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1.
The main goal of this research was to analyse in vitro compatibility of Nile tilapia (Oreochromis niloticus) digestive proteinases and enzymes recovered from species comprising fishery waste as Pleoticus muelleri, Artemesia longinaris and Patagonotothen ramsayi. It evaluated the use of exogenous enzymes as feed supplements to increase digestion efficiency in tilapia fingerlings and juveniles (3.5 ± 0.11 g and 11.6 ± 1.5 g, respectively). We successfully have obtained proteinases from fishery waste as source of exogenous enzymes. P. muelleri and A. longinaris enzymes had more activity of acid and alkaline proteinases than P. ramsayi. SDS‐PAGE gels demonstrated that Nile tilapia digestive proteinases keep their activity when combined with each exogenous proteinase. Exogenous enzymes varied in their ability to enhance hydrolysis of different feed ingredients. P. muelleri by‐products are the best candidates to be employed as feed supplements for tilapia juveniles. Enzymes from this by‐product did not affect the activity and integrity of fish digestive enzymes, improved the hydrolysis of different protein sources (fish meal, squid meal, shrimp meal and soybean meal), and maintained its activity after being exposed to high temperatures and acid pHs. Our findings are applicable to other places where O. niloticus is raised utilizing local fishery waste, and also to different cultured species.  相似文献   

2.
Digestive enzymes of Cirrhinus mrigala (Ham.) were studied during ontogenic development. Specific amylase activity was detected in first feeding fish. The enzyme activity decreased up to day‐18 and then it increased with the age of fish to reach the highest level on day‐34. Protease activity was 28.61 ± 8.90 mU mg protein?1 min?1 on day‐4 and increased with the age throughout the study period. Trypsin activity was 31.86 ± 1.12 mU mg protein?1 min?1 on day‐4. The activity decreased up to day‐10 and from day‐12 onwards increased up to day‐26. Chymotrypsin activity was 14.56 ± 2.74 mU mg protein?1 min?1on day‐4 and constantly increased up to day‐26. A significant increase in lipase activity was observed between days‐24 and 34. SDS‐PAGE and substrate SDS‐PAGE showed the diversity of protein (17.4–127.8 kDa) and protease activity bands (16.6–88.8 kDa) during ontogenesis. Soybean trypsin inhibitor, phenyl methyl sulphonyl fluoride, N‐α‐p‐tosyl‐l ‐lysine chloromethylketone and N‐tosyl‐l ‐phenylalanine chloromethylketone inhibited the protease activity up to 79.72–97.21, 65.55–94.83, 45.41–75.31 and 40.78–64.72%, respectively. Inhibition study in substrate SDS‐PAGE revealed the abundance of serine proteases and the presence of isoforms of trypsin and chymotrypsin. Ethylenediamine‐tetraacetate showed 5.56–22.78% inhibition of metal ion‐specific enzyme activity.  相似文献   

3.
Digestive enzymes of freshwater fish Labeo rohita (family Cyprinidae; class Actinopterygii; infraclass Teleostei; order Cypriniformes) were studied during ontogenic development. Amylase, protease and lipase activities showed a polynomial relationship with age of rohu, whereas trypsin, chymotrypsin and lipase activities exhibited exponential trends with the age of rohu larvae. SDS‐PAGE of crude enzyme extract revealed that the proteases of higher molecular weight (MW) appeared during early ontogeny, whereas low MW proteases were observed at a later stage. Substrate SDS‐PAGE supported the quantitative study of protease activities as evidenced with the increasing number and intensity of activity bands with the age of fish. The number of protease activity bands observed in 4, 10, 12 and 24 DAH (days after hatching) larvae were 5, 7, 8 and 9, respectively. Inhibition of protease activities with soybean trypsin inhibitor (SBTI) (58.6–81.2%), phenyl methyl sulphonyl fluoride (PMSF) (55.6–70%), N‐α‐p‐tosyl‐l ‐lysine chloromethyl ketone (TLCK) (41.1–52.3%) and N‐tosyl‐l ‐phenylalanine chloromethyl ketone (TPCK) (27.9–44.5%) indicated the presence of serine proteases, trypsin‐ and chymotrypsin‐like enzymes in rohu larvae. Inhibition with SBTI and TPCK showed power, TLCK and ethylenediaminetetraacetic acid showed exponential, whereas PMSF showed polynomial relationships during the study period.  相似文献   

4.
Characteristics and functional efficacy of digestive proteases of Catla catla, catla, Labeo rohita, rohu and Hypophthalmichthys molitrix, silver carp were studied. Total protease activity was significantly (P < 0.05) higher in rohu (1.219 ± 0.059 U mg protein−1 min−1) followed by silver carp (1.084 ± 0.061 U mg  protein−1 min−1), and catla (0.193 ± 0.006 U mg  protein−1 min−1). Trypsin activity of silver carp and rohu was 89–91% higher than catla. Chymotrypsin activity was significantly (P < 0.05) higher in silver carp compared with rohu and catla. The protease activity of rohu and silver carp displayed bell‐shaped curves with maximum activity at pH 9; whereas in catla, maximum activity was found between pH 8 and 11. Inhibition of protease activity with soybean trypsin inhibitor (SBTI), phenylmethylsulfonyl fluoride (PMSF) revealed the presence of serine proteases and inhibition of activity with N‐α‐p‐tosyl‐L‐lysine‐chloromethyl ketone (TLCK) and N‐tosyl‐L‐phenylalanychloromethane (TPCK) indicated the presence of trypsin‐like and chymotrypsin‐like enzymes in all these three carps. SDS‐PAGE showed the presence of several protein bands ranging from 15.3 to 121.9 kDa in enzyme extracts of catla, rohu and silver carp. The substrate SDS‐PAGE evidenced the presence of various protease activity bands ranging from 21.6–93.7, 21.6–63.8 and 26.7–98.5 kDa for catla, rohu and silver carp respectively. In pH‐stat hydrolysis of Chilean fishmeal showed significantly (P < 0.05) higher degree of hydrolysis compared with soybean meal, silver cup (a commercial fish feed of Mexico) and wheat flour, with enzyme preparations of three fishes. The rate of hydrolysis was significantly (P < 0.05) higher in silver carp compared with others.  相似文献   

5.
Redclaw aquaculture has developed at a rapid pace during the past few years, yet no specialized diet for the species has been developed. The present study was designed to evaluate whether soybean‐based diets containing either fishmeal (FM), poultry by‐product meal (PBM), ground peameal (GPM) or distillers dried grains with solubles (DDGS) meal as a protein source are suitable for redclaw aquaculture. Juvenile redclaw crayfish Cherax quadricarinatus (0.125±0.025 g) were stocked into 20 rectangular tanks at a stocking density of 12.5 m?2 for 8 weeks. Crayfish in four replicate tanks were maintained on one of five diets formulated to contain 35% crude protein and 7.1% lipids. Each 100 g of diet contained 25 g of protein from soybean meal (SBM) and 10 g of protein from the alternative protein sources mentioned above. A fifth treatment that did not receive feed was included to account for growth from natural productivity. There were no significant differences in survival (86–90%), growth (3.84–4.98 g animal?1) or feed conversion ratio (2.10–2.79) of crayfish among the four treatments (P>0.05). Survival and growth of crayfish in the treatment that received no supplementary feed were significantly less than those in treatments offered experimental diets. Results of the present experiment suggest that SBM‐based diets with PBM, FM, DDGS or GPM have similar effects on growth performance and survival of juvenile redclaw, C. quadricarinatus.  相似文献   

6.
Juvenile redclaw crayfish, Cherax quadricarinatus, show non‐selective feeding behaviour when commencing feeding, but change to a dietary preference for plant material as they grow. This study aimed to find any ontogenic changes in digestive enzymes that might be associated with the change of diet. The enzymes in the digestive tract initially showed high levels of proteases and low levels of carbohydrases, but as the crayfish grew, the levels of proteases fell and the levels of the carbohydrases generally rose, in parallel with the increased preference for plant food. Cellulase was present in all free‐living stages. Dietary requirements of juvenile redclaw are probably quite similar to those of larger specimens.  相似文献   

7.
Partial characterization of digestive proteases in the three‐spot cichlid Cichlasoma trimaculatum juveniles was conducted. It was determined that there is higher alkaline proteases activity (3.95 ± 0.32 IU mg?1 protein) compared to acidic proteases (2.01 ± 0.57 IU mg?1 protein). Optimal temperature for alkaline proteases is 60 °C which resulted in more thermostability to temperature changes. On the other hand, optimal temperature for acidic proteases is 50 °C. Optimal pH for acidic proteases was pH 2, while for alkaline proteases, it was pH 10, which resulted in more stability in relation to pH changes than acidic protease. The use of specific inhibitors and the SDS‐PAGE electrophoresis analysis revealed seven types of bands for alkaline proteases, which make evident the main presence of serine proteases. In acidic proteases, more than 98 g kg?1 of the activity was inhibited with pepstatin A inhibitor. Therefore, it is evident that C. trimaculatum digestion is composed by acidic and alkaline proteases; thus, it should be considered an omnivorous fish.  相似文献   

8.
The effect of three crude dietary lipid levels (4, 8, and 12%) on egg quality of the female redclaw crayfish, Cherax quadricarinatus, was evaluated. Three replicate groups of 40 crayfish (mean initial weight ± SD, 23.0 ± 3.0 g) per diet treatment were stocked in 15,000 ‐L tanks, at 28 C. After 75 d of culture, there were significant differences (P < 0.001) in egg area (3.52–3.90 mm2), volume (34.0–39.3 mm3), weight (4.81–5.36 mg), and diameter (2.15–2.27 mm) in response to dietary lipids in the diet. Using the quadratic equation, the maximum responses of these parameters corresponded to lipid levels from 8.60–8.84%. There were no significant differences (P > 0.05) in survival (82.1–86.9%), final weight (41.0–43.7 g), number of spawning females (18.12–27.38%), and fecundity (9.07–10.31 eggs/g female). Additionally, there were no significant differences (P > 0.05) in egg mean protein (2306.87 ± 440.08 μg/egg), lipid (441.95 ± 83.23 μg/egg) and carbohydrate (74.81 ± 10.12 μg/egg) contents, and energy (13.49 ± 2.25 kcal/egg). The optimum overall response was obtained for dietary lipid content of 8.7%, which we recommend to optimize egg quality of this freshwater crayfish.  相似文献   

9.
Feeding trials with five levels of crude protein (CP) (22%, 27%, 33%, 39%, and 45%) and digestible energy ranging from 14.32 to 15.21 kJ g?1 were carried out to determine optimum dietary protein for the growth and production of pre‐adult freshwater crayfish Cherax quadricarinatus. Triplicate groups of males (ω=23.1±0.58 g) and females (ω=21.8±0.33 g) were fed diets for 70 days in monosex culture. Optimal digestible protein (DP) was estimated for males by fitting mean weight gain to a quadratic model (y=27.86+0.839x–0.0216x2, r2=0.945), and a linear model for females (y=32.84–0.0745x, r2=0.959). Optimal male growth occurred with 25.6% CP, equivalent to 19.4% DP. Male crayfish attained significantly higher final weights and absolute growth rate than females fed the 22%, 27%, and 33% CP diets. The results indicated that a 19.4% DP diet, with a P/E ratio of 11.4 mg kJ?1, produces an optimum growth response for pre‐adult C. quadricarinatus under experimental conditions.  相似文献   

10.
In vitro assays used porcine or bovine trypsin as models of exogenous enzymes to determine functioning in the presence of enzymatic extracts from the digestive gland of whiteleg shrimp Penaeus vannamei. Using electrophoresis and zymograms, when enzymes from the shrimp were mixed in the absence of protein substrate, they hydrolysed the trypsin from bovine or porcine origin. Porcine or bovine trypsin, when mixed with shrimp enzymes in pH‐stat assays in the presence of shrimp commercial feed, fish meal, or casein, there was added activity to hydrolyse the protein substrate. Hydrolysis of protein substrate was twofold to threefold stronger if exogenous enzymes were added. Results suggest that porcine or bovine trypsin could be used as feed supplements for whiteleg shrimp P. vannamei to enhance hydrolysis of proteins in feeds, because the commercial enzymes contributed to the hydrolysis of the protein in the three substrates in the presence of shrimp enzymes.  相似文献   

11.
Precipitation techniques play a vital role in the industrial extraction of enzymes. The present study aimed to extract the proteases from the viscera of Labeo rohita (commonly called Rohu) and to compare the precipitation techniques for the isolation and purification of the enzyme. The enzyme is usually discarded as tons of waste during processing. Hence, a trial has been carried out to isolate the protease enzymes from viscera of the freshwater fish Labeo rohita. The proteases were precipitated with ammonium sulfate, ethanol, and acetone. Acetone precipitation was found to be the best option for the recovery of enzymes (54%) from the viscera of Rohu, and two caseinolytic protease bands were shown in the zymogram. The precipitates with highest proteolytic activity were further subjected to dialysis, and their molecular weight was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).  相似文献   

12.
This study determined the effect of different dietary protein and lipid levels on growth and survival of juvenile redclaw Cherax quadricarinatus. Nine practical test diets were formulated to contain three crude protein (CP) levels [260, 310 and 360 g kg?1, equivalent to 225, 260 and 296 g kg?1 digestible protein (DP) respectively] at three crude lipid (CL) levels (40, 80 and 120 g kg?1, equivalent to 38, 67 and 103 digestible lipids respectively), with digestible protein : digestible energy (DP : DE) ranging from 14.6 to 22.6 mg protein kJ g?1. Three replicate groups of 15 crayfish (initial weight mean ± SD, 0.71 ± 0.13 g) per diet treatment were stocked in 40 L tanks, at 28 °C for 60 days. The highest mean weight, specific growth rate and biomass, with values of 7.0 g, 3.67% day?1, and 370.2 g m?2, respectively, were achieved by feeding a diet with P : L = 310 : 80 (P < 0.05). The treatments resulted in a survival rate of 80–91%, feed conversion ratio of 1.08–1.61 and protein efficiency ratio of 2.24–3.08. Results indicated that a diet containing 270 g kg?1 DP (equivalent to 320 g kg?1 CP), 75 g kg?1 digestible lipid (DL) with a DP/DE of 18.4 mg protein kJ?1, and 0.031 g protein per animal per day was optimum for juvenile C. quadricarinatus under the tested experimental conditions.  相似文献   

13.
Quantification of proteases activities and their class structure have been studied in a cladoceran, Daphnia carinata. Protease activity ranged from 0.28 to 0.55 Unit mg−1 protein min−1 with an average value of 0.42±0.06 Unit mg−1 protein min−1. Chymotrypsin activity was more than twofold higher (0.49±0.09 Unit mg−1 protein min−1) than the trypsin activity (0.21±0.02 Unit mg−1 protein min−1). Protease activity and reduction of activity in bands of samples treated with specific inhibitors were documented in photometric assay and substrate SDS–PAGE. Proteinase activity against azocasein was inhibited (91.4±1.5%) with SBTI. PMSF reduced the enzyme activity by 53.1±6.5%, and the azocasein hydrolysis was reduced up to 64.6±3.8% by the specific inhibitor of trypsin, TLCK. In the present investigation, the molecular weight of various activity bands ranged from 16.3 to 51.1 kDa. The molecular weights of several protein bands are similar to protease activity zones. The knowledge of digestive enzyme profiles of fish food organisms generated in the present study may assist in the formulation of age-specific feed.  相似文献   

14.
Idiopathic lesions observed in redclaw freshwater crayfish, Cherax quadricarinatus (von Martens), from farms in northern Queensland, Australia, over a 4‐year period are described. Various idiopathic lesions were observed in the exoskeleton, antennal gland, mandibular organ, haemolymph vessel endothelium and enteric tissues of C. quadricarinatus collected for histopathological surveys, investigations of chronic mortalities or during other activities. The need for an increased use of histopathology in crayfish disease diagnosis is highlighted.  相似文献   

15.
Four commercially available proteolytic enzymes active over a wide pH range were assessed as shrimp feed additives. Feeds were formulated to contain 1% of the test enzymes by weight. A feed of the same composition with no enzyme addition served as a control. In vitro feed protein digestibility of the feeds was determined using a modified pepsin digestibility method. In vitro protein digestibility results indicated that commercially available proteases added to a shrimp feed formulation and then hot pelleted/dried did retain enzyme activity. Feeds exhibited low weight losses through leaching (≤ 10.9%) and low or no losses of protein content (≤ 2.7%), indicating that the test feeds were available in their entirety to the shrimp. An 8-week feeding trial using the enzyme-supplemented feeds was conducted to evaluate shrimp growth. Mean shrimp survival was high for all treatments (> 90%). One-way analysis of variance did not detect significant differences (P > 0.05) among feeds in terms of survival, weight gain, specific growth rates and feed–weight gain ratios. Although in vitro digestibility indicated the presence of active enzymes in the feed, the feeding trial revealed that shrimp growth was not enhanced by the inclusion of proteases in these feeds. This study provides sufficient proof to emphasize the importance of in vivo investigations before enzyme-supplemented feeds are used with shrimp.  相似文献   

16.
应用实时荧光定量PCR技术,结合生物酶和代谢产物测定,研究了氨氮急性胁迫对红螯光壳螯虾幼虾代谢及免疫系统的毒性影响及其毒后恢复情况。实验首先进行3 d的氨氮胁迫,取样后剩余虾移入曝气自来水进行7 d的毒后恢复实验。结果表明,3 d氨氮胁迫后,肌肉ACP、AKP、SOD活性表达均受到显著影响,随着氨氮浓度的升高酶活性分别降低,最高浓度组(16 mg/L)降低为对照组的76%、68%和62%。线粒体MnSOD、胞外Cu/ZnSOD的mRNA表达量也随着氨氮浓度增加而下降,最高浓度组降低至对照组的69%和68%。CAT、GPX活性以及GPX和GST的mRNA表达量变化不显著。肝胰腺中可溶性蛋白和甘油三酯含量随着氨氮浓度升高而降低,最高浓度组分别降低至对照组的72%和59%,AST活性在12 mg/L浓度组显著升高至对照组的134%。7 d恢复期过后,ACP和AKP活性以及各代谢指标恢复到正常水平;而SOD和GPX活性高于对照组。各抗氧化基因的表达量都不同程度高于对照组。实验表明,高浓度氨氮胁迫能抑制部分免疫相关酶的活性及基因表达,对免疫系统造成损害。氨氮胁迫下,红螯光壳螯虾动员蛋白质和脂肪来供能应对胁迫。7 d的恢复时间不足以让红螯光壳螯虾从胁迫中完全恢复,其肌肉仍处于轻度氧化应激状态。  相似文献   

17.
Growth of juvenile redclaw Cherax quadricarinatus fed 400 g kg?1 crude protein isoproteic diets substituting fish meal with soya bean meal at various levels (250, 500, 750 and 1000 g kg?1) was evaluated under laboratory conditions. Juvenile crayfish were reared individually in 1‐L plastic containers for 56 days. Total survival was recorded for all treatments at the end of the experiment. There were no significant differences (P > 0.05) for the specific growth rate among the treatments, although the animals fed with the control diet (100% fish meal) obtained the highest mean daily weight gain (0.055 g day?1), individual final weight (4.12 g), molts per individual (0.04 molts day?1) and molting frequency (23.57 days) values at the end of the experiment. Dietary inclusion of soya bean meal at the tested levels hampered the C. quadricarinatus juvenile growth.  相似文献   

18.
Five dietary lipid sources (fish oil, soybean oil, palm oil, rapeseed oil and linseed oil) were evaluated in juvenile red claw crayfish, Cherax quadricarinatus, based on the response of growth, antioxidant capacity, intestine histology, whole‐body composition, fatty acid nutrition and lipid metabolism. Crayfish were fed in quadruplicate net cages for 8 weeks. Crayfish fed diets with fish oil, soybean oil and linseed oil obtained significantly higher weight gain and specific growth rate than those fed the other two diets. Survival, condition factor and hepatosomatic index were not significantly affected by lipid sources. Lipid sources also do not affect the whole‐body composition of crayfish. Serum SOD, T‐AOC and GSH‐PX activities of crayfish fed the palm oil and rapeseed oil diets had a significantly lower value than those fed other diets. The minimum concentrations of MDA have been observed in crayfish fed the soybean oil diet. The activity of ACC in the hepatopancreas of crayfish fed the linseed oil diet showed the highest value, and the CPT‐1 activity was not significantly affected by different lipid sources. Crayfish fed the soybean oil diet showed significantly higher TC and TG contents in hepatopancreas than those fed other diets. Crayfish fed linseed oil diet had a significantly higher percentage of EPA, C18:3n?3 and Σn?3 PUFA in muscle than those fed other treatments. Most of the fatty acid compositions in the hepatopancreas had a close correlation to fatty acid compositions in diets. All findings in this study indicate that soybean oil is the advantageous lipid source for juvenile C. quadricarinatus which can reflect in satisfactory growth performance, antioxidant capacity and fatty acid nutrition of edible tissues.  相似文献   

19.
The activity of different protease classes was monitored in developing discus (Symphysodon spp.) larvae using a combination of biochemical assays and substrate SDS–PAGE techniques. Results showed the presence of alkaline proteases of serine proteases such as trypsin with a significant increase in activity levels detected beginning 3 days after hatching. Other alkaline proteases such as metallo‐proteases and chymotrypsin, a type of serine protease, were only detected in older larval stages, at around 20–30 days after hatching. Acidic protease activity was very low during the first 20–25 days of development before showing a significant (P < 0.01) rise. This is despite the formation of a stomach observed 10 days after hatching. Based on the development of the protein digestive system observed, the use of microdiets to replace Artemia should be considered 25 days after hatching.  相似文献   

20.
  1. Crayfish are invasive polytrophic keystone species, which are phylogenetically unique on the African continent. The Australian redclaw crayfish Cherax quadricarinatus is invasive in southern African freshwater systems including the Zambezi River Basin. Surveys conducted across the Zambezi Basin (Botswana, Zimbabwe, Namibia and Zambia) between 2017 and 2019 showed that C. quadricarinatus is broadly distributed across the Upper and Middle Zambezi and is rapidly spreading through natural and human-mediated means across several ecoregions.
  2. The probability of capture (Pcapture), catch per unit effort (CPUE) and population characteristics of C. quadricarinatus from the recent Barotse floodplain invasion were compared with older invasions from Lake Kariba and Kafue River.
  3. The Pcapture and CPUE of C. quadricarinatus in the recently invaded region of the Barotse floodplain were similar to those of the older invasions. Mass and carapace length of C. quadricarinatus from the Barotse floodplain were significantly lower than those of C. quadricarinatus from the older invasions. Sex ratios differed significantly between the three invasive populations. The Barotse floodplain population had a disproportionate investment in females (65.3%) and intersex individuals (8.4%). No crayfish were detected in the Zambezian Headwaters or the Okavango Floodplains ecoregions, but current spread rates are 49 km yr−1 downstream and 12 km yr−1 upstream.
  4. Investment in population management and the prevention of spread will have high conservation value across the invaded regions in order to restrict crayfish ecological impacts via direct predation and competition. In areas where crayfish are in high abundance, existing fisheries are affected through damage to nets, leading to increased net abandonment and ghost gear pollution in invaded regions.
  5. The emphasis should be on developing cohesive transboundary biosecurity policies in southern Africa to limit further spread that will threaten the integrity of freshwater ecosystems. However, long-term monitoring is needed to gauge invasion risk to sensitive areas such as the Okavango Floodplains ecoregion and determine field-based ecological impacts.
  相似文献   

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