29 results on '"silver carp (Hypophthalmichthys molitrix)"'
Search Results
2. A Thermostable Type I Collagen from Swim Bladder of Silver Carp (Hypophthalmichthys molitrix).
- Author
-
Jiang, Honghui, Kong, Yuanyuan, Song, Lili, Liu, Jing, and Wang, Zhihong
- Abstract
As a major component of the extracellular matrix, collagen has been used as a biomaterial for many purposes including tissue engineering. Commercial collagen derived from mammals is associated with a risk of prion diseases and religious restrictions, while fish-derived collagen can avoid such issues. In addition, fish-derived collagen is widely available and low-cost; however, it often suffers from poor thermal stability, which limits its biomedical application. In this study, collagen with a high thermal stability was successfully extracted from the swim bladder of silver carp (Hypophthalmichthys molitrix) (SCC). The results demonstrated that it was a type I collagen with high purity and well-preserved triple-helix structure. Amino acid composition assay showed that the amounts of threonine, methionine, isoleucine and phenylalanine in the collagen of swim bladder of silver carp were higher than those of bovine pericardium. After adding salt solution, swim-bladder-derived collagen could form fine and dense collagen fibers. In particular, SCC exhibited a higher thermal denaturation temperature (40.08 °C) compared with collagens from the swim bladder of grass carp (Ctenopharyngodon idellus) (GCC, 34.40 °C), bovine pericardium (BPC, 34.47 °C) and mouse tail (MTC, 37.11 °C). Furthermore, SCC also showed DPPH radical scavenging ability and reducing power. These results indicate that SCC presents a promising alternative source of mammalian collagen for pharmaceutical and biomedical applications. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
3. Improvement of Flesh Quality of Farmed Silver Carp (Hypophthalmichthys molitrix) by Short-Term Stocked in Natural Water.
- Author
-
Miao, Xiaomin, Guo, Hui, Song, Yong, Du, Chunying, Feng, Jingyun, Tao, Yixi, Xu, Hao, and Li, Yun
- Subjects
- *
SILVER carp , *DIGESTIVE enzymes , *PONDS , *ESSENTIAL amino acids , *FOOD composition , *FATTY acids - Abstract
To investigate the effects of different raising environments on the flesh quality, the study set up three groups of silver carp (Hypophthalmichthys molitrix), that is, pond-farmed; short-term stocked; and ranched, grown in a natural water ranch. This study compared the differences in muscle proximate composition and amino acid composition among three groups. The results showed that there was no difference between the ranched and the stocked groups in content of crude protein, ash, total amino acids (TAA), essential amino acids (EAA), and umami-flavored amino acids (UAA), with both groups being significantly higher than the farmed group. This shows that, in terms of flesh quality, the stocked group was closer to the ranched group and better than the farmed group after being stocked in natural water for 30 days. The results suggest that the flesh quality of farmed fish was improved by short-term stocking in natural water. To elucidate the reasons of flesh quality change in the stocked group, this study compared the food composition and intestinal digestive enzyme activity in three groups, demonstrating that the ranched and stocked groups had similar food composition, with their detritus proportion lower than that of the farmed group, consuming easily digestible phytoplankton diatoms rich in amino acids and fatty acids. No significant difference was seen between the ranched group and the stocked group, for which amylase, lipase, and trypsin activities were lower than that of the farmed group. It is hypothesized that differences in environmental and food composition resulted in lower digestive enzyme activity in the ranched group and the stocked group. In summary, the short-term stocking of farmed fish in a natural water ranch can effectively improve the nutrient content, quality and flavor, and promote value of fish products. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
4. Effect of pH-Shifting Process on the Cathepsin Activity, Muddy Off-Odor Compounds' Content and Gelling Properties of Isolated Protein from Silver Carp.
- Author
-
Guo, Weidan, Zhan, Miao, Liu, Hui, Fu, Xiangjin, and Wu, Wei
- Subjects
SILVER carp ,GELATION ,ODORS ,PROTEIN crosslinking ,ELASTIC modulus ,MOLECULAR weights - Abstract
Silver carp (Hypophthalmichthys molitrix) is a potential source for making surimi products. However, it has the disadvantages of bony structures, high level of cathepsines and muddy off-odor which is mainly caused by geosmin (GEO) and 2-methylisoborneol (MIB). These disadvantages make the conventional water washing process of surimi inefficient (low protein recovery rate, and high residual muddy off-odor). Thus, the effect of the pH-shifting process (acid-isolating process and alkali-isolating process) on the cathepsins activity, GEO content, MIB content, and gelling properties of the isolated proteins (IPs) was investigated, comparing it with surimi obtained through the conventional cold water washing process (WM). The alkali-isolating process greatly boosted the protein recovery rate from 28.8% to 40.9% (p < 0.05). In addition, it removed 84% GEO and 90% MIB. The acid-isolating process removed about 77% GEO and 83% MIB. The acid-isolated protein (AC) displayed the lowest elastic modulus (G′), the highest TCA-peptide content (90.89 ± 4.65 mg/g) and the highest cathepsin L activity (65.43 ± 4.91 U/g). The AC modori (60 °C for 30 min) gel also demonstrated the lowest breaking force (226.2 ± 19.5 g) and breaking deformation (8.3 ± 0.4 mm), indicating that proteolysis caused by the cathepsin deteriorated the gel quality of AC. The setting (40 °C for 30 min) considerably increased the breaking force (386.4 ± 15.7 g) and breaking deformation (11.6 ± 0.2 mm) of the gel made from the alkali-isolated protein (AK) (p < 0.05). In AC and AK gel, a clearly visible cross-linking protein band with a molecular weight greater than MHC was seen, demonstrating the presence of endogenous trans-glutaminase (TGase) activity, that improved the gel quality of AK. In conclusion, the alkali-isolating process was an effective alternative method for making water-washed surimi from silver carp. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
5. The impacts of cold plasma on the taste and odor formation of dried silver carp products.
- Author
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Zhai, Yueying, Luan, Aonan, Yang, Zhimeng, Rong, Zhixing, Liu, Yongle, Wang, Faxiang, and Li, Xianghong
- Subjects
- *
LOW temperature plasmas , *SILVER carp , *FLAVOR , *ODORS , *GIBBS' free energy , *ADENINE nucleotides , *TASTE - Abstract
This study investigated non-thermal pretreatment (cold plasma, CP) on the flavor (taste and odor) profiles of dried fish products. CP treatment of 5 min contributed to accumulation of umami nucleotides adenosine 5′-monophosphate (AMP) from 30.96 to 40.82 μg/g and inosine 5′-monophosphate (IMP) from 2009.29 to 2132.23 μg/g, and significant reduction of bitter hypoxanthine ribonucleoside (HxR) and hypoxanthine (Hx), respectively (P < 0.05) in the dried fish products. A noticeable enhancement in sweet glycine (from 429.41 to 490.03 mg/100 g) and umami glutamic acid (from 55.68 to 67.76 mg/100 g) accompanied with the CP treatment (P < 0.05) based on taste activity value (TAV > 1). And the characteristic odor volatiles (nonanal, hexanal and 1-octen-3-ol) were strengthened 2.13-, 2.16- and 2.17- folds, respectively (P < 0.05). The results of equivalent umami concentration and Gibbs free energy calculation, combining with the correlation analysis, indicate that nucleotides and free amino acids synergically enhanced the taste improvement of dried fish products. Moderate lipids oxidation favored the formation of characteristic volatiles. The CP pretreatment offered new strategies for enhancing flavor of dried fish products. Fig. 1. Schematic illustration of the design of this study. [Display omitted] • Non-thermal pretreatment (cold plasma, CP) enhanced the flavor of dried fish samples. • CP treatment increased umami AMP and IMP levels and reduced bitter HxR and Hx. • CP treatment induced a noticeable promotion in sweet Gly and umami Glu (TAV > 1). • Nucleotides and FAAs synergistically improved the taste of the samples. • Correlation analysis and ΔG calculation revealed the flavor formation pathway. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
6. A Thermostable Type I Collagen from Swim Bladder of Silver Carp (Hypophthalmichthys molitrix)
- Author
-
Honghui Jiang, Yuanyuan Kong, Lili Song, Jing Liu, and Zhihong Wang
- Subjects
swim bladder ,thermal stability ,collagen ,antioxidant activity ,silver carp (Hypophthalmichthys molitrix) ,Biology (General) ,QH301-705.5 - Abstract
As a major component of the extracellular matrix, collagen has been used as a biomaterial for many purposes including tissue engineering. Commercial collagen derived from mammals is associated with a risk of prion diseases and religious restrictions, while fish-derived collagen can avoid such issues. In addition, fish-derived collagen is widely available and low-cost; however, it often suffers from poor thermal stability, which limits its biomedical application. In this study, collagen with a high thermal stability was successfully extracted from the swim bladder of silver carp (Hypophthalmichthys molitrix) (SCC). The results demonstrated that it was a type I collagen with high purity and well-preserved triple-helix structure. Amino acid composition assay showed that the amounts of threonine, methionine, isoleucine and phenylalanine in the collagen of swim bladder of silver carp were higher than those of bovine pericardium. After adding salt solution, swim-bladder-derived collagen could form fine and dense collagen fibers. In particular, SCC exhibited a higher thermal denaturation temperature (40.08 °C) compared with collagens from the swim bladder of grass carp (Ctenopharyngodon idellus) (GCC, 34.40 °C), bovine pericardium (BPC, 34.47 °C) and mouse tail (MTC, 37.11 °C). Furthermore, SCC also showed DPPH radical scavenging ability and reducing power. These results indicate that SCC presents a promising alternative source of mammalian collagen for pharmaceutical and biomedical applications.
- Published
- 2023
- Full Text
- View/download PDF
7. Changes in Myosin from Silver Carp (Hypophthalmichthys molitrix) under Microwave-Assisted Water Bath Heating on a Multiscale.
- Author
-
Cong, Haihua, Lyu, He, Liang, Wenwen, Zhang, Ziwei, and Chen, Xiaodong
- Subjects
SILVER carp ,HYDRONICS ,MYOSIN ,PARTICLE size distribution ,MICROWAVE heating ,SULFHYDRYL group - Abstract
To further prove the advantages of microwave-assisted water bath heating (MWH) in low-value fish processing, the effects of different heating methods (two heating stage method, high temperature section respectively using MWH1, MWH2, MWH3, WH—water heating, MH—microwave heating) on secondary and tertiary myosin structures, SDS-PAGE, surface morphology, scanning electron microscopy (SEM), and particle size distribution were compared and analyzed. The findings revealed that MH and MWH aided in the production of gel formations by promoting myosin aggregation. Myosin from silver carps demonstrated enhanced sulfhydryl group and surface hydrophobicity after MWH treatment, as well as a dense network structure. The distribution of micropores becomes more uniform when the microwave time is increased. Actually, the total effect of microwave time on myosin is not substantially different. The correlation between particle size distribution and protein aggregation was also studied, in terms of time savings, the MWH of short microwave action is preferable since it not only promotes myosin aggregation but also avoids the drawbacks of a rapid warming rate. These discoveries give a theoretical foundation for understanding silver carp myosin under microwave modification, which is critical in the food industry. [ABSTRACT FROM AUTHOR]
- Published
- 2022
- Full Text
- View/download PDF
8. A Pilot Study Assessing a Concentration of 100 mg/L Alizarin Complexone (ALC) to Mark Calcified Structures in Hypophthalmichthys molitrix.
- Author
-
Zhu, Yahua, Jiang, Tao, Chen, Xiubao, Liu, Hongbo, Phelps, Quinton, and Yang, Jian
- Subjects
- *
SILVER carp , *ALIZARIN , *OTOLITHS , *PILOT projects , *LIGHT sources , *FISH anatomy , *FLUORESCENCE microscopy - Abstract
The effectiveness of chemical compounds for marking hard tissues in juvenile silver carp (Hypophthalmichthys molitrix) is not well known. We analyzed the use of alizarin complexone (ALC) as a fluorescent marker to mark the various hard structures of juvenile silver carp. Experimental fish (~2 months old) were randomly assigned to either control or marking groups, which were immersed in 0 or 100 mg/L ALC solutions, respectively, for 2 days. The otoliths, fin rays, and scales of the fish were then sampled, visualized using fluorescence microscopy, and evaluated after 10 days. The ALC treatment was effective for marking certain hard structures and the marking color was affected by the light source. There were no obvious differences in the marking efficiency of rays from pectoral, dorsal, ventral, anal, and caudal fins, but the lapilli and lateral line scales were marked most effectively from the sampled otolith and scale types, respectively. Our findings indicate that ALC immersion and fin ray and scale sampling can be used for the effective marking and non-lethal evaluation of hard structures in juvenile silver carp. [ABSTRACT FROM AUTHOR]
- Published
- 2022
- Full Text
- View/download PDF
9. Effect of pH-Shifting Process on the Cathepsin Activity, Muddy Off-Odor Compounds’ Content and Gelling Properties of Isolated Protein from Silver Carp
- Author
-
Weidan Guo, Miao Zhan, Hui Liu, Xiangjin Fu, and Wei Wu
- Subjects
silver carp (Hypophthalmichthys molitrix) ,pH-shifting process ,muddy off-odor ,gelling properties ,cathepsin ,Chemical technology ,TP1-1185 - Abstract
Silver carp (Hypophthalmichthys molitrix) is a potential source for making surimi products. However, it has the disadvantages of bony structures, high level of cathepsines and muddy off-odor which is mainly caused by geosmin (GEO) and 2-methylisoborneol (MIB). These disadvantages make the conventional water washing process of surimi inefficient (low protein recovery rate, and high residual muddy off-odor). Thus, the effect of the pH-shifting process (acid-isolating process and alkali-isolating process) on the cathepsins activity, GEO content, MIB content, and gelling properties of the isolated proteins (IPs) was investigated, comparing it with surimi obtained through the conventional cold water washing process (WM). The alkali-isolating process greatly boosted the protein recovery rate from 28.8% to 40.9% (p < 0.05). In addition, it removed 84% GEO and 90% MIB. The acid-isolating process removed about 77% GEO and 83% MIB. The acid-isolated protein (AC) displayed the lowest elastic modulus (G′), the highest TCA-peptide content (90.89 ± 4.65 mg/g) and the highest cathepsin L activity (65.43 ± 4.91 U/g). The AC modori (60 °C for 30 min) gel also demonstrated the lowest breaking force (226.2 ± 19.5 g) and breaking deformation (8.3 ± 0.4 mm), indicating that proteolysis caused by the cathepsin deteriorated the gel quality of AC. The setting (40 °C for 30 min) considerably increased the breaking force (386.4 ± 15.7 g) and breaking deformation (11.6 ± 0.2 mm) of the gel made from the alkali-isolated protein (AK) (p < 0.05). In AC and AK gel, a clearly visible cross-linking protein band with a molecular weight greater than MHC was seen, demonstrating the presence of endogenous trans-glutaminase (TGase) activity, that improved the gel quality of AK. In conclusion, the alkali-isolating process was an effective alternative method for making water-washed surimi from silver carp.
- Published
- 2023
- Full Text
- View/download PDF
10. Improvement of Flesh Quality of Farmed Silver Carp (Hypophthalmichthys molitrix) by Short-Term Stocked in Natural Water
- Author
-
Xiaomin Miao, Hui Guo, Yong Song, Chunying Du, Jingyun Feng, Yixi Tao, Hao Xu, and Yun Li
- Subjects
silver carp (Hypophthalmichthys molitrix) ,flesh quality ,food composition ,digestive enzyme activity ,Biology (General) ,QH301-705.5 ,Genetics ,QH426-470 - Abstract
To investigate the effects of different raising environments on the flesh quality, the study set up three groups of silver carp (Hypophthalmichthys molitrix), that is, pond-farmed; short-term stocked; and ranched, grown in a natural water ranch. This study compared the differences in muscle proximate composition and amino acid composition among three groups. The results showed that there was no difference between the ranched and the stocked groups in content of crude protein, ash, total amino acids (TAA), essential amino acids (EAA), and umami-flavored amino acids (UAA), with both groups being significantly higher than the farmed group. This shows that, in terms of flesh quality, the stocked group was closer to the ranched group and better than the farmed group after being stocked in natural water for 30 days. The results suggest that the flesh quality of farmed fish was improved by short-term stocking in natural water. To elucidate the reasons of flesh quality change in the stocked group, this study compared the food composition and intestinal digestive enzyme activity in three groups, demonstrating that the ranched and stocked groups had similar food composition, with their detritus proportion lower than that of the farmed group, consuming easily digestible phytoplankton diatoms rich in amino acids and fatty acids. No significant difference was seen between the ranched group and the stocked group, for which amylase, lipase, and trypsin activities were lower than that of the farmed group. It is hypothesized that differences in environmental and food composition resulted in lower digestive enzyme activity in the ranched group and the stocked group. In summary, the short-term stocking of farmed fish in a natural water ranch can effectively improve the nutrient content, quality and flavor, and promote value of fish products.
- Published
- 2023
- Full Text
- View/download PDF
11. Changes in Myosin from Silver Carp (Hypophthalmichthys molitrix) under Microwave-Assisted Water Bath Heating on a Multiscale
- Author
-
Haihua Cong, He Lyu, Wenwen Liang, Ziwei Zhang, and Xiaodong Chen
- Subjects
silver carp (Hypophthalmichthys molitrix) ,myosin ,heating ,protein structure ,protein aggregation behavior ,Chemical technology ,TP1-1185 - Abstract
To further prove the advantages of microwave-assisted water bath heating (MWH) in low-value fish processing, the effects of different heating methods (two heating stage method, high temperature section respectively using MWH1, MWH2, MWH3, WH—water heating, MH—microwave heating) on secondary and tertiary myosin structures, SDS-PAGE, surface morphology, scanning electron microscopy (SEM), and particle size distribution were compared and analyzed. The findings revealed that MH and MWH aided in the production of gel formations by promoting myosin aggregation. Myosin from silver carps demonstrated enhanced sulfhydryl group and surface hydrophobicity after MWH treatment, as well as a dense network structure. The distribution of micropores becomes more uniform when the microwave time is increased. Actually, the total effect of microwave time on myosin is not substantially different. The correlation between particle size distribution and protein aggregation was also studied, in terms of time savings, the MWH of short microwave action is preferable since it not only promotes myosin aggregation but also avoids the drawbacks of a rapid warming rate. These discoveries give a theoretical foundation for understanding silver carp myosin under microwave modification, which is critical in the food industry.
- Published
- 2022
- Full Text
- View/download PDF
12. THE GENETIC STRUCTURE OF DIFFERENT AGE GROUPS OF SILVER (HYPOPHTHALMICHTHYS MOLITRIX) AND BIGHEAD (ARISTICHTHYS NOBILIS) CARPS FROM FISH FARM LIMANSKE
- Author
-
Т. Nagorniuk, I. Hrytsyniak, and N. Borysenko
- Subjects
silver carp (Hypophthalmichthys molitrix) ,bighead carp (Aristichthys nobilis) ,age-1 ,age-1+ ,age-2 ,genetic-biochemical markers ,genetic structure ,alleles ,genotype ,heterozygosity ,Aquaculture. Fisheries. Angling ,SH1-691 - Abstract
Purpose. Studying the peculiarities of the genetic structure of different age groups of silver and bighead carps from fish farm Limanske with the use of genetic-biochemical markers. Methodology. The methods of vertical polyacrylamide and horizontal starch electrophoresis with our own modifications have been used for the study. Sampling of the biological material and histochemical staining of gel plates were carried out using the generally accepted methods. Statistical analysis of the obtained data were performed in "Biosys-I". Findings. We analyzed the genetic structure of age-1, age-1+ and age-2 of silver and bighead carps with the use of genetic-biochemical markers - Рralb, EST, MDH, МЕ, СА. We demonstrated the peculiarities of allele frequencies and genotypic composition of protein system loci. A high level of heterozygosity in the examined was detected loci for age-1 silver carp – 66.7-88.9% and age-1 bighead carp – 65.4-77.8%. A significantly higher level of the observed average heterozygosity compared to the expected heterozygosity of age-1 as silver carp (Ho = 75.9%, He = 49,6%), and bighead carp (Ho = 73,6%, He = 47,9%) was observed. This fact indicated on the necessity for performing a stabilization of their genetic structure. The calculated Wright's F index in different age groups showed a predominance of the observed level of the average heterozygosity over the expected one for 22.7–53% in silver carp and 24.5-53.7% in bighead carp. A stabilization of the genetic structure in age-2 silver carp (Ho = 57.8%; He = 47.1%) by the heterogeneity level was observed. Originality. For the first time we analyzed the genetic structure and the level of heterogeneity of the breeding groups of age-1, age-1+, age-2 silver and bighead carps from fish farm Limanske with the use of genetic-biochemical markers. Practical value. The results of the study can be used in selection and breeding works aimed at creating the breeding stocks of silver and bighead carps to control the level of interspecies genetic variability. The obtained experimental data of the genetic structure analysis of silver and bighead carps of different age groups will be used for the monitoring of the genetic structure changes during their adaptation to new environmental conditions.
- Published
- 2015
- Full Text
- View/download PDF
13. THE EFFECTS OF WATER TEMPERATURE REGIME FLUCTUATIONS ON THE EMBRYONIC DEVELOPMENT OF SILVER CARP (HYPOPHTHALMICHTHYS MOLITRIX)
- Author
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А. Vodyanitskyi, А. Potrokhov, and О. Zinkovskyi
- Subjects
embryonic development ,silver carp (Hypophthalmichthys molitrix) ,fluctuations of temperature regime in water bodies ,enzymatic activity ,Aquaculture. Fisheries. Angling ,SH1-691 - Abstract
Purpose. To determine the effect of temperature regime fluctuations on the development of silver carp embryos, as well as the activity of enzymatic reactions in fish eggs. Methodology. The studies were conducted at the experimental station of the Institute of Hydrobiology of Bila Tserkov, Ukrainian National Academy of Sciences, from June to July. The biological materials were silver carp eggs, embryos and larvae. The dissolved oxygen content was determined using the Winkler method at four o’clock in the morning. Alkalinity phosphatase and LDG activity were determined using a set of reagents «Alkalinity phosphatase» and «LDG» (Phyllis diagnosis, Ukraine). SDH activity was determined by Vexy. The activity of Na, K-Mg-dependent-activated ATPase was determined as growth of inorganic phosphorus in the incubation medium by Kindratova M.N. et al. Protease activity was determined using immune enzymatic method of Tyurina et al. The obtained results were processed statistically in Statistica 5.5, Epaprobit analysis was used for calculating LC/EC values (Version 1.5). Findings The results showed that a delay of embryonic stages of development occur, the number of abnormal embryos increases, and the reproduction efficiency of fish reduces with an increase in water temperature and decrease in the dissolved oxygen content in water. The temperature factor had a significant effect on the activity of key enzymes, in particular the energetic metabolism changed from aerobic to anaerobic. Originality. It was found a negative effect of abiotic factors of water medium and drastic fluctuations in water temperature and gas regime of water bodies on the course of embryogenesis of silver carp that is especially important in the conditions of climate change. Practical value. The obtained results showed that the level of optimum and unfavorable environmental factors during the change of embryonic stages in embryonic and larval fish can be established based on the activity of lactate dehydrogenase, succinate dehydrogenase, ATPase and protease.
- Published
- 2015
- Full Text
- View/download PDF
14. INFORMATIONAL CONTENT OF MICROSATELLITE LOCI FOR ANALYSIS OF GENETIC STRUCTURE OF SILVER (HYPOPHTHALMICHTHYS MOLITRIX) AND BIGHEAD (HYPOPHTHALMICHTHYS NOBILIS) CARPS
- Author
-
N. Borysenko, I. Hrytsyniak, and S. Tarasjuk
- Subjects
silver carp (Hypophthalmichthys molitrix) ,bighead (Hypophthalmichthys nobilis) carp ,microsatellite loci ,Aquaculture. Fisheries. Angling ,SH1-691 - Abstract
Purpose. The goal of this work was the selection of microsatellite markers to study the genetic structure of populations of silver (Hypophthalmichthys molitrix) and bighead (Hypophthalmichthys nobilis) carps of fish farm JSC "Donrybkombinat" of Donetsk region. Methodology. Products of amplification were separated in 2 % agarose gel in 1×TBE buffer. Visualization was performed with help of transilluminator in UV-region of the spectrum and photographing of electrophoregrams by digital camera. Processing and analysis of gels were performed using program TotalLab v2.01. The frequency of each amplicon by particular locus was determined as a percentage from the total number of amplicons at this locus. Statistical analysis of results was carried out using programs Excel, «Biosys-1». Findings. At the results of investigations of populations silver carp and bighead carp were analyzed genotypes of individuals by using three microsatellite DNA loci: MFW 15, MFW 23, MFW 06. Originality. It has been first shown that to investigate genetic structure of Ukrainian silver and bighead carps of JSC "Donrybkombinat" the most informative microsatellite loci were MFW 15 and MFW 23. Practical value. Microsatellite loci MFW 15 and MFW 23 used in research were suitable for interspecies differentiation of Ukrainian silver and bighead carps and can be applied in population-genetic investigations of fishes.
- Published
- 2013
- Full Text
- View/download PDF
15. Cloning, Expression, Characterization, and Tissue Distribution of Cystatin C from Silver Carp (Hypophthalmichthys molitrix)
- Author
-
Song Li, Xiaoqian Tan, Ran Li, Shulei Li, Yu Jin, Per E. J. Saris, Timo M. Takala, Shuhong Li, Department of Microbiology, and Antimicrobials, probiotics and fermented food
- Subjects
0106 biological sciences ,Proteases ,protein quantification ,tissue distribution ,cloning ,AMINO-ACID-SEQUENCE ,silver carp (Hypophthalmichthys molitrix) ,Molecular cloning ,01 natural sciences ,chemistry.chemical_compound ,EGG-WHITE ,Affinity chromatography ,cystatin C ,transcription level ,Polyacrylamide gel electrophoresis ,GENE-EXPRESSION ,chemistry.chemical_classification ,Silver carp ,MOLECULAR-CLONING ,CATHEPSIN-B ,PURIFICATION ,biology ,OVARIAN CYSTATIN ,IDENTIFICATION ,010401 analytical chemistry ,General Chemistry ,MUSCLE ,biology.organism_classification ,3. Good health ,0104 chemical sciences ,Amino acid ,Papain ,Cystatin C ,Biochemistry ,chemistry ,416 Food Science ,biology.protein ,1182 Biochemistry, cell and molecular biology ,CYSTEINE PROTEINASE-INHIBITOR ,General Agricultural and Biological Sciences ,010606 plant biology & botany - Abstract
Cystatins are proteins, which inhibit cysteine proteases, such as papain. In this study, the 336-bp cystatin C gene (family II, HmCysC) of silver carp (Hypophthalmichthys molitrix) was cloned and expressed in Escherichia coli BL21 (DE3). HmCysC encodes the mature peptide of cystatin C (HmCystatin C), with 111 amino acids. A typical QXXXG motif was found in HmCystatin C and it formed a cluster with Cyprinus carpio and Danio rerio cystatin C in the phylogenetic tree. Quantitative real-time polymerase chain reaction analysis indicated that HmCysC was transcribed at different levels in five tested tissues of silver carp. Following purification with Ni2+– nitrilotriacetic acid agarose affinity chromatography, HmCystatin C displayed a molecular weight of 20 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis. Purified HmCystatin C had strong inhibitory effects toward the proteolytic activity of papain. Immunochemical staining with anti-HmCystatin C antibody showed that HmCystatin C was widely distributed in silver carp tissues. These results collectively demonstrated the properties of HmCystatin C, providing information for further studies of cystatins from fish organisms.
- Published
- 2021
16. A Pilot Study Assessing a Concentration of 100 mg/L Alizarin Complexone (ALC) to Mark Calcified Structures in Hypophthalmichthys molitrix
- Author
-
Yahua Zhu, Tao Jiang, Xiubao Chen, Hongbo Liu, Quinton Phelps, and Jian Yang
- Subjects
Ecology ,marking ,alizarin complexone ,hard tissue ,fin ray ,otolith ,scale ,silver carp (Hypophthalmichthys molitrix) ,sense organs ,Aquatic Science ,Ecology, Evolution, Behavior and Systematics - Abstract
The effectiveness of chemical compounds for marking hard tissues in juvenile silver carp (Hypophthalmichthys molitrix) is not well known. We analyzed the use of alizarin complexone (ALC) as a fluorescent marker to mark the various hard structures of juvenile silver carp. Experimental fish (~2 months old) were randomly assigned to either control or marking groups, which were immersed in 0 or 100 mg/L ALC solutions, respectively, for 2 days. The otoliths, fin rays, and scales of the fish were then sampled, visualized using fluorescence microscopy, and evaluated after 10 days. The ALC treatment was effective for marking certain hard structures and the marking color was affected by the light source. There were no obvious differences in the marking efficiency of rays from pectoral, dorsal, ventral, anal, and caudal fins, but the lapilli and lateral line scales were marked most effectively from the sampled otolith and scale types, respectively. Our findings indicate that ALC immersion and fin ray and scale sampling can be used for the effective marking and non-lethal evaluation of hard structures in juvenile silver carp.
- Published
- 2022
- Full Text
- View/download PDF
17. Investigation on Structural Changes of Myofibrillar Proteins from Silver Carp (Hypophthalmichthys molitrix) during Frozen Storage.
- Author
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Ren LINA, XU YANSHUN, Jiang QIXING, Xia WENSHUI, and Qiu CHUNJIANG
- Abstract
The effect of frozen storage at -20°C and -50°C on the structure of myofibrillar proteins (MP) from silver carp was determined using several analytical methods. Differential scanning calorimetry thermo-grams demonstrated that the thermal stability of MP decreased and some structural changes occurred particularly in the rod-like tail portion of myosin. The trend of surface hydrophobicity was first an increase and then a decrease, indicating that the unfolding of protein molecules and exposure of buried hydrophobic residues occurred firstly, followed by the aggregation of MP through hydrophobic interactions, and this was also supported by intrinsic fluorescence and Raman. Circular dichroism and Raman data indicated that there was a decrease in α-helix content with a concomitant increase in β-sheet during storage. All the experimental data confirmed that the MP stored at -20°C suffered more pronounced structural alteration than those stored at -50°C. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
18. Effect of high hydrostatic pressure (HHP) on myofibril-bound serine proteinases and myofibrillar protein in silver carp (Hypophthalmichthys molitrix).
- Author
-
Qiu, Chunjiang, Xia, Wenshui, and Jiang, Qixing
- Subjects
- *
HYDROSTATIC pressure , *MYOFIBRILS , *SERINE proteinases , *SILVER carp , *FRESHWATER fishes , *SURIMI , *SEAFOOD , *FOOD texture - Abstract
Abstract: Silver carp (Hypophthalmichthys molitrix) is a popular cultured freshwater fish in China, and becomes a potential raw material for surimi product. However, silver carp surimi exhibits considerable gel softening (modori) and myofibril-bound serine proteinases (MBSP) involved in the degradation of myofibrillar proteins have been assumed to be crucial to the modori phenomenon. High-pressure technology has demonstrated its potential application to control enzyme-related seafood texture deterioration. Herefrom, it could be further applied to the freshwater fish processing and the effect of high pressure treatments on the activity of MBSP implicated in texture deterioration in myofibrils and crude enzyme extracts was evaluated. The inactivation kinetics of MBSP pressured from 200MPa to 500MPa at room temperature (20°C) were fitted first-order kinetics, The extent of enzyme inactivation was lower in silver carp myofibrils in comparison with the crude enzyme extracts from myofibrils showing a protective effect against the high pressure treatment. The effect of HHP on MBSP myofibrillar degradation shown by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was associated with a corresponding decrease coincided with the elevated high pressure levels in intensity of myosin heavy chain (MHC). Dynamic rheological measurements indicated that high-pressure treatment exerted considerable influence on MBSP activities as well as proteins (mainly myofibrils), resulting in structural modifications and texture changes. In this study, 300MPa (≥10min) was the process condition (pressure and time) that caused apparent MBSP inactivation derived from the first order kinetic model and had the efficacy in controlling silver carp texture deterioration both verified by SDS-PAGE and dynamic rheological measurements. [Copyright &y& Elsevier]
- Published
- 2013
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19. Purification and characterisation of cathepsin L2 from dorsal muscle of silver carp (Hypophthalmichthys molitrix)
- Author
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Li, ShuHong, Zhou, XiaoQiu, Zhang, Nan, Liu, Huan, and Ma, ChangWei
- Subjects
- *
SILVER carp , *CHROMATOGRAPHIC analysis , *HYDROLYSIS , *POLYPEPTIDES - Abstract
Abstract: Cathepsin L2 was purified to homogeneity from silver carp muscle using an array of chromatography methods. The enzyme showed affinity to con A-sepharose. Although it appeared to be 78kDa on non-reducing SDS–PAGE and gel–substrate-activity SDS–PAGE, it completely degraded into 31kDa and 26kDa sub-units, as well as some small polypeptides on reducing SDS–PAGE. The optimum pH and temperature of cathepsin L2 for hydrolysis of Z-Phe-Arg-MCA were pH 4.5–5.5 and 45°C, respectively. It was stable at pH 5.5 and below 40°C, but almost inactivated at pH 7.0 and 60°C. Substrate specificity analysis indicated that it could hydrolyse Z-Phe-Arg-MCA but not Z-Arg-Arg-MCA or l-Arg-MCA. Cathepsin L2 was efficiently activated by Cys, DTT and β-ME, but was completely inhibited by E-64. and Cl− have inhibitory effects on its activity. Cathepsin L2 showed a high K m value of 9.5μmol/l, but extremely low K cat and K cat/K m values of 0.8s−1 and 84.2s−1 mM−1, respectively. Except for under optimum conditions (pH 5.0, 35°C), silver carp cathepsin L2 could also hydrolyse myosin heavy chain at softening temperatures ranging from 50 to 60°C and at surimi pH of 6.5. [Copyright &y& Elsevier]
- Published
- 2008
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20. Effect of mixed feeding schedules with varying dietary protein levels on the growth of sutchi catfish,Pangasius hypophthalmus(Sauvage) with silver carp,Hypophthalmichthys molitrix(Valenciennes) in ponds.
- Author
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Ali, Md Zulfikar, Hossain, Md Arshad, and Mazid, Md Abdul
- Subjects
- *
CATFISHES , *FISH feeds , *LOW-protein diet , *SILVER carp , *FISH ponds - Abstract
A 6-month feeding trial was conducted in field condition using 10 farm ponds (400–600 m2) to investigate the effect of mixed feeding schedules on the growth of sutchi catfish,Pangasius hypophthalmuswith silver carp,Hypophthalmichthys molitrix. Fish were stocked at a ratio of 80:20 (sutchi catfish, 4.9±0.5 g: silver carp, 12.0±0.8 g) at the total rate of 25 000 ha−1. Two diets of high protein (30%, HP) and low protein (16%, LP) were prepared using locally available feed ingredients. Five different feeding schedules of high-protein diet continuously (HP), low-protein diet continuously (LP), 1-day low–protein/1-day high-protein diet (1LP/1HP), 7 days low–protein/7 days high-protein diet (7LP/7HP) and 14 days low–protein/14 days high-protein diet (14LP/14HP) were tested. The fish were fed twice daily at the rate of 15%, 10%, 8% and 5% of their body weight for first, second, third month and rest of experimental period respectively. Feeding rate was calculated only on the basis of sutchi catfish weight only and was adjusted every 2 weeks according to weight gain. Fish fed LP and HP on alternate day (1LP/1HP) resulted in significantly (P<0.05) higher growth rate, feed utilization and production among the treatments. However, there were no significant differences (P>0.05) between the growth rates and production of fish fed HP regularly and fish fed 7 days LP followed by 7 days HP (7LP/7HP). Fish maintained on LP grew the least. The feed conversion ratio (FCR) values for sutchi catfish ranged between 2.04 and 2.79 with feeding schedule 1LP/1HP showing the best FCR. The total production of fish (including silver carp) ranged between 8310 and 12 422 kg ha−1 6 months−1 with 1LP/1HP feeding schedule resulting in the highest production and net profit. The study demonstrated that feeding fish continuously with HP is less economical. Thus, for profitable sutchi catfish culture with silver carp, farmers can use the mixed feeding schedule of alternate day feeding of LP and HP as a means of reducing feed costs. [ABSTRACT FROM AUTHOR]
- Published
- 2005
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21. THE GENETIC STRUCTURE OF DIFFERENT AGE GROUPS OF SILVER (HYPOPHTHALMICHTHYS MOLITRIX) AND BIGHEAD (ARISTICHTHYS NOBILIS) CARPS FROM FISH FARM LIMANSKE
- Author
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I. Hrytsyniak, Т. Nagorniuk, and N. Borysenko
- Subjects
lcsh:SH1-691 ,Hypophthalmichthys ,biology ,genotype ,Fish farming ,age-1+ ,Aristichthys nobilis ,silver carp (Hypophthalmichthys molitrix) ,genetic-biochemical markers ,bighead carp (Aristichthys nobilis) ,biology.organism_classification ,age-2 ,lcsh:Aquaculture. Fisheries. Angling ,age-1 ,Fishery ,Age groups ,alleles ,Genetic structure ,genetic structure ,heterozygosity ,sense organs - Abstract
Purpose. Studying the peculiarities of the genetic structure of different age groups of silver and bighead carps from fish farm Limanske with the use of genetic-biochemical markers. Methodology. The methods of vertical polyacrylamide and horizontal starch electrophoresis with our own modifications have been used for the study. Sampling of the biological material and histochemical staining of gel plates were carried out using the generally accepted methods. Statistical analysis of the obtained data were performed in "Biosys-I". Findings. We analyzed the genetic structure of age-1, age-1+ and age-2 of silver and bighead carps with the use of genetic-biochemical markers - Рralb, EST, MDH, МЕ, СА. We demonstrated the peculiarities of allele frequencies and genotypic composition of protein system loci. A high level of heterozygosity in the examined was detected loci for age-1 silver carp – 66.7-88.9% and age-1 bighead carp – 65.4-77.8%. A significantly higher level of the observed average heterozygosity compared to the expected heterozygosity of age-1 as silver carp (Ho = 75.9%, He = 49,6%), and bighead carp (Ho = 73,6%, He = 47,9%) was observed. This fact indicated on the necessity for performing a stabilization of their genetic structure. The calculated Wright's F index in different age groups showed a predominance of the observed level of the average heterozygosity over the expected one for 22.7–53% in silver carp and 24.5-53.7% in bighead carp. A stabilization of the genetic structure in age-2 silver carp (Ho = 57.8%; He = 47.1%) by the heterogeneity level was observed. Originality. For the first time we analyzed the genetic structure and the level of heterogeneity of the breeding groups of age-1, age-1+, age-2 silver and bighead carps from fish farm Limanske with the use of genetic-biochemical markers. Practical value. The results of the study can be used in selection and breeding works aimed at creating the breeding stocks of silver and bighead carps to control the level of interspecies genetic variability. The obtained experimental data of the genetic structure analysis of silver and bighead carps of different age groups will be used for the monitoring of the genetic structure changes during their adaptation to new environmental conditions.
- Published
- 2015
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22. THE EFFECTS OF WATER TEMPERATURE REGIME FLUCTUATIONS ON THE EMBRYONIC DEVELOPMENT OF SILVER CARP (HYPOPHTHALMICHTHYS MOLITRIX)
- Author
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О. Zinkovskyi, А. Vodyanitskyi, and А. Potrokhov
- Subjects
lcsh:SH1-691 ,Silver carp ,Hypophthalmichthys ,biology ,Chemistry ,fluctuations of temperature regime in water bodies ,enzymatic activity ,Embryogenesis ,silver carp (Hypophthalmichthys molitrix) ,biology.organism_classification ,lcsh:Aquaculture. Fisheries. Angling ,Water temperature ,embryonic development ,Food science - Abstract
Purpose. To determine the effect of temperature regime fluctuations on the development of silver carp embryos, as well as the activity of enzymatic reactions in fish eggs. Methodology. The studies were conducted at the experimental station of the Institute of Hydrobiology of Bila Tserkov, Ukrainian National Academy of Sciences, from June to July. The biological materials were silver carp eggs, embryos and larvae. The dissolved oxygen content was determined using the Winkler method at four o’clock in the morning. Alkalinity phosphatase and LDG activity were determined using a set of reagents «Alkalinity phosphatase» and «LDG» (Phyllis diagnosis, Ukraine). SDH activity was determined by Vexy. The activity of Na, K-Mg-dependent-activated ATPase was determined as growth of inorganic phosphorus in the incubation medium by Kindratova M.N. et al. Protease activity was determined using immune enzymatic method of Tyurina et al. The obtained results were processed statistically in Statistica 5.5, Epaprobit analysis was used for calculating LC/EC values (Version 1.5). Findings The results showed that a delay of embryonic stages of development occur, the number of abnormal embryos increases, and the reproduction efficiency of fish reduces with an increase in water temperature and decrease in the dissolved oxygen content in water. The temperature factor had a significant effect on the activity of key enzymes, in particular the energetic metabolism changed from aerobic to anaerobic. Originality. It was found a negative effect of abiotic factors of water medium and drastic fluctuations in water temperature and gas regime of water bodies on the course of embryogenesis of silver carp that is especially important in the conditions of climate change. Practical value. The obtained results showed that the level of optimum and unfavorable environmental factors during the change of embryonic stages in embryonic and larval fish can be established based on the activity of lactate dehydrogenase, succinate dehydrogenase, ATPase and protease.
- Published
- 2015
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23. Relationship between genetic risk and stock enhancement of the silver carp (Hypophthalmichthys molitrix) in the Yangtze River.
- Author
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Fang, Di-an, Luo, Yu-Ting, Xu, Dong-po, Yang, Xi-wen, and Wang, Xiao-hao
- Subjects
- *
SILVER carp , *MICROSATELLITE repeats , *FISHERY resources , *LIVESTOCK breeding , *GERMPLASM - Abstract
• SSR labeled-release-recapture routines were established to evaluate stock enhancement. • Silver carps were facing population depression and genetic structure homogeny. • Potential genetic risk existed between recaptured populations by constant SE. • Urgent scientific SE breeding programs and traceability system should be constructed. Hatchery release of silver carp (Hypophthalmichthys molitrix) is an efficient strategy that has been carried out for years, and its scales expand year by year in the lower reaches of the Yangtze River (LRYR) in China. As the cultured juveniles are released into self-recruiting wild population to increase fisheries resources yields, they may potentially come along with negative impacts such as intra-inter specific completion, loss of genetic diversity and fitness, even causing genetic risk. To assess the relationship between genetic risk and stock enhancement (SE) of the silver carp in the Yangtze River (YR); microsatellite loci amplification, parentage assignment, genetic diversity, genetic differentiation and structure were conducted and testified by the help of eleven fluorescence-labeled microsatellite markers method. Results showed that silver carp populations in the YR had relatively higher genetic diversity (UHe), inbreeding coefficient (Fis), and effective population size (Ne) than breeding stock populations from eight hatcheries. By using the analysis of molecular variance (AMOVA), results showed that the percentage of variance among populations and within populations were 4.15 % and 95.85 %, respectively, which indicated that genetic structure of YR populations possibly had been changed by SE. Therefore, the present study suggests that the large-scale SE possibly resulted in negative genetic impact on silver carp population. Most importantly, an evaluation system in fisheries resource recruitment and genetic risks was established for SE and management. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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24. Cloning, Expression, Characterization, and Tissue Distribution of Cystatin C from Silver Carp ( Hypophthalmichthys molitrix ).
- Author
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Li R, Tan X, Li S, Jin Y, Li S, Li S, Takala TM, and Saris PEJ
- Subjects
- Animals, Cloning, Molecular, Cystatin C, Phylogeny, Tissue Distribution, Carps genetics, Cystatins genetics
- Abstract
Cystatins are proteins, which inhibit cysteine proteases, such as papain. In this study, the 336-bp cystatin C gene (family II, Hm CysC ) of silver carp ( Hypophthalmichthys molitrix ) was cloned and expressed in Escherichia coli BL21 (DE3). Hm CysC encodes the mature peptide of cystatin C (HmCystatin C), with 111 amino acids. A typical QXXXG motif was found in HmCystatin C and it formed a cluster with Cyprinus carpio and Danio rerio cystatin C in the phylogenetic tree. Quantitative real-time polymerase chain reaction analysis indicated that Hm CysC was transcribed at different levels in five tested tissues of silver carp. Following purification with Ni
2+ - nitrilotriacetic acid agarose affinity chromatography, HmCystatin C displayed a molecular weight of 20 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis. Purified HmCystatin C had strong inhibitory effects toward the proteolytic activity of papain. Immunochemical staining with anti-HmCystatin C antibody showed that HmCystatin C was widely distributed in silver carp tissues. These results collectively demonstrated the properties of HmCystatin C, providing information for further studies of cystatins from fish organisms.- Published
- 2021
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25. Pressure-induced changes of silver carp (Hypophthalmichthys molitrix) myofibrillar protein structure
- Author
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Qiu, Chunjiang, Xia, Wenshui, and Jiang, Qixing
- Published
- 2014
- Full Text
- View/download PDF
26. Development of 201 tri- and tetra-nucleotide repeat microsatellites for bighead carp (Aristichthys nobilis)
- Author
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Zhu, Chuankun, Yu, Xiaomu, Fu, B., and Tong, Jingou
- Published
- 2013
- Full Text
- View/download PDF
27. Enclosure experiments on and lacustrine practice for eliminatingMicrocystis bloom
- Author
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Jian-kang, Liu and Ping, Xie
- Published
- 2002
- Full Text
- View/download PDF
28. 2011 In-Water Testing of Aquatic Nuisance Species Dispersal Barriers IIA and IIB with Increased Voltage and Frequency Operating Parameters
- Author
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ENGINEER RESEARCH AND DEVELOPMENT CENTER CHAMPAIGN IL CONSTRUCTION ENGINEERING RESEARCH LAB, McInerney, Michael K, Aubin, Brianna S, Trovillion, Jonathan C, Baxter, Carey L, Trovillion, Ethan T, Hock, Jr , Vincent F, Weir, David M, ENGINEER RESEARCH AND DEVELOPMENT CENTER CHAMPAIGN IL CONSTRUCTION ENGINEERING RESEARCH LAB, McInerney, Michael K, Aubin, Brianna S, Trovillion, Jonathan C, Baxter, Carey L, Trovillion, Ethan T, Hock, Jr , Vincent F, and Weir, David M
- Abstract
US Army Engineer District - Chicago operates an electric field-based aquatic nuisance species dispersal barrier system in the Chicago Sanitary and Ship Canal (CSSC), Romeoville, IL. The barriers were constructed to prevent the movement of invasive species, such as Asian big-head carp (Hypophthalmichthys nobilis) and silver carp (Hypophthalmichthys molitrix) between the Mississippi River and Great Lakes basins. The objective of this project was to perform a series of in-water tests on the barrier addressing field-strength mapping, sparking potential during barge fleeting and collision, voltage potentials between barges traversing the barriers, personnel in-water shock potential, stray-current corrosion potential, and optimal settings for the parasitic barrier system. Test results and analysis indicate there is no significant risk of personnel shock hazard in the fleeting area during barrier operations for any operating configuration. Also, while some operational scenarios were found to increase sparking risk if barges collide with each other or separate metal objects, analysis indicates that concerns about coal dust explosion hazard from sparking are not supported by the technical literature. A detailed set of data, analysis, conclusions, and recommendations is provided in the report text and four appendices., Project title: CSSC Dispersal Barrier II, ERDC-CERL Safety Studies . The original document contains color images.
- Published
- 2011
29. Preparation and identification of peptides and their zinc complexes with antimicrobial activities from silver carp (Hypophthalmichthys molitrix) protein hydrolysates.
- Author
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Jiang L, Wang B, Li B, Wang C, and Luo Y
- Abstract
The antibacterial activities of peptide-zinc complexes (PZCs) prepared from silver carp (Hypophthalmichthys molitrix) protein hydrolysates treated by four enzymes were evaluated. PZCs prepared from flavourzyme hydrolysates exhibited the highest antibacterial activities. The peptides that were able to produce PZCs with high antibacterial activity were isolated from flavourzyme hydrolysates using chromatographic procedures. Eleven peptides were identified through nano-liquid chromatography-electrospray ionization mass spectrometry. Five of these identified peptides were synthesized. The PZCs obtained were characterized and tested for antimicrobial activities. Results suggested that all five PZCs present strong antibacterial activities but no antifungal activities. Peptides with more acidic amino acid residues in their sequences could bind more zinc ions than peptides with fewer acidic amino acid residues. Zinc complexes of acidic amino acid-richer peptides showed stronger antibacterial activities. Results confirmed the antibacterial activities of PZCs from silver carp hydrolysates, and suggested the feasibility of PZCs as a provider of natural zinc ligands for antibacterial agents in the food and feed industries., (Copyright © 2014 Elsevier Ltd. All rights reserved.)
- Published
- 2014
- Full Text
- View/download PDF
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