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1.
ABSTRACT:   The post-mortem changes of type I and V collagens in Japanese flounder muscle during chilled storage were examined. The muscle softened significantly within 12 h under chilled storage. Both type I and V collagens isolated by ion-exchange chromatographies showed no remarkable changes in band patterns even after 24 h of storage, suggesting that degraded collagen molecules may be removed from the collagen fraction by a conventional preparation method. In contrast, type I and V collagen molecules were detected by Western blotting with each specific antibody in a saline extract and gradually increased during chilled storage. These results suggest that both type I and V collagens may participate in the post-mortem softening of fish muscle.  相似文献   

2.
Acid soluble collagen (ASC) and pepsin soluble collagen (PSC) were isolated from rohu skin with the yield of 64.2 and 6.8% (dry weight basis), respectively. Both collagens had glycine as the major amino acid with imino acid content of 196–202 residues/1,000 residues and were characterized as type I collagen with molecular composition of (α1)2α2-heterotrimer. Fourier transform infrared spectra of both collagens were similar, with no shift in wavenumber of all amide bands. The Tmax value of ASC and PSC was 36.40 and 35.48°C, respectively. The zero surface net charge of ASC and PSC was found at pH 5.9 and 5.3, respectively.  相似文献   

3.
ABSTRACT

Skin collagen of six discarded fish species was analyzed and compared. Acid soluble collagen (ASC) was extracted; a characteristic sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) profile for type I collagen was obtained, except for Chimaera mostrosa. Contents of collagen calculated from HPro (31.85% average) were higher than those determined from ASC extracts (17.75% average), with Galeus spp. being the species with the higher percentage. Amino acid analysis revealed the typical composition of collagen, with very few differences among species. Specific profiles were obtained after protease digestion. Denaturation temperature of ASC correlated well with imino and hydroxyproline contents.

Results demonstrate the feasibility of using the obtained collagens in different industrial applications.  相似文献   

4.

Pepsin-solubilized collagen (PSC) was prepared from the dermis of sea cucumber Apostichopus japonicus (green type) by performing pepsin digestion to collagen fiber pretreated with disaggregating solution (0.1 M Tris–HCl, pH 8.0, containing 0.5 M NaCl, 0.05 M ethylenediaminetetraacetic acid (EDTA), and 0.2 M 2-mercaptoethanol) and 0.1 M NaOH. On sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), the PSC clearly showed two alpha bands under phosphate buffer system in the presence of 3.5 M urea. An antiserum was raised against chromatographically purified major molecular species in the PSC, and immunoblot analyses were performed for the soluble fractions at 4 M guanidine hydrochloride treatment and disaggregation as well as the collagen fiber before and after treatment. These fractions and collagen fibers showed quite similar band patterns to that of the PSC, showing mainly two alpha bands. These combined results suggest that the major molecular species of collagen contains at least two distinct alpha components and that the effect of pepsin digestion is relatively small on the structure of this collagen type.

  相似文献   

5.
The mantle muscles of five cephalopod species, Todarodes pacificus, Photololigo edulis, Sepioteuthis lessoniana, Sepia esculenta, and Sepia longipes, were extracted with 0.1 M NaOH to prepare crude collagen fiber, called ‘residue after alkali extraction’ (RS-AL). Solubility of the collagens in water was examined in the temperature range 20–90°C. The collagens showed a similar tendency in solubility, which gradually increased depending on the treating temperature, and the values at 40–90°C were constantly less than 47.0% for all the species examined. In addition, the collagens were estimated to denature in the approximate temperature range of 37.5–42.5°C. These results suggest that the collagens in RS-AL from these species may have relatively high resistance to hot-water extraction even after their denaturation.  相似文献   

6.
KEN  TOUHATA  YUKI  TOKUDA  MORIHIKO  SAKAGUCHI  HARUHIKO  TOYOHARA 《Fisheries Science》2002,68(5):1118-1123
We previously cloned cDNA of type V/XI collagen α1 chain (ColVa1) gene from cultured cells derived from red sea bream embryo. We raised an antibody against the deduced C-telopeptide of ColVa1 in order to detect the translation products of this cDNA and their degradation products in red sea bream muscle. To improve its specificity, the antibody was purified from rabbit antiserum by use of an affinity column cross-linked with recombinant C-terminal peptide of ColVa1 produced by Epicurian coli. The purified antibody recognized a band corresponding to the α chain of type V/XI collagen in western blot analysis of the extract of cultured cells. The antibody also recognized two bands in acid-soluble and pepsin-solubilized collagens, indicating that the translation products of the ColVa1 gene are present in muscle and that bands correspond to α and β chains of type V/XI collagen. A band corresponding to a molecular weight of approximately 65 k was detected in the NaOH extracts of muscle, suggesting that type V/XI collagen α1 chain is restrictedly digested in red sea bream muscle.  相似文献   

7.
为增加暗纹东方鲀附加值,考察不同处理方法对提取暗纹东方鲀鱼皮胶原蛋白的影响,实验以暗纹东方鲀鱼皮为对象,通过热水法、酸法和胃蛋白酶、木瓜蛋白酶、无花果蛋白酶处理鱼皮提取胶原蛋白。通过对不同胶原蛋白的提取率、氨基酸组成、傅里叶红外光谱(FTIR)、紫外光谱(UV)、扫描电镜、十二烷基硫酸钠—聚丙烯酰胺凝胶电泳(SDS-PAGE)进行研究,对暗纹东方鲀鱼皮胶原蛋白特性进行表征和比较。结果显示,用胃蛋白酶得到的胶原蛋白提取率最高;氨基酸组成相似但含量不同;3种酶制备的胶原蛋白中,脯氨酸含量显著低于酸和热水制备的胶原蛋白;FTIR扫描结果表明,5种处理方法得到的胶原蛋白都存在Amide A、Amide B、AmideⅠ、AmideⅡ和AmideⅢ,均保持了胶原蛋白三螺旋结构;紫外光谱显示在235 nm左右有强吸收峰,结合FTIR确定其为典型的胶原蛋白,经过SDS-PAGE分析,确定暗纹东方鲀鱼皮胶原蛋白为Ⅰ型胶原蛋白。酸法和胃蛋白酶较好地保留了胶原蛋白的β、α1和α2链,木瓜蛋白酶作用化学键比其他酶广泛,得到小分子量的胶原蛋白分子;扫描电镜结果显示,酸法提取的胶原蛋白最适合应用在生物医学材料上运载药物。由此可见,不同处理方法提取的胶原蛋白理化特性存在一定差异,不同的酶制备的胶原蛋白分子量分布会产生明显差别,可根据研究需要选用不同处理方法开发胶原蛋白产品。  相似文献   

8.
In order to study the similarities and differences of physical-chemical properties of collagens from different parts of the fish body, acid-soluble collagens and pepsin-soluble collagens were isolated from skin, scale, and bone of grass carp (Ctenopharyngodon idellus), with yields ranging from 0.7 to 25.5% on the basis of dry weight. The content of acid-soluble collagen in skin and scale was much higher than that in bone according to the data of extraction yield. Collagens from grass carp have similar amino acid composition, protein patterns, and sequences of primary structure. Thermal transition temperatures of collagens from different parts of grass carp body decreased in the order of bone > skin > scale. The pepsin-soluble collagens showed more resistance to collagenase, compared to acid-soluble collagens extracted from the same part of the grass carp body. All grass carp collagens did not induce a significant cytotoxic effect according to the results of in vitro cytotoxicity test.  相似文献   

9.
Little is known about molecular mechanisms involved in gonad differentiation in sturgeon species. In non‐mammalian vertebrates, sox9, dmrt1, cyp17a1 and ar are male specific genes in testicular differentiation and are highly conserved. In order to understand the mechanism underlying testicular development in sturgeon, we investigated sex steroids level of 11‐ketotestosterone (KT) and testosterone (T) and relative expression of sox9, dmrt1, cyp17a1 and ar in great sturgeon gonads during different stages of sexual maturity. The results showed all studied genes had dimorphic patterns in males. In immature gonads (stage I), only cyp17a1 expressed in male gonads, while other genes did not express, and KT and T levels were low. Dmrt1 showed dimorphic expression pattern in male gonads at maturity stage II, III and IV. Furthermore, sox9 and ar mRNA presented significant dimorphic expression pattern in male gonads only at maturity stage 4. Plasma androgens levels were significantly higher in males compared to females during maturity stages II, III and IV. The results showed that among these four genes, only cyp17a1 expressed in male gonads at maturity stage I (immature), suggesting that this gene may be applicable as a sex marker in recently differentiated male great sturgeon. Sexually dimorphic patterns in other studied genes (sox9, dmrt1 and ar) suggesting that these genes may be important for testicular development and differentiation in premature great sturgeon. The obtained results provide a foundation for further research on sex differentiation and developing strategies for the sexing of sturgeon for aquaculture.  相似文献   

10.
The influence of three different initial stocking densities (low stocking density [LSD] = 1.5 kg/m2; medium stocking density [MSD] = 3 kg/m2; and high stocking density [HSD] = 6 kg/m2) in flow‐through systems was evaluated on growth and welfare in beluga, Huso huso, and ship sturgeon, Acipenser nudiventris, juveniles for 2 mo. Fish were kept in 18 concrete square tanks (2.0 × 1.0 × 1.2 m3) at 22.3 ± 0.4 C and under a natural photoperiod. In both species, the growth performance in terms of final body weight, weight gain, specific growth rate, and feed intake significantly decreased with increasing stocking density (P < 0.05). In both species, the percent of neutrophils increased after 60 d of trial (P < 0.05). Moreover, hematocrit and white blood cell counts increased after 60 d of trial in ship sturgeon (P < 0.05). Plasma immunoglobulin significantly decreased with increasing stocking density in both species. Plasma insulin‐like growth factor I decreased with increasing stocking density in beluga; however, it was not affected in ship sturgeon. Overall, these results showed that the LSD group in both species demonstrated more homogeneous and higher growth rate than the MSD and HSD groups.  相似文献   

11.
在葛洲坝下中华鲟(Acipenser sinensis)产卵场连续两年未监测到自然繁殖的情况下,2015年6月在江苏溆浦段发现15尾疑似中华鲟幼鱼。本研究通过形态学和分子生物学方法对该15尾幼鲟进行鉴定。形态学度量结果显示,除1尾个体外,新发现的14尾幼鲟与历史记载的中华鲟野生幼鱼生物学性状完全一致,但其体长和体重低于历史同期出现在该区域的中华鲟。同时采用11个多态微卫星位点,对15尾幼鲟样本和分布于我国的3种鲟鱼,包括30尾中华鲟(A.sinensis)、39尾达氏鲟(A.dabryanus)和33尾施氏鲟(A.dabryanus)进行遗传分析,发现这15尾幼鲟与中华鲟的遗传距离最近(F_(st)=0.039,P0.05),其次为达氏鲟(F_(st)=0.108,P0.05),最远为施氏鲟(F_(st)=0.209,P0.05),从遗传和进化的角度判定该批幼鱼为中华鲟,该结果暗示中华鲟在葛洲坝下游极有可能找到了新的产卵场,为中华鲟的繁殖群体和幼鱼的种群监测提供了依据。遗传多样性分析显示,该批鱼的遗传多样性(SW=1.998)略低于野生中华鲟(SW=1.886),暗示中华鲟幼鱼遗传多样性水平降低,野生亲本的数量仍不容乐观。  相似文献   

12.
Two studies were conducted using 5- to 6-month-old juvenile (mean weight: 3-4 g) white sturgeon, Acipenser transmontanus, to examine the effects of density and other stressors on manifestation of disease caused by the white sturgeon iridovirus (WSIV). In the first study, replicate groups of Snake River white sturgeon were stocked at three densities (953, 1,907, and 3,178 fish/m3) in 0.31-m3 fiberglass aquaria with spring water flow through of 30.0 L/minute. No significant (P > 0.05) differences were observed among the three groups during the initial 6-week period. However, the high density group cumulative mortality was significantly (P < 0.05) greater than the mortality detected in the other groups at 59 days. Pathognomonic signs of WSIV were detected in moribund fish from each group by histological evaluation; however, no virus was isolated. These results suggested that maintaining low sturgeon densities in fish younger than I year may be a prudent strategy for minimizing mortality caused by WSIV. In the second study, triplicate or replicate groups of Kootenai River white sturgeon that had been transported for approximately 14 hours at 2-6°C, prior to acclimation at lS°C, were stocked at high and low densities previously tested. No signs of WSIV or abnormal mortality were observed during the initial 2-week period. However, after 36 days the mortality increased to 10-18% and 14-18% in the low and high density groups, respectively, and signs of WSIV disease were detected in moribund fish examined from each treatment by histological evaluation. The presence of the virus was further confirmed by electron microscopy of gill tissue. Although fish density did not appear to affect the occurrence of disease or cumulative mortality, the results suggested that a stressor (e.g., handling, transport, and temperature) in subyearling sturgeon may enhance clinical manifestation of WSIV.  相似文献   

13.
草鱼鱼鳞胶原蛋白的提取及其部分生物学性能   总被引:5,自引:2,他引:3  
以草鱼鱼鳞为原料, 分别提取鱼鳞中的酸溶性胶原蛋白(ASC)和酶溶性胶原蛋白(PSC), 着重开展了其包括热稳定性、体外酶降解性以及胶原海绵材料特性在内的相关研究, 并与哺乳动物来源的猪皮胶原(PC)相比较。实验结果表明, 制备所得的3种胶原蛋白均为典型的Ⅰ型胶原并具有完整的三螺旋结构; PC的热变性温度(41.6 ℃)明显高于ASC(34.8 ℃)和PSC(35.2 ℃); 3种胶原蛋白的体外酶降解性能受水解酶的种类、胶原蛋白提取方法、胶原蛋白来源、胶原蛋白受热历史以及蛋白的自组装程度影响。胶原蛋白酶、胰蛋白酶和木瓜蛋白酶对淡水鱼胶原均具有不同程度的降解能力, 但胶原蛋白酶的降解能力最强; 相同条件下, 3种胶原蛋白体外酶降解率依次为ASC>PSC>PC; 经热变性处理后胶原蛋白的体外酶降解率明显提高而经体外自组装处理后其体外酶降解率均出现不同程度的降低; 3种胶原样品冻干后得到的胶原海绵材料具有不同的机械性能和组织结构, ASC和PSC海绵是一种多孔但拉伸承受力较弱的海绵材料, 而PC则与之相反。  相似文献   

14.
The issue of deep hooking is of concern in white sturgeon, Acipenser transmontanus Richardson, fisheries because nearly all anglers use bait with a stationary presentation on the river bottom to catch them, and bait fishing is often associated with higher instances of deep hooking and hooking mortality. Deep hooking rates, landing success and catch rates were investigated for anglers bait fishing for white sturgeon using circle and J hooks with inline and offset alignments fished with both active and passive hook‐setting methods. Anglers hooked 578 white sturgeon and landed 508 fish, ranging in size from 60 to 316 cm total length (mean = 137 cm). Deep hooking rates averaged 0.6% and did not differ between hook types, hook alignments or hook‐setting methods. Landing success (the proportion of hooked sturgeon that were successfully landed) and catch rates were also equivalent between hook types, hook alignments and hook‐setting methods; landing success averaged 88% and catch rates averaged 0.27 fish/hr. Results of this study indicate that deep hooking is rare when angling for white sturgeon using standard bait‐fishing gear regardless of hook‐setting method or whether circle or J hooks were used; regulations restricting hook type in sturgeon bait fisheries are therefore unwarranted.  相似文献   

15.
A study was conducted to evaluate tissue storage and mobilisation of L-ascorbic acid (AA) in the Siberian sturgeon (Acipenser baeri) fed three different experimental diets. The three treatments consisted of a diet devoid of vitamin C (diet A0) and two diets supplemented with equivalent of 300 mg AA kg–1 in the form of either silicone-coated ascorbic acid (diet SC) or of ascorbyl-2-polyphosphate (diet AP). During the first phase (4 months) of the trial, six batches of 130 Siberian sturgeon (initial body weight: 25.5±0.5 g) each were fed one of the three diets in duplicate. During the second phase (3 months), fish from groups SC and AP were switched to diet A0 and those fed diet A0 during the first phase were switched to diet SC. Irrespective of the dietary treatment, growth rates were not significantly different from each other. At the end of phase I, in all tissues studied, total ascorbic acid (TAA) concentrations were higher in Siberian sturgeon fed diet AP than in the other two groups. During phase II, tissue ascorbate depletion was also higher in the AP group than in the other two groups. Transfer of the AA-free diet fed group onto a diet supplemented with 300 mg AA kg–1 (diet SC) led to a slight increase in the TAA concentrations in all tissues. Blood plasma tyrosine concentrations were not significantly different between the three groups. Whole-body collagen levels were affected by dietary AA levels or forms at the end of phase I; the differences were not significant at the end of phase II. Muscle collagen levels were slightly affected. L-Gulonolactone oxidase activity was found in the kidney of Siberian sturgeon, but not in the liver. The ascorbyl-2-polyphosphate appears to be either better utilised by Siberian sturgeon, like in many other teleosts, or more stable than the silicone-coated AA during food processing and storage. Presence of L-gulonolactone oxidase activity in Siberian sturgeon kidney combined with the absence of gross scorbutic signs in AA-free diet fed groups expressing very good growth rates suggested no need of dietary AA byA. baeri.  相似文献   

16.
River–floodplain complexes represent some of the most variable and diverse habitats on earth, yet they are among our planet's most threatened ecosystems. Use of these habitats by large‐bodied fishes is especially poorly understood, particularly in temperate regions. To provide insight into the factors that affect floodplain assemblages and migration, we sampled large‐bodied fishes with a fyke trap for 7 years in the Yolo Bypass, the primary flood basin of the Sacramento River, California. We collected a total of 18,336 individual fish comprised of 27 species, only 41% of which were native. Year‐round resident species white catfish Ameiurus catus, channel catfish Ictalurus punctatus and common carp Cyprinus carpio (all alien species) were the most abundant and comprised 74% of the total catch. Splittail Pogonichthys macrolepidotus (3.8%), white sturgeon Acipenser transmontanus (2.3%) and Sacramento sucker Catostomus occidentalis (1.1%) were the primary native species. We found that seasonal variation in water temperature and flood stage were important factors affecting the fish assemblage structure and the presence of migratory species. American shad Alosa sapidissima, an alien species, showed highest abundance during the early summer upstream migration, when temperatures were warmer. For native species, the abundances of white sturgeon, splittail, Sacramento pikeminnow Ptychocheilus grandis and Sacramento sucker were all highest during flood pulses. While our results suggest that flow alone is not sufficient to control alien species, the strong linkage between native fish migration and flow pulses highlights the importance of river–floodplain connectivity for the conservation of native fishes.  相似文献   

17.
Fish skin collagen solutions (0.3%, w/v) were treated using N-hydroxysuccinimide activated adipic acid (NHS-AA) as a cross-linking agent. Partial important physicochemical properties such as the molecular weight, the morphologies, the interaction between molecules, and especially the thermal stability of the cross-linked collagens were examined by various methods. Results from sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) indicated the formation of cross-links, and thus the molecular weight of collagen was increased. Atomic force microscopy (AFM) suggested that the fiber diameter of cross-linked collagens was larger than that of collagen control. Scanning electron microscopy (SEM) showed that the number of pores from collagen sponges was reduced, accompanied by the rise in their pore sizes. According to Fourier transform infrared spectroscopy (FTIR) spectra, the value of amide III/amide I increased as a small amount of NHS-AA was used. Then it decreased and remained at a relatively constant level as NHS-AA was further added, which might indicate the altered interaction between collagen molecules due to cross-linking. Most important is the improved thermal stability of the cross-linked collagens as determined by differential scanning calorimetry (DSC). The reason for this improvement was further exposed by two-dimensional (2D) FTIR with temperature as perturbation.  相似文献   

18.
Acid-solubilized collagen (ASC) and pepsin-solubilized collagen (PSC) from golden pompano skins were extracted and characterized. The molecular weight of ASC was about 130 kDa for α1 and 115 kDa for α2, which were slightly higher than those of PSC. Similar amino acid composition and Fourier transform infrared (FTIR) spectra were observed in both collagens, but slight differences were found in the peptide maps of collagen digested by V8 protease and trypsin. The denaturation temperatures (Tds) of ASC and PSC calculated from the reduced viscosity were 31.8 and 30.0°C, while the transition temperature (Tm) of ASC and PSC analyzed by DSC were 33.0 and 32.0°C, respectively. ASC has a lag phase, a growth phase, and a plateau phase in the turbidity–time curve, while PSC does not have similar phenomenon. It was found that the fibril gel of ASC could be formed at 25°C, leading to improved thermal stability.  相似文献   

19.
Iridoviridae are known to cause disease in sturgeons in North America. Here, histological and molecular methods were used to screen for this family of virus in sturgeons from various European farms with low‐to‐high morbidity. Some histological samples revealed basophilic cells in the gill and labial epithelia, strongly suggesting the accumulation of iridovirus particles. Newly developed generic PCR tests targeting the major capsid protein (MCP) gene of sturgeon iridoviruses identified in North America, namely the white sturgeon iridovirus and the Namao virus (NV), produced positive signals in most samples from four sturgeon species: Russian (Acipenser gueldenstaedtii), Siberian (A. baerii), Adriatic (A. naccarii) and beluga (Huso huso). The sequences of the PCR products were generally highly similar one another, with nucleotide identities greater than 98%. They were also related to (74–88%), although distinct from, American sturgeon iridoviruses. These European viruses were thus considered variants of a single new virus, provisionally named Acipenser iridovirus‐European (AcIV‐E). Moreover, three samples infected with AcIV‐E showed genetic heterogeneity, with the co‐existence of two sequences differing by five nucleotides. One of our European samples carried a virus distinct from AcIV‐E, but closely related to NV identified in Canada (95%). This study demonstrates the presence of two distinct sturgeon iridoviruses in Europe: a new genotype AcIV‐E and an NV‐related virus.  相似文献   

20.
The effects of dopamine on the secretion of two sturgeon gonadotropins (stGTH I and stGTH II) in sexually mature male white sturgeon (Acipenser transmontanus) were evaluated. In Experiment I, sturgeon were given intraperitoneal injections of physiological saline (PS), dopamine (100 mg kg−1), the gonadotropin releasing hormone analog D-Ala6-des-Gly10-GnRH ethylamide (GnRHA) (10 μg/kg−1), and a combination of GnRHa and dopamine. Fish receiving only GnRHa had significantly higher (p < 0.05) concentrations of plasma stGTH I and stGTH II compared to fish receiving PS, dopamine, or a combination of GnRHa and dopamine. Two hours following its administration, dopamine was effective in decreasing plasma concentrations of both stGTHs that were previously elevated by GnRHa. Dopamine or PS administered by themselves did not alter plasma concentrations of either stGTH. In Experiment II, sturgeon injected intraperitoneally with a combination of GnRHa and pimozide had significantly higher (p < 0.05) concentrations of plasma stGTH I and stGTH II compared to males receiving GnRHa or pimozide alone. While this effect of GnRHAa + pimozide was observed in the spring, no such potentiation was seen in these fish during the summer (Experiment III). These results represent the first evidence of dopaminergic inhibition of GnRH-induced pituitary gonadotropin secretion in Chondrostean fish.  相似文献   

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