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EDITORIAL BOARD

2014

KNOWLEDGE, ATTITUDE AND PRACTICES OF LOCAL POPULATIONS OF UNION COUNCIL KOAZ BAHRAM DEHRI PAKISTAN TOWARDS GREEN TEA (CAMELLIA SINESIS)

By Tauseef Ahmad*1, Haroon2 , Arif Jan2 , Sikander Khan Sherwani3 , Sana Ullah4 , Naseer Ullah5 , Muhammad Ayub Jadoon1 , Muhammad Waqar6 , Akbar Hussain1 , Abdul Majid1 , Afzal Khan1 , Naiz Ali7 , Tanwe

VOL-11 NO-(2)

Abstract

After water the tea is the biggest beverage use worldwide. This study is aimed to find out the knowledge, attitude and practices of local population of Union Council Koaz Bahram Dehri District Charsada, Khyber Pakhtunkhwa Pakistan towards green tea. A descriptive observational study was design. Data were collected from 150 subjects through a self design questionnaire. The majority of participants were males belonging to different age groups. The ratio of education was reported low. The local populations have sufficient knowledge about green and also used it for different purposes.

EFFECT OF INDOL-3 ACETIC ACID ON THE BIOCHEMICAL PARAMETERS OF Achoria grisella HEMOLYMPH AND Apanteles galleriae LARVA

By Fevzi Ukan, Havva Kbra Soydaba?* Rabia zbek

VOL-11 NO-(2)

Abstract

Biochemical structures such as lipid, protein, sugar and glycogen are known to play a pivotal role on the relationship between host and its parasitoid. Any changes in these parameters may have potential to alter the balance of the host-parasitoid relation. Taking this into account, the effects of plant growth regulator, indol-3 acetic acid (IAA) on the biochemical parameters of the host and parasitoid were investigated. Achoria grisella Fabricus (Lepidoptera: Pyralidae) is a serious pest and causes harmful impacts on honeycomb. Endoparasitoid Apanteles galleriae Wilkinson (Hymenoptera: Braconidae) feeds on the hemolymph of the A. grisella larva and finally causes mortality of the host. Different concentrations (2, 5, 10, 50, 100, 200, 500, and 1,000ppm) of IAA were added to the synthetic diet of host larvae. Protein, lipid, sugar, and glycogen contents in hemolymph of host and in total parasitoid larvae were determined by Bradford, vanillin-phosphoric acid, and hot anthrone tests using UV-visible spectrophotometer, respectively. Protein level in host hemolymph increased upon supplement of each doses of IAA except for 10ppm. IAA application enhanced the level of sugar at 100 and 200ppm whereas a decrease was observed in lipid at 5, 10, 200, and 1,000ppm doses in host. All doses were effective on the parasitoid larvae. Nevertheless, the most effective dose was 50ppm, which increased glycogen but decreased lipid amount. Similarly, trearment of 1000ppm IAA increased protein level and 100ppm reduced level of sugar. Our study indicated that application of IAA resulted in different effects on the amount of biochemical structures associated with the hemolymph of pest species and its natural enemy. Therefore, results showed that not only the target but also the non-target organisms exposed to widely-used plant growth regulators may be affected and this, in turn may also change the host-parasitoid interaction.

POTENTIAL OF SOME NATIVE BACILLUS STRAINS TO PROMOTE GROWTH OF TOMATO

By Sana Hanif1 , Tehmina Anjum1 , Sabin Fatima1 , Amna Ali1 ,Asrar Mahboob2 and Waheed Akram1*

VOL-11 NO-(2)

Abstract

Plant growth promoting bacteria are soil inhabiting around or on the root surface and are involved in plant growth promotion and development via direct or indirect mechanisms. Some native strains of Bacillus were evaluated for their ability to promote growth of tomato. Different in vitro biochemical assays indicated capabilities of these bacterial agents for producing Indole acetic acid, siderophores and phosphate solubilization. Pot experiments indicated ability of B. megaterium FCBP520 strain to induce maximum significant increase in growth parameters like root length, shoot length and total biomass in tomato plants. B. subtillis FCBP170 was second best performer to induce growth promotion in tomato plants. Calorimetric assays were performed to quantify changes in plant growth related biochemicals. Both these strains induced plants for significantly higher production of total chlorophyll, anthocyanin and carotenoid contents. Overall this study presents the potential of B.megaterium FCBP520 and B.subtillis FCBP 170 to promote growth of tomato and the mechanism behind growth promotion with respect to both bacterial inducer and tomato plants. Our results may be the first to demonstrate plant growth promotion by these two bacterial strains during vegetation growth of tomato. This study will also be functionally relevant for future studies directed towards better tomato growth and ultimately better yields in agriculture system of Pakistan

ESTIMATES OF HERITABILITY AND EXPECTED RESPONSE FOR MATURITY AND GRAIN YIELD RELATED TRAITS IN HALF-SIB RECURRENT FAMILIES OF MAIZE

By Muhammad Ishaq1,2* , Ghulam Hassan1 , Hidayatur Rahman1 , Muhammad Iqbal2 , Ibni Amin Khalil1 , Sabaz Ali Khan3 , Sheraz Ahmad Khan1 , Rafiullah1 and Jamal Hussain3

VOL-11 NO-(2)

Abstract

Genetic improvement of the segregating populations is an important aspect of the maize breeding programs. This research was undertaken to estimate heritability and expected responses for maturity and grain yield related traits in maize. For this purpose, one hundred and forty-four half-sib families that were derived from Sarhad white maize population were used. These half-sib families were statistically evaluated in a partially balanced 12x12 lattice design. The heritability estimates varied from 0.28 to 0.88 and were high for days to tasseling days to silking, days to pollen shedding, ear height, kernels row-1 , grain yield, medium plant height, 1000 kernel weight and low for kernels ear-1 . The genetic and environmental coefficients of variation varied from 1.85 to 16.13 and 1.64 to 13.85, respectively. The index of variation ranged from 0.44 to 1.92 among the traits. Based on the values of the heritability and selection differential the highest expected response was observed for grain yield (900.99 kg ha-1 ) followed by 1000 kernel weight (2.51 g), plant height (3.26 cm) and ear height (2.72 cm). These high heritability and high index of variation estimates (I.V >1) for various important traits indicates greater heterogeneity and genotypic differences among the tested half-sib families. We have shown for the first time that this improved recurrent population Sarhad white has the potential to recurrent selection and could be used in future maize breeding programs. Moreover, the improved recurrent population might be used by poor farmers to save production cost and to use the seeds for future farming.

THERMOSTABLE RECOMBINANT ESTERASE PRODUCTION IN 3-L STIRRED TANK BIOREACTOR, PURIFICATION AND CHARACTERIZATION

By Nadia A. Soliman*a , Yasser R. Abdel-Fattaha , Samar M. Yousef a , Ehab R. El-Helowb

VOL-11 NO-(2)

Abstract

Thermophilic lipases/esterases are currently attracting enormous attention because of their biotechnological potential. It was thus aimed in the present study to optimize enzyme on large scale production, to purify and characterize the produced enzyme. The est fragment of Geobacillus sp. AZ1 (ac: KM823656) was cloned directly by PCR into the pCYTEXP1 expression vector under the control of lambda promoter. Good intracellular expression of the studied gene was obtained in Escherichia coli DH5?. Optimization of the recombinant protein production was carried out using a 3L bench-top bioreactor with a stirred tank. The results showed that the fermenter condition and induction by shifting temperature from 37C to 42C at OD. 0.7 resulted in a 2-fold increase. The expressed protein was fused C-terminally with 6x-his tag to allow one step purification using IMAC (Immobilized Metal Affinity Chromatography). A 28kD protein was achieved. The kinetic characterization of the purified enzyme exhibited maximum activity at 50C and pH 7.4. The enzyme had a considerable thermal stability. The percentage of remained activity after 1h exposure to 50, 55 and 60C reached to 50, 42 and 27%, and retained more than 60% of its original activity at 65C for 15min. However, exposure of crude enzyme to these conditions showed a complete stability and more than 90% of activity. The enzyme was also highly stable in a pH range of 3-10 for 24h. The enzyme activity was promoted in the presence of Co+2, K+1, Ca+2 and Fe+2 and was inhibited by Mn+2, Mg+2, Cu+2, Hg+2 , Zn+2. Diethyl-ether, and acetone but hexane enhanced the activity. On the contrary nbutanol, DMSO, ethanol, isopropanol, glycerol, methanol and chloroform, reduce the enzyme activity. EGTA, DTT, EDTA, PMSF and SDS decreased the enzyme activity, whereas the presence of urea, oxidizing and reducing agents, some non-ionic surfactants increased the enzyme activity. The values of Km and Vmax as calculated from the Lineweaver- Burk plot were 12.66 mM and 333.33 U/mg protein respectively.

MOLECULAR DIVERSITY IN SOME A-GENOME WHEAT AMPHIPLOIDS (2n=6x=42; BBAAAA)

By Sania Ahmed1 , Hadi Bux2* , Alvina Gul-Kazi3 , Abdul Wajid Channa2 , Sadaf Tabasum Qureshi2 , Aijaz Ahmed Soomro4 ,Mahboob Ali Sial5 , Abdul Rauf1 and Abdul Mujeeb-Kazi6

VOL-11 NO-(2)

Abstract

Wild relatives of wheat have defying capability against detrimental conditions as they possess rich reservoirs of valuable genes. Through hybridization, many desired traits have been successfully introgressed from wild relatives to cultivars for various stress tolerances for wheat improvement. Wheat amphiploids (BBAAAA) have been created from diploid resources ( Triticum monococum (AA), Triticucm urartu (AA), and Triticum bioeticum (AA) and Triticum turgidum the tetraploid durum (BBAA) wheat cultivars through bridge crossing. These amphiploids possess enormous variability for biotic and abiotic stresses. In current study, molecular characterization of a collection of 79 amphiploids (2n=6x=42, BBAAA) by 25 SSR primers have been carried out. The molecular scanning produced 58 polymorphic bands and all were polymorphic showing 100% polymorphism. Dendrogram based on Nei and Lis similarity coefficient, clearly distinguished the genotypes in the clusters showing abundant diversity. The genetically diverse germplasm identified through genetic similarity and cluster analysis in current study are accession 13, 16, 42, 52 and 50. These amphiploids received the A genome from diploid Triticum bioeticum. The selected collection should be used for the genetic improvement of wheat and the selected collection needs further studies to reveal the hidden desirable variability of agricultural utility.

PLASMID PROFILING AND RAPD MARKER ANALYSIS OF PAKISTANI GROWTH PROMOTING BACTERIA

By Sadaf Tabasum. Qureshi1 *, Hadi Bux1 , Syed Habib Ahmed Naqvi2 , Muhammed Rafiq2 , Aneela Naz Soomro3 , Mahjabeen Panwahar1

VOL-11 NO-(2)

Abstract

Nineteen chickpea (Cicer arietinum L.) and eleven green pea (Pisum sativum L.) root nodule rhizobia collected from different localities of Pakistan were characterized for plasmid profiles and genomic diversity. Plasmid profile analysis and RAPD finger printing revealed genetic diversity among studied chickpea and green pea rhizobia. Cluster analysis of plasmid categorized chickpea in four groups and green pea into five groups at the distance of 0.3 and 1.5 respectively. Plasmid numbers varied from 1 to 3, while size of plasmid varied from 2.3kb to 33kb for chickpea isolates. Plasmid number for green pea rhizobia varied from 1 to 2 and size from 9.4kb to 33kb. Soil isolates nodulating both crops showed more variations for plasmid size than plasmid number. Chickpea rhizobia were less diverse for plasmid number and size than green pea rhizobia. Cluster analysis of RAPD banding patterns revealed three clusters for chickpea and six clusters for green pea. Therefore RAPD analysis is an efficient discriminatory method for genetic diversity estimation of Rhizobium strains.

INCIDENCE OF A BRIGHTER VARIANT OF VIBRIO HARVEYI STRAIN N2 UNDER CONTINUOUS LAB CULTURING

By Aisha Nawaz* and Nuzhat Ahmed

VOL-11 NO-(2)

Abstract

A brighter variant of a lab cultured strain of Vibrio harveyi was observed under routine lab culturing procedures and was investigated for differences in luminescence characteristics and protein profile as compared to the wild type parent. A lysate of the variant and wild type strain was subjected to BCA protein analysis and proteins were separated on PAGE to elucidate differences. Luminescence studies revealed a faster growing, 20% brighter variant that had a clear difference in protein profiles when subjected to quantitative and qualitative protein analyses. The bright variant of N2 had 0.21gm/l more total protein content as compared to the wild type and exhibited darker protein bands for some of the proteins separated on PAGE.

EFFECTIVY OF THYROXINE HORMONE SUPLEMENTATION IN THE OVARIAN MATURATION OF FEMALES MUD CRAB (SCYLLA SERRATA)

By Heppi Iromo1 , M. Zairin Junior1 , M. Agus. S1 ., Wasmen Manalu2 .

VOL-11 NO-(2)

Abstract

The aim of this research was to study the effectiveity of thyroxine hormone to increase ovarian maturition of broodstock mud crab (scylla serrata). The thyroxine hormone has been tested to increase survival and growth of larvae fish but have yet on crab. The weight of broodstock mub crab was 350-400 g. The thyroxine hormone used in this research of levothyroxine sodium tablets production by NV organon, Oss, The Netherland and each tablet have contains 100 g of thyroxine. This study used treatment with more doses of the thyroxine hormone supplementation were; with doses 0 (control), 0.05ug/BW (low dose); and 0.5ug/BW (high dose). The analysis of variance that the suplementation thyroxine hormone low dose of broodstock mud crab were significant effect (P

ELIMINATION OF HYDROGEN SULFIDE FROM SOUR GAS IN CSTR BIOREACTOR USING NATIVE ISOLATED STRAIN OF SULFUR OXIDIZING BACTERIA

By Hamid Heydarzadeh1 , Ghasem D. Najafpour*1 , Aliasghar Ghoreishi1 and Habibollah Younesi

VOL-11 NO-(2)

Abstract

Biodesulfurization of sour gas has been carried out in a continuous stirred tank bioreactor with culture volume of 3.5 L. Pure culture of sulfur oxidized bacteria was isolated from Ramsar (Iran) hot spring. Bioreactor performance was evaluated in terms of elimination capacity (EC, mg/L.h) and removal efficiency (RE, %). The highest RE% of H2S at high concentration of sour gas was determined to be 99%. The effect of agitation rate on the removal efficiency was evaluated at four different gas flow rates ranged in 0.5 to 2mL/min. The optimum gas flow rate was achieved 0.5mL/min. In addition, the achieved optimal dilution rate and cell productivity were 0.01 1/h and 0.018 g/L/h, respectively. Furthermore, the limitation of dilution rates in the bioreactor and wash-out phenomena were considered. The biodegradation kinetic values obtained from Michaelis Menten model. The proposed model was satisfactorily fitted with the experimental data with three agitation rates in the bioreactor operation for the purpose of biodesulfurization of natural gas.

ISOLATION, OPTIMIZATION AND PRODUCTION OF BIOPOLYMER (POLY 3-HYDROXY BUTYRATE) FROM MARINE BACTERIA

By Muthezhilan R*1 , C.P. Kaarthikeyan1,3 , M. Jayaprakashvel1 , A. Jaffar Hussain2

VOL-11 NO-(2)

Abstract

The synthetic polymer plastics become an integral part of contemporary life. Excess use of plastics and indiscriminate dumping of it in soil and water is polluting the environment and other living organisms. To overcome this problem, the production and applications of eco-friendly biodegradable products (such as bioplastics) from microbes are becoming inevitable from the last decade and also good alternatives of synthetic polymers. Keeping this point in mind the present study aimed at isolating and identifying the poly 3-hydroxybutyrate (PHB) producing bacteria from marine sources that can be effectively utilized for the synthesis of bioplastics. For isolation of poly 3-hydroxybutyrate producing bacteria, spread plate technique was followed using E2 mineral medium. After incubation, based on the morphological characteristics 32 strains were isolated and identified from the sand dunes plants of rhizosphere vegetation of Chennai coast. While staining with Sudan Black, six strains viz., AMET 5103, AMET 5111, AMET 5113, AMET 5121, AMET 5124 and AMET 5128 were identified as poly 3-hydroxybutyrate producing bacterial strains. Based on the dry weight of total biopolymer content, the strain AMET 5111 showed the maximum accumulation and was selected for optimizing at different pH, temperature, salinity, carbon and nitrogen source, incubation period and peak time of poly 3- hydroxybutyrate accumulation. Based on the biochemical tests, the strain AMET 5111 was found to be Pseudomonas spp. The optimum pH, temperature and salinity were found to be 7.0, 30oC and 5%. The peak time of poly 3-hydroxybutyrate accumulation was found to be 36hrs. The best carbon and nitrogen sources were found to be sucrose and (NH4)2HPO4. The selected strain was massively cultured using the optimized media and poly 3-hydroxybutyrate was extracted by solvent extraction.

KNOWLEDGE, ATTITUDE AND PRACTICES OF LOCAL POPULATIONS OF UNION COUNCIL KOAZ BAHRAM DEHRI PAKISTAN TOWARDS GREEN TEA (CAMELLIA SINESIS)

By Tauseef Ahmad*1, Haroon2 , Arif Jan2 , Sikander Khan Sherwani3 , Sana Ullah4 , Naseer Ullah4 , Muhammad Ayub Jadoon1 , Muhammad Waqar5 , Akbar Hussain1 , Abdul Majid1 , Afzal Khan1 , Naiz Ali6 , Tanwe

VOL-11 NO-(2)

Abstract

After water the tea is the biggest beverage use worldwide. This study is aimed to find out the knowledge, attitude and practices of local population of Union Council Koaz Bahram Dehri District Charsada, Khyber Pakhtunkhwa Pakistan towards green tea. A descriptive observational study was design. Data were collected from 150 subjects through a self design questionnaire. The majority of participants were males belonging to different age groups. The ratio of education was reported low. The local populations have sufficient knowledge about green and also used it for different purposes.

OPTIMIZATION OF SARDINE (SARDINELLASP.) OIL REFINING USING RESPONSE SURFACE METHOD (RSM)

By Sugeng Heri Suseno1 , Jeny Tambunan, Bustami Ibrahim, Saraswati, Sri Hayati and Ayu Fitri Izaki

VOL-11 NO-(1)

Abstract

RSM (Response Surface Method) is a set of mathematical techniques and statistics that are useful to analyze the problems so that the level of response was influenced by several variables. This research aimed to determine the optimal treatment of sardine (Sardinella sp.) oil refining through centrifugation and adsorption. Based on the results, it can be concluded that the optimum treatment was centrifugation at 9494.89 rpm for 31.48 minutes with addition of combined adsorbents attapulgite and bentonite (1:1) 3%. The optimum point resulted a peroxide value at 2.97 meq/Kg, p-anisidine value at 0.81 meq/Kg, totox value at 6.32 meq/kg, and free fatty acid at 4.07%

DETERMINATION OF FATTY ACIDS AND ELEMENTS FROM COCONUT (COCOS NUCIFERA) SHELL

By Nasreen K. Thebo1 , Altaf A. Simair2 *, Wazir A. Sheikh1 , Sher M. Mangrio1 , Pirkash L. Nagni3 , Sughra G. Mangrio2 and Hassan M. Nizamani4

VOL-11 NO-(1)

Abstract

GCMS analysis showed twenty eight fatty acids, 12 n-octanoate, dodecanotic acid, n-hexadecanoic acid, n-heptaecenoate, tridecatrienoate acid methyl-2-tridecynote and essential elements were isolated from the shell of Cocos nucifera viz., along with calcium, cadmium, chromium, copper, iron, lead, potassium, magnesium, manganese, nickel and zinc. Present study revealeds that accumulation of tetradeconic acid was higher among saturated fatty acids among metals magnesium and iron (593.06 42.17, 997.9 52.25) respectively were highest following by low accumulation of lead (1.46 0.21) was recorded in coconut shell.

THE OIL EXTRACTION FROM EEL (ANGUILLA BICOLOR BICOLOR) BONE AS BY-PRODUCT FROM KABAYAKI PROCESSING INDUSTRY

By Irfan Syahbana and Sugeng Heri Suseno*

VOL-11 NO-(1)

Abstract

Fish oils have been recognized as good sources of polyunsaturated fatty acids (PUFA) which are widely used for pharmaceutical purposes and as food supplements. In this study, fish oil from eels (Angguilla bicolor bicolor) bone were extracted using a Bligh and Dyer methods. The fatty acid composition of the oil was analyzed and quantified using gas chromatography. Results showed that the total yield of by-product of Eel (Anguilla bicolor bicolor) reached 26,38 %, the lipid content of eels bone was 17.33 0.58 g/100 g. Yield of eel bone oil extracted by Bligh and Dyer method was 17.12%. In the fatty acid analysis of eel bone oil, it was discovered that SFA was 19.87%, MUFA was 25.84%, and PUFA was 13.84%. The major fatty acids in the oil from the bone were palmitic acid (13.58%), oleic acid (20.94%), linoleic acid (4.01%), EPA (1.57%), and DHA (4.84%).

IN VITRO REGENERABILITY OF DIFFERENT SUGARCANE (SACCHARUM OFFICINARUM L.) VARIETIES THROUGH SHOOT TIP CULTURE

By Mangrio G. Sughra, Simair A. Altaf1*, Rind M. Rafique , Mangrio S. Muhammad2 , Shereen N. Rind Balouch3 and Dahot M. Umar

VOL-11 NO-(1)

Abstract

An experiment was conducted to develop an efficient protocol for micropropagation of sugarcane using the method of growing shoot tips of three varieties of sugarcane viz. BL-4, Thatta-10, and Larkana-2001. A protocol for the regeneration of direct shoot without the intervention of the callus phase was developed using shoot tip culture in a basic medium of Murashige and Skoog (MS) supplemented with different concentrations of auxin (NAA) and cytokinin (BAP). The analysis of variance due to varieties, concentration and variety x concentration interaction was significant for all characters. In general, the best results were seen from the BL-4 and Thatta-10 varieties with 1.0 mg/L BAP and 3.0 mg/L NAA for various parameters. Different concentrations of BAP for shoot initiation and multiplication were used and 1.0 mg/L BAP showed the most effective concentration for induction and shoot multiplication, while MS medium supplemented with different concentrations of NAA for in vitro formation of roots from proliferated shoots, the maximum root formation on MS medium supplemented with 3.0 mg/L NAA was recorded. However, BL-4 had high power (85.3 %) of regenerating from explants took minimum (11.00) days for shoot induction and gave highest number of shootlets (6.50) with maximum length of shootlets (5.50 cm) by 1.0 mg/L BAP. For root induction BL-4 variety produced higher number of roots per shootlet (6.80) after minimum 9 days with maximum length of the roots (2.50 cm) at 3.0 mg/L NAA.

BATCH AND CONTINUOUS PRODUCTION OF LACTIC ACID USING LACTOBACILLUS BULGARICUS (ATCC 8001)

By Hamidreza Ghafouri Taleghania , Ghasem D. Najafpoura* , Ali Asghar Ghoreyshib

VOL-11 NO-(1)

Abstract

In this work lactic acid was produced in batch and continuous fermentation processes. The objective of this work was to investigate the effects of initial substrate concentration (30-120 g/l) and hydraulic retention time (HRT: 10-40 h) on cell growth, lactose utilization and lactic acid production. Whey lactose was used in batch and continuous systems; obtained results were compared. Substrate and product inhibitions were investigated at high initial substrate concentration. The results of continuous system indicate that production of lactic acid has increased with an increase in HRT along with increasing substrate concentration. The optimum initial lactose concentration for maximal lactic acid production in batch and continuous fermentation were found to be 90 and 120 g/l, respectively. The maximum concentrations of lactic acid in batch and continuous cultures were 32.1 and 42.9 g/l, respectively. The results showed that optimal lactic acid production was obtained at HRT of 30 hours. Mathematical projection model and kinetic studies for lactic acid production in batch cultures were performed. It was found that initial lactose concentration of 60g/l resulted in maximum specific growth rate of 0.215h-1 .

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, 2014 to December 31, 2014 for Vol. 11 No. 1 and 2, 2014.
Prof. Dr. Alexandre SemenovMoscow, Russia
Prof. Dr. Atef S. SadikTaif, Saudi Arabia
Prof. Dr. Sher Muhammed MangrioUni. Of Sindh, Jamshoro, Pakistan
Prof. Dr. Jian He XuShanghai, China
Prof. Dr. Muhammed Umar Dahot Uni. Of Sindh, Jamshoro, Pakistan
Dr. S. A. Anitha ChristyHouston, USA
Dr. Mostafa RahimnejadUniv. Babool, Iran
Dr. Muhammed RafiqUni. Of Sindh, Jamshoro, Pakistan
Dr. Muhammed Yakoub KhanUni. Of Sindh, Jamshoro, Pakistan
Dr. S. Habib Ahmed NaqviUni. Of Sindh, Jamshoro, Pakistan
Dr. Imtiaz Ahmed KhanNIA Tando Jam, Pakistan
Dr. Mehboob Ali sialiNIA Tando Jam, Pakistan