Agricultural Research Management Information System

  • Home
  • Research Summary
    • All
    • Government Organization
      • Agriculture Training Institute, Ishwardi, Pabna
      • Bangabandhu academy for poverty alleviation and rural development (BAPARD)
      • Bangabandhu Sheikh Mujibur Rahman Science & Technology University
      • Bangladesh Bureau of Statistics
      • Bangladesh Institute of Health Sciences
      • Bangladesh Institute of Tropical & Infections Diseases (BITID)
      • Bangladesh Meteorological Department
      • Bangladesh National Herbarium
      • Bangladesh Space Research and Remote Sensing Organization
      • Bangladesh Technical Educational Board
      • Barind Multipurpose Development Authority
      • Central Cattle Breeding Station
      • Department of Agriculture Extension
      • Department of Fisheries
      • Department of Livestock Services
      • Department of Youth Development
      • Dhaka Medical College
      • Geological Survey of Bangladesh
      • Institute of Epidemiology, Disease Control & Research
      • Jatiya Kabi Kazi Nazrul Islam University
      • Khulna Govt. Women College
      • Livestock Training Institute
      • Local Government Engineering Department
      • Ministry of Agriculture
      • Ministry of Environment and forest
      • Ministry of Fisheries and Livestock
      • Ministry of Labour & Employement
      • Ministry of Land
      • Ministry of Public Administration
      • Ministry of Textiles and Jute
      • Ministry of Water Resources
      • Ministry of Youth and Sports
      • National Agricultural Training Academy
      • National institute of preventive and social medicine
      • National Mushroom Development and Extension Centre
      • Pabna University of Science and Technology
      • Seed Certification Agency
      • Shaheed Suhrawardy Medical College
      • Sheikh Hasina University
      • University Grants Commission
      • Youth Training Centre
    • Autonomous/Semi-gov Org
      • Bangladesh Academy for Rural Development
      • Bangladesh Agricultural Development Corporation
      • Bangladesh Atomic Energy Commission
      • Bangladesh Council of Scientific and Industrial Research
      • Bangladesh Fisheries Development Corporation
      • Bangladesh Institute of Development Studies
      • Bangladesh Institute of Management
      • Bangladesh Milk Producers Cooperative Union Limited
      • Bangladesh Water Development Board
      • BIRDEM
      • Center for Environmental and Geographic Information Services
      • Hortex Foundation
      • Institute of Water Modeling
      • National Institute of Biotechnology
      • River Research Institute
      • Rural Development Academy
    • NARS
      • Bangladesh Agricultural Research Council
      • Bangladesh Agricultural Research Institute
      • Bangladesh Fisheries Research Institute
      • Bangladesh Forest Research Institute
      • Bangladesh Institute of Nuclear Agriculture
      • Bangladesh Jute Research Institute
      • Bangladesh Livestock Research Institute
      • Bangladesh Rice Research Institute
      • Bangladesh Sericulture Research and Training Institute
      • Bangladesh Sugarcrop Research Institute
      • Bangladesh Tea Research Institute
      • Bangladesh Wheat and Maize Research Institute
      • Cotton Development Board
      • Soil Resource Development Institute
    • Public University
      • Ahsanullah University of Science and Technology
      • Bangabandhu Sheikh Mujibur Rahman Agricultural University
      • Bangamata Sheikh Fojilatunnesa Mujib Science and Technology University
      • Bangladesh Agricultural University
      • Bangladesh Open University
      • Bangladesh University of Engineering and Technology
      • Bangladesh University of Professionals
      • Bangladesh University of Textiles
      • Barisal Government Veterinary College
      • Begum Rokeya University
      • Chittagong University of Engineering and Technology
      • Chittagong Veterinary and Animal Science University
      • Comilla University
      • Dhaka University of Engineering & Technology
      • Dinajpur Government Veterinary College, Dinajpur
      • Gono Bishwabidyalay
      • Hajee Mohammad Danesh Science and Technology University
      • Islamic University, Kushtia
      • Jagannath University
      • Jahangirnagar University
      • Jessore University of Science and Technology
      • Jhenaidha Government Veterinary College
      • Khulna Agricultural University
      • Khulna University
      • Khulna University of Engineering & Technology
      • Mawlana Bhashani Science and Technology University
      • Millitary Institute of Science and Technology
      • National University
      • Noakhali Science and Technology University
      • Patuakhali Science and Technology University
      • Rajshahi University of Engineering and Technology
      • Shahjalal University of Science & Technology
      • Sher-e-Bangla Agricultural University
      • Sylhet Agricultural University
      • Sylhet Government Veterinary College
      • University of Barisal
      • University of Chittagong
      • University of Dhaka
      • University of Rajshahi
    • Private University
      • Asian University of Bangladesh
      • Atish Dipankar University of Science and Technology
      • BGC Trust University Bangladesh
      • BGMEA University of Fashion & Technology (BUFT)
      • BRAC University
      • City University
      • Daffodil International University
      • East West University
      • Exim Bank Agricultural University
      • Gana Bishwabiddalaya
      • Hamdard University
      • Independent University, Bangladesh
      • International Islamic University Chittagong
      • International University of Business Agriculture and Technology
      • Islamic University of Technology
      • Leading University, Sylhet
      • North South University
      • Premier University
      • Primeasia University
      • Private University
      • SOAS, University of London
      • Southeast University
      • Stamford University
      • State University of Bangladesh
      • The Millenium University
      • University of Asia Pacific
      • University of Development Alternative
      • University of Information Technology and Sciences
      • University of Liberal Arts Bangladesh
      • University of Science and Technology, Chittagong
      • World University
    • INGO/IO/NGO/Private Org
      • ACI Limited
      • Agricultural Advisory Society (AAS)
      • Apex Organic Industries Limited
      • Arannayk Foundation
      • Bangladesh Academy of Sciences
      • Bangladesh Centre for Advanced Studies
      • Bangladesh Institute of Social Research
      • Bangladesh Science Foundation
      • Bangladesh Unnayan Parishad
      • BAPA
      • BRAC
      • CARE Bangladesh
      • CARITAS
      • Centre for Environmental Geographical Information System
      • Centre for Policy Dialogue (CPD)
      • Creative Conservation Alliance
      • Dhaka Ahsania Mission
      • Dwip Unnayan Sangstha
      • EMBASSY OF DENMARK, BANGLADESH
      • Energypac Limited Bangladesh
      • FAO- Bangladesh
      • FIVDB
      • ICDDRB, Mohakhali, Dhaka-1212
      • iDE Bangladesh
      • Innovision Consulting Private Ltd.
      • International Center for Climate Change and Development
      • International Centre for Integrated Mountain Development
      • International Development Research Centre
      • International Fertilizer Development Center, Bangladesh
      • International Food Policy Research Institute
      • International Maize and Wheat Improvement Centre
      • International Potato Center
      • IRRI- Bangladesh
      • IRRI-Philippines
      • Ispahani Agro LTD
      • IUCN, Bangladesh
      • Krishi Gobeshina Foundation
      • Lal Teer
      • Mennonite Central Committee
      • Metal (Pvt.) Ltd
      • Modern Herbal Group
      • Palli Karma-Sahayak Foundation
      • Practical Action Bangladesh
      • Proshika
      • RDRS Bangladesh
      • RIRI-Philippines
      • Rothamsted Research
      • SAARC Agricultural Centre
      • SAARC Meteorological Research Centre
      • Social Upliftment Society
      • South Asia Enterprise Development Facility
      • Square Pharmaceuticals Ltd.
      • Supreme Seed
      • Transparency International Bangladesh
      • Unnayan Onneshan
      • USAID
      • Water Resources Planning Organization
      • Winrock International
      • World Bank
      • World Food Program
      • World Vegetable Center
      • WorldFish Centre, Bangladesh
    • Foreign University
      • Asian Institute of Technology
      • Auckland University of Technology
      • Australian National University
      • Bidhan Chandra Krishi Viswavidyalaya
      • BOKU-University of Natural Resources and Applied Life Sciences
      • Cranfield University
      • Curtin University
      • Foreign University/ Institute
      • Hiroshima University
      • Hokkaido University
      • Huazhong Agricultural University
      • International Islamic University, Malaysia
      • Kagawa University
      • Kangwon National University
      • Kochi University
      • Kyoto University
      • Kyushu University
      • Ladoke Akintola University of Technology
      • Murdoch University
      • Nagoya University
      • NOAA-CREST, CCNY
      • Royal Veterinary and Agricultural University
      • San Diego State University
      • Shinshu University
      • Tottori University
      • United Nations University
      • University Malaysia Kelantan
      • University Malaysia Pahang
      • University Nova de Lisboa
      • University of Alberta
      • University of Bremen
      • University of Bremen
      • University of Calgary
      • University of california
      • University of Greenwich
      • University of Hamburg, Hamburg
      • University of Hannover
      • University of Hawaii
      • University of Helsinki, Finland
      • University of Kalyani
      • University of Leeds
      • University of Liverpool
      • University of Malaya
      • University of Milan
      • University of New England
      • University of Philippines
      • University of Plymouth
      • University of Queensland
      • University of Reading
      • University of Southampton
      • University of Texas
      • University of the Punjab
      • University of Tokyo
      • University of Toronto
      • University of Wales
      • University of Washington
      • University of Wollongong
      • University Putra Malaysia
      • University Sains Malaysia
  • Search
    • Search by Keyword
    • Search by Organization
    • Search by Program Area
    • Search by Commodity/Non-commodity
    • Search by Funding Source
    • Search by Researcher
    • Custom Search
    • On-going Research
  • About Us
    • ARMIS
    • Brochure
  • Contact Us
    • BARC Personnel
    • ARMIS Personnel
    • Feedback
  • Report
    • All
    • By Organization
      • Bangladesh Agricultural Research Council
      • Bangladesh Agricultural Research Institute
      • Bangladesh Fisheries Research Institute
      • Bangladesh Forest Research Institute
      • Bangladesh Institute of Nuclear Agriculture
      • Bangladesh Jute Research Institute
      • Bangladesh Livestock Research Institute
      • Bangladesh Rice Research Institute
      • Bangladesh Sericulture Research and Training Institute
      • Bangladesh Sugarcrop Research Institute
      • Bangladesh Tea Research Institute
      • Bangladesh Wheat and Maize Research Institute
      • Cotton Development Board
      • Soil Resource Development Institute
    • Research Trend Analysis
  • User Request
  • Data Input
  • Help
    • Operation Manual
      • PDF
      • Video
    • Program Area & Commodity
  • We have reached 37600 number of research entries at this moment.
    • Logout

Research Detail

  1. Home
  2. Research
  3. Detail
Silvia Ferdous
Fisheries and Marine Resource Technology Discipline, Khulna University, Khulna 9208, Bangladesh

Md. Shamim Ahmed
Fisheries and Marine Resource Technology Discipline, Khulna University, Khulna 9208, Bangladesh

Zerin Sultana
Fisheries and Marine Resource Technology Discipline, Khulna University, Khulna 9208, Bangladesh

Md. Monirul Islam
Fisheries and Marine Resource Technology Discipline, Khulna University, Khulna 9208, Bangladesh

Md. Ayaz Hasan Chisty*
Fisheries and Marine Resource Technology Discipline, Khulna University, Khulna 9208, Bangladesh

The study was carried out to identify muscle protein patterns and gut protease activity of rohu (Labeo rohita), catla (Catla catla), climbing perch (Anabas testudineus) and spotted snakehead (Channa punctatus). Twelve percent Polyacryleamide substrate SDS-PAGE was done to visualize the activity of the enzyme after casein hydrolysis and protein pattern of the species. Three alkaline proteases of different molecular weight were observed in rohu, two in catla, five in climbing perch and four in spotted Snakehead from the zymogram. This revealed that the existence of alkaline protein in the alimentary tract of respective species. Fourteen distinct protein bands in rohu, fifteen in catla, sixteen in climbing perch and nineteen in soptted snakehead were visualized from the electrophoretic patterns of fish muscle extract. From the result, it is assumed that heavy molecular weight myosin and low molecular weight actin were prominent in all the experimental species.

  Casein, Enzyme, Protein patterns, Actin, Myosin
  Fisheries and Marine Resource Technology Discipline, Khulna University, Khulna 9208, Bangladesh
  
  
  Quality and Nutrition
  Rohu, Catla, Koi/Climbing

As protein is the most costly nutrient in diets and the enzymes that are involved in protein digestion might vary from species to species, it is important to get the muscular protein pattern and the proteases present along with the digestive tract of the species to help selecting the feed ingredients as well as to maintain the protein levels in feed. Therefore, the present study was designed to identify gut protease activity and muscular protein pattern of different commercially valuable species.

To observe the muscular protein pattern and the gut protease activity of the valuable fish species, different procedures and protocols were followed:

Sample collection: Four different commercial valuable fish species were selected as follows:  Live fish (viz. rohu, catla, climbing perch and spotted snakehead) were purchased from local market (Gollamari bazaar) and kept in aquaria (climbing perch and spotted snakehead) and ponds (ruhu and catla) until needed. Upon the arrival of the fish, they were acclimatized by commercial feed (protein level 40%) in the aquaria and in the ponds. Later they were sacrificed for muscle protein and gut enzyme collection, according to Garcia-Carreno et al., (1994).

Preparation of enzyme extract: Live fish were caught from the aquaria and pond and sacrificed by dissection to collect the guts of the species. The guts of the same species were pooled together, kept at chilled condition (≤4°C) and weighed using electronic balance (Electric balance, Model-AND GF-300, Japan) and homogenized in a Potter Thomas tissue grinder with a Teflon pestle at cool temperature (≤4°C) by keeping the tissue grinder on to ice and diluted with cool distilled water (4°C) at a ratio of 1:10 W/V. The homogenate were poured into 1.5ml microfuge tubes (previously kept into ice and marked) and immediately centrifuged at 12000×g for 15 minutes at 4°C in a refrigerated centrifuge (Hettich Zentrifugen, model- D 78532, Mikro 22 R, Germany). The upper lipid layer of the supernatant, after centrifuging, was discarded. The aqueous supernatant was collected in previously cooled microfuge tubes, frozen and stored at -20°C until use (Chisty, 2005).

Preparation of protein extract: Live fish were sacrificed by dissection to collect the muscle just beneath the 1st and or 2nd dorsal fin, but above the lateral line of the respective species. The muscle of the same species were pooled together, kept at chilled condition (≤4°C) and weighted. Later, the muscle was homogenized in a Potter Thomas tissue grinder with a Teflon pestle at cool temperature (≤4°C) by keeping the tissue grinder on to ice and diluted with cool distilled water (4°C) at a ratio of 1:10 w/v. The homogenate were poured into 1.5ml microfuge tube and centrifuged at 12000×g for 15-20 minutes in a centrifuge (Eppendorf, model-5415D, Germany). The aqueous supernatant was taken in previously cooled another microfuge tube, after centrifuging. Immediately after collection of the muscle protein extract of electrophoresis was done to identify the protein pattern according to Bollag and Edelstein (2000).

Sample preparation for enzyme activity: Ten µl crude enzyme extract and 10µl sample buffer was taken, vortexed (Rocking on rotary sucker, Type 16700 Mixer, Thermoclyne) for 30 second, after vortex, the sample was ready for electrophoresis. The sample buffer was prepared according to Bollag and Edelstein (2000).

Sample preparation for protein pattern: Ten µl crude protein extract and 10µl sample buffer was taken. It was then vortexed for 30 second and heated 3-5 min at 95°C temperature in water bath (Grant instruments Ltd., Model- Hearts SG86PZ, England). Again it was vortexed and centrifuge at 12000×g for 15-20 min in a centrifuge (Eppendorf, Model- 5415 D, Germany) and was taken in a previously cool microfuge tube and was ready for electrophoresis.

Sodium Dodecyle Sulfate Polyaccrylamide Gel Electrophoresis: SDS-PAGE was done to identify the enzyme zymogram and protein pattern. Bio-Rad Mini -Gel Apparatus (Bio-Rad, MiniPROTEIN 3 cell) was used for this purpose. In brief, Twelve percent polyaccrylamide was used to prepare the linear slab gel at a dimension of 10 cm × 8 cm × 0.75 mm. In brief, the gel was prepared by pouring the separating gel in the glass slab. Stacking gel was poured onto separating gel, just after the polymerization of the separating gel. Comb (0.75 mm thickness and with 10 teeth) was inserted on it. Electrophoretic buffer was poured properly according to Bollag and Edelstein, (2000) and 10 µl sample solution was loaded onto each well. The tub was placed in a bowl containing ice to maintain the temperature below 10°C. For running the gel, 120V constant power supply was ensured (Bio-Rad, Powerpack 1000). Electrophoresis continued until the dye front migrated about to 1 cm above the bottom of the gel (around 1 hr) then Spacer (0.75 mm) was removed carefully by inserting the spacer in one corner between the plates and the gel plates were apart gently. To obtain the enzyme zymogram, the gel was soaked in 2% caesine solution (w/v) by using caesine and Tris HCl buffer, (pH 7.8) and kept at 4°C for 30 min that helped to absorb casein into the gel. Later, keeping the gel into the same solution; it was incubated for 1.5 hrs at room temperature. To obtain the protein zymogram, soaking in caesine solution was omitted. Now the gel was subjected for staining and distaining.

Gel staining: The gel was picked up and transferred to a small container, containing a small amount of coomassie gel staining solution by using coomassie Blue R-250, methanol, distilled water and glacial acetic acid and the gel, keeping into the staining solution, was agitated gently for 1-2 hrs. After staining of the gel, it was taken out from the solution, washed a few times with distilled water and was subjected to destaining.

Destaining of gel: The gel was soaked in destaining solution and kept over night with a gentle stirring. In brief the destaining solution was prepared by addition of methanol, distilled water and glacial acetic acid. Finally the gel was taken out from the solution, washed with distilled water and preserved for the zymogram.

  Khulna University Studies Volume 10 (1&2) : 161-166 : June-December 2010
  
Funding Source:
1.   Budget:  
  

The result of this study as three alkaline proteases of different molecular weight were observed in rohu (L. rohita), two in catla (C. catla), five in climbing perch (A. testudineus) and four in spotted snakehead (C. punctatus). Muscular protein pattern of different fish species was visualized after electrophoresis. Fourteen distinct bands in rohu were visualized from the electrophoretic patterns of fish muscle extract. And that of fifteen in catla, sixteen in climbing perch and nineteen in spotted snakehead fish ware visualized from SDS-PAGE. So, the findings of the research would be useful tools for to identify gut protease activity and muscular protein pattern of different experimental species.

  Journal
  


Copyright © 2025. Bangladesh Agricultural Research Council.