Vol. 32, June, 2009.

1/32 CYTOLOGICAL STUDIES AND SPERMODERM ORNAMENTATIONS IN SOME TAXA OF ERYTHRINA, L. (LEGUMINOSAE- PAPILIONOIDEAE) IN EGYPT

Wael Taha Kasem and   Mahmoud Mansour

Botany and Microbiology Department, Faculty of Science,

Al-Azhar University, Egypt

Cytological characters (nucleic acid and mitotic index), seedshape, as well as seed topography were studiedcomparativelyfor11taxarepresenting 7 speciesof Erythrina, L. (Leguminosae-Papilionoideae) cultivated inEgypt. These taxa were Erythrina caffra, E. corallodendrum, E. spinosa, E.crista-gall, E. speciosa and E. variegata. Twenty binary characters were subjected to the numerical analysis by using UPGMA program (unweighted pair-group method with arithmetic means). Fivetypesofbasicornamentationpatternwererecognized:reticulate(E. variegata & E. speciosa), pitted (E.poppigiana & E. caffra), granulate (E.crista-gall), undulate (E. corallodendrum) and stellate (E.spinosa). The present studies showed great correlation in seed morphological and cytological characters between the different taxa especially the three accessions of E. corallodendrum, two accessions of E.crista-gall and the two accessions of E. variegate. Data of seed topography by electron microscopywereusedinthe construction of an indented key to identify the studied taxa.

2/32 EFFECT OF BIOCOMPOSTING ON THE GROWTH AND PRODUCTION OF SUNFLOWER AND CORN CULTIVATED IN DESERT SOIL

Mahmoud M. Amer*, Seham M. Shash*, Bouthina F. Abdel Ghani**

and Amal M. Omar**

*Botany Department, Faculty of Science, Benha University, Benha, Egypt.

**Desert Research Center, Cairo, Egypt.

The highly active cellulolytic strains (Kocuria rosea, Micrococcus luteus and Stachybotrys sp.) were used in composting some organic wastes such as rice straw and garbage manure. Bio composting organic wastes supplemented with chemical accelerator and mixed with biofertilizer (Azotobacter chroococcum and Bacillus spp.) supported better chemical characteristics of the final products free from coliform groups, Salmonella, Shigilla and Proteus and rich in humuscontent.Highly significant increase in microbial growth, plant parameters and oil content of sunflower and corn plants were recorded by amending alkaline calcareous soil of with 5 ton/ feddan of rice straw or garbage manure compost applied with biofertilizer in presence of full ~ half recommended field doses of inorganic nitrogen fertilizer.

3/32 Seed protein Electrophoresis AND ITS TAXONOMIC

SIGNIFICANCE on some taxa of Cruciferae in Egypt

Amaal H. Mohamed, Fareida M. Al Saied *, Azza A. F. Khafagi and Dalia G. Gabr

Botany Department, Faculty of Science (Girls Branch), Al-Azhar University,

Cairo, Egypt

*Desert Research Center-Matariya, Cairo, Egypt

On the basis of SDS – PAGE electrophoresis, seed protein profiles of 22 taxa belong to16 genera of Cruciferae were valuable in the study of their phylogenetic relationships. The seed proteins banding showed that there were no institute bands in all examined taxa. Based on obtained dendrograms the indices of distance were calculated. The dendrograms were constructed using PRIMER analysis used Euclidean distance measure similarity and the SPSS version 16 program used Agglomeration Schedule method with Euclidean distance measure average linkage distance(between group).The clustering of investigated taxa showed that the taxa were grouped into two major clusters including groups and subgroups. The results of this investigation could be useful as an attempt to clarify taxonomic relationships of these taxa. The seed protein electrophoresis characters are important in the delimitation of clades within Cruciferae.

4/32 EFFECT OF POTASSIUM FOLIAR APPLICATION ON GROWTH AND NUTRITIONAL STATUS OF ALFALFA (MEDICAGO SATIVA L.) UNDER SALINITY CONDITIONS.

Samya Mohamed Zayied Sohsah

Botany Department, Faculty of Science, Al-Azhar University (Girls).

Alfalfa plant (Medicago sativa L.) was grown in greenhouse experiment to study the interactive effects of different salinity levels (0, 2500, 500 and 7500 ppm NaCl) in the irrigation water and additional K rates (0, 0.5, l and 1.5 %) as foliar application on growth parameters and mineral compostion at three sampling times. Results revealed that growth parameters including plant height, shoot fresh and dry weights, leaf area and number of branches / plant have been adversely affected by salinity levels. However, K foliar application mitigates the harmful effect of salinity and induced a pronounced stimulative effect on all growth parameters throughout the growth period. Concerning mineral composition, salinity increased total ash, Na , Cl- as well as Ca and decreased K and N contents in alfalfa shoots. However, no obvious trend was observed concerning Mg content. K foliar application relatively increased their concentrations except Na content which was markedly decreased as compared with unsprayed plants under salinized and non salinized conditions.

5/32 ISOLATION AND CHARACTERIZATION OF BACTERIA CAPABLE OF DEGRADING BENZENE AND TOLUENE FROM PETROLEUM-CONTAMINATED SOILS IN SAUDI ARABIA

Majdah M.Y. Al-Tuwaijri

Botany Department, Faculty of Applied Science (Girls Branch) Umm Al Qura University, Makkah Almukarramah, Saudi Arabia.

A vast number of bacterial strains have been isolated from petroleum-contaminated soils at the Eastern region in Saudi Arabia. The isolates were screened for their capacity to utilize benzene and toluene as sole source of carbon and energy. Two isolates identified as Bacillus subtilis (Strain No. Bs2) and Bacillus mycoides (Strain No. Bm7) exhibited the highest benzene and toluene degrading potentials. Both isolates proved capable of utilizing the two petroleum derivatives for their growth with a concomitant production of acidic metabolic products which resulted in a shift in the pH of the medium towards acidity. Supplemental addition of peptone (at a concentration of 0.05% w/v) to the mineral salt basal medium appeared to enhance growth of both isolates but had a slight impact on the drift of pH of the medium.

6/32 GROWTH AND CERTAIN METABOLIC ACTIVITIES AS TOOLS FOR EARLY DIAGNOSIS OF GRAFT COMPATIBILITY OR INCOMPATIBILITY IN SOME FRUIT TREES

Abd El-Monem M. Sharaf*, Mohamed M. Ali**, Ibrahim I. Farghal*

and Mohamed S. Mohamed**

Faculty of Science, Al-Azhar Univ. and Horticulture Res. Instit., Cairo, Egypt

Growth and some biochemical measurements were studied at early stages of growth after grafting process using Local apricot and Nemagaurd peach as rootstocks and different cultivars (Okinawa peach, Sour almond, Hansen peach, Florida prince peach and Canino apricot) as scions. Compatible cultivars recorded significant increases in the scion length, number of leaves and rootstock thickness. However, grafting area thickness as wall as scion thickness were decreased. At the early stages of growth, the contents of photosynthetic pigments, gibberellic acid (GA3) and total indols were significantly increased in compatible cultivars. Different responses were also detected concerning contents of carotenoids, total phenols and abscisic acid (ABA) in compatible and incompatible cultivars.

7/32 THERMOTOLERANCE AND ANTIOXIDANT SYSTEMS IN CHICKPEA (CICER ARIETINUM L.) SEEDLINGS: INVOLVEMENT OF HEAT SHOCK AND SALICYLIC ACID

Dina Z. Abdel-Kader, Amal A.H. SalehandAmr M. El Elish

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

The present work was carried out to study the role of heat shock and salicylic acid in antioxidative system in chickpea (Cicer arietinum L.) plant. 15-day chickpea seedlings were divided into 6 groups. The first one kept in open air then harvested after 25 and 32 day post germination (negative control).The second was exposed to 45°C for 3 hours (heat stress) and then harvested after 10 days (positive control). The 3rd and 4th groups were exposed to 2 different heat shock temperature (35°C and 40°C) for 1, 1.5 and 2 hours. The fifth and sixth groups were sprayed with two different salicylic acid (SA) concentrations (0.5 or 1 mM). The groups from third to sixth were exposed to heat stress (45°C) for 3 hours either directly or after week after treatment with heat shock or salicylic acid. The seedlings were harvested 10 days post heat stress. The results showed that, high temperature stress increased lipid peroxidation, electrolyte leakage and hydrogen peroxidase level. High temperature stress decreased catalase, peroxidase, ascorbic peroxidase activities and glutathione content. It was also found that high temperature stress induced a significant increase in endogenous SA concentration and superoxide dismutase activity.These deteriorative symptoms in the chickpea seedlings were ameliorated by heat shock treatments or SA application by decreasing lipid peroxidation and increasing catalase, peroxidase, ascorbic peroxidase, superoxide dismutase activities and glutathione content. Based on these results, it was concluded that the stress protection caused by heat shock treatments or SA application contributes to some extent to the enhanced activity of the free-radical scavenging systems.


8/32 PRODUCTION, PURIFICATION AND IDENTIFICATION OF GLYCOLIPID BIOSURFACTANT FROM HYDROCARBONS UTILIZING PSEUDOMONAS PSEUDOALCALIGENES-R 41

Reda A. Bayoumi and Magdy M. Muharram

Botany & Microbiology Department, Faculty of Science (Boys), Al-Azhar University, Cairo, Egypt.

Biosurfactants are a unique class of compounds that have been shown to have a variety of potential applications in the remediation of organic- and metal-contaminated environments applications in enhancing bacterial, oil recovery, as cosmetic additives, and in biological control. However, little is known about the distribution of biosurfactant-producing bacteria in the environment. The goal of this study is to isolate and purify biosurfactants produced by bacteria and its characterization. Five potent biosurfactant producing bacterial isolates were selected from twenty five isolates isolated from five petrochemical polluted and petroleum sludge samples and well identified two bacterial strains Bacillus subtilis NCTC10400 and Pseudomonas aeruginosa TCC10415ATCC10415 were used for this study. These bacterial isolates were tested for their ability to grow in presence of 0.2 % (w/v or v/v) concentration of each phenanthrene, cyclohexane, toluene, anthracene, naphthalene, salicylate, nitrophenol, α- and β-naphthol as only source of carbon and energy on two types of media. Five potent hydrocarbon utilizing bacterial isolates were tested for their ability for biosurfactant biosynthesis using hemolytic activity and oil spread techniques. These five unknown bacterial isolates could be identified on the basis of cell morphology, physiological and biochemical characteristics as Bacillus subtilis R2, Bacillus coagulans R4, Bacillus amyloliqueficans R5, Acinetobacter calcoaceticus R13 and Pseudomonas pseudoalcaligenes R41. Bacillus amyloliqueficans R5 and Pseudomonas pseudoalcaligenes R41 exhibited high biosurfactant biosynthesis strains were tested for their ability of biosynthesis on different vegetable oils and different carbohydrate sources. The biosurfactant produced by Pseudomonas pseudoalcaligenes R41 was characterized by Ultraviolet (UV), Infra red (IR), amino acid, fatty acid profile, gas liquid –mass spectrometry (GC-MS), high performance liquid chromatography (HPLC) and nuclear magnetic resonance (NMR) as glycolipid. This biosurfactant may have a potential for application in bioremediation of crude oil contamination.

9/31 EFFECTS OF HEAVY METALS (Cd or Pb), GIBBERELLIC ACID AND THEIR INTERACTIONS ON GROWTH, YIELD AND LEVELS OF CERTAIN ENDOGENOUS HORMONES IN BROAD BEAN AND LUPIN PLANTS

Abd El-Monem M. Sharaf, Ibrahim I. Farghaland Mahmoud R. Sofy

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

Effects of heavy metals (20 ppm Cd or 100 ppm Pb) and gibberellic acid (100 ppm GA3) and their interactions on growth, yield and certain endogenous growth hormones in broad bean and lupin plants were studied. Treatment with either Cd or Pb caused significant reduction in most of the tested growth characteristics. Significant decreases in levels of endogenous GA3, indole acetic acid (IAA) and significant increases in levels of abscisic acid (ABA) were recorded in both broad bean and lupin plants in response to the treatment with either Cd or Pb. Application of GA3, alone and combined with either Cd or Pb, markedly stimulated most of the growth and yield characters as well as the levels of endogenous GA3 and IAA of both broad bean and lupin plants. Levels of ABA in both plants were decreased in response to the treatments with GA3 singly or combined with either Cd or Pb.

 

10/32 THE BENEFICIAL USE OF BIOFERTILIZERS ON GROWTH AND YIELD OF WHEAT PLANTS GROWN ON SANDY SOIL WITH OR WITHOUT NITROGEN FERTILIZATION

Rawhia A.M. Arafa*, Bouthaina F. Abd El-Ghany**, Nagwa M. Sidkey*

and Mona M. El-Shazly**

*Fac. of Science (Girls) Al-Azhar Univ., Cairo, Egypt. ** Desert Research Center, Cairo, Egypt.

Biofertilizers are the most advanced applied biotechnology, necessary to support developing organic agriculture, They have been considered as a promising alternative for mineral fertilizers especially for desert development .They are effective in decreasing agricultural costs, maximizing crop yield and reducing environmental pollution .A comparative study using three Azotobacter chroococcumisolates (MGO2, MOM2 and MKZ1) were used either individually or as a mixture .Chemical nitrogen fertilizer was either omitted (0N) or added with recommended full dose (N) or half dose (1/2N). A greenhouse pot experiment was carried out using sandy soil supplementd by 1% sheep manure as organic fertilizer and Ca, N, P and K as inorganic fertilizers. The total number of bacteria in the rhizospere was evaluated. Wheat growth createrea were measured.Generally inoculation of wheat grain with Az.chroococcum isolates, increased the total microbial counts and total Azotobacter counts in rhizosphere, which in all cases, resulted in a significant increase in all growth and yield parameters. Which resulted in an increase in nitrogen content of wheat plants by about 1.50 fold in N content of the control and lead to increase in plant biomass, root length and grain yield compared with the control .The increase in all plant parameters was more pronounced in plants treated with bacterial mixture, than in the control especially grain yield about (1.5- 1.65 folds). Meanwhile, the highest dose of inorganic N fertilizer was approximetly not associated with any significant increase in values of all the tested plant parameters compared with the 1/2N dose. Thus, inoculation with Az.chroococcum could save half of the re-commended dose of inorganic N biofertilizer without any significant reduction in wheat yield.

11/32 GAMMA-IRRADIATION FOR STIMULATION OF SEED GERMINATION OF MAIZE UNDER SALINITY-INDUCED STRESS CONDITIONS

Ibrahim M. Zeid and Amira M. Abd El-Sattar

Botany Department, Faculty of Science, Helwan University, Cairo 11792, Egypt

The response of maize grown under salinity-induced stress conditions to gamma-irradiation was studied during seed germination of two differentially tolerant varieties of maize (Giza 2 and TCH 352). The results indicated that Giza 2 variety was more tolerant to salt stress than TCH 352, since it showed a higher germination percentage, enzymatic activity, and higher content of total soluble sugars and proteins under salinity stress, while the lipid-peroxidation and relative permeability of the root membranes were lower. Application of gamma-irradiation, particularly at 1Krad dose stimulated seed germination and the metabolic activities of salt-stressed and unstressed seeds, while the lipid-peroxidation and relative permeability of the root membranes were reduced.Protein banding patterns exhibited a great variations in dry seeds of both varieties before sowing in response to γ-irradiation.

12/32 CYTOLOGICAL EFFECTS OF TWO DRY MIX ANTIBIOTICS IN ALLIUM CEPA ROOT ASSAY SYSTEM AND THEIR MONITORING BY PLANT EXTRACTS

Abdel-Reheem Qenawy

Botany Department, Faculty of Science Qena, South Valley University, Egypt.

The present work aims to investigating the cytogenetic effects of the two antibiotics amoxicillin-clavulanic acid (AC) and amoxicillin-trihydrate (AT) in root tip cells of A. cepa L. The concentrations used of the two antibiotics were; 0.25, 0.5 and 1% for 8 hours followed by recovery for 24 hours. The two antibiotics induced cytotoxic effects, which include the induction of chromosomal aberrations and reduction in mitotic activity. An increase in the ratio of abnormal mitosis was dose-dependent. AC blocked DNA synthesis, as adjudged by the content of DNA in non-dividing cells. The present study also aims to evaluate the ability of extracts of the medicinal plants; Cumin, Spearmint and Anise to counteract the undesirable effects of these antibiotics. The results showed highly positive ability of some of these extracts to alleviate the cytotoxic actions of these semi synthetic antibiotics.

13/32 ALUM WASTE AND ITS EFFECT ON PHYSICO-CHEMICAL PROPERTIES OF WATER AND SOIL AND AQUATIC PLANTS OF ABU-ZA'ABAL WETLAND

Ahmed A. Khalafallah

Botany Department, Women's Collage for Arts, Sci. and Edu., Ain Shams University.

The present study aimed to evaluate alum waste thrown on shores of Abu-Za'abal 3rd depression shores and to study its effect on the physico-chemical properties of water and soil, in addition to response of the native plants. Water, soil and plants (Najas marina and Phragmites australis) samples were collected from the littoral zone of the 1st, 2nd and 3rd depressions. The analysis indicated that alum waste contains 15% dissolved constituents mainly Al3 and SO42- which dissolved in water with high concentrations. Dissolved alum in water increased the acidity of water and soil and reduced P availability. The high concentration of Al3 and SO42- in 3rd depression reduced dry biomass and photosynthetic pigments of Najas marina, while those of Phragmites australis didn't affected. Uptake of K, Ca and Mg by Najas marina plants was significantly decreased compared to Phragmites australis plants, in addition, phosphate availability decreased to plants by increasing Al3 and SO42- concentrations. The present study showed that alum waste had detrimental effect on the physico-chemical properties of Abu-Za'abal Wetland. Phragmites australis plants accumulated high amount of Al3 in their rhizomes, so it can be used as phytoremediator to aluminum.


14/32 CYTOLOGICAL AND PHYSIOLOGICAL RESPONSES OF VICIA FABA TO A GLYPHOSATE-BASED HERBICIDE TREATMENTS

Abdel-Reheem Qenawy, Mohamed Atya El-Tayeb and Hoida Zaki

Botany Department, Faculty of Science Qena, South Valley University, Egypt.

The cytological and physiological effects of various treatments of the Round up herbicide on Vicia faba (cv. Assuit 25) have been studied. Mitotic activity, expressed as MI, decreased and frequency of chromosomal aberrations increased in root tip meristems as the concentration of the herbicide increased and the time of treatment prolonged. The reported chromosomal anomalies included chromosomal bridges and breaks that have been regarded as an indication of a mutagenic potential caused by the herbicide. Seedling growth, expressed as fresh and dry weight, was adversely affected as the concentration of Round up increased and the duration of treatments prolonged. A dose-dependent reduction was also observed for the amount of soluble protein and total free amino acids in the roots and shoots of seedlings exposed to the herbicide. Similar results were also found for the amount of photosynthetic pigments content in the leaves. In contrast, soluble sugar and proline contents were highly raised as the concentration of the herbicide increased and the time of treatment prolonged particularly at the highest concentration (10-4%) and the longest duration used (24 hrs). Meanwhile, the peroxidase activity decreased by increasing the concentration of herbicide and duration of treatment the activities of catalase and ascorbate peroxidase increased.

15/32 PURIFICATION AND CHARACTERIZATION OF EXTRACELLULAR CHOLESTEROL OXIDASE FROM STREPTOMYCES CORCHORUSII CX-3.

El-Sayed, A. El-Sayed, Mohamed F. Ghaly, Ibrahim A. Abou-Elkhair

and Amira I. Abou-saty

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

The objective of this study is to obtain an abundant source of cholesterol oxidase for industrial and medicinal needs. Five Streptomycete isolates that express high level of inducible extracellular cholesterol oxidase could be isolated from cultivated soil. One of these isolates, named CX-3, related to Streptomyces corchorusii, was found to produce the highest level of cholesterol oxidase. Cholesterol oxidase from strain CX-3 was precipitated from the culture supernatant using 80% ammonium sulphate fraction. The precipitate was subjected for purification using Sephadex G-100 gel filtration. The purified enzyme revealed a single polypeptide band on SDS-PAGE with an estimated molecular weight of about 58 KDa, and exhibited maximum absorption at 240 nm. The enzyme was stability active in a wide range of pH and temperature with an optimal pH and temperature of 7.5 and 40ºC, respectively. Moreover, the thermal stability of the enzyme exhibited high stability at 40ºC in the first 24 h.

16/32 ALLELIC DETECTION AT THE MICROSATELLITE XGWM261 LOCUS LINKED TO THE RHT8 DWARFING GENE IN WHEAT

Khaled F.M. Salem* and Magdy Z. Mattar**

* Plant Biotechnology Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), P.O. Box, 79, Sadat City, Menoufia University, Egypt.

** Botany Department, Faculty of Science, Menoufia University, Egypt.

Wheat (Triticum aestivum L.) varieties with greater coleoptiles elongation are preferred, but the presence of gibblerellin (GA3) insensitive dwarfing genes tends to restrict coleoptile elongation. The agronomic value of Rht8 and the discovery of its diagnostic microsatellite marker, Xgwm261, have accelerated breeders interest in Rht8 as an alternative dwarfing gene. A wheat microsatellite locus, Xgwm 261, whose 192-bp allele closely linked to the dwarfing gene Rht8, on chromosome 2DS, was used to screen 16 wheat varieties from nine countries to identify the reduced height (Rht) or semi-dwarfing genes at this locus Xgwm261 in the presence and absence of GA3. Screening of this wheat varieties showed that a 174-bp allele and a 192-bp allele were the most frequent. Seven wheat varieties possessed a 192-bp allele specific to Rht8; three wheat varieties from UK; three from Egypt in addition one Russian variety. The allele frequency ranged from 6-45%. The highest allele frequency was observed for a 192-bp fragment (45%) followed by a 174-bp fragment (31%). The lowest mean values of coleoptile elongation and seedling length in response of GA3 (2.28 and 5.63 cm) were detected in varieties have 192-bp allele, while the highest mean values (2.67, 2.78, 3.00, 3.22, 3.40 cm for coleoptile elongation and 10.72, 11.41, 12.67, 12.95, 14.40 cm for seedling length) were observed in varieties have another allele than 192-bp at Xgwm261 locus.

17/32 CHARACTERIZATION OF SOFT ROTTING ISOLATES OF ERWINIA ISOLATED FROM DIFFERENT online casino VEGETABLES IN EGYPT

Hoda H. El-Hendawy, Salah A. Moustafa*, Nahla G. Mohamady

Botany and Microbiology Department, Faculty of Science, Helwan University, Ein Helwan, Helwan1, Egypt.

*Agricultural Genetic Engineering Institute, Agricultural Research Center, Giza, Egypt.

Comparison of the cultural, biochemical and physiological characteristics of a soft rotting bacterial isolate Le1 isolated from diseased lettuce leaf with those reported for fifteen local isolates of Erwinia spp. revealed that this isolate belongs to Erwinia carotovora subsp. carotovora. Also, the ability of isolate Le1 to utilize 95 different carbon sources was examined by the biolog microplate system. The obtained results confirmed the identification of the pathogen as Erwinia carotovora subsp. carotovora. Analysis of cellular fatty acid methyl ester of this isolate showed that the fatty acid profile is similar but not identical to that of most Ecc isolates tested. However, analysis of total cellular protien of all Ecc and Eca isolates showed that similar protien banding profile was detected for Eca isolates wherease 5 different protien banding profiles were observed among Ecc isolates which might suggest that Ecc isolates represent 5 different strains. Furthermore, PCR primers deduced from published recA gene sequences of Erwinia carotovora were used for identification of the tested isolates. 730 bp DNA fragment was obtained from each of the 16 Erwinia isolates tested.

18/32 EVALUATION OF GENETIC DIVERSITY IN EGYPTIAN WHEAT VAREITIES USING MICROSATELLITE MARKERS

Khaled F. M. Salem* and Magdy Z. Mattar**

* Plant Biotechnology Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), P.O. Box, 79, Sadat City, Menoufia University, Egypt.

** Botany Department, Faculty of Science, Menoufia University, Egypt.

This study was conducted to assess the genetic diversity in some Egyptian bread wheat (Triticum aestivum L.) varieties. A collection of twelve Egyptian bread wheat varieties representing four wheat breeding programs performed at Sakha, Gemmiza, Giza and Seds was analyzed using eleven wheat microsatellite markers. A total of 39 alleles were detected with an average value of 3.55 alleles per locus. Allele number per locus ranged from 2 to 6. Gene diversity ranged from 0.260 for Xgwm631 to 0.754 for Xgwm165, with a mean value of 0.617. The largest number of alleles per locus (15) was found in the A and B genomes, compared to only 9 for the D genome. Varieties collected from the same breeding program were pooled and the number of alleles and gene diversity were calculated over the twelve simple sequence repeats. The number of alleles was 31, 28, 25 and 24 for Seds, Giza, Gemmiza and Sakha, respectively. PIC value was 0.727, 0.698, 0.594 and 0.497 in Giza, Seds, Gemmiza and Sakha, respectively. Genetic similarity coefficients were estimated for all possible six pairs of breeding program comparisons and ranged from 0.229 to 0.365. Cluster analysis based on UPGMA method provided a clear separation of the four breeding program into two main cluster.

19/32 EVALUATION OF PHENOLIC COMPOUNDS OF VIRGIN OLIVE OILS CULTIVARS AND THEIR POMACE AS NATURAL ANTIOXIDANTS

Mona M. Abd El Magied*, Salah H. Abu Raya*, Laila El-Mahydi**

and Rania M. El Sayed**

*Department of Food Sci., Of Agric, Cairo Univ., Giza, Egypt.

**Food Tech. Res. Inst., Agric. Res Center, Giza, Egypt.

The phenolic compounds in virgin olive oil are considered as an important factor to its shelf life for their antioxidative properties. The virgin olive oils under examination included Kronakii and Coratina cultivars. The findings indicate that both cultivars contained 280 and 297 ppm total polyphenols, respectively. Moreover, the residual Olive pomace from both cultivars contained 105 and 112 ppm, respectively. The phenolic compounds were analyzed by HPLC. The major phenolic compound were found to be tyrosol, hydroxytyrosol, p.Coumaric, vanillic acid ,cinnamic acid and gallic acid. The virgin olive oils of both cultivars contained higher amount of phenolic acids than their corresponding pomaces. The induction period of sunflower oil (SFO) with different concentrations of extracted polyphenols from olive pomaces was determined. The highest induction period (hr at 100˚C) was shown for SFO with 600 ppm polyphenols of Coratina olive pomace (12.86 hr and 71.46 %). The antimicrobial activity of virgin olive oils and their pomaces were evaluated, virgin olive oil and the phenolic compounds extracted from their pomaces had higher antimicrobial activity on the gram positive bacteria than those of gram negative bacteria. Also, extracted phenolic compounds of olive pomace as well as the original virgin olive oil were evaluated as antioxidant agent through the biological evaluation tests. The phenolic compounds were examined at the levels of 600, 1200 and 2400 ppm and the butylated hydroxytoluene (BHT) 200 ppm through the daily ingestion of albino rats for 6 weeks. Meanwhile, the virgin olive oils were separately mixed with the diet of rats at the level of 10 %. The content of total polyphenolic compounds presence in extra virgin olive oils had strong effect on the level of total cholesterol and LDL that decreased them.


20/32 RISK ASSESSMENT OF FOOD COLORING AGENTS ON DNA DAMAGE USING RAPD MARKERS

Hala M. Abdelmigid

Botany Dept, Faculty of Science, Mansoura University, Egypt

       In this study, faba bean (Vicia faba) seedlings were used as bioindicator to determine genotoxic effect ofsynthetic dyes currentlyused as food color additives in many countries.   Novel short-term assays are required to substantiate the batteryof assessment methods for evaluating the genotoxicity of candidate substances. Therefore, an attempt has been made to evaluate randomlyamplified polymorphic DNA (RAPD) analysis for its potentialto establish genotoxic effect of colored food. For the preliminary assessment, this study compared the effects occurring at molecular levels in Vicia faba exposed to colored food at concentrations inthe range of 0.2% to 18.2% . The qualitative modifications arising in random amplified polymorphic DNA (RAPD)profiles as a measure of DNA effects were compared with control. Results suggested thattreatments of the above test food samples reflectingchanges in RAPD profiles. Changes in RAPD patterns included variation in band intensity; loss of normal bands and appearance of new bands compared with control. These results indicated that DNA polymorphisms detected by RAPD analysis could be used as an investigation tool for environmental toxicology and as a useful biomarker assay for the detection of genotoxic effects of food dyes. In conclusion, the measurementof parameters at molecular levels is valuablefor investigating the specific effects of agents interactingwith DNA. Ultimately, this methodology may allow the ecotoxicologicalexamination of the link between molecular alterations and measurableadverse effects at higher levels of biological organization.

21/32 DESTRUCTION OF AFLATOXIN B1 DURING ACID HYDROLYSIS OF CONTAMINATED CORN GLUTEN

Soher E. Aly and Amal S. Hathout

Toxicology & Food Contaminant Department, National Research Centre, Dokki,

Cairo, Egypt

Aflatoxins are a group of mycotoxins that cause serious chronic disease outbreak and contaminates many food products such as corn and their by product corn gluten. Therefore, the aim of this study is to evaluate the effect of HCl on aflatoxin B1 destruction during the production of hydrolyzed vegetable protein. Different HCl concentrations (1, 3, and 5N), hydrolysis temperatures (25oC, 100oC, and 110oC) and different hydrolysis time (2, 4, 6, and 8 hours) were used. Aflatoxin B1 was destructed by 27.6% after 4 hours of hydrolysis and reached 42.5% after 8 hours in the presence of 1N HCl at 110oC. On increasing HCl normality from 1N to 3N at 25oC, percentage of aflatoxin B1 destruction increased from 9.3% to 59.9% after 8 hours of hydrolysis, meanwhile in the presence of 5N HCl, percentage of destruction reached 77.6%. During the 8 hours of hydrolysis, the percentage of aflatoxin B1 destruction recorded 50.4%, 60.3%, 71.6% and 79% after 2, 4, 6 and 8 hours respectively at 100oC in the presence of 3N HCl. Complete aflatoxin destruction (100%) was recorded in the presence of 5N HCl, at 110oC after 4 hours of hydrolysis. The amino acid profile for the hydrolyzed vegetable protein was evaluated and compared to those of commercial soy sauce. The dicarboxylic acid group was moderately present at 32.28%, meanwhile the non-essential amino acid, glutamic acid (24.27%) was found to be higher than the other hydrolyzed vegetable protein. It could be concluded that the manufacture of hydrolyzed vegetable protein is considered a suitable method for decontamination of aflatoxin without any residual hazard or toxicity on human and animal health.

22/32 MONITORING OF ANTIMICROBIAL ACTIVITY OF ESSENTIAL OILS USING MOLECULAR MARKERS

Hend A. Hamedo

El Arish, Faculty of Education, Suez Canal University, Egypt

Technological application of essential oils, as natural antimicrobial agents, to reduce the effect of pathogenic microorganisms, requires the a new methods of detection. The present work evaluated the parameters of antimicrobial activity of the essential oils of rosemary (Rosmarinus officinalis) on two pathogenic strains Escherichia coli and Staphylococcus aureus. The MBC and MIC values were of 2.5, 25 μl ml-1, and values of 1.25 and 5 μl ml-1 for the two strains respectively. In this study, an attempt has been made to evaluate randomlyamplified polymorphic DNA (RAPD) analysis for its potentialto establish antimicrobial effect of rosemary essential oil. For the preliminary assessment, this study compared the effects occurring at molecular levels in E. coli and Staph. aureus exposed to rosemary essential oil at the MIC concentrations for the two organisms. The qualitative modifications arising in random amplified polymorphic DNA (RAPD)profiles as a measure of DNA effects were compared with control which showed many differences. In conclusion, the measurementof parameters at molecular levels is valuablefor investigating the specific effects of agents interactingwith DNA.

23/32 BIOMARKER CHANGES IN ERUCA SATIVA AS A RESULT OF APPLICATION OF BIOFERTILIZER TRICHODERMA HARZIANUM IN METALS CONTAMINATED SOIL

Tarek M. Abd El -Ghany,   Eman M. El – Taher* andMona S. Ashour*

FROM

Botany and Microbiology Department, Faculty of Science, Al-Azhar University,

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

Eruca sativa plants cultivated in soil inoculated with T. harzianum as a biofertilizer and were exposed to different concentrations (50,100 and 200 ppm) of copper and zinc mixture as sulfate salts. The fatty acid composition of Eruca sativa could be used as an additional biomarker of soil contamination by metals mixture. Saturated (palmitic and stearic acid) and unsaturated (oleic acid) fatty acid contents could be detected in the plant cultivated in soil at different treatments with T. harzianum in presence of different mixtures of copper and zinc concentrations, while other fatty acids were found to be affected by metals in the soil. Behenic acid was detected only at high concentration (100 and 200 ppm) in absence of T. harzianum inoculation. Stearic, arachidic and lignoceric acids could be detected at high concentrations with T. harzianum inoculation. Inoculation of T. harzianum in the soil improved the plant growth and minimize the toxic effect of metals mixture. At the same time an increase in the content of chlorophyll (a) and (b) could be detected. Addition of T. harzianum to cultivated soil minimize the absorbed   amount of metals in plant , where the percentage of copper was 6.04 and 4.69 % with uninoculation and inoculation of T. harzianum respectively. On the other hand, copper and zinc mixture decreased the percentage of Mg 2 metal in plant and the addition of T. harzianum increased their percentage, where their percentage was 4.92 % in presence of 100 ppm of metal mixture without T. harzianum and becoming 7.08 % with T. harzianum Exposure of Eruca sativa plant to high concentrations of metals mixture (100 ppm) without T. harzianum induced a significant increase catalase and peroxidase enzymes activities.  

24/32 RESPONSE OF MAIZE TO GAMMA-IRRADIATION AND MYCORRHIZA UNDER SALINITY STRESS

Ibrahim M. Zeid and Amira M. Abd El-Sattar

Botany Department, Faculty of Science, Helwan University, Cairo, Egypt

Maize grains were pre-sowing exposed to gamma rays and grown under salinity-induced stress conditions in presence and absence of mycorrhiza, as a biofertilizer, for increasing the plant resistance to salinity stress.The results indicated that application of gamma-radiation, particularly at 1 Krad dose stimulated plant growth, enzymatic activity, content of sugars, soluble proteins, chlorophyll content, and activity of Hill-reaction of salt-stressed and unstressed plants, Mycorrhiza treatment markedly increased root growth, metabolic activity and the mineral ions content of the leaves, as well as the K /Na ratio under salinity stress indicating alleviation of the adverse effects of salinity.Protein banding patterns showed a great variation in the leaves of mature plants, during the fruiting stage, in response to γ-irradiation, as well as mycorrhiza treatment, indicating a response at the gene expression level.

25/32 TREHALOSE AS OSMOPROTECTANT FOR MAIZE UNDER SALINITY-INDUCED STRESS

Ibrahim M. Zeid

Botany Department, Faculty of Science, Helwan University, Ain Helwan,

Cairo, Egypt.

Response of maize grains (Giza 2) to the pre-soaking treatment with 10 mM trehalose was studied under salinity-induced stress conditions. Trehalose treatment induced growth of salt-stressed and unstressed plants. Trehalose pretreatment alleviated the adverse effects of salinity stress on the metabolic activity of maize seedlings. Hill-reaction activity, photosynthetic pigments and nucleic acids content increased in response to trehalose application. Organic solutes e.g., sugars, soluble proteins and proline content increased under salinity stress. This increment was associated with increased hydrolytic activity of amylase and protease enzymes. Trehalose treatment may be ameliorate salinity stress through stabilization of the plasma membranes, since it decreased the rate of ion leakage and the rate of lipid peroxidation of the root cells, and increased the ratio of K/Na ions in the leaves of maize seedlings. Electrophoretic protein banding patterns were also investigated. Trehalose treatment reduced salt expression.


26/32 FACTORS AFFECTING BIODEGRADATION OF LOW CONCENTRATION 2,4-DICHLOROPHENOXYACETIC ACID (2,4-D) IN NILE WATER BY THE PSEUDOMONAS SP.MA-100
Mohamed A. Ramadan, Alaa M. Shawky, *Aml E. Saafan and *Eman A. Elgebaly
Department of Microbiology, Faculty of Pharmacy, Cairo University and Faculty of Pharmacy, *Beni-Suef University.
The study was conducted to determine the potential capacity of the local isolate Pseudomonas sp MA-100 to degrade 2,4-D at low concentrations in Nile water samples and how to enhance the degradation process. Pseudomonas sp has the capacity to degrade 2,4-D at a concentration of 1 µg ml-1 in salt solution.The bacterium was able to bring about biodegradation of 2,4-D at a concentration of 1 µg ml-1 in Nile water under the same conditions. About 70% of 1 μg/ml 2,4-D disappeared within 4 hours of incubation in non-sterile Nile water inoculated with 3 ×107 cfu ml-1 of the tested isolate. The bacterium has the capacity to degrade very low concentrations of 2,4-D up to 100 ng ml-1 where complete disappearance of 100 ng ml-1 occurred within 5 hours of incubation. Increasing the inoculum density up to 3×109 cfu ml-1 enhanced the degradation rate since the substrate completely disappeared from the medium within 3 hours of incubation. Addition of inorganic nutrients to Nile water samples at a concentration of (10 µg ml-1 NHNO3 and 10 µg ml-1 KH2PO4 & 20 µg ml-1 K2HPO4) dramatically enhanced the degradation rate where 2,4-D was completely degraded after one hour of incubation. Addition of second carbon sources as oxalate, citrate and phenol at a level of 5 µg ml-1, enhanced degradation where complete disappearance of 2,4-D occurred within 3-4 hours for oxalate, citrate and phenol compared to 6 hours for unsupplemented sample. Immobilization of isolate MA-100 on activated carbon induced complete disappearance of 2,4-D within 4 hours of incubation compared to 6 hours for the free cells under the same conditions. Pseudomonas sp was able to bring about degradation of 2,4-D at low concentrations up to 100 ng ml-1 in Nile water samples and the extent of degradation could be enhanced by increasing the inoculum size, amendments of Nile water samples with inorganic nutrients and second carbon sources. Immobilized Pseudomonas sp cells also enhanced the degradation process. The results presented here show that the Pseudomonas sp MA-100 could be used in bioremediation of low concentrations of chlorinated aromatic herbicides in polluted aquatic ecosystems such as the river Nile in Egypt since it contains residual amounts of 2,4-D. 

27/32 ANTIBACTERIAL EFFECTS OF CRUDE ETHANOLIC EXTRACTS OF ENTEROMORPHA AND ULVA ON PSEUDOMONAS AERUGINOSA

Alaa A. Said

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

The present study was mainly intended to study the antimicrobial effect of crude ethanolic extracts of Enteromorpha and Ulva species, green algae on pathogenic bacteria Pseudomonas aeruginosa isolated from Benghazi children hospital that was not treated with single-drug therapy. A total of 5 species of Enteromorpha and 7 species of Ulva were collected and identified from Benghazi coasts during the period from April 2007 and October 2008. The crude ethanolic extracts of Enteromorpha species was significantly (p<0.05) more effective than those of Ulva species (99.69% and 73.88%, respectively), as compared with the antibiotic, Chloramphenicol. The algal ethanolic extracts also had significant antibacterial effect on P. aeruginosa (p<0.05). It was noticed the high efficiency of Enteromorpha species (117.73% and 105.77%) at 400 and 200 mg/dry weight, respectively. The aim of the work is studying of the antibacterial effect of crude ethanol extracts of Enteromorpha and Ulva on Pseudomonas aeruginosa.

 

28/32 EFFECT OF NACL AND SUCROSE ON IN VITRO PRODUCTION OF CHAMOMILE ESSENTIAL OILS.

Nomaan S. Hassan, Karima H.A. Salama and Mohamed M.F. Mansour

Department of Biology, Faculty of Science, Taif University, Taif, Saudi Arabia

Seeds of chamomile (Chamaemelum nobile L.) were surface sterilized and inoculated on the surface of sterilized Murashige and Skoog medium supplemented with 2.0 mg/L kinetin, 0.5 mg/L 2,4-D and 30g/L sucrose. The pH of the medium was adjusted to 5.8 0.1 prior to the addition of 0.8% bacteriological agar. The cultures were incubated at 27oC 1 and 16 h photoperiod. Callus was produced after four weeks of incubation. This callus was subcultured successfully on the same medium composition. After three passages (four weeks each), part of this callus line (control) was transferred to media containing 100 or 150 mM NaCl. Another part was transferred to media containing 9 or 18% sucrose (90 or 180 g/L). After incubation for another four weeks at the same conditions, essential oils were extracted from the control, treated callus and seeds and identified using the gas liquid chromatography. Eight essential oils were identified in the seed: the greatest of which was transpinocarveol (42.83%) followed by amyl-angelate (12.96%), transpinocarvone (7.38%), amyl-tiglate (8.85%), isoamyl-tiglate (6.17%) and hexyl-tiglate (5.65%). Farnesol and 1,8-cineole were minor fractions. In the callus, the eight essential oils were detected with different relative distributions: the greatest was amyl-tiglate (44.46%) while transpinocarveol dropped to 3.31%. Isoamyl-tiglate and hexyl-tiglate contents were higher in the control callus compared with those of the seeds. Two new essential oils, α-pinene and nerolidol, were identified in the control callus. Farnesol and 1,8-cineole were not detected in the control callus.Exposing chamomile callus to salinity stress (100 and 150 mM NaCl) and sucrose (90 and 180 g/L) resulted in the production of eleven essential oils. The major essential oils under salinity were amyl-tiglate, hexyl-tiglate and 1,8-cineole. The major essential oils were Hexyl-tiglate (38.11%) and amyl- angelate (15.18%) under 90 g/L sucrose, and amyl-tiglate (44.04%) and Hexyl-tiglate under 180 g/L sucrose.The results indicated that exposing callus tissues to salinity and sucrose resulted in the greater production of already present essential oils as well as newly synthesized of other essential oils, e.g., ά-pinene, nerolidol and hexyl-angelate. It is concluded that in vitro cultures of chamomile seeds may provideessential oils, which may have medicinal or industrial importance.


29/32 STUDIES ON STRELITZIA REGINAE, AIT BY MICROPROPAGATION METHOD

BY

Mamdouh A. El-Shamy, Boshra B. Rezk Alla* and Ali Nabih

FROM

Botanical Garden Res. Dept Hort. Res. Inst. (ARC), Giza, Egypt

*Ornamental plants Res. Dept Hort. Res. Inst. (ARC), Giza, Egypt

Strelitzia reginae, Ait is one of world’s most popular, economic and important cut flowers plant. The study was carried out at Plant Tissue Culture Laboratory at El-Zohria Botanical Garden, Horticulture Research Institute, ARC, during 2007 – 2009 years. It is intended to investigate the possibility of propagating this very attractive plant by using tissue culture technique and to detect the most suitable methodology in this respect. Strelitzia reginae buds and seeds were tried as explants. All explants of Strelitzia reginae were effectively surface sterilized with a mixture of mercuric chloride (HgCl2) and Clorox (commercial bleach) at 40 % Clorox plus 0.7 mg/L HgCl2 and at 40 or 50 % Clorox plus 0.7 mg/L HgCl2, respectively, for sterilization explants, in which the best result (100%) survival and free contamination of buds and seeds explants were recorded. The supplementation of the culture medium with activated charcoal was the best for the initial establishment stage. In the multiplication stage of Strelitzia reginae, 6.0 mg/L Kin formed not only the highest number of leaves but also the highest extension length of shoots. BA was better than Kin during the multiplication stage of shoot number. The most suitable medium for rooting stage was MS 4 mg/L IBA. In vitro, rooted shoots could be transferred with great success for acclimatization to greenhouse conditions. The survival percentage was 50.    

30/32 FORTIFICATION OF SOME BAKERY PRODUCTS BY USING OF ORANGE FRUIT PEELS

Mohamed H. Abdeldaiem and Hoda G. Mohamed

Atomic Energy Authority, Nuclear Research Center, Egypt

This study aims to prepare orange dietary fiber concentrate from Balady orange (Citrus sinensis L.) waste fruits as waste of orange juice industry.One such recent trend is to increase the fiber content in food products to overcome health problems such as hypertension, diabetes, and colon cancer, among others. The results showed that the contents of orange dietary fiber concentrate for moisture 6.79, protein7.52, fat 3.40, ash2.64, crude fiber61.27, total carbohydrates and total phenols as gallic acid were, and 0.97%.Vitamin C and trace elements of iron, zinc and manganese were 152.6 mg/100g and 63.89, 19.21 & 5.05 mg/100, respectively. Also, amino acids composition of orange dietary fiber samples was determined. Biscuits were prepared from blends of 72% wheat flour containing 0, 5, 10, 15 and 20% of orange dietary fiber concentrate. While, cakes were prepared from 72% wheat flour mixed by   0, 10, 20, 30 and 40% of orange dietary fiber concentrate. On the other hand, the results of sensory evaluation illustrated that highly acceptable biscuits and cakes could be obtained by incorporating 5% and 10% of orange dietary fiber in the formulation, respectively. Meanwhile, finely ground orange dietary fiber was incorporated in wheat flour at 0, 5 and 10% levels and studied for rheological characteristics.Water absorption increased from 58.3% to 65.8 and 67.4% with increase in orange fiber from 0% to 5 and 10%. Dough stability increased and weakening of the dough decreased with increasing mixtures of 5 and 10% of orange fibers. Thus, it can be recommended that orange waste fruits as an important source of dietary fibers and nutritional valueas well as bioactive components (such as flavonoids, polyphenols and carotenes) and have possible uses in the enrichment of foods.

31/32 POSSIBLE APPLICATION OF MICROBIAL BIOREMEDIATION OF DIBENZOTHIOPHENE AND DURSBAN POLLUTED ENVIRONMENTS

Reda A. Bayoumi; Ezz El-Deen M. Mohamed; Samir S. El-Louboudy

and Abd-Allah H. Hendawy

Botany & Microbiology Department, Faculty of Science (Boys), Al-Azhar University, Cairo, Egypt,

This study concerned with the biodesulfurization of organic sulfur heterocyclic compounds and organophosphate pesticide chlorpyrifos (Dursban) by using microorganisms. Twenty eight and sixteen microbial isolates capable of growing on dibenzothiophene (DBT) and Dursban as a secondary substrate were isolated from six crude oil and agricultural pesticide polluted soil samples respectively. The two most potent yeast isolates capable of utilizing dibenzothiophene (DBT) as the only sole source of sulfur were selected and identified as Candida parapsilosis S1-Y1 and Cryptococcus humicolus S1-Y2. The most potent two bacterial isolates capable of utilizing Dursban as only source of carbon and energy were identified as Pseudomonas stutzeri S7-B4 & Flavobacterium balustinum S8-B6. These four isolates were investigated in some environmental and nutritional conditions controlling the biodesulfurization of dibenzothiophene and biodegradation or detoxification of Dursban. These conditions included incubation period, DBT or Dursban concentrations, inoculum size, incubation temperature, pH, introducing carbon sources, nitrogen sources, amino acids, vitamins and incubation conditions. Different three groups containing nine treatments were inoculated under all optimal environmental and cultural conditions by their consortia after 15 days incubation period at 35˚C. By gas chromatography- mass spectrometry analysis, desulfurized DBT metabolites and dursban biodegradation indicated that the concentration of DBT decreased and reached up to 0.0% in some cases. These results indicated that strains C. parapsilosis and Cr. humicolus in case of DBT& sulfur-containing crude petroleum oil and P. stutzeri & F. balustinum in case of Dursban are able to utilize the organic sulfur of DBT and Dursban in application to large variety of hydrocarbons that occur extensively in commercial fuel oils in three tested environments. Therefore possible C. parapsilosis and Cr. humicolus in case of DBT& sulfur-containing crude petroleum oil and P. stutzeri & F. balustinum in case of Dursban are considered a good potential candidates for industrial and environmental applications for the biodesulfurization of DBT and dursban degradation in different fields.

32/32 BIOLOGICAL BEHAVIOR OF VARICELLA- ZOSTER VIRUS RECOVERED FROM EGYPTIAN CHILDREN IN CELL CULTURE AS A USEFUL TOOL FOR ANTIVIRAL SCREENING ASSAYS

Rania I. Shbel*, Amal E. Ali**, Ali F. Mohamed* and Magdi A. Amin**.

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

*The Holding Company for Biological Products and Vaccines (VACSERA)

       The biological behavior of four Varicella-Zoster virus (VZV) isolates recovered from clinically diagnosed children with varicella, were studied in cell culture. Following isolation and identification, isolates were propagated in african green monkey kidney cells (Vero cells). Propagation of the isolates showed 90% cytopathic effect (CPE) on the 12th – 16th day. On further passage, they showed a progressive growth activity; where the time to detect CPE had been shortened to 48-72 hours in the 5th passage. Studying the growth kinetics of these isolates using endpoint of CPE (CCID50) assay revealed significant rise in the viral infectivity titer relatively to time reaching 104.5,103. 8,103. 5 and 105 for isolates no.1, 2, 3 and 4 respectively seven days post inoculation. The in vitro cytotoxicity and antiviral activity of liquorice powder extract containing glycrrhizin, acyclovir and interferon alpha 2a against one isolate were evaluated respectively by crystal violet uptake assay and by reduction in virus infectivity titer. Liquorice powder extract containing glycrrhizin (125 μg/ml) showed slight antiviral activity with o.5 log unit decrease in virus infectivity titer. Comparatively, acyclovir (250 μg/ml) elicited higher antiviral activity, and resulted in 4 log unit decrease in virus infectivity titer. On further dilution of acyclovir, the 50% inhibitory concentration (IC50) was 7.8 μg/ml. Diluted interferon alpha 2a (10ng/ml) showed complete suppression of the virus infectivity and had the most potent antiviral activity among the tested drugs. These results enable biological characterization of local VZV isolates in cell culture and allow definitive identification of the isolates and their susceptibilities to antiviral drugs.
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