Indian Journal of Experimental Biology

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VOLUME 51

NUMBER 4

APRIL 2013

CODEN: IJEB (A6) 51 (4) 269-342 (2013)

ISSN: 0019-5189 (Print); 0975-1009 (Online)

 

CONTENTS

 

 

Mini Review

 

 

 

Population genetic structure of malaria vector Anopheles stephensi Liston (Diptera:Culicidae)

273

S K Gakhar, Richa Sharma & Arvind Sharma

 

 

 

Papers

 

 

 

Evaluation of vaccine candidate potential of ΔaroA, ΔhtrA and ΔarohtrA mutants of Salmonella enterica subspecies enterica serovar Abortusequi in guinea pigs

280

 

 

Bhoj Raj Singh, Mudit Chandra, Dhananjoy Hansda, Javed Alam4, Narayanan Babu, Mehtab Z Siddiqui, Ravi K Agrawal & Gautam Sharma

 

 

 

Effect of Vanillin on lipid profile in a model of hyperlipidemia, a preliminary study

288

Yogesh Belagali, Sheetal D Ullal, Ahsan Shoeb, Vani Bhagwath, Ramya K, Rakshitha Maskeri

 

 

 

Effect of thymoquinone on ethanol and high fat diet induced chronic pancreatitis—a dose response study in rats

292

 

 

Periyanayagam Suguna, Arumugam Geetha, Ravikumar Aruna & Ganesan Vijaiyan Siva

 

 

 

Comparative effect of cane syrup and natural honey on abdominal viscera of growing male and female rats

303

Abdulwahid Ajibola, Joseph P Chamunorwa & Kennedy H Erlwanger

 

 

 

Effect of chinese traditional herb Epimedium grandiflorum C.Morren and its extraction Icariin on osteoarthritis via suppressing NF-kB pathway

313

Zhang Wenliang, Li Rongheng, Wang Shumei, Mu Fangzheng & Jia Ping

 

 

Statistical and evolutionary optimization for enhanced production of an anti-leukemic enzyme,
L-asparaginase, in a protease-deficient Bacillus aryabhattai ITBHU02 isolated from the soil contaminated with hospital waste

322

Yogendra Singh & S K Srivastava

 

 

 

Statistical medium optimization of an alkaline protease from Pseudomonas aeruginosa MTCC 10501, its characterization and application in leather processing

336

 

 

Naidu Ramachandra Boopathy, Devadas Indhuja, Krishnan Srinivasan, Mani Uthirappan, Rishikesh Gupta, Kamini Numbi Ramudu & Rose Chellan

 

 

_______________________

 

Announcement

 

Indian Council of Medical Research

 

ICMR International Fellowships

(2013-14)

 

The Indian Council of Medical Research (ICMR) invites applications from Indian biomedical scientists for the international fellowships for the year 2013-14.

Number of fellowships: Young scientists: 12; Senior scientists: 06

Duration: 3-6 months for Young scientist; 10-15 days for senior scientist

 

The last date of receipt of applications is 15 April 2013.

 

            For further details and format of application visit ICMR website: http://www.icmr.nin.in

 

____________________

 

 

Editor’s Note

The Indian Journal of Experimental Biology is covered by the following international abstracting and indexing services:

 

Science Citation Index ExpandedTM

PubMed (http://www.ncbi.nim.nih.gov/)

MEDLINE

BIOSIS

Chemical Abstracts Service

Excerpta Medica

Informascience

Refrativnyi Zhurnal

Zoological Records

 

Author Index

Agrawal Ravi K

280

Ajibola Abdulwahid

303

Alam Javed

280

Aruna Ravikumar

292

 

 

Babu Narayanan

280

Belagali Yogesh

288

Bhagwath Vani

288

 

 

Chamunorwa Joseph P

303

Chandra Mudit

280

Chellan Rose

336

 

 

Erlwanger Kennedy H

303

 

 

Fangzheng Mu

313

 

 

Gakhar S K

273

Geetha Arumugam

292

Gupta Rishikesh

336

 

 

Hansda Dhananjoy

280

 

 

Indhuja Devadas

336

 

 

K Ramya

288

 

 

Maskeri Rakshitha

288

 

 

Numbi Ramudu Kamini

336

 

 

Ping Jia

313

 

 

Ramachandra Boopathy Naidu

336

Rongheng Li

313

 

 

Sharma Arvind

273

Sharma Gautam

280

Sharma Richa

273

Shoeb Ahsan

288

Shumei Wang

313

Siddiqui Mehtab Z

280

Singh Bhoj Raj

280

Singh Yogendra

322

Srinivasan Krishnan

336

Srivastava S K

322

Suguna Periyanayagam

292

 

 

Ullal Sheetal D

288

Uthirappan Mani

336

 

 

Vijaiyan Siva Ganesan

292

 

 

Wenliang Zhang

313

 

Keyword Index

16S rRNA gene

322

 

 

Abdominal viscera

303

Anopheles stephensi

273

Antioxidants

292

 

 

Bacillus aryabhattai

322

 

 

Cane syrup

303

 

 

Dehairing

336

 

 

Epimedium grandiflorum

313

Ethanol

292

 

 

FAME analysis

322

 

 

Gastrointestinal tract

303

Genetic markers

273

Growing rats

303

Guinea pig

280

 

 

High fat diet

292

Honey

303

Hypertriglyceridemia

288

 

 

Icariin

313

Inflammation

292

ITBHU02

322

 

 

L-asparaginase

322

Lipid profile

288

 

 

Malaria

273

Microsatellite

273

Mucosal

280

 

 

NF-κB (P65)

313

 

 

Osteoarthritis

313

Pancreatitis

292

Population genetics

273

Protease

336

Protease-deficient

322

Pseudomonas aeruginosa

336

 

 

Response surface     methodology

336

 

 

Salmonella Abortusequi

280

 

 

Thymoquinone

292

 

 

uPA

313

 

 

Vaccine

280

Vanillin

288

 

 

ΔaroA

280

ΔhtrA

280

 

            Correspondent author is marked by *

 

 

Indian Journal of Experimental Biology

Vol. 51, April 2013, pp. 273-279

 

 

Population genetic structure of malaria vector Anopheles stephensi Liston (Diptera: Culicidae)

S K Gakhar*, Richa Sharma & Arvind Sharma

Centre for Biotechnology, Maharshi Dayanand University, Rohtak 124 001, India

Malaria is a complex disease that afflicts human today. Malaria epidemiology is associated with drug resistance in parasite and differential distribution and insecticide resistance in vector. Efforts are being made to eradicate malaria but burden of malaria is still increasing. Vector control is essential for malaria prevention strategies. Knowledge of population genetic structure is pre-requisite for determining prevention strategies, particularly using transgenic mosquitoes. Population genetic study can predict level of gene flow between different populations. Anopheles stephensi Liston is urban vector of malaria in Indo-Pakistan subcontinent. About 12% of malaria cases of malaria in India are contributed by A. stephensi. Studies conducted on population genetics of A. stephensi using various markers in different parts of the world are discussed in this communication.

 

 

Indian Journal of Experimental Biology

Vol. 51, April 2013, pp. 280-287

 

 

Evaluation of vaccine candidate potential of ΔaroA, ΔhtrA and ΔarohtrA mutants of Salmonella enterica subspecies enterica serovar Abortusequi
in guinea pigs

 

Bhoj Raj Singh1*, Mudit Chandra2, Dhananjoy Hansda3, Javed Alam4, Narayanan Babu5, Mehtab Z Siddiqui6,
Ravi K Agrawal1 & Gautam Sharma5

National Salmonella Centre (Vet)

Centre for Animal Disease Research and Diagnosis, Indian Veterinary Research Institute (IVRI), Izatnagar 243 122, India

Received 17 August 2012; revised 10 January 2013

Salmonella enterica subspecies enterica serovar Abortusequi (S. Abortusequi), a host adapted Salmonella causes abortions, still births and foal mortality in equids. Though known since more than 100 years, it is still a problem in many of the developing countries including India. There is dearth of really good vaccine affording immunity lasting at least for one full gestation. In search of a potential vaccine candidate, three defined deletion mutants (ΔaroA, ΔhtrA and ΔarohtrA) of S. Abortusequi were tested in guinea pig model for attenuation, safety, immunogenicity, humoral immune response, protective efficacy and persistence in host. The ΔhtrA and ΔarohtrA mutants were found to be safe on oral inoculation in doses as high as 4.2×109 cfu/animal. Also through subcutaneous inoculation ΔarohtrA mutant did not induce any abortion in pregnant guinea pigs. All the three mutants did not induce any illness or death in 1-2 week-old baby guinea pigs except ΔhtrA mutant which caused mortality on intraperitoneal inoculation. Inoculation with mutants protected against challenge and increased breeding efficiency of guinea pigs. After >4.5 months of mutant inoculation, guinea pigs were protected against abortifacient dose of wild type S. Abortusequi and mother guinea pigs also conferred resistance to their babies to the similar challenge. Early humoral immune response of S. Abortusequi mutants was characteristic. Faecal excretion of ΔaroA and htrA mutants was detected up to 45 days of inoculation in guinea pigs while ΔarohtrA mutant could not be detected after 21 days of inoculation. The results indicated that the double deletion mutant (ΔarohtrA) was the most effective and safe candidate for vaccination against S. Abortusequi through mucosal route of inoculation.

 

 

Indian Journal of Experimental Biology

Vol. 51, April 2013, pp. 288-291

 

 

Effect of Vanillin on lipid profile in a model of hyperlipidemia,
a preliminary study

Yogesh Belagali, Sheetal D Ullal*, Ahsan Shoeb, Vani Bhagwath, Ramya K, Rakshitha Maskeri

Department of Pharmacology, Kasturba Medical College, Manipal University, Mangalore 575 001, India

Received 3 August 2012; revised 23 January 2013

To evaluate the effect of vanillin on the lipid profile of high fat diet induced hyperlipidemia in rats, the hyperlipidemia was induced by feeding cholesterol-rich high fat diet for 45 days in wistar rats of either sex. The reduction in the triglycerides and VLDL-C was significant at 200 & 400 mg/kg dose of vanillin compared to atorvastatin group. Reduction in total cholesterol was significant at 200 and 400 mg/kg doses compared to hyperlipidemic control. The results demonstrate that vanillin at a dose of 200 and 400 mg/kg body weight lowers the serum triglyceride, VLDL-C and total cholesterol level significantly in high fat diet induced hyperlipidemic rats. However there was no significant effect on the lipid profile at 100 mg/kg dose. There were no statistically significant changes in the HDL-C and LDL-C levels at any of the given doses.

 

 

Indian Journal of Experimental Biology

Vol. 51, April 2013, pp. 292-302

 

 

Effect of thymoquinone on ethanol and high fat diet induced chronic pancreatitis—a dose response study in rats

Periyanayagam Suguna1, Arumugam Geetha1*, Ravikumar Aruna1 & Ganesan Vijaiyan Siva2

1Department of Biochemistry, Bharathi Women’s College, Affiliated to University of Madras, Chennai 600 108, India

2Department of Biotechnology, University of Madras, Chennai 600 025, India

Received 31 July 2012; revised 6 December 2012

A significant increase in serum lipase, amylase, capase-1 and myeloperoxidase activities, oxidative stress index (OSI),
IL-1β and IL-18 was observed in rats receiving ethanol (EtOH) and high fat diet (HFD). Thymoquinone (TQ) supplementation along with EtOH and HFD significantly decreased the levels of serum lipase, amylase, capase-1, myeloperoxidase, OSI and maintained the antioxidant status when compared to untreated EtOH and HFD fed rats. Among the 4 doses, 100 mg of TQ/kg body weight was found to provide optimum protective effect on pancreas against EtOH and HFD induced abnormal changes. Histological observations added more evidence for the anti-inflammatory effect of TQ.

 

 

Indian Journal of Experimental Biology

Vol. 51, April 2013, pp. 303-312

 

 

Comparative effect of cane syrup and natural honey on abdominal viscera of growing male and female rats

Abdulwahid Ajibola1,3*, Joseph P Chamunorwa2 & Kennedy H Erlwanger3

1Department of Veterinary Physiology & Pharmacology, Faculty of Veterinary Medicine,
University of Ilorin, P.M.B 1515, Ilorin, Kwara State, Nigeria

2Department of Veterinary Anatomy and Physiology, Faculty of Veterinary Science,
University of Pretoria, Onderstepoort 0110, South Africa

3School of Physiology, Faculty of Health Sciences, University of the Witwatersrand,
7 York Road, Parktown 2193, South Africa

Received 10 September 2012; revised 1 January 2013

The high intake of refined sugars, mainly fructose has been implicated in the epidemiology of metabolic diseases in adults and children. With an aim to determine whether honey can substitute refined sugars without adverse effect, the long-term effects of natural honey and cane syrup have been compared on visceral morphology in growing rats fed from neonatal age. Honey increased the caecum and pancreas weights in male rats, which could enhance enzymatic activities of pancreas and digestive functions by intestinal microflora of caecum. Unlike honey, cane syrup caused fatty degenerations in the liver of both male and female rats. Honey enhanced intestinal villi growth, and did not cause pathology in the rodents’ abdominal viscera, suggesting potential nutritional benefit as substitution for refined sugars in animal feed.

 

 

Indian Journal of Experimental Biology

Vol. 51, April 2013, pp. 313-321

 

 

Effect of chinese traditional herb Epimedium grandiflorum C.Morren and its extract Icariin on osteoarthritis via suppressing NF-kB pathway

Zhang Wenliang1,2, Li Rongheng1*, Wang Shumei3*, Mu Fangzheng1 & Jia Ping1

1Department of Traditional Chinese and Western Medicine, the First Affiliated Hospital, Chongqing Medical University,
Chongqing, 400016, P.R.China

2Chongqing Electric Power Hospital , Chongqing, 400030, P.R.China

3College of Traditional Chinese Medicine, Chongqing Medical University, Chongqing, 401331, P.R.China

Received 19 July 2011; revised 21 December 2012

Osteoarthritis (OA), which is also called degenerative arthritis, is the leading cause of disabilities in the old people. The Chinese traditional herb Epimedium grandiflorum had long been found to attenuate osteoarthritis process, but the detailed mechanism was not clear. To study the mechanisms of E. grandiflorum in the treatment of osteoarthritis, rabbit osteoarthritis model combined with D-galactose was used. After different treatments for 10 weeks, cartilage sections were analyzed by immunohistochemistry for uPA, uPAR and PAI expression level. E. grandiflorum could significantly attenuate OA condition and decrease uPA, uPAR and PAI expression. The extract of E. grandiflorum, icariin also had a similar effect when compared with E. grandiflorum treatment alone. Rabbit chondrocytes were further isolated to be  stimulated by TNFα combined with different reagents treatment. Here, icariin treatment significantly reduced nuclear factor kappa B NF-κB (P65) activity, decreased uPA expression level and increased IκBα protein level. The results indicated that E. grandiflorum and its extract icariin could attenuate OA condition, reduce the expression of uPA and uPAR and increase PAI in experimental rabbit model and this effect may be conducted by suppressing NF-kB activity by increasing IκBα level.

 

 

Indian Journal of Experimental Biology

Vol. 51, April 2013, pp. 322-335

 

 

Statistical and evolutionary optimization for enhanced production of an anti-leukemic enzyme, L-asparaginase, in a protease-deficient Bacillus aryabhattai ITBHU02 isolated from the soil contaminated with hospital waste

Yogendra Singh & S K Srivastava*

School of Biochemical Engineering, Indian Institute of Technology, Banaras Hindu University, Varanasi 221005, India

Received 17 August 2012; revised 19 December 2012

Over the past few decades, L-asparaginase has emerged as an excellent anti-neoplastic agent. In present study, a new strain ITBHU02, isolated from soil site near degrading hospital waste, was investigated for the production of extracellular
L-asparaginase. Further, it was renamed as Bacillus aryabhattai ITBHU02 based on its phenotypical features, biochemical characteristics, fatty acid methyl ester (FAME) profile and phylogenetic similarity of 16S rDNA sequences. The strain was found protease-deficient and its optimal growth occurred at 37 °C and pH 7.5. The strain was capable of producing enzyme L-asparaginase with maximum specific activity of 3.02±0.3 Umg-1 protein, when grown in un-optimized medium composition and physical parameters. In order to improve the production of L-asparaginase by the isolate, response surface methodology (RSM) and genetic algorithm (GA) based techniques were implemented. The data achieved through the statistical design matrix were used for regression analysis and analysis of variance studies. Furthermore, GA was implemented utilizing polynomial regression equation as a fitness function. Maximum average L-asparaginase productivity of 6.35 Umg-1 was found at GA optimized concentrations of 4.07, 0.82, 4.91, and 5.2 gL‑1 for KH2PO4, MgSO4.7H2O, L-asparagine, and glucose respectively. The GA optimized yield of the enzyme was 7.8% higher in comparison to the yield obtained through RSM based optimization.

 

 

Indian Journal of Experimental Biology

Vol. 51, April 2013, pp. 336-342

 

 

Statistical medium optimization of an alkaline protease from Pseudomonas aeruginosa MTCC 10501, its characterization and application in leather processing

Naidu Ramachandra Boopathy*, Devadas Indhuja, Krishnan Srinivasan, Mani Uthirappan,
Rishikesh Gupta, Kamini Numbi Ramudu & Rose Chellan

Department of Biotechnology, CSIR-Central Leather Research Institute (CLRI), Adyar Chennai 600 020, India

Received 23 July 2012; revised 26 December 2012

Proteases are shown to have greener mode of application in leather processing for dehairing of goat skins and cow hides. Production of protease by submerged fermentation with potent activity is reported using a new isolate P. aeruginosa MTCC 10501. The production parameters were optimized by statistical methods such as Plackett-Burman and response surface methodology. The optimized production medium contained (g/L); tryptone, 2.5; yeast extract, 3.0; skim milk 30.0; dextrose 1.0; inoculum concentration 4%: initial pH 6.0; incubation temperature 30 °C and optimum production at 48 h with protease activity of 7.6 U/mL. The protease had the following characteristics: pH optima, 9.0; temperature optima 50 °C; pH stability between 5.0-10.0 and temperature stability between 10-40 °C. The protease was observed to have high potential for dehairing of goat skins in the pre- tanning process comparable to that of the chemical process as evidenced by histology. The method offers cleaner processing using enzyme only instead of toxic chemicals in the pre-tanning process of leather manufacture.