Research Journal of Biotechnology

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Research Journal of Biotechnology





Effects of Phosphate on Texture and Water Retention Capacity of Pork-Egg Sausage

Feng Cheng-Cheng, Ma Mei-Hu, Jin Yong-Guo and Wei Shuai

Page No: 1-6

Abstract: Texture and water retention are important properties of pork-egg sausage made from egg and pork. The effects of sodium tripolyphosphate (STPP), tetrasodium pyrophosphate (TSPP), sodium hexametaphosphate (SHMP) on texture and water retention properties of pork-egg sausage were studied. The results showed that texture and water retention of pork-egg sausage were effectively improved by adding three kinds of phosphates. Compared with STPP and SHMP, TSPP was more effective in improving the hardness of the sausage.

Based on the single factor experiment, orthogonal experiment was done. When a combination of STPP (0.101%), SHMP (0.211%) and TSPP (0.117%) was added, the hardness, elasticity and water retention capacity of pork-egg were increased by 221.915g, 0.11 and 10.08% respectively compared with those without phosphate which provided a high gel quality of pork-egg sausage.

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High-frequency multiple shoot induction from nodal segments and assessment of genetic homogeneity of micropropagated Atriplex halimus L.

Kumar Vinod, AlMomin Sabah, Shajan Anisha, Al-Aqeel Hamed, Al-Salameen Fadila and Nair Sindhu

Page No: 7-15

Abstract: Atriplex halimus L. is a potentially useful plant in the arid region for animal feed, soil conservation and phytoremediation. Propagation of this plant through seeds is hindered by seed dormancy and a short viability period. Development of a robust tissue culture-based regeneration method would greatly facilitate rapid multiplication. In order to develop an efficient micropropagation method, we evaluated the effects of various growth regulators in the synthetic culture media. For initiation of the shoot multiplication process, surface-sterilized nodal segments were cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of growth hormones such as 6-benzyladenine (BA) or thidiazuron (TDZ) alone or in combination with indole-3-acetic acid (IAA). The optimum response for shoot bud multiplication was observed on MS medium supplemented with both BA and TDZ. The shoot buds were then allowed to elongate and root.

The plants were acclimatized, transplanted to soil and the regenerated plants were subjected to molecular analysis to determine their genetic homogeneity. Jaccard’s coefficient and Nei and Li coefficient indicated high degree of genetic identity between the mother and the tissue-cultured plants. The results suggest suitability of this propagation technique for large-scale multiplication of A. halimus L.

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Morphological, genetic and virulence diversity of Rhizoctonia solani isolates from different rice growing regions of Southern India

Nagaraj B.T., Gururaj Sunkad, Pramesh D., Naik Manjunath K., Patil Mahanthesh B., Yadav Manoj K. and Patil Naveenkumar B.

Page No: 16-23

Abstract: Rice sheath blight disease caused by Rhizoctonia solani is an economically important disease of rice in India. To characterize the R. solani population structure, sixty-five isolates were collected from rice and rice associated weed species from different agroclimatic zones of Karnataka, India. All 65 isolates were characterized for their morphological, genetic and virulence diversity. All isolates showed wide morphological variation in terms of size of sclerotia (0.13 mm to 9.44 mm), colour and texture of sclerotia ranging from dark brown to light brown with rough to smooth in texture and abundance (0-1054). The genetic diversity of R. solani populations was dissected based on anastomosis grouping. Anastomosis group (AG) was identified by restriction fragment length polymorphism (RFLP) of internal transcribed spacer (ITS) sequences. We identified that 65 isolates of R. solani belonged to four different anastomosis groups such as AG1-IA (40 isolates, contributed to 61 per cent of the total isolates characterized), AG1-IB (4 isolates), AG1-IC (15 isolates) and AG2-1 (6 isolates).

The virulence of all 65 isolates of R. solani was analyzed based on relative lesion height on four different rice varieties such as TN1 (Susceptible), BPT 5204 (Susceptible), IR 64 (Moderately Susceptible) and Tetep (Moderately Resistance) by artificial inoculation. Out of 65 isolates, 11 isolates were categorized as highly virulent, 23 isolates as moderately virulent and 31 isolates were less in virulence. The relative lesion length on rice varieties varied with respect to isolate and cultivar combination. Present investigation provides the existence of diversity in the population of R. solani causing rice sheath blight disease in Karnataka, Southern India.

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Comparison of antimicrobial activity of bio-synthesized silver and zinc oxide nanoparticles using Lavandula stoechas leaf extract

Dilek Demirezen Yilmaz, Koca Fatih Doğan and Nurhan Ertaş Onmaz

Page No: 24-29

Abstract: In this study, the extract of Lavandula stoechas for production of silver and zinc oxide nanoparticle without use of any chemical agent was investigated. The silver (AgNPs) and zinc oxide nanoparticles (ZnONPs) showed strong antibacterial activity against both tested Escherichia coli O157:H7 (Gram negative) and Staphylococcus aureus (Gram positive) bacteria. The antibacterial activity of bio-synthesized nanoparticles against two pathogens was assessed by minimal inhibitory concentration (MIC) assays. The MIC values of AgNPs and ZnONPs of 10.63 µg/mL and 12.5 µg/mL for E.coli O157:H7 AgNPs and Staphylococcus aureus against ZnONPs were at 3.32 µg/mL and 6.25 µg/mL respectively.

Biologically synthesized nanoparticles were characterized by Scanning Electron Microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), UV-Vis spectroscopy, X-ray diffraction (XRD) and Zeta potential analysis. This study concludes that the bio- synthesized AgNPs and ZnONPs may be used for effective antimicrobial activity, so can be projected as future generation antimicrobial agents and designing newer drugs.

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Prevalence of clumping factor associated genes (clfA and clfB) in isolates of Staphylococcus aureus obtained from clinical mastitis in cattles

Choudhary Sunita, Diwakar, Bhati Taruna and Kataria Anil Kumar

Page No: 30-35

Abstract: The present study was conducted to type Staphylococcus aureus obtained from clinical mastitis in cattle on the basis of clfA and clfB genes, both responsible for producing clumping factors. In the present investigation, S. aureus isolates were isolated as per standard protocols. Typing of S. aureus was carried out by detection of clfA and clfB gene by polymerase chain reaction.

In the present study, 20 (66.66%) isolates produced amplicons of 1000 bp indicating presence of clfA gene and 27 (90%) isolates produced amplicons of 205 bp indicating presence of clfB gene and three isolates were non-typable. Twenty (66.66%) isolates were typable for both of the genes. No polymorphism was observed for both of the genes. The genes for clumping factors were present in most of the isolates indicating their pathogenic potential.

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Interaction between Serratia fonticola and Chryseobacterium jejuense on tomato phylloplane influences the cellular protein profile

Bhadauria Neha, Shilpi Shilpi, Sharma P.D. and Paul P.K.

Page No: 36-43

Abstract:Phylloplane is colonized by an array of microbes including those where natural habitats are not plants. Human enteric pathogens (HEPs) are one of them which can colonize plant surfaces. During the last two decades, the numerous gastrointestinal disorder outbreaks were reported worldwide which were caused due to the consumption of raw vegetables contaminated with human enteric pathogens. The present study aims to understand the molecular changes in plant during Plant-HEP interaction. The study reveals HEPs could cause changes in cytoplasmic protein profile of plant tissues either singly or in combination. Aseptically grown tomato plants were inoculated with HEPs i.e. Serratia fonticola and Chryseobacterium jejuense either singly or in combination. Cytoplasmic proteins were extracted from in vivo crosslinked leaves and SDS PAGE separated proteins were studied.

Enhanced concentrations of proteins were observed after 48 hours to 96 hours post inoculation. Electrophoretic separation demonstrated the crosslinking of proteins in Serratia fonticola, Chryseobacterium jejuense and the combination of S. fonticola + C. jejuense inoculated plants where changes were observed predominantly at 72 hours post inoculation. The study shall help to understand the impact of these HEP on molecular physiology of host which if disrupted could lead to reduction of HEP load on plants.

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Genetic diversity and phylogenetic profiling of Fusarium sp., the causing storage rot of ginger (Zingiber officinale) in Himachal Pradesh and their potential environmental eco-friendly management strategies

Sharma Sushma, Veerubommu Shanmugam, Brar Gaganpreet Singh, Thakur Neelam, Thakur Sapna, Thakur Priyanka, Phurailatpam Sumitra and Yadav Ajar Nath

Page No: 44-54

Abstract:With an advance of modern molecular markers technology, it becomes easy for the rapid identification, characterization and detection of some important plant fungi. In present investigation, different Fusarium sp. were isolated from ginger and were identified using ITS amplification. The identities of different Fusarium sp. causing storage rot of ginger which were recovered from infected rhizomes from different locations established by PCR amplification and sequencing of ITS gene sequences. Genetic diversity study of isolated Fusarium sp. was carried out by using four ISSR primers.

Four primers selected from a group of 5 primers generated excellent results and were used in this investigation. The 90.9 % genetic diversity was present in ISSR-1, ISSR-2 and ISSR-3 and 100 per cent genetic diversity was present in all the isolates of Fusarium sp. when ISSR-4 primer was used. In management studies, cow urine and neemazal were found most effective and resulted in minimum disease incidence with highest rhizome recovery.

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Statistical optimization of culture conditions for endopolygalacturonase production by Aspergillus niger using solid orange waste

Benmalek Nabila and Benchabane Ahmed

Page No: 55-66

Abstract: Scanning electron microscopy analysis of Aspergillus niger growth on orange residue showed that the microorganism grows well on the medium by synthesizing the pectolytic enzymes. A two-level full factorial design was used for screening the most important factors for the production of endopectinase (endo PG) from dry orange waste by Aspergillus niger among dry orange waste pellet size, concentration of ammonium sulphate, concentration of glucose and moisture content. Three significant factors influencing endo PG production were identified: concentration of ammonium sulphate, moisture content and glucose concentration. The particle size of the dry orange waste may not affect the process bat its interaction with moisture has significant effect.

The effects of the significant factors were further investigated using a design of rotation-regression-orthogonal combination. The optimised medium according to response surface methodology (RSM) analysis contained 1.98% ammonium sulphate 1.83% glucose and 54,63 of distilled water. Under the optimised conditions, the model predicted 25,57 U/ml in crude extract, whereas validation experiments produced an endo PG enzymatic activity of 32.41 U/ml. This study is innovatively a fermentation medium to use inexpensive agro-industrial wastes to produce a high yield of pectinase.

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Role of various signaling pathways for glycerol synthesis in appessoria of Magnaporthe oryzae under Climate Change Scenario

Rajput L.S., Sharma Taru, Madhusudhan P., Kumar Sanjeev and Sinha P.

Page No: 67-73

Abstract: Temperature rise leads to change infection process of M. oryzae causal agent of rice blast pathogen. Glycerol synthesis is significantly changed by enhanced temperature of M. oryzae which play a key role in formation of infection structure such as appressoria. Glycerol synthesis was enhanced up to 27°C and later on reduced drastically with high temperature (32°C). At the time of appressoria production, high temperature leads to delay the glycerol synthesis process by 2 h which indicated that higher temperature leads to partially inhibit or reverse in glycerol synthesis mechanism. Inhibition of glycerol synthesis by specific signalling inhibitors viz. K252a (cAMP dependent PKA pathway inhibitor) and EGTA (calcineurin pathway inhibitor) revealed that MAPK pathway and cAMP dependent PKA signalling pathways have major role in glycerol production rather than calcineurin signalling at both germinated spore as well as production of appressoria.

Metabolic control analysis of glycerol biosynthesis synthesis pathways depicted that dihydroxyacetone phosphate is key compound to control glycerol biosynthesis. However, experimentally it was seen that cAMP dependent PKA concentration has direct control on glycerol formation. Glycerol synthesis inhibition leads to possibility of disease management without any biocidal use and it may usher a new era of disease management without harming environment.

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Anticancer activity of camptothecin producing endophytes isolated from Chonemorpha fragrans (Moon) Alston. (Apocynaceae)

Prince Clarance, Jebalsy Lalitha, Jerin Sales, Ameer Khusro and Paul Agastian

Page No: 74-82

Abstract: Chonemorpha fragrans (Moon) Alston is an endangered medicinal herb used in the indigenous system of medicine belonging to the family Apocynaceae. The present study was aimed to isolate and characterize Camptothecin (CPT) producing endophytic fungi and bacteria from C. fragrans, a novel plant source for endophytes. The presence of CPT was identified in both fungi and bacteria by RP-HPLC and ESI–Q-TOF analysis of the extract.

The fungal extract was subjected to Topoisomerase I inhibitory assay and found partially inhibiting. One of the fungal isolates as well as bacterial isolates showed the presence of 9-methoxy-CPT in the cultures along with CPT. The production of CPT from bacterial strain was estimated 16-fold higher than that of fungal strain. The anticancer activity of the lyophilized endophytic fungal extract was confirmed using an MTT reduction assay. The sub-culturing of the endophytic bacteria up to 4th generation brought forth attenuation in the culture. The gene level research in endophytic fungus and bacteria will expectantly bring their application for the enhanced production of camptothecin and 9-methoxy camptothecin.

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Mango intra-varietal hybrid confirmation through SPAR markers

Srivastava Navin, Rajan Shailendra, Muthukumar M., Srivastava Manoj K. and Bajpai Anju

Page No: 83-88

Abstract:Genetic studies in mango can increase pace of precision breeding due to development of true mapping populations endorsed by molecular markers. Concerns over the suitability of single dominant marker system for determining parentage have arisen due to appearance of nonparental bands in offspring of known pedigrees as well as heterozygosity at the marker loci. Twenty-one mango (Mangifera indica L.) hybrid progenies developed from inravarietal crossing could be established as true hybrids using RAPD, ISSR and DAMD markers. UPGMA tree showed clustering of the hybrids with the male parents i.e. Vanraj and Janardan Pasand respectively. Amrapali being common parent is equidistant from all hybrids and is isolated as a separate shoot confirming the pedigree.

This may be attributed to the bearing habits as Amrapali is a regular bearer and late maturating variety, while Vanraj and Janardan Pasand are biennial bearers and peel colour donors. The results of the present study indicated that three markers together serve as a candidate molecular tool to correlate parent-hybrid relationship.

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Bactericidal activity of green synthesized silver nanoparticles (AgNPs) against Escherichia coli and Staphylococcus aureus

Das Sarita, Prasanti Y., Naik Priyanka, Padhy Janaki Kumari and Panigrahi Suchasmita

Page No: 89-96

Abstract:Antimicrobial activity of silver nanoparticles is gaining importance due to its broad spectrum of targets in cells in comparison to the conventional antimicrobial agents. In this present study, silver nanoparticles (AgNPs) were synthesized by using a cost effective and ecofriendly green synthesis method (using mango kernel powder). Plant extracts act as reducing as well as capping agent during AgNPs synthesis. The antimicrobial activity of silver nanoparticles against Escherichia coli and Staphylococcus aureus was measured by disc diffusion, agar well diffusion and modified agar well diffusion method. Cfu/ml was determined using serial dilution followed by spread plate method or taking OD600nm (absorbance) value. Green synthesized AgNPs showed better inhibitory activity against E. coli than S. aureus in agar well diffusion and modified agar well diffusion method. It was found that in disc diffusion method, S. aureus was more sensitive than E. coli.

In serial dilution method and absorbance method, cfu/ml was found to be reduced in a dose dependent manner in the AgNPs treated bacteria. The results of this investigation showed significant bactericidal activity of AgNPs against the most common gram negative and gram-positive pathogenic bacteria. However, further research can elucidate the mechanism of bactericidal activity of AgNPs and their applications in preventing different opportunistic infections.

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Genetic diversity and molecular characterization of Ganoderma spp. affecting plantation crops and forest tree plants using ITS-PCR

Raju J., Naik S.T., Jayalakshmi K., Sonavane Priti and Giri Madhu S.

Page No: 97-102

Abstract:Basal stem rot caused by Ganoderma lucidum is the most serious and destructive disease in coconut, arecanut gardens and forest tree plants. Fifty-one Ganoderma isolates were collected from different parts of southern India and were characterized using ITS-PCR. In this study, the primers Gan1 and Gan2 produced a product of 167 bp in size for all the Ganoderma isolates were tested. Simultaneously, ITS 1 and ITS 4 primers amplified a fragment of be 550 bp in the Ganoderma isolates.

In addition, the phylogenetic analysis showed that all studied isolates separated into two groups: Groups 1 and 2. Within group 1, there was further formation of sub-group between isolate ARS-8, PUN-1 with bootstrap value of 65 per cent which showed that they are related to each other whereas in group 2 there is closely related grouping of three isolates viz. SRS-1, COMB-1 and DWD-2 with bootstrap value of 100 per cent. As expected, branching of Amerouderma rogosum was altogether different having wide divergence with separate branch in dendrogram. It indicated existence of variability between the two groups of isolates.

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In Vitro Mass Propagation System in Banana Cv. Kamalapur Red (AAA)

Devalakere Prakasha, Gurunath Ramya, Ryavalad Shivayogi, Kustagi Gajanana and Krishnappa Kattimani

Page No: 103-113

Abstract:This study was taken up for finding out effect of various factors affecting in vitro and to develop an in vitro mass propagation system in Banana cv. Kamlapur red. Healthy sword suckers were collected, washed, cleaned, outer layers removed sterilized using different sequences involving sodium hypochlorite mercuric chloride and ethanol and then inoculated. There was significantly high of 77.23 per cent clean culture established using the treatment sequence having bavistin (0.1%-60mins), mercuric chloride (0.1%-15mins), sodium hypochlorite (4%-15mins) and ethanol (70%-60 sec) and three water wash after every chemical treatment. The explants cultured on the MS medium containing 5mgL- BAP and 1mgL- IAA have showed the significantly average highest of 8.53 shoots per explants whereas significantly average highest shoot height of 3.98cm was obtained on 3mgL- BAP.

In the primary and secondary hardening, the highest (89.33%, 97.17% respectively) and lowest (31.33%, 90.33% respectively) number of plants survived in low headed tunnel under shade house and shade house respectively. There was remarkable improvement in plantlet survivability with reduced mortality rate in secondary hardening even under shade net. A system for mass propagation was developed by combining best treatment in all the experiments and it was evaluated for eight passages for its responses. It showed an average multiplication ratio, number of shoots per explants and shoot height of 1.87, 7.01 and 3.49cm respectively. A sucker initiated by using the protocol will yield 1373 in vitro rooted plantlets after eight passages. This protocol would be useful in production of large-scale planting material in banana cv. Kamalapaur Red.

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Enhanced Production of Diffusible Eumelanin from Streptomyces Rochei isolated from Marine Sources

Periyasamy Satheesh and Kathirvel Preethi

Page No: 114-122

Abstract:In the present study, 39 pure cultures of actinomycetes were isolated from different marine samples collected around Vishakhapatnam and Chennai coast and screened for melanin production. The isolate that yielded highest quantity of diffusible pigment production was identified by morphological, 16S rRNA sequencing and phylogenetic analysis as Streptomyces rochei. Melanin pigments isolated were purified by precipitation of the cell free supernatant using 6N HCl, digested with papain and vacuum dried.

The yield of melanin from Streptomyces rochei was increased nearly 3-fold to 1.41 g/L by optimizing 19 fermentation parameters using Plackett burman design and face centered composite design. The critical parameters were optimized and fixed at Sodium Chloride (1 mg/mL), Temperature (30°C) and incubation period (14 days).

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Xenobiotic hepatotoxicity

Priya Josson Akkara and Evan Prince Sabina

Page No: 123-138

Abstract:Drug induced and environmental toxin induced hepatotoxicity is of major concern among xenobiotic hepatotoxicities in today’s world. The potential of drug-based or environmental hepatotoxins to generate liver cell injury results from a complex interaction of cellular processes and is based on direct or indirect reactions of the toxins with basic hepatocyte constituents such as proteins, lipids, RNA, or DNA. This is a review of xenobiotics which causes hepatotoxicity, the various in vitro models to study hepatotoxicity and the biochemical reactions leading to biotransformation of xenobiotics which include different drugs as well as environmental pollutants. The effect on mitochondrial enzymes, mitochondrial dysfunction, oxidative stress leading to liver damage is dealt.

Molecular physiology of inflammation in hepatotoxicity, lipid peroxidation and its consequences, the role of various cytokines and chemokines in toxicity of liver have been discussed. The biochemical markers of hepatotoxicity and the enzymes and proteins involved have been elaborated. Bromobenzene has been taken as a candidate for environment pollutant causing hepatotoxicity. The biochemical reactions leading to the breakdown of bromobenzene into its metabolites and its correlation with pathophysiological conditions have been discussed henceforth. The latest methods in understanding xenobiotic hepatotoxicity at the trancriptome level and the contribution of proteomics and toxico-metabolomics in detecting chemically induced hepatotoxicity would facilitate the generation of new hypotheses on the cellular mechanism.

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