail to respond to conventional therapy.

 

15/6 EFFECT OF AMINOGLYCOSIDE AND Β-LACTAM ANTIBIOTICS ON THE PRODUCTION OF GENTAMICIN BYMICROMONOSPORA PURPUREA DSM 43036

S.M. Kheira* and M.M.A. El-Sokkary

Department of Microbiology, Faculty of Pharmacy, Mansoura University,

Mansoura 35516, Egypt.

Different mutant strains ofMicromonospora purpurea DSM 43036 were selected by growing the organism on a solid medium containing gradually increased concentrations of different aminoglycoside antibiotics namely gentamicin (GM), streptomycin and amikacin up to one mg/ml and β-lactam antibiotics namely ampicillin (400 mg/ml) and cephradine (20 mg/ml). GM activity was determined by agar diffusion method and its production by different mutants was followed by daily sampling from shake culture flasks containing casein glucose medium grown at 32oC. Results of investigations revealed that GM produced by either amikacin or gentamicin-resistant mutants was insignificantly increased or unaffected when compared with the original culture. However, GM production was increased 1.45 times with streptomycin-resistant mutant. A noticeable increase of GM production was observed when using ampicillin resistant mutant (from 38 to 105 mg/ml= 2.76 times) and 1.37 times (from 38 to 52 mg/ml) by using cephalosporin resistant one. Determination of the dry weight of cells revealed that such increase in antibiotic production was unrelated to the biomass production by the three mutants. It was suggested that the increased GM production by β-lactam mutants might be due to the formation of mutants having an altered cell wall murein composition that led to increased transport of the antibiotic outside the cell. Separate addition of sub-bacteriostatic concentrations of the above mentioned antibiotics to the original culture during the whole process of production was accompanied by a slight insignificant decrease which was related to a decrease in mass produced.

16/6 ISOLATION, CHARACTERIZATION AND 16S rRNA SEQUENCE OF A CELLULOLYTIC BACTERIUM,

BACILLUS PUMILUS

A.M. Ibrahim and S. Karita*

Genetic Engineering and Biotechnology Research Institute (GEBRI), Menoufiya University,Sadat City, Egypt. and *Department of Sustainable Resource Science,

Mie University, Japan.

A native aerobic, mesophilic, spore-forming bacterium is described. Carboxymethylcellulose (CMC) is digested within 24h around colonies formed in CMC agar plates. Cells stain Gram positive and are short rods. Cellulose, locust bean gum, xylan, cellobiose, glucose, maltose, galactose, sucrose, lactose, and mannose serve as substrates for growth. The optimum temperature for growth is 37oC and optimum pH is 7.0. The 16S rRNA of this bacterium was partially sequenced.

17/6 SURVEILLANCE OF YEAST INFECTION IN IMMUNOCOMPROMISED EGYPTIAN HOSTS: A MYCOLOGICAL, MOLECULAR AND SEROTYPING STUDY.

Y.M. Shetaya; M. Mohamed; A. Karam El-Din; L. El-Shawarby*; Y.A. Youssef*.

Microbiology and Biochemistry department, Faculty of Science, Ain Shams University

Clinical pathology department, Faculty of Science, Ain Shams University

Traditional culture as well as serological methods (ELISA, immunofluoresence and western blotting) were used for detecting yeast infection in immunocompromised hosts (suffering from Leukemia and Cancer). Out of 300 diagnosed cases, 158 were positive. One hundred and forty nine were infected with Candida species (C. albicans, C. glabrata, C. tropicalis, C. guilliermondii, C. parapsilosis), 6 with Trichosporon beigelii, and 3 with Rhodotorula rubra. In the present study, the sensitivities and specifities were 64.8 and 83.3 % for the detection of IgM, 85.9 and 41.7% for the detection of IgG by ELISA technique, and 91.4 and 62.5% for immunofluoresence antibody assay respectively. For detection of Candida species, T. beigelii, and R. rubra antigens in patient's sera by immunoblotting technique, the sensitivities and specifities were 87.5 and 100%. However, the specific rotation of cell wall mannan of yeast as well as RAPD techniques in candidiasis are promising for the identification of subtypes of C. albicans. Data of the present study indicated that, immunoblotting of serum protein antigens was most specific (100%), while immunofluoresence technique was most sensitive method (91.4%) among all collected samples regardless of source of culture.

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