Vol 8, July, 2000.

1/8 BIOSYNTHESIS OF A CONSTITUTIVE THERMOSTABLE PROTEASE BY HUMICOLA GRISEA VAR. THERMOIDEA.

E.F. Sharaf

Bot. Dept, Fac. of ScL, Cairo Univ., Cairo, Egypt.

Humicola grisea var. thermoidea, a thermophilic fungus exhibited a remarkable ability to biosynthesize constitutive thermostable protease (s) in the absence of the inducing substrate. Optimal parameters controlling the enzyme productivity \vere found to be: incubation temperature, 55°C; incubation period, 24 h; inoculum size, one disc (5.83x 10s spores/disc); inoculum age, 4 days; pH , 7; C-source, starch; N-source, NaNOj; amino acids, L-alanine and vitamins, pyridoxine.

2/8 CEMENT KILN DUST INDUCEDCHROMOSOMAL ABERRATIONS IN ALLIUM CEP A

A.I. Shehata

Botany Departmen, Faculty of Science, Zagazig University, Zagazig, Egypt

Cement Kiln dust (CKD showed mitodepressive effect on the Mitotic index of Alii urn cepa and it was inversely proportional to the concentrations of the CKD, while the frequency of abnormality, Chromosome!, breakage, stickness, chromosome, bridge lagging chromosomes clumping, C-metaphase, c-anapha.se, binucleate telophase were observed in different stages of mitosis in treated, cells. The harmful effect of environmental pollution by CKD proved that it may act as a strong mutagen.

3/8 SCANNING ELECTRON MICROSCOPIC STUDIES ON SEED COAT OF SOME SPECIES OF PAPAVER (L.) (PAPAVERACEAE)

M.E. Tantawy* and S.H. Rabie**

*Botany Dept. Fac. of Sci. Ain Shams Univ. Cairo, Egypt.

**Flora & Phytotaxonomy Researches; H.R.I.; A.R.C. Giza, Egypt.

The seed coat of 15 species of Papaver was studied by LM and SEMfor a sharp differentiation. All the taxa were found to have sunken hilum and a reticulation pattern, which were considered constant at the generic level. Intera-specific variations were recorded for both the anticlinal and periclinal walls. The characters revealed by SEMand LMwere used in the construction of an identificatory key for the studied species.

4/8 TAXONOMIC SIGNIFICANCE OF SEED CHARACTERS IN SOME SPECIES OFRUMEXL. (POLYGONACEAE) WITH SPECIAL REFERENCE TO THE SEED COAT SCULPTURE

M. E. Tantawy Botany Dept, Faculty of Science, Ain Shams Univ.

Seed characters of12 species of Rumex L. were studied. The shape, dimensions, form of the hilum and elevation of seeds as well as details of seed sculpture prove their taxonomic value for the separation between the species. A key with the seed characters of the investigated species as well as SEM micrographs of such characters are provided.


5/8 MICROBIAL DEGRADATION OF AROMATIC SUBSTANCES BY LOCAL BACTERIAL ISOLATES

II - KINETICS OF DEGRADATION OF PARA-NITROPHENOL BY PSEUDOMONAS PUTIDA

O.M. El-Tayeb, F.H. Ali*, S.A. Megahed and M. El-Azizi

Department of Microbiology and the Microbial Biotechnology Center,

Faculty of Pharmacy, Cairo University and *National Research Center, Cairo,

Egypt.

Degradation ofp-nitrophenol (PNP) as a sole source of carbon by a locally isolated strain of Ps. putida in both shake flasks and in a 7 I fermentor is affected by: PNP concentration, inoculum size, agitation, aeration, temperature, and additional carbon sources. The parameters followed were: PNP concentration, pH, dissolved oxygen (DO) and biomass. High concentrations of PNP were inhibitory to growth but repeated incremental addition allowed the use of higher concentrations and the reaction appeared faster and open ended when the pH was maintained at 6.9. The reaction was shown to be inducible and stopped in the presence of amikacin, which inhibits de novo protein synthesis. The linearized form of the integral method "Walker plot" (a Monod model with no growth) fits the degradation data. A linear least square regression delineated the parameter values Vmax and Km. The zero, first order, Lineweaver-Burk equations did not fit the data satisfactorily. Application of the Walker model to the data indicates an enzymatic growth-independent reaction. It is proposed that disappearance of the PNP may be controlled by an initial enzymatic step, which requires oxygen and energy.

6/8 DESALINATION OF SEA WATER USING A BIOLOGICALLY CONTROLLED MICROBIAL SYSTEM

M.S. Ammar; M.S. Azab and M.M. Muharram

Botany and Microbiology Dept, Fac. of Sci., Al - Azhar Univ.,

Medinet Nasr, Cairo, Egypt

More than 73.5% of the world water on earth is a saline in nature which could be a good source of fresh water. Day after day, fresh water represents a global problem and becomes rare at many places all over the world. The costs of the desalination process-is-the main challenge of using sea waters as a source of fresh water. The present study is concerned with the desalination of sea water using a biologically controlled microbial system. It is a closed system consisted of three stages connected to each other. Data indicated that the desalination percentage (D%) reached up to 80.04% in case of Ca++ using A. fumigatus, 1-27B with the presence of 0.5% sucrose and 0.25% yeast extract at 30°C for 6 days incubation. In case ofMg++ the (D%) was 53.35% using A. Jumigatus 1-5., in case of Ct it was by the fungus isolate I-(I) since it reached up to 41.64% at the end of the third stage of sea water recycling, sodium Na+ recorded with I-(50) since it recorded to 77.87% and K* recorded 80.40% at the end of the third stage using the fungus I - KP/o .Furthermore, the tested fungal isolates showed high abilities to remove various heavy metals parallel to their ability of sea water desalination.


7/8 THE POSSIBLE APPLICATION OF MICROBIOLOGICALLY DESALINATED SEA WATER IN CROP IRRIGATION IN EGYPT.

M.S. Ammar; M.S. Azab and M.M. Muharram

Botany and Microbiology Dept., Fac. of Sci., Al-Azhar Univ.

Madinet Nasr, Cairo, Egypt.

The application of the microbiologically desalinated sea water in the irrigation of an Egyptian crop was clearly emphasized. The redesalinated sea water collected at the end of the third stage of the biologically controlled multi-redesalination system was passed, firstly through ion cation exchanger and secondly on an anion exchanger resins for the sake of removing the remnant ions ofZn++, tC, A/g++ and CT which were still represented at a desalination percentage (D%) of 78.67, 99.59, 82.77and87.16respectively. After passing process, the desalination percentage jumped to 100% for each ion except CT that was still at 94.28% after passage process. In a trial to test the redesalinated sea water for the irrigation of Zea mays Giza 2, experimental pots were seeded and irrigated with the third stage product of the desalinated sea water whereas the control was irrigated by the ordinary tap water. After 12 days of irrigation the plants (shoots + roots) were measured and their weights were recorded. Also, chlorophyll a, b, and carotenoidpigments were spectrophotometrically measured. The recorded results confirmed that the effect of the fungal desalinated sea water did not markedly vary from that of the tap water. Finally, the redesalinated sea water was tested for its content ofMycotoxins (extracellular) using thin layer chromatography. The results gave negative tests for all the five used isolates which emphasizes the effectiveness of the present introduced biologically controlled multi-desalination system.

8/8 EFFECT OF MAGNETIC FIELD ON GROWTH CRITERIA

AND YIELD, GROWTH REGULATING SUBSTANCES, PHOTOSYNTHETIC PIGMENTS AND METABOLISM OF WHEAT

A.A. Aly Botany

Department, Faculty of Science, Ain Shams University, Cairo

In a pot trial, it was found that exposing wheat grains to both stationary and pulsating magnetic fields of 200, 150, 100 & 50 gaus for 24 hours increased growth criteria (shoot length, root length, number and area of leaves, fresh and dry weights of plant as well as the number of seeds/spike) of the treated plants. There was an increase in auxin and gibberellic acid contents and a decrease in abscisic acid content concomitant with the increase in growth criteria of the treated plants. It was observed that, most of the magnetic fidd treatments increased the photosynthetic pigment contents, carbohydrates and protein levels of wheat. Meanwhile, magnetic field treatments induced an increase in stomatal resistance of the treated wheat plants as well as a reduction in transpiration rate.

9/8 GROWTH RESPONSES OF SUDAN GRASS TO SOIL AMENDMENT WITH THE CONDITIONER RAPG AND CONCOMITANT CHANGES IN GROWTH REGULATORS AND SOME RELATED ENZYMES

A.A. Aly and J.G. Sadek Botany Department, Faculty of Science, Ain Shams University, Cairo.

The present study highlights the effect of different concentrations of the soil conditioner reclaimer -ameliorator polymeric gel (RAPG) on growth parametres, the activity levels of endogenous auxins, growth inhibitors, gibberellins and cytokinins as well as some related enzymes namely indole acetic oxidase (IAA-O),catalase, peroxidase (P-O) and polyphenol oxidase (PP-O). The data obtained revealed an increased growth(shoot length, root length, number of leaves/plant, area of leaf/plant and fresh and dry weights of root & shoot)and yield (number of seeds/plant and fresh and dry weights of seeds /plant) of Sudan grass plants cultivated in soil amended with the low and moderate rates of the conditioner. A similar increase in the activity of growth substances (auxins .gibberellins and cytokinins) as well as a corresponding enhancement of the activity levels ofcatalase and peroxidase and a decrease of IAA-O and PP-O were observed with the same treatments. On the other hand, the higher rate of the conditioner caused reverse changes to those of the previous results. The trends and magnitude of effectiveness of the plants cultivated in normal clay soil were more or less similar to those of the plants cultivated in the soil supplemented with the moderate rate of the conditioner.

10/8 PURIFICATION AND CHARACTERIZATION OF CONSTITUTIVE THERMOSTABLE THREE ISO-PROTEASES PRODUCED BY HUMICOLA   GRISEA VAR. THERMOIDEA AT 55°C.

E.F. Sharaf* and M.S. Ammar ** Botany Dept. Fac. of Sci. Cairo Univ* and Al-Azhar Univ. **

Three thermophilic iso-proteases secreted by Humicola grisea var. thermoidea were purified through ammonium sulphate precipitation (60% saturation), dialysis and then passing through gel filtration on sephadex 0200 followed by sephadex giqo columns. The three proteases (A, B&C) showed obvious thermal stabilities within 20-80 °C with a maximum one at 60 *C. pHstability was remarkable within 5.8- 8 with a maximum value at pH 7for proteases A& Band 7.4 for protease C. The optimal substrate concentration was 0.25% for protease A corresponding to l%for other proteases (B& C). In general, there was a linear relationship between enzyme concentration and its activity up to a fixed concentration beyond which no increase in enzyme activity was detected. Although Ba+* acclerated the activity of protease A, yet it nullified the activity of protease B and exhibited no effect against protease C. In contrast, both Mg*+ and Zn++ exerted stimulatory effects on both proteases B&C with little inhibition against protease A. EDTA exhibited no effect against protease A with little inhibitory effect against protease B and stimulatory one on protease C. Ca++ stimulated only protease C. However, Cu++, Li+ and glutamic acid inhibited the three proteases at various degrees. Data emphasized the fact that the three proteases are Iso-enzymes produced by the same strain under the same specified production conditions at 55 f.

11/8 MICRO-BIOCONTACTOR (MBC) AS A NEW TECHNIQUE FOR WASTEWATER TREATMENT

M.S. Azab

Botany and Microbiology Department, Fac. Sci, Al-Azhar Univ. Madinet Nasr, Cairo, Egypt. P.N. 11884 .

The industrial wastewater (WW) of potato-chips factory is characterized by its high biological oxygen demand (BOD) and chemical oxygen demand (COD), in addition to a medium concentration of oil & grease (O&G), total dissolved slats (TDS) and total suspended solids (TSS). A new technique for wastewater treatment has been applied by using bio-mixture of selected strains ofAspergillus terreus or Rhizopus sexualis in addition to the natural flora of rice-husks in the form of mobile micro-biocontactor (MBC). Different kinds of composted rice-husks (bio-mixture) were used as a microbial carrier, support and source of nutrients and enzymes to enhance the wastewater treatment process; in order to improve the quality of treated wastewater and resulting sludge. The parameters of treated wastewater in terms of BOD, COD, O&G, TDS and TSS were greatly improved by 55.6 - 85.0, 65.7 - 79.0, 61.8- 82.7, 67.5 - 74.6 and 76.8 -87.7%respectively, in correlation to the retention time and kind of tested materials. The 14 days microbial-treated (composted) rice-husks by A. terreus, or R. sexualis as (MBC) exhibited the highest enzymes contents and were the most efficient materials for the wastewater treatment process. Furthermore, the retention time of the treatment process could be reduced to 4 h only. Finally, the resulting sludge(s) of (MBC) were easy to separate (in 5-10 min.)from wastewater. The sludge, according the chemical analysis, can be used in agriculture as an organic fertilizer and soil conditioner. In addition, different kinds of resulting sludge have been tested as biosorbents and exhibited high ability to remove chromium (89.1 -99.3%), nickel (84.3 - 98.0%) and zinc (85.6 - 97.7%) from the industrial wastewater. Data indicated the possibility of magnifying the introduced (MBC) as a new technique for the treatment ofwastewater in general and that of industrial waste in particular.

12/8  NEW APPROACH FOR ASSESSMENT AND FOLLOW UP OF PULMONARY TUBERCULOSIS

H.A. Abdel-Hafeez, A.M. Ibrahim and M. Abd El-Hady* Departments of Internal Medicine and Microbiology*, Faculty of Medicine,

Al-Azhar University.

Thirty five patients presented with pulmonary tuberculosis and 10 apparently healthy controls were studied for quantitative detection of circulating intercellular adhesion molecule - 1 (ICAM-1) and Clq-containing circulating immune complexes (Clq-ClCs). The patients were divided into 3 groups, group A included 11 untreated patients with mild to moderate lesions, group B 12 untreated patients with advanced lesion and group C included 12 patients receiving anti-tuberculous therapy for 2-3 months since diagnosis of their advanced lesions. Serum levels of 1CAM-1 and Clq-ClCs were significantly increased in all patient groups as compared with the control group (P<0.001). Moreover, both parameters were significantly higher in untreated groups A and B than the treated group C (P<0.01). Also, the presence of significant positive correlations between serum levels of 1C AM-1 and Clq-CICs in control and patient groups were proved (r = 0.99, 0.99, 0.99 and 0.93; respectively - P<0.0001). This means that both parameters are in close association and they tend to be high or low together which clarifies their significance in the clinical evaluation. In conclusion, ICAM-1 and Clq-CICs reflect the immuno-pathologic state of pulmonary tuberculosis and they can be reliably used as markers of the disease activity and/or its therapeutic response.


13/8 PURIFICATION AND RHEOLOGICAL PROPERTIES OF AN EXTRACELLULAR POLYSACCHARIDE SYNTHESIZED BY STREPTOMYCES VIRIDOCHROMOGENES

W.A. El-Shouny, A.R. El-Shanshouiy, O.Y. Mady* and N. G. Allam

Botany Department, Faculty of Science, Tanta University, Egypt.

*Faculty of Pharmacy, Tanta University, Egypt.

An extracellular polysaccharide (EPS) was isolated from culture broth of Streptomyces viridochromogenes by successive fractional ethanol precipitation. A completely deproteinized EPS yield (1.7 g/l) was recovered at the end of the purification process. The polymer had no thixotropy. The viscosity increased throughout a range of increasing shear rates from 0 to 250 s'1. Such property referred to a dilatant behaviour of EPS. The versatile effect of different tested salts on viscosity did not change the dilatancy nature of the purified polymer.

14/8  INFLUENCE OF SALT STRESS ON THE AMINO ACIDS AND

LIPID COMPOSITION OF THE PLASMA MEMBRANE OF

CLADOSPORIUM CLADOSPORIOIDES

A.A. El-Mougith

Botany Department, Faculty of Sciences, Zagazig University, Zagazig, Egypt.

When the salt-tolerant isolate of Cladosporium cladosporioides was grown in the presence of 15% sodium chloride the growth was strongly inhibited and many significant changes occurred in amino acids and lipids composition of the mycelium and plasma membrane of the fungus under study. The most important changes in amino acids due to salt stress were the significant increase ofproline, arginine, serine and isoleucine in both mycelium and plasma membrane. Salt stress also, was accompanied by an increase in the total lipids of the mycelium to 6 times more than that of the control, whereas it was accompanied by a decrease in the total lipids of plasma membrane by 32 %. Salinity enhanced the total protein (by 42 and 51 %) and total sterols (by 223 and 73 %) in both mycelium and plasma membrane, respectively. Important quantitative changes in fatty acids of the mycelium and plasma membrane were induced by salt stress, which leads to a decrease in the unsaturation index by 37 and 54 %, respectively. The ratio of sterols to phospholipid increased by 4 times more than control mycelium and to 2.75 times of control plasma membrane, whereas the ratio of phospholipid to protein in mycelium and in plasma membrane was decreased by 17 and 70 %, respectively. Also salt stress caused significant quantitative variations in phospholipid fractions and in neutral lipid components in both mycelium and plasma membrane.

15/8 HORMONAL REGULATION OF DORMANCY AND SWEETENING IN DIFFERENT CULTIVARS OF POTATO

TUBERS

K. Khattab

Botany Department, Faculty of Science, Ain Shams University

Potato tubers (Solanum tuberosum L) are dormant at the time of harvest. The dormancy period influenced by cultivar and temperature. Potato tubers from four different cultivars (cv. Diamant, cv. Desiree, cv. Gigant and cv. Agria) were exposed to two different storage temperatures (4 and 20°C). The tubers differ in the length of the dormancy period and the response to cold-induced sweetening. Studies of the regulation of dormancy and sweetening in potato tubers are reviewed with reference to the content of abscisic acid (ABA), gibberellic acid (GAj) andfree-indole acetic acid (IAA). Changes in, free- amino acids, reducing sugars and starch contents were also, examined Amylases and phosphorylases were detected by - polyacrylamide gel electrophoresis prepared with 0.5% soluble starch. This study revealed that, there was a decline in ABA level which seems to be associated with potato dormancy. The gibberellic acid (GAj) remained at low level during dormancy but increased near the resumption of sprout growth. Thefree-indole acetic acid (IAA) content declined concomitantly with sprouting. There was a net loss of starch content which concomitant with high levels of reducing sugars. Storage of potato tubers for several weeks results in accumulation of free amino acids and reducing sugars which was associated with the unacceptable quality of fried potato products. The function and the pattern of the major enzymes including amylases andphosphorylases involved in starch breakdown and control of breakdown through hormonal regulation were discussed in this investigation.

16/8  EFFECT OF NICARDIPINE, VERAPAMIL, AND RESERPINE

ON THE SENSITIVITY OF EFFLUX MEDIATED MULTIDRUG RESISTANTSALMONALLA PARATYPH1B MUTANT SELECTED

BY CIPROFLOXACIN

W. Faisal, N.A. Hassouna and M.A.M. Yassien

Department of Microbiology and Immunology, Faculty of Pharmacy,

Ain-Shams University

A spontaneous single step active efflux multi-drug resistant (MDR) Salmonella paratyphi B mutant, selected on ciprofloxacin, showed cross resistance to norjloxacin, ft-lactam antibiotics (cefoperazone and cefotriaxone), and to aztreonam. The development of multi-drug resistance in the selected mutant was associated with overproduction of 55 KDa outer membrane protein. In the presence of the transporter inhibitors (0.5 ug/ml nicardipine, 0.75 mg/ml verapamil, or 15 ug/ml reserpine), the MICs value of fluoroquinolones (ciprofloxacin and norfloxacin), B-lactam antibiotics (cefoperazone and ceftriaxone), and aztreonam were reduced by 2-32, 2-8, and 2-8 times as compared to that of the control (untreated mutant), respectively. Since, nicardipine is the only transporter inhibitor that is effective at concentration smaller than its maximum plasma concentration, it was selected for further studies. In the presence of 20 ug/ml nicardipine, the ciprofloxacin uptake by selected MDR S. paratyphi B mutant showed sevenfold increase in the intracellular concentration as compared to that of the control (without nicardipine). These data suggest that nicardipine may be useful to increase the efficacy of the antimicrobial therapy against active efflux multi-drug resistance infections.

17/8  IN VITRO ANTAGONISTIC ACTION OF PHYLLOPLANE INHABITING BACTERIA TO SOME PARASITIC FUNGI

N.M. Atef

Department of Botany, Faculty of Science, Cairo University

Bacteria were isolated from the phylloplane of two vegetable plants cultivated in Egypt. Thirty one bacteria were isolated from the leaf surface of Solatium ntelongena L, and other twenty from the phylloplane qflpomoea batatas cv. "Mabrouka ". These isolates were screened for their antagonistic behavior against some foliar and soil-borne phytopathogens. The data derived from the 51 isolates were subjected to cluster analysis. Two isolates from the phylloplane ofS. ntelongena and two isolates from L batatas were chosen for further identification as they showed the greatest inhibition to all the five phytopathogens tested. These four isolates were identified as: Envinia herbicola, from the two plants Pseudomonas fluorescens and Bacillus subtilis.

18/8 IMPROVEMENT OF CITRIC ACID PRODUCTION BY

ASPERGILLUS NIGER G-7 FROM SWEAT POTATO UNDER

SOLID-STATE FERMENTATION

M.A. Khalaf and A.A. Hassan

Microbiol. Dept. NCRRT, PO. Box: 29, Nasr City, Cairo, Egypt

The production of citric acid from sweat potato tubers by Aspergillus niger G-7 (gamma irradiated citric acid producer isolate) by solid state fermentation was studied. The experiments showed that sweat potato was an excellent substrate for citric acid production under SSF. The maximal citric acid production (I42.5g/kg wet substrate) and biomass dry weight (31.6 g/kg wet substrate) were obtained from raw substrate supplemented with 5% (wAv) methanol at an initial moisture content 70%; inoculum level l(f spores/50g, temperature of3Q°C andpHoj"5 after8 days of"fermentation. Akinetic analysis of citrate production showed an overall reactor productivity ofO. 003 g citrate/kg wet substrate per hour, with a conversion of 58%.

19/8 EFFECT OF BRASSINOLIDE ON GROWTH, METABOLITE CONTENT AND YIELD OF SALINIZED VICIA FABA

L.M. Zaky

Faculty of Education, Ain ShamsUniversity.

The effect of priming Vicia faba seeds with 400, 800 and 1600 mg/L ofNaClfor 12 hours is presented. In addition, the application ofbrassinolide on the growing plants was studied. It has been found that, salinity at 1600 mg/L NaCl reduced shoot length, fresh and dry weights and number of leaflets per plant, total chlorophylls, total carbohydrates, hormonal contents and yield of Vicia faba plants while 400 and 800 mg/L of the same salt increased these contents. Spraying the salinized plants with 0.05 ppm brassinolide counteracted the toxic effects of salinity by increasing IAA and decreasing ABA levels which resulted on higher rates in the accumulation of the metabolites led to higher growth and yield. SDS-PAGE electrophoretic patterns of water soluble protein fraction extracted from Vicia faba seeds of the harvested plants which were previously soaked in different salt concentrations (400, 800 & 1600 mg/L) and those which were sprayed with BR after soaking in different salts treatments were studied. Anatomical studies of root cross section showed that cortex area may be used as diagnostic criteria for salt sensitive genotypes in Vicia faba plants.


20/8  OPTIMIZATION OF CULTURE CONDITIONS FOR THE

PRODUCTION OF INTRACELLULAR URICASE BY

ASPERGILLUS SYDOWU (BAINIER & SARTORY)

THOM & CHURCH.

MM. Nour El-Dein, H.S. Hamdy* and M. Ghareib

Botany Department, Faculty of Science, Damietta, Mansoura University (Damietta

Branch).

*Biology Department, Faculty of Education, Ain Shams University

Aspergillus sydowii synthesized intracellular uricase (E.C. 1.7.3.3) most actively after four days of rotary shaking (150 rev min1) in a medium containing sucrose, 20.0: uric acid, 2.0; KH2PO4, 1.0; KCl, 0.5; MgSO4.7H2O, 0.5 andFeSO4. 5H2O, 0.01 gLf, giving greatest yield of the enzyme amounting to 2.19 U/50 ml at initialpH6.0 and 3Q°C, while biomass formation was better produced at pH 6.5. It is worthy of mentioning that no correlation was established between uricase production and biomass formation. Yeast extract (0.1%, W/V) was the most potent nutritional additive that enhanced enzyme production amounting to 2.36 U/50 ml while malt extract (0.1 %, W/V) similarly did with regard to biomass formation (0.49 g/50 ml). On the other hand, beef extract, retarded enzyme synthesis while fish meal although slightly enhanced enzyme production, yet, its presence caused a decrease in biomass formation.

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