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EDITORIAL BOARD

2016

MOLECULAR DIAGNOSTIC OF ESCHERICHIA COLI AMONG URINARY TRACT INFECTIONS PATIENTS USING POLYMERASE CHAIN REACTION (PCR)

By Israa Adnan Ibraheam*, Ishraq Abdul Amir Saleh*, Hawraa Jawad Kadhim*, Zainab Khudhur Ahmad Al-Mahdi **

Vol-13 No-4

Abstract

Infection with E. coli represented the highest causes of Urinary tract infections among community. Molecular techniques such as polymerase chain reaction (PCR) have become most important technique for more rapid and accurate for detection bacterial causal organisms in species level. E. coli based on detecting gene targets lac Z which encode for the enzyme ?-galactosidase is important target gene, this gene is responsible for energy production through break down of disaccharide lactose to the glucose and galactose. The urine samples were collected from UTI’s patients initially diagnosed according to the presence of pyuria, a classical culture methods for detection of E. coli was recorded the presence of E. coli in 20 out of 30 UTI’s samples (66%) while in molecular method by using polymerase chain reaction (PCR) for detection lac Z gene, E. coli was detect in 18 out of 30 UTI’s samples (60%), this difference between classical culture and PCR for identification of E. coli was non-significant (p=0.05), and this may be due to the presence of evolved beta-galactosidase lac Y in some strain of E. coli which have the same galactose permease activity as lac Z galactosidase. So the present work strongly encouraged using PCR technique as a novel, perfect and fast test for E. coli diagnosis on molecular level. Further study for developed multiplex for both lac Z and lac Y galactosidase is highly recommended.

PRODUCTION OF BIODIESEL FROM FRESHWATER ALGAE

By Syed Adnan Hasan1*, Ambreen Aziz2, Khalid Hassan2, Syed Imran Ali1, Suraiya Jabeen3

Vol-13 No-4

Abstract

The production of biodiesel from micro algal lipids is studied by using newly isolated fresh water algal strains. Four different strains of Chlorococcum, Deasonia, Chlorella and Botryococcus species were isolated. The selection of the fastest growing specie was made through the results of growth kinetic studies when grown under optimum conditions. Chlorella sp. strain S-3 was grown in a large quantity of synthetic algal growth medium. This strain produced a large amount of oil (up to 22% of biomass), which was efficiently extracted by using (i) solvent extraction method through Soxhelet extractor with a 1:1 mixture of n-hexane and diethyl ether and (ii) by centrifugation of sonicated algal cells in liquid medium. By alkaline transesterification of algal lipids, biodiesel was formed as the main product with glycerol as by-product. The physicochemical quality of algal biodiesel was found comparable to conventional diesel.

EVALUATION OF PHYTOCHEMICAL CONSTITUENTS AND ANTIBACTERIAL POTENTIAL OF AVICENNIA MARINA AND RHIZOPHORA MUCRONATA FROM INDUS DELTA OF PAKISTAN

By A.M. Mangrio1, M. Rafiq2*, S.H.A. Naqvi2, S.A. Junejo3, S.M. Mangrio1, N.A. Rind2

Vol-13 No-4

Abstract

In the present study, phytoconstituents and antibacterial response of mangroves Avicennia marina and Rhizophora mucronata were evaluated. Various parts of both plants were collected from Indus delta, Thatta. Qualitatively test results confirmed that alkaloids, flavonoids, tannins, terpenoids, saponins and sterols are present in crude extracts of different parts of both plants. Total phenol contents, flavonoids and antioxidant activity were determined by spectrophotometer. Total phenol contents were varied from 10.7?0.5 to 29.85?1.2 mg/ml; total flavonoids from 2.37?0.42 to 16.53?0.19 mg/ml and antioxidants from 2.4?0.5 to 14.4?0.17 mg/ml in different parts of both plants. Leaf extracts of both mangrove species showed the highest amounts of total phenol, total flavonoids and antioxidant activity as 29.85?1.2 mg/ml, 16.53?0.19 mg/ml and 14.4?0.17 mg/ml respectively, while root extracts of both plants accumulated the lowest amount of total phenol, flavonoids and antioxidant activity. Crude ethanol extracts of A. marina and R. mucronata were also tested against three pathogenic bacterial species. T1 preparations (10% ethanol extracts) of leaves of both species showed a wide range of growth inhibitions 5.5?0.7 – 13.5?0.4 mm while T2 preparations (5% ethanol extract) showed poor response 3.0?0.65 - 7.6?0.2 mm against E. coli, K. pneumoniae, S. aureus.

COMPARISON OF EFFECTIVENESS BETWEEN BEES-CARNAUBA MIXED WAX AND CHITOSAN WHEN COMBINATION WITH PHENYLLACTIC ACID ON POSTHARVEST QUALITIES OF ORANGE FRUIT CV. CANH DURING LOW TEMPERATURE STORAGE

By Huyen Nguyen Thu1,2, and Tanachai P.1

Vol-13 No-4

Abstract

Comparison of effectiveness between bees-carnauba mixed wax (MW) and chitosan when combination with phenyl lactic acid (PLA) on postharvest qualities of orange fruit cv. Canh during low temperature storage was studied, by soaking fruit in 2.5% PLA before coating with 8% MW and 2% chitosan solution, and then storing at 5 ± 1°C for 60 days. While, untreated fruits were used as control. Percentage of weight loss and fruit decay, total sugar content, titratable acidity content, total soluble solids (TSS) content, ascorbic acid content, and total microorganisms were monitored during the preservative time. The results showed that 2.5% PLA in combination with 8% MW had a higher preservative effectiveness when compared to 2.5% PLA in combination with 2% chitosan and control fruits, which was expressed as lower percentage of weight loss and fruit decay, and lower total microorganisms for 60 days in storage. Moreover, fruits maintained nutrient ingredients expressed as total sugars, titratable acidity, TSS and ascorbic acid content.

ANABOLIC ANDROGENIC STEROIDS AND LIVER DYSFUNCTION IN MALE ADULT MICE

By Shaima Obied Abd-Allh and Shyma Ahmed Rahiem

Vol-13 No-4

Abstract

Liver is one of the most important members of the body that play a fundamental role in the metabolism and secretion xenobiotics which makes it highly influences to their adverse and toxic effects. Liver infection or liver dysfunction caused by different drugs and toxic chemicals or their reactive metabolites (hepatotoxicants) is known as hepatotoxicity. The current study is designed to know the effect anabolic androgenic steroids in liver function, histological structure and liver biomarkers. Experimental animals used in the study (30 Swiss mice) were allocated to three groups as general control group, and test groups (N=10). The test groups received therapeutic dose 5 mg/kg of body weight and high dose 10 mg/kg of nandrolone decanoate (ND) by subcutaneous injection, the dose was given each 48 hours, but control groups received the same volume of normal saline. The duration of the experiment was 4-weeks. The collection of blood samples, two days after the end of the period experimental from the control and treated groups. The serum samples were assayed for liver biomarkers (alanine aminotransferase (ALT), aspartate aminotransferase (AST), total proteins, alkaline phosphatase (ALP) and total bilirubin). It has been sacrificed animals in each group after the end of the experiment and the extirpation of liver for histopathological study. Results revealed that all ND-treated animals exhibited a significant increase of ALT, AST, ALP, total protein and total bilirubin. Also, our results of this study indicate that anabolic steroids at supraphysiological and high doses exerts histopathological changes in liver and might lead to toxic liver of males. The abuse of such drug by youth and adolescents requires taking steps represented by programs of awareness and guidance known health risk of these materials to public health to curb or at least minimize the spread of communities.

INFLUENCE OF GAMMA RADIATION ON THE PHYSIOCHEMICAL PROPERTIES OF IN VITRO TRIPLOID AND TETRAPLOID BANANA SPECIES

By Mehnaz Qamar1*, Sadaf Tabassum Qureshi1, Imtiaz Ahmed Khan2, Sajjad Ahmed Memon3, Zeenat Bano1 and Sardar Khatoon Solangi1

Vol-13 No-4

Abstract

Resistance to different stresses of banana are much needed, therefore, introductions of new and healthy clones are required. Applications of in vitro mutagenesis are considered as the reliable tool to create variations. For that purpose, in vitro cultured multiple shoots of three Musa clones: Giant Cavendish Tissue Culture variant (GCTCV215), Yangambi KM-5 and FHIA-23 were exposed to different gamma doses (0, 10, 20, 30, 40 and 50 Gy). Radiosensitivity of in vitro shoots was assessed by counting data on survival % after 40 days of culture, a rate of shoot multiplication, average shoot, fresh weight (g), root number and root length (cm). Different level of radiation sensitivity was viewed in all Musa varieties. Lower dosage 10 Gy produced raising effect compared to control. The present study may be helpful to introduce new Musa germplasm in our region.

BREAST CANCER DIAGONOSIS THROUGH ANALYSIS OF BRCA GENE USING MACHINE LEARNING ALGORITHMS

By J. Sumitha* and T. Devi

Vol-13 No-4

Abstract

Worldwide, the breast cancer is the second leading cancer type which leads to death among women. Breast cancer exists due to the mutation in the normal growth of Breast Cancer Gene (BRCA) under certain circumstances. In this paper, we proposed the existing machine learning algorithms for finding the disease – causing BRCA gene. These existing machine learning algorithms are compared with each other to determine the efficiency in detecting the diseases from gene expression value. The results proved that the Hybrid Radial Bias Neural Network (HRBFNN) performs better than Divide and Conquer Kernel Solving Support Vector Machines (DCKSVM) and other classification algorithms than ever before.

THE METHOD OF Agrobacterium Tumefaciens-MEDIATED MmCu/Zn-SOD GENE TRANSFORMATION IN THE RED SEAWEED Kappaphycus alvarezii

By St. Hidayah Triana*1,2,3, Alimuddin*1, Utut Widyastuti3,4 , Suharsono3,4, Emma Suryati5 and Andi Parenrengi5

Vol-13 No-4

Abstract

Superoxide dismutase (SOD) has an important role in the defense system of the body, especially the activity of reactive oxygen compounds that can cause stress. MmCu/Zn-SOD gene successfully regenerated in N. benthamiana and N. tabacum, rice, and Jatropha transgenic plants. The assembling of transgenic K. alvarezii has been previously performed with other genes, however, the successful introduction of genes was still low. The objective of this study was to optimize a method for the Agrobacterium mediated MmCu/Zn-SOD gene transformation based on the optical density (OD), the duration of inoculation and co-cultivation period factors to increase the effectiveness of transgenesis in K. alvarezii. The experimental method was divided into the three stages: 1) the optimization of MmCu/Zn-SOD gene transformation in Kappaphycus alvarezii based on optical density (OD), inoculation duration and co-cultivation period; 2) the analysis of the existence of MmCu/Zn-SOD genes in Kappaphycus alvarezii using PCR and 3) the challenge test with salinity treatments (15, 30 and 45g/L). Six levels of A. tumefaciens OD600 (0.3, 0.4, 0.5, 0.6, 0.7 and 0.8), four duration of inoculation (10, 20, 30 and 60minutes) and two periods of cocultivation (3 and 4days) were tested. The results demonstrated that the highest putative bud (100%) and transformation (100%) efficiencies were obtained by using inoculation duration of 60minutes and 3days of co-cultivation with OD600 of 0.5. However, regeneration efficiency of explants (6.67%) was lower than the non-transgenic control (33.33%). The results of PCR analyses demonstrated the presence of MmCu/Zn-SOD gene in the transgenic explants, whereas the nontransgenic explants seaweed showed no amplification product. In conclusion, transgenic K. alvarezii could be produced and optimized by the Agrobacterium tumefaciens-mediated method based on optical density (OD), duration of inoculation and co-cultivation period factors. Transgenic explants that carry encoding genes of MmCu/ Zn-SOD tolerant to salinity of 15g /L and 45g/L so all were survive, while all the non-transgenic were died.

VIDEO CONTENT RETRIEVAL USING IMAGE FEATURE SELECTION

By D. Saravanan

Vol-13 No-3

Abstract

ABSTRACT Multimedia is any combination of content written in text, drawing, and sound, moving picture, delivered to you by computer or other electronic or digitally manipulated means. Through the rapid growth of multimedia technology, multimedia content can be created, shared and distributed easily. The amount of available digital resources is continuously increasing, promoted by a growing interest of users and by the development of new technology for the ubiquitous enjoyment of digital resources. The acquisition and storage of video data is an easy task but retrieval of information from video data is challenging .The general method of representing each video segment is shot that is a sequence of key frame(s) where those frames contained the ‘meaningful’ frames also the frame contained the important contents of the shot. The key frame(s) based shot method is specifically assisted for searching the video content as clients provided image query/search where an image will be matched with the indexed key frames with assist of resemblance distance. As a result, the key frames selection is most significant and several methods are used to automate the process. This paper proposes a new technique for key frame selection. The proposed method show significant ally good and the experiments prove the above statement.

PROTOCOL OPTIMIZATION FOR EFFECTIVE IN VITRO ROOT FORMATION OF SUGAR CANE SOMA CLONES IN NIA-2012, NIA-105 AND GULABI-95 VARIETIES

By Sardar Khatoon Solangi1*, Sadaf Tabasum Qureshi1, Mukhtiar khatoon Solangi2, Altaf Hussain Solangi2

Vol-13 No-3

Abstract

ABSTRACT The aim of this study is to develop an efficient In vitro regeneration and root formation procedure from callus. Present work was conducted in 2015 at Nuclear Institute of Agriculture, Tando jam. Study relevant with three sugarcane varieties of NIA like NIA-2012, NIA-105 and GULABI-95. Root formation of plantlets was compared under different concentration of auxins IBA (1.0, 2.0, 2.5 mg1-1 and sucrose 2, 3 and 4 % respectively. Highly significant (p<0.05) variations were observed for all parameters of root formation. Interactive effect of variety treatment and concentration was non-significant for the number of root and length of root formation. Auxins 3% and sucrose 4% mgl-1 were highly effective for rooting. In vitro root formation for all genotypes of sugarcane showed the maximum root and highest length in ½ MS media supplemented with 2.5 mg1-1 IBA and 4% sucrose, therefore this concentration of growth regulators was used for future in vitro culture of sugarcane.

GRAIN YIELD, PHOSPHORUS CONTENT AND UPTAKE OF WHEAT (TRITICUM AESTIVUM L.) AS AFFECTED BY PHOSPHORUS FERTIGATION

By Mukhtiar Khatoon Solangi*1, SardarKhtoon Solangi2, Nusrat Khatoon Solangi1

Vol-13 No-3

Abstract

ABSTRACT Most of the Pakistani soils are alkaline and calcareous in nature; therefore the efficiency of P-uptake by wheat is low. Among the agronomic practices that influence the efficiency of applied fertilizer, time and method of application are critically important. Fertigation is one of the techniques which enable the application of water soluble fertilizers and other chemicals along with irrigation water to the plant, uniformly and more efficiently. Two years field studies on wheat was undertaken to compare the response of P applied in the irrigation water with broadcast method. The treatments include: control (no P applied), P applied through broadcast method (farmers practice) and P applied with irrigation water (Fertigate P). The experiment was conducted in four replications using randomized complete block design. Generally, fertigated applied P increase grain yield, P content and P-uptake in wheat as compared to broadcast method. In wheat, fertigated P increased grain yield by 7-8%, P content by 8-16% and P uptake by 15 to 23% during two cropping years as compared to broadcast method. In conclusion, the use of P fertilizer through fertigation could be useful technique to increase grain yield and P uptake in cereals.

EFFECT OF PHYTOHORMONES ON SHOOT AND ROOT REGENERATION IN ROSE UNDER IN VITRO CONDITIONS

By Nighat Seema Soomro1, Imtiaz Ahmed Khan1, Shahla Baloch2, Ghulam Shah Nizamani1 Shafquat Yasmeen1 and Muhammad Tahir Khan1

Vol-13 No-3

Abstract

Abstract In vitro propagation of rose has played a very important role in rapid multiplication of cultivars with desirable traits and production of healthy and disease free plants. Murashige and Skoog medium supplemented with various concentrations and combinations of Indole-3-butyric acid (IBA), Naphthylacetic acid (NAA), Benzylaminopurine (BAP), and Indole-3- acetic acid (IAA) were employed for shoot regeneration in this study, whereas MS½ and full strength medium containing different amounts of IBA (0.5 and 1mg/l) were used for root induction. The highest multiple shoot formation was recorded in MS medium containing IAA (1 mg/l) and BAP (4 mg/l) with 30 g/l sugar. Cytokinin (BAP) in addition to auxin (IBA) caused the maximum bulbus formation which turned into elongated shoots, while the NAA was observed to have antagonistic effect on shoot formation. Highest numbers of roots were produced on MS½ strength + IBA 0.5 mg/l. The aim of this study was to determine growth and development of rose on different growth media, to shorten the multiplication time of virus-free plant material.

OPTIMIZATION OF CULTURE CONDITIONS FOR THE PRODUCTION OF PROTEASE BY Pleurotus eryngii

By Safia Bano, M. Umar Dahot and S.H.A. Naqvi

Vol-13 No-3

Abstract

ABSTRACT Present study focuses to optimize the protease enzyme production by edible mushrooms. For the specified goal to achieve the growth of Pleurotus eryngii cultured on potato dextrose agar medium incubating at temperature 24ºC, pH 5.6 for 7 days. The maintained culture was processed for the production on fermentation media, i.e., glucose 20 g/L, peptone 10 g/L, corn steep 3.0 g/L and ammonium nitrate 1.0 g/L on pH 6.5. The best time for maximum production was observed to be four days from incubation. After selecting the optimum time for the enzyme production, the optimum nutrient contents were checked. Sucrose (0.5%) and casein (1%) were found to be the best carbon and nitrogen sources, respectively. The maximum protease production was achieved at pH 6.5 and the maximum enzyme yield was found at 22ºC,

GENETIC TRANSFORMATION OF POTATO (Solanum tuberosum L. ) Cv. NOOKSACK WITH FBPase/ClRan1 GENES MEDIATED BY Agrobacterium tumefaciens.

By Fatahillah1, Suharsono2, UtutWidyastuti2

Vol-13 No-3

Abstract

ABSTRACT Nooksack is one of potential cultivar to be used as french fries material. Fructosa 1.6-bisphosphatase (FBPase) and Citrullus lanatus Ran1(ClRan1)genes play a very important role by increasing potato productivity. Overexpression of FBPase gene is able to accelerate photosynthesis whereas overexpression of ClRan1 genesis to develop the stolon. This study aimed to introduce FBPase/ClRan1 genes into the genome of potato cv. Nooksack used Agrobacterium tumefaciens. Leaf disc and internode of potato was inoculated with A. tumefaciens EHA105 bringing FBPase/ClRan1 genes under control of rbsS3C promoter for FBPase gene and 35SCaMV promoter for ClRan1 gene. Leaf discs have a transformation efficiency (83.33%) and regeneration efficiency (16%) better than internode with transformation efficiency (50%) and regeneration efficiency (6.66%). PCR analysis indicated that FBPase/ClRan1 genes has been integrated into potato with size of the FBPase/CLRan1 genes was 1047 bp. Data analysis used SPSS 16 with one way ANOVA method on the level of 95%. FBPase/ClRan1 gene was significantly different from the photosynthesis rate, leaf number, plant height, stem diameter, the number of bulbs and tuber weight. Whereas, the wet weight and dry weight of shoots were not significantly different.

THE EFFECT OF CITRIC ACID AND SODIUM CHLORIDE (NACL) TO QUALITY OF SARDINE OIL (Sardinella Sp.)

By Sugeng Heri Suseno*, Agoes M. Jacoeb, Stephanie Bija, Nadia Fitriana, Nilam Puspa Ruspatti

Vol-13 No-3

Abstract

ABSTRACT The amount of sardine oil as by-product from a fish-meal processing industry is large and still contains high amount of omega-3 , but its quality is still for feed. Purification of fish oil is very important to improve the quality of fish oils for human consumption. This study aimed to determine citric acid and Sodium chloride (NaCl) treatment in the degumming process to improve the quality of fish oil. The Citric acid and NaCl used were 0.2% and 40%, respectively. The results showed that reduction of peroxide value, free fatty acid, p-ansidine, and TOTOX were 72.42-92.37%, 77.95-85.83%, 5.43-51.90%, and 49.59-76.99% respectively while transmission percentage range was 48.61-63.63%.

EXTRACTION OF BIOACTIVE COMPOUND CURCUMIN FROM TURMERIC (CURCUMA LONGA L.) VIA DIFFERENT ROUTES: A COMPARATIVE STUDY

By Foozie Sahne1, Maedeh Mohammadi1, Ghasem D. Najafpour1*and Ali Akbar Moghadamnia2

Vol-13 No-3

Abstract

ABSTRACT In this study, extraction of curcumin, the bioactive compound of turmeric (Curcuma longa L.), through different extraction methods was investigated. For this purpose, microwave-assisted, ultrasound-assisted and enzyme-assisted extractions were used as modern extraction routes to extract curcumin from turmeric powder and some of their essential extraction parameters were optimized; the results were compared to those obtained from Soxhlet extraction as the traditional and reference method. The presence of curcumin in the extracted samples was confirmed by UV-vis spectroscopy and quantification of curcumin was carried out by high performance liquid chromatography (HPLC) analysis. Result showed that the curcumin extraction yield using Soxhlet method (6.9%) was considerably higher than those obtained from microwave-assisted (3.72%), ultrasound-assisted (3.92%) and enzyme-assisted (4.1%) extractions; however, the highlighted features of advance extraction methods including cost-effectiveness (due to much saving in time and energy consumption) and environmentally benign nature make them more favorable extraction methods.

EFFECT OF BRASSIN-LIKE SUBSTANCE ON THE QUALITY OF EARLY GERMINATED ARABICA COFFEE BEAN (Coffea arabica L.)

By Primluck Praphutphitthaya1*, Chantalak Tiyayon1, Thaneeya Chetiyanukornkul2, Tanachai Pankasemsuk1

Vol-13 No-3

Abstract

ABSTRACT Arabica coffee beans were monitored to evaluate the changes of chemical compositions during early germination period after soaking with brassin-like substance (BS). The experiment was designed as 4×5 factorial in completely randomized design (CRD) by soaking coffee seeds into BS at 4 different concentrations (0 (control), 0.5, 1.0 and 2.0 mg/L), which considered as factor 1. Chemical compositions were determined at 5 germination times (before soaking, day 2, 4, 6 and 8 after soaking), which considered as factor 2. Three replications were made on each combination and each replication consisted of 100 coffee seeds. The results showed that chemical compositions and antioxidant activity were clearly effected by BS and germination time. Caffeine content significantly declined upon BS concentrations. Furthermore, the content also gradually decreased during early germination. Interestingly, soaking coffee seeds in 2.0 mg/L BS could enhanced the greatest amount of chlorogenic acid content within 4 days after soaking. The compound, however, sharply declined afterward. BS could decrease total phenolic compounds in germinated coffee seeds. BS applications at concentrations of 1.0 and 2.0 mg/L also showed greater antioxidant activity than those seeds applied by BS 0.5 mg/L and the control. Moreover, radical scavenging activity of coffee seed crude extracts also indicated that the activity tended to increase during early germination.

TRANSFORMATION OF KAPPA (?)-CARRAGEENASE GENE FROM Pseudoalteromonas IN SEAWEED Kappaphycus alvarezii [DOTY]

By Muh. Alias L. Rajamuddin1, Alimuddin2, Utut Widyastuti3 and Enang Harris2

Vol-13 No-3

Abstract

ABSTRACT Foreign gene transformation techniques have been applied to improve the valuable traits of farmed organism. As the first step towards increasing of kappa (?)-carrageenan content, in this study ?-carrageenase gene (?car) from Pseudoalteromonas was transformed into K. alvarezii mediated by Agrobacterium tumefaciens method. The pMSH-?car binary plasmid construct was controlled by 35S CaMV promoter and Nos terminator. DNA analysis of transformants was conducted by polymerase chain reaction (PCR) method using two sets of primer. The result showed that 36% transformed explants were sprouted and 100% of them were having the ?car gene. The size of amplified DNA fragment from K. alvarezii transformants using 35S-F and tNos-R set primers was 2,000 bp, the same size as PCR product of the positive control of pMSH-?car plasmid, while no amplification product in non-transformated (negative control) was found. Furthermore, PCR analysis using 35S-F and 35S-R set primers showed amplification product of 300 bp, the same size as 35S CaMV promoter sequence. Thus, K. alvarezii transformants carrying pMSH-?car had been generated.

SOLID STATE CULTIVATION AND APPLICATION OF XYLANASE

By Kianoush Khosarvi-Darani1 and Dina Karamad2

VOL-13 NO-2

Abstract

Research for xylanase biosynthesis is an interesting area due to its important industrial application. This review paper serves as an overview of xylanase bioproduction and application as well as its producing microorganisms, substrates and process variables, to consider the future prospects of xylanases in biotechnological applications. Several approaches should be applied to overcome main limitations which inhibit widespread commercial and industrial application of this enzyme; low production yield and the high total cost.

STUDY OF ANTIMICROBIAL SUSCEPTIBILITY PATTERN OF ESCHERICHIA COLI ISOLATED FROM NOSOCOMIAL INFECTIONS IN HILLAH CITY, IRAQ

By Tsahel Hamid Kadhim Al-Dulaimi

VOL-13 NO-2

Abstract

Escherichia coli is a Gram-negative , rod-shaped organism it present as commensal in the gut of human’s alsoits isolates from many different types of animals as well as this kind of bacteria presence in water provides, so it’s considered as an index of stool contamination .in addition this organism can causes diseases in the alimentary tract ,urinary tract includes urethra, bladder, otitis media and respiratory system. The current study aims to identify of antimicrobial agents use in the treatment E. coli that it’s acquired by nosocomial infections. The E. coli has been isolated and then exposure to different classes of antibiotics by using Vitek2 technique method. The total number of E. coli was isolated is 45. the findings have appeared so resistance to Ticarcillin with in 45 (100%), Piperacillin 45 (100%), Ceftazidime 42 (93.4%), Cefepime and Aztreonam 43 (95.6%). On the other hand the samples was showed high sensitivity with Amikacin 44 (97.8%), Imipenem 43 (95.6%), Meropenem 43 (95.6% ) and Gentamicin 37 (82.2%) , while of Piperacillin\Tazobactam 12 (26.7 %) and Tobramycin 7 (15.5%) has intermediate susceptibility to antibiotics study .finally found the high frequency of resistance to Ticarcillin, Piperacillin,Ceftazidime,Cefeoime and Aztreonam was seen among E. coli isolates. As a result they were not recommended the last drugs used for treatment of E. coli infections as optimal therapy. Keywords : E. coli, Antimicrobial susceptibility, Nosocomial infections, Vitek2 technique.

A SENSITIVE ELECTROCHEMICAL SENSOR BASED ON GOLD NANOPARTICLES DROPLET DEPOSITION ON GLASSY CARBON ELECTRODE FOR BISPHENOL A DETECTION

By Hoda Ezoji1 , Mostafa Rahimnejad1 *, Maryam Asghary2

VOL-13 NO-2

Abstract

A convenient, rapid and extremely sensitive electrochemically detection of bisphenol A (BPA) was proposed in phosphate buffer (pH =7) on a gold nanoparticles modified glassy carbon electrode (GNs/GCE). Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were applied for characterization of GNs/GCE. Differential pulse voltammetry (DPV) was employed for measuring BPA. The studies exhibited lower over potentials, improved current response, high reproducibility and lower detection limit at the GNs/GCE versus the bare electrode. Therefore, GNs displays excellent electroactivity toward BPA. Within the concentration range of 1×10-8 to 6×10-6M. The fabricated electrochemical sensor demonstrated linear response to BPA with the detection limit of (1.5×10-9M).

GENETIC TRANSFORMATION OF TOBACCO PLANT USING INHIBITOR OF MERISTEM ACTIVITY (IMA) GENES

By Asri Pirade Paserang 1,2,3*, Aris Tjahjoleksono 3,4, Utut Widyastuti 3,4 , Suharsono 3,4

VOL-13 NO-2

Abstract

Inhibitor of Meristem activity (IMA) gene encoded a Mini Zinc Finger protein (MIF). IMA gene regulates flowering and ovule development. It controls WUSCHEL gene in meristem center and determined nucellus during the development of ovule. IMA gene is activated by the gene expression of type-D inhibited the proliferation of cell during floral termination, controlled the amount of carpels during flower development, and has a role in the primordial ovule initiation. IMA gene plays crucial roles during shoot apical meristem organization which associated with cell division, differentiation, and growth hormone. IMA gene has been successfully constructed into vector pGWB402 and introduced into Agrobacterium tumefaciens LB4404 through electroporation means. IMA gene is transferred to Nicotiana tabacum by using cotyledons in vitro culture. The transformed plant tissue is selected by addition of kanamycin in culture medium. The validation of transgene is detected by PCR using CaMV 35 S forward primer and IMA gene reverse specific primer. Transgenic plants show band size 500 bp indicating IMA gene has been integrated into the plant genome of N. tabacum.

INDUCTION OF SYSTEMIC RESISTANCE IN TOMATO BY PSEUDOMONAS AERUGINOSA PM12 AGAINST FUSARIUM WILT

By Sabin Fatima* and Tehmina Anjum

VOL-13 NO-2

Abstract

This study was carried out to check the potential of a bacterial strain Pseudomonas aeruginosa PM12 for inducing systemic resistance in tomato against Fusarium oxysporum f. sp. lycopersici. Significant reduction in disease index of 14.91% was observed under the influence of this bacterial strain. P. aeruginosa was found to be involved in induction of systemic resistance through upregulation of defense related compounds like phenolics, polyphenol oxidase (PPO), peroxidase (PO), and phenyl ammonia lyase (PAL). Greenhouse study revealed that P. aeruginosa significantly increased growth parameters like biomass, root and shoot length in tomato plants whereas calorimetric assays showed the ability of this strain to produce growth related biochemicals (total soluble sugars, cholorophyll and carotenoid content) efficiently. Therefore it can be concluded from this study that P. aeruginosa can be used for the management of fusarium wilt of tomato and to promote sustainable agriculture in Pakistan. This study will open new ways for the use of indigenous biocontrol agents against the pathogens to obtain higher yields.

CHARACTERIZATION OF BACTERIAL SOFT ROT STRAINS AND THEIR SPECIFIC PHAGES ISOLATED FROM SOIL AT TAIF

By Sonya H. Mohameda,b , Gado E.A.a , Gomaa H.a and Sadik A.S.a,c*

VOL-13 NO-2

Abstract

In this study some bacteriophage(s) specific to some plant pathogenic bacteria (the causal agent of soft rot diseases), were isolated and characterized from soil of Taif, KSA. Soil samples were randomly collected from some rhizosphere soils cultivated with various plant species including potato. Ten bacterial isolates were obtained from soil and used as hosts for enrichment and isolation of the virulent bacteriophages. The spot test and turbidity tests were used to detect the presence of the phage of interest in the suspension. The area of clear zones were represented by threes levels, i.e., weak lysis (+) (three isolates), moderate lysis (++) (one isolate) and high lysis (+++) (two isolates). The phage(s) was propagated and partially purified for determining the morphology of viral particles via electron microscopy. Sperm shape virus-like particles with long tail and icosahedral head were shown in the electron micrographs of partially purified phages specific to the two selected bacterial isolates (# 08 and # 10). These two bacterial isolates were then biologically and molecularly identified. The nucleotide sequences of 16S rRNA gene of the two bacterial isolates was determined and final sequences of 942 and 940 nts for the 16S rRNA gene of two soft rot bacteria (# 08 and # 10), respectively were recorded. Data of the phylogentic trees show that the two bacterial isolates (# 08 and # 10) could be strains of Pseudomonas stutzeri (LC053456.1) and Bacillus pumilus (LC053854.1).

PRODUCTION AND OPTIMIZATION OF ?-AMYLASE FROM ASPERGILLUS NIGER USING POTATO PEEL AS SUBSTRATE

By 1Shahzad Mahmood, 1Memuna Ghafoor Shahid, 2Muhammad Nadeem, 2,3Muhammad Irfan*, 2Quratulain Syed

VOL-13 NO-2

Abstract

The present study is concerned with the production and characterization of ?-amylase by Aspergillus niger in solid-state fermentation using food waste as substrate. Various cultural conditions such as incubation period, incubation temperature, pH of the medium, moisture level and inoculum size were optimized for maximum ?- amylase yield. The maximum activity of enzyme (1262.27 ± 2.11 U/g) was recorded after 72 h of incubation at 30°C temperature, pH 5 with 5% moisture level and inoculum size. Among different nitrogen and carbon sources evaluated, peptone (1.5%), NH4NO3 (0.75%) and soluble starch (1.25%) gave maximum ?-amylase production under optimized conditions. Under all the optimized culture conditions, the maximum enzyme production was 1298.12 ± 2.14 U/g.

SCREENING OF POTENTIAL BACTERIAL BIO-CONTROL AGENTS AGAINST THE HELMINTHOSPORIUM ORYZAE IN-VITRO CONDITIONS. CAUSED BY BROWN SPOT OF RICE

By Ghulam Hussain Jatoi1,5* , Manzoor Ali Abro1**, Javed Asghar Tariq3**, Shabana Memon2** , Naimtullah Mangi**2, Azhar-ul-Din Keerio4 , Muswar Ali Gadhi1 , Shahid Hussain1 and Dewen Qiu5

VOL-13 NO-2

Abstract

Brown spot of rice caused by Helminthosporium oryzae is one of the most destructive diseases of rice worldwide. This disease causing rotting of seeds that ultimately reduce yield by 16-43%. This study was aimed to evaluate bio-control agents under in vitro conditions against H. oryzae. The method followed was based on survey and sampling of rizhospheric soils samples from Larkana district and surrounding areas. Rhizobacteria were isolated from soil-rice plants and 25 isolates were studied for their antagonistic effect on the linear colony growth of H. oryzae by using PDA medium as for growth. Results of the present study showed that the use of bio-control agent (Rhizobacteria) against the tested fungus showed that, four isolates out of 24 bacterial isolates has inhibited the linear colony growth of H. oryzae up to 1-2 mm with different levels (-, +, ++, +++).. The use of bio-control agent Rhizobacteria like (Agrobacterium and Pseudomonas) isolates also showed antagonistic activity against the tested fungus (H. oryzae). This study could be helpful for researchers and farming community in future for better management of this disease.

GENETIC VARIABILITY AND RELATIONSHIP AMONG DURIAN CULTIVARS (Durio zibethinus Murr) IN THEKAMPAR, INDONESIA ASSESSED BY RAPD MARKERS

By Rosmaina1 , Joko Warino2 , Suhaida3 , Zulfahmi1*

VOL-13 NO-2

Abstract

Durian (Durio zibethinus Murr) is an important tropical fruit tree economically and ecologically. This research aims to observe the genetic variability and relationship among durian cultivars based on RAPD(Random Amplified Polymorphic DNA) markers. The results of this study obtained a total of 49DNA fragments of which 34 fragments (69.39%) were polymorphic. Overall size of the PCR amplified product bands ranged from 200 bp (OPO-11) to 1800 bp (OPO-05). Six primers (OPT-09, OPO-05 OPY-16, OPY-15, OPD-08, and OPY-14) can distinguish among five durian cultivars e.g. Bakul, Ome Kampar, Tembaga, Sijantung and Keong Mas. The genetic distance among durian cultivars ranged from 0.276-0.477. Based on UPGMA dendrogram, seven durian cultivars were divided into four groups. The first cluster consisted of Sijantung cultivar, second cluster consisted of Bakul cultivar, third cluster consisted of Keong Mas and Tembaga cultivars, and fourth cluster consisted of Empu Kunyit, Ome Kampar, and Montong cultivars. The results of this study will helpthe breeders toformulate the breeding strategy for the durian improvementin future.

COMPARATIVE STUDIES ON CITRIC ACID PRODUCTION FROM CHEESE WHY BY SUBMERGED AND IMMOBILIZED ASPERGILLUS NIGER

By Sepideh A. Ghanbartabar, Ghasem D. Najafpour*, Maedeh Mohammadi

VOL-13 NO-2

Abstract

Citric acid production from cheese whey using submerged culture and immobilized cell of Aspergillus niger was investigated. In this work, effect of substrate (lactose) and product (citric acid) concentrations, medium pH, sesame oil on production of citric acid was evaluated. In addition, the cell dry weight was measured during the fermentation process. For submerged culture of A. niger, maximum citric acid concentration of 4.45 g/l was obtained at pH value of 4, lactose concentration of 50 g/l, and 0.1% (v/v) sesame oil for incubation of 6 days. For incubation of 5 days, in immobilized cell operation at optimal pH value of 3 and lactose concentration of 70 g/l, maximum citric acid concentration of 8.4 g/l was achieved. Immobilized cells of A. niger had a significant impact on improvement of citric acid production and increased the yield up to 62.6%. The immobilized cells had about 3 folds increase in citric acid productivities in comparison with free cells of A. niger. The productivity of citric acid in immobilized cell system was 4 times greater than the free cells.

RESPONSE OF DIFFERENT DOSES OF PHOSPHATE FERTILIZER AND PHOSPHATE SOLUBILIZING_BACTERIA AND MYCORRHIZA IN MUNGBEAN (VIGNA RADIATA L.)

By Niaz Ali Sial, S.A Abro, Javed Ahmed Shah and M.Y. Memon,

VOL-13 NO-(1)

Abstract

A field experiment was conducted in randomized complete block design (RCBD) with seven treatments in four replications at Nuclear Institute of Agriculture (NIA), Tando Jam. Five plants were selected from middle rows of the plot for yield parameters. Yield and yield components were recorded such as plant height (cm) number of pods/plant, single plant biomass weight (g), number of branches/plant, number of panicles/plant, single plant grain yield (g) biological yield kg/ha, grain yield (kg/ha), days to flowering, days to maturity of the crop, and 100- grain weight (g). Bacterial enumeration after sowing of one week and after harvesting of the crop were recorded. The results revealed that the all yield and yield components were highest in T2 (80 kg P/ha), whereas, the control was the lowest. In treatment Bacillus megatherium and 40 kg P/ha were identical with each other. T5 (Bacillus megatherium+40 kg P/ha), T6 (Mycorrhiza) and T7 (Mycorrhiza + 40 kg P/ha) were similar with each other but lower than T2, T3, T4 and higher than control. Biological yield and grain yield were highest in T2 (2541 kg/ha and 883 kg/ha). Same trend was found for bacterial enumeration after sowing of one week the crop and after harvesting of the crop at the depth of 0-15 and 15-30 cm in one gram of soil. It was observed that the addition of Bacillus megatherium increased the yield and other yield associated traits of mungbean. The results showed that the P solubilizing bacterial inocula alone may be used for obtaining reasonable yield in mungbean.

ASSESSMENT OF INTERNAL QUALITY OF JAPANESE PUMPKIN (CUCURBITA MAXIMA) USING NEAR INFRARED SPECTROSCOPY

By Muenmanee, N.1,2* , A. Joomwong4 , J. Natwichai5 and D. Boonyakiat1,2,3

VOL-13 NO-(1)

Abstract

Near infrared spectroscopy (NIRS) is a non-destructive technique for fruit quality assessment. This study applied NIRS to assess internal quality of Japanese pumpkin fruit during maturation. Japanese pumpkin fruit (Cucurbita maxima) were harvested at three stages of fruit maturation, namely, immature, mature and overmature, i.e., 40, 50 and 60 days after fruit set (DAFS). It was found that there was a possibility to use short wavelength region of near infrared radiation (700-1100 nm) to determine quality of Japanese pumpkin in terms of total soluble solids (TSS). The spectra were measured by NIRSystem 6500. Partial least squares regression (PLSR) was introduced to develop prediction models for the aforementioned parameters. The best PLS model for quality assessment of Japanese pumpkin fruit was found in TSS, resulting the coefficient of determination (R2 ) of 0.77. The prediction model also provided low values of SEC, SEP and bias which were 0.62%, 0.96% and 0.13%, respectively.

EFFECT OF INOCULATION DENSITY ON POTATO MICROPROPAGATION

By Tufail Shakeel+, Zafar Jamal* and Ihsan Ullah**

VOL-13 NO-(1)

Abstract

The present study was conducted to examine the effect of inoculation density on potato micropropagation var. Desiree and investigate the nutrients competition among potato micro plants. The different growth factors were examined, i.e., shoot length, numbers of roots and numbers of nodes of each microplant. A number of 1-5 treatments of micro plants was examined with 10 repetitions of each treatment. In this experiment 1, 2, 3, 4 and 5 nodal cuttings were transferred in each test tube. The morphology of the plantlets during this experiment was observed, and remained same whether single inoculated plantlets or five nodal cuttings were inserted in a test tube. Post two weeks of culture certain numbers of nodes/microplant, roots/micro plant and shoot length were obtained from each culture tube. The numbers of nodes and roots were more in T5 as compared to other treatments. During this experiment it had been observed that the inoculation density did not affect the normal growth of the plantlets. So the inoculation density effect did not affect (i.e., p=0.05) the plantlets. Four week post explant culture the plantlets showed maximum growth under artificial condition of growth chamber. So, this experiment showed that the inoculation density has no significant effect (i.e., p=0.05) on potato microplants.

ISOLATION OF FUNGI FROM VARIOUS AGRICULATURAL FIELDS AS A FUNCTION OF SOIL DEPTH AND SEASONAL VARIABILITY

By Abdul Hameed1* , Muddasir Asrar1 , Abdul Aziz2 and Hidayatullah3

VOL-13 NO-(1)

Abstract

Study of population and species occurrence of fungi as a function of soil depth and seasons was conducted during 2009 and 10. The samples were collected from ten agricultural fields at Quetta district. Soil sampling was conducted during four seasons such as February, 2009 and 10 (S1 and S3) and August, 2009 and 10 (S2 and S4) across soil depth of 0-06 and 06-12 inches. The serial dilution plate technique was used for isolation of fungi which were significantly affected by seasonal variability and soil depth. The results exhibited higher fungal (18.30 cfu g -1soil) population in location 9 at upper depth in S1 and lower (6.50 cfu g -1soil) in location 8 at lower depth. From each location 18 fungal species were isolated and Aspergillus niger was found dominant fungal species in all locations. From this study it was suggested that less perturbed soil like orchards showed more O.M. contents indicating higher available nutrients and high fungal population than field crops which were subjected to more perturbation.

ISOLATION AND CHARACTERIZATION OF FREE-NITROGEN FIXER BACTERIAL STRAINS (Azotobacter sp.) AND THEIR PHAGES FROM MAIZE RHIZOSPHERE-SOIL AT TAIF

By Sadik A.S.a,b* , Noof A. El-Khamasha and Sonya H. Mohameda,c .

VOL-13 NO-(1)

Abstract

We are focusing on isolation of bacteriophage(s) specific to some free N2-fixer bacteria from rhizosphere and non-rhizosphere soils of Taif region of KSA as a first record for such study in Taif area (Makkah, Jeddah and Taif). A number of 10 bacterial isolates grown on the nitrogen-free specific medium (Waksman base No.77 Medium) were purified and separately used as hosts for bacteriophage(s) isolation. The spot test and turbidity tests were used to detect the presence of the phage of interest in the prepared phage suspensions. The level of lysis revealed the presence of turbid zones (plaques), as weak (+) lysis (for isolates # 07) and (# 09); moderate lysis (++) (for isolate # 05), and high lysis (+++) (for isolates # 01 and # 03). The phage(s) was propagated and partially purified for determining the morphology of viral particles via electron microscopy. Sperm shape virus-like particles with long tail and icosahedral head were shown in the electron micrographs of partially purified phages specific to the two selected bacterial isolates ((# 01 and # 03). These two bacterial isolates were then morphologically and molecularly identified. The nucleotide sequences of 16S rRNA gene of the two bacterial isolates was determined and final sequences of 927 and 873 nts for the 16S rRNA gene of two isolates of this study ((# 01 and # 03), respectively, were recorded. Data of the phylogentic trees show that the two bacterial isolates (# 01 and # 03) could be strains of Azotobacter sp. (LC053456.1 and LC053854).

EXPRESSION OF RUBBER ELONGATION FACTOR (REF) AND SMALL RUBBER PARTICLE PROTEIN (SRPP) RELATES TO DRY RUBBER YIELD OF CLONAL VARIETIES

By Auksorn Klaewklad1,3 , Korakot Nakkanong1 , Chatchamon Daengkanit Nathaworn2 and Charassri Nualsri1*

VOL-13 NO-(1)

Abstract

REF and SRPP are necessary for a functional role in rubber polymerization. It involved in the final polymerization step of natural rubber biosynthesis in Hevea. The aims of this research were to relate the REF and SRPP gene expressions and latex yield, in order to use these genes as markers in selection criteria of rubber tree breeding programs. Dry rubber yield and expressions of the two genes were determined for four selected clones (SK1, SK3, NK1 and T2) and the by location paired controls (RRIM 600 clone). Results from cDNA sequencing indicated the REF amino acid sequence of lower yield clone present one amino acid, which is different from Proline (CCC) as Serine at site 113 (CCT). The SRPP amino acid sequences, Glutamic acid are absence at site 6 of RRIM 600. One amino acid at site 67 of RRIM 600 clone was different from other clone resulting in Threonine (ACT) instead of Alanine (GCT). In NK1 clone, at site 178, different in one protein was recorded different code protein from Threonine (ACT) to Valine (GTT). The annual dry rubber yields of the four clones exceeded that of RRIM 600. Quantitative RT-PCR showed a positive relationship between latex yield and expression levels of REF and SRPP genes. In seedling leaves, REF and SRPP genes were expressed at low level, but both genes were abundantly expressed in bark. These results suggest good potential in the early selection of rubber trees, in breeding programs for yield improvement.

EFFICACY OF SELECTED FUNGICIDES ON THE LINEAR COLONY GROWTH OF THE HELMINTHOSPORIUM ORYZAE CAUSED BY BROWN SPOT DISEASE OF RICE .

By Ghulam Hussain Jatoi1*, Manzoor Ali Abro1**, Javad Asghar Tariq4**, Shabana Memon2**, Naimatullah Mangi2 , sultan Ahmed Maitlo1 , Azhar-ul-Din Kerio3 , Shahid Hussain1 and Abdul Salam Mengal1 ,

VOL-13 NO-(1)

Abstract

Brown spot of rice caused by Helminthosporium oryzae, attacks the plant at any growth stage and it appears on the foliage as scattered brown spot that coalesces and result in withering and yellowing of the leaves. This fungal disease lowers the seed germination, causing rotting of seeds and roots that ultimately reduce yield by 16% to 43%. The method followed was based on survey and sampling of diseased seed samples from Larkana. Isolation and purification of fungi from these samples were conducted and performed pathogenicity test of the most predominant isolated fungus (H. Oryzae). So, this study was aimed to evaluate different fungicides. In vitro study of four fungicides efficacy revealed that Mencozeb and Thiomal completely inhibited growth of the test fungus. H. oryzae failed to grow at 150 and 200 ppm, (00. mm) as compared to control (36.62 mm) respectively. Whereas, Bavistin was moderately effective (10.50 mm) and Melody duo was less effective (12.87 mm) as compared to control (36.62 mm). So, one can conclude that Mencozeb and Thiomal was the most effective fungicides. These studies could be helpful for researchers and farming community in future for better management of this disease.

BIODEGRADATION OF PETROLEUM INDUSTRY OILY SLUDGE BY BACTERIAL CONSORTIUM AND ITS APPLICATION IN LAND FARMING

By Anwaar Arafa1 , Soha Farag2* , Manal A. Mahdaly1 , Mai M. Badr1

VOL-13 NO-(1)

Abstract

Oily sludge is considered the most important solid waste resulted from crude oil storage tanks. Conventional oily sludge treatment methods (landfill and incineration) are high in cost and need specialized equipment and skilled personnel. Therefore, bioremediation provides a technique for cleaning up oily sludge, totally or mostly, by enhancing the natural biodegradation processes. The three bacterial isolate Bacillus cereus sp. M1, Bacillus cereus sp. M7 and Pseudomonas mendocina sp. M10, which were isolated from a soil sample from SidiKerir branch, have a high capability to degrade oil sludge. A consortium of isolates (M1, M7 and M10) and sp48 (Pseudomonas sp.) was applied for bioremediation of 1% of oily sludge with different inoculum amounts and at different time periods, where it resulted in higher biodegradation rate of total petroleum hydrocarbons, reaching 89% after 12 days. This consortium was used in bioaugmentation as land farming for oily sludge degradation which achieved 99.2% removal (biodegradation rate) of total petroleum hydrocarbons and 93.7% of oil and grease. Results obtained suggest that the bacterial consortium for oil sludge degradation may be better than each bacterium unaccompanied.

Acknowledgment of Reviewers

The Editor, Pak. J. Biotechnol. is very grateful to the following scientists who dedicated their considerable time and expertise to the journal by serving as reviewers from January 1, 2016 to December 31, 2016 for Vol. 13 No. 1 to , 20164.
Prof. Dr. Alexandre SemenovMoscow, Russia
Prof. Dr. Atef S. SadikTaif, Saudi Arabia
Prof. Dr. Sher Muhammed MangrioUni. Of Sindh, Jamshoro, Pakistan
Prof. Dr. Wei MaShanghai, China
Prof. Dr. Jian He Xu Shanghai, China
Prof. Dr. Ghasem D. NajafpourUniv. Babool, Iran
Prof. Dr. Ozlem OksoyKocaeli, Turkey
Prof. Dr. Muhammed Umar DahoUni. Of Sindh, Jamshoro, Pakistan
Prof. Dr. Muhammed Hanif NoomrioHyderabad, Pakistan
Prof. Dr. M. R. ZamaniNIGEB, Tehran, Iran
Dr. Nisa Rachmania MubarikBogor Agriculture University, Indonesia
Prof. Dr. P. VenkatachalamPeriyar University, Salem-636011, TN, India
Prof. Dr. Hong Kyoon NoCatholic University of Daegu, Gyeongsan, South Korea
Prof. Dr. Ismael KhatabKafrelsheikh University, Egypt
Dr. Sedky H. A. HassanAssiut University, Egypt
Dr. Mostafa RahimnejadUniv. Babool, Iran
Dr. Dang T.M. QuyenDanang city - Vietnam
Dr. Muhammed RafiqUni. Of Sindh, Jamshoro, Pakistan
Dr. Altaf Ahmed SemiarShanghai, China
Dr. Afsheen AmanUniv. Karachi, Pakistan
Dr. S. Habib Ahmed NaqviUniv. Of Sindh, Jamshoro, Pakistan
Dr. Ameer KhusroChenni, India
Dr. Bilal Haider AbbasiIslamabad, Pakistan
Dr. Mahdev MaliKunming, China
Dr. S. RajalakshmiBotswana Intl. Univ. of Sci. and Technol. Botswana
Dr. Mehboob Ali sialiNIA Tando Jam, Pakistan