Purification and properties of β-galactosidase from Tilapia intestine: Digestive enzyme of Tilapia-X

ABSTRACT:   β-galactosidase of the intestine of Tilapia nilotica was purified by ammonium sulfate precipitation, followed by PAPTG-Sepharose 4B affinity chromatography, ethylenediamineetetraacetic acid ion-exchange chromatography, polyexchanger PBE 94 chromatofocusing, and Sephadex G-100 gel filtration. β-galactosidase was found to be a single band when examined by poly-acrylamide gel electrophoresis. The purifications of β-galactosidase were 27-fold from the crude extract. β-galactosidase showed optimum activity at pH 5.0 at 40°C, and was specifically found to be able to hydrolyze p -nitrophenyl β-galactopyranoside. It degrades galactan and agarose, and produces galactose. β-galactosidase was strongly inhibited by Hg²⁺ and PCMB. β-galactosidase is considered to be secreted by the upper and middle parts of the intestine and most of the activity was detected in the intestinal juice.     

罗非鱼鱼种肠炎病的病理组织学及病原体的研究

本文从病理组织学的角度,研究了罗非鱼鱼种肠炎病的发生与发展,为其诊断与防治奠定了理论基础。结果表明,该病病原体为产气单胞菌。     

利用变温促进罗非鱼生长的研究

在实验室内不同温度条件(恒温28℃、30℃和周期性变温28±4℃、30±4℃)下,分四组饲养罗非鱼,测定其增重、耗氧率、并推算能量同化率和组织生长效率(K_2)。结果表明,变温组均比相应恒温组生长迅速。其中以变温28±4℃组日增重最快,增重百分率最高,其次为变温30±4℃组,恒温28℃组日增重最慢,增重百分率最低。能量同化率和组织生长效率也以变温28±4℃组最高;30±4℃组能量同化率虽较高,但组织生长效率却最低。本文对变温促进罗非鱼生长的机理和其在生产实践中的意义进行了分析与探讨。     

Behavioral responses of tilapia (Oreochromis niloticus) to acute fluctuations in dissolved oxygen levels as monitored by computer vision

Behavioral and ventilatory parameters have the possibility of predicting the stress state of fish in vivo and in situ. This paper presents a new image-processing algorithm for quantifying the average swimming speed of a fish school in an aquarium. This method is based on the alteration in projected area caused by the movement of individual fish during frame sequences captured at given time intervals. The image enhancement method increases the contrast between fish and background, and is thus suitable for use in turbid aquaculture water. Behavioral parameters (swimming activity and distribution parameters) and changes in ventilation frequency (VF) of tilapia (Oreochromis niloticus) responded to acute fluctuations in dissolved oxygen (DO) which were monitored continuously in the course of normoxia, falling DO level, maintenance of hypoxia (three levels of 1.5, 0.8 and 0.3 mg l⁻¹) and subsequent recovery to normoxia. These parameters responded sensitively to acute variations in DO level; they displayed significant changes (P < 0.05) during severe hypoxia (0.8 and 0.3 mg l⁻¹ level) compared with normoxic condition, but there was no significant difference under conditions of mild hypoxia (1.5 mg l⁻¹ level). There was no significant difference in VF between two levels of severe hypoxia 0.8 and 0.3 mg l⁻¹ level during the low DO condition. The activity and distribution parameters displayed distinguishable differences between the 0.8 and 0.3 mg l⁻¹ levels. The behavioral parameters are thus capable of distinguishing between different degrees of severe hypoxia, though there were relatively large fluctuations.     

Effects of dietary cottonseed meal protein level on growth, gonad development and plasma sex steroid hormones of tropical fish tilapia Oreochromis sp

Five experimental diets containing increasing proportions of cottonseedmeal (CSM) protein (0, 25, 50, 75 and 100%; diets 1 to 5, respectively) toreplace fish meal (FM) protein were formulated for intensive culture of tilapiaOreochromis sp. Each diet was fed to three replicategroupsof fish (mean weight ± SE = 11.3 ± 3.9 g) in30L aquaria connected as a closed recirculating-water system andmaintained at 27 ± 1 °C. Fish were fed three times adayby hand at a rate of 3% of body weight during four weeks, after which thefeeding rate was gradually decreased to reach 1.5% at 16 weeks. Thesubstitutionof 75 and 100% of FM proteins by CSM proteins resulted in significantly lowerbody weights in both sexes. In both sexes, gonadosomatic indexes and plasmaconcentrations of sex steroids (testosterone, 11-ketotestoterone,estradiol-17 and 17,20-dihydroxy-4-pregnen-3-one) were notsignificantly different among dietary treatments. The concentration ofgossypol,an antifertility agent contained in CSM, was measured in reproductive tissues.The total gossypol concentration in the testis was consistently lower than thatmeasured in the ovaries of the same group. Moreover, in both sexes, theconcentration of the (+)isomer of gossypol was always higher than that of(–)isomer. The total gossypol concentrations in testes increasedsignificantly with the increase of CSM in the diet. The highest levels of the(+)isomer (7.64 ± 0.62 g g^–1)were found in the testes of fish fed diet 4, whereas the (–)isomerreached its highest values in the testes of fish fed diet 5. The highest levelsof both enantiomers of gossypol were found in the ovaries of fish fed diet 4(14.2 ± 2.7 and 5.6 ± 1.5 g g^–1for (+) and (–)isomers, respectively). In both sexes, thehistological analysis of the gonads did not reveal differences among the fishfed different levels of CSM. Although CSM at any levels did not affect thereproductive parameters examined in this study, it cannot be used to substitutemore than 50% of FM since at higher levels growth of tilapia was compromised.     

Determining design parameters for recovery of aquaculture wastewater using sand beds

Design information for the use of sand beds to remove suspended solids from wastewater discharged from recirculating aquaculture systems (RAS) was developed. Wastewater from a commercial RAS tilapia farm with 2% total solids and 1.6% total suspended solids (TSS) was applied to sand columns to determine infiltration rates and phosphorus capture. Various hydraulic loading rates and drying periods between application events were evaluated. Infiltration rates stabilized after five application events to 3.5 cm/day (S.D.=1.7). Practically, all suspended solids were captured at the top of the columns, creating the primary resistance to infiltration. Concrete sand removed approximately 93% of the soluble phosphorous in the wastewater and wollastonite, an economical aggregate alternative to sand, removed at least 98%. A modified Darcy equation is presented to predict infiltration based upon TSS and the number of sequential applications.     

Genetic improvement of farmed tilapias: Biochemical characterization of strain differences in Nile tilapia

Four African wild strains (Egypt, Ghana, Senegal and Kenya) and four established Asian farmed strains of Nile tilapia, Oreochromis niloticus (popularly known in the Philippines as Taiwan, Thailand, Singapore and Israel) were analysed electrophoretically at 30 protein loci to estimate genetic differences among the strains. All strains shared alleles at 14 monomorphic and 16 variable loci. Among the African strains, characteristic allele frequency differences were observed at AAT-1 ^* 46 for Ghana and Senegal, ADH ^* 83 for Kenya, ADH ^* 120 for Senegal, G3PDH-2 ^* 300 for Egypt, IDDH ^* 67 for Senegal, sMDH-1 ^* 120 for Kenya and SOD ^* 150 for Senegal. Genetic distance values among the strains revealed a clustering of the farmed strains with Egypt and Ghana O. niloticus, a slight separation of the Senegal strain and a larger separation of the Kenya strain. This profile may reflect the origins of the few founder populations of this species previously introduced to Asia. It also confirms the wider genetic divergence of the Kenya strain (O. niloticus vulcani) from the others studied here, which are all O. n. niloticus. Observed heterozygosities of the strains ranged from 0.026 to 0.071, with the African wild strains the lower values (mean Ho = 0.036) and the farmed strains the higher ones (mean Ho = 0.056). The implications of these results to the ongoing tilapia genetic improvement programme in the Philippines are discussed.     

雄性化罗非鱼精养高产试验

１９９４年进行了主养雄性化罗非鱼高产试验，经过１０９天养成，每亩产鱼１１８０ｋｇ。其中罗非鱼１０３０ｋｆ，产值１２３００元，纯利４３００元。专家鉴定认为，实际产量，技术水平达到山东省先进水平。     

氨基酸螯合盐对罗非鱼促生长作用的研究

本文报道了氨基酸螫合盐与无机盐添加剂在罗非鱼饲养中的对比试验结果，以及在螯合盐中添加稀土对罗非鱼生长的影响，测定了各组试验鱼的耗氧率以及肌肉中蛋白质、脂肪和水分的含量，结果表明，添加螯合盐的四个试验组鱼的增重效果均优于添加无机盐的对照组，四个试验组分别比对照组多增重７５．７％、８６．５％、１０８．５％．８９．０％，饵料系数下降率分别为２９．２％．３３．４％，４２．５％，３３．７％。ｔ检验表明，四个试验组与对照组差异显著。     

Ontogeny of protease, amylase and lipase in the alimentary tract of hybrid Juvenile tilapia (Oreochromis niloticus × Oreochromis aureus)

Juvenile tilapias (Oreochromis niloticus × O. aureus), with average body weights of groups I, II and III of 55.14, 122.82, and 225.68 g respectively were used to study changes in the activity of protease, amylase, and lipase in different organs and sections of the alimentary tract. Additionally, tract containing digesta, which is closer to physiological conditions, may help reveal the mechanisms of whole chemical digestion process. Another group of tilapias (mean body weight of 111.81 ± 0.14 g) was used to analyze changes in activity of protease, lipase, and amylase in response to varying pH and temperatures. The activity of enzymes in all digestive organs increased when body weight increased from 55 to 122 g, and the activity further increased when the weights increased from 122 to 225 g. The rates of increase in digestive enzyme activity were significantly greater than the rates of growth of the fish. When the activity was compared in different sections of the tract, a similar regularity was found for the three enzymes in all fish. The highest amounts of activity were seen in the foregut and the lowest in the rectum, with the middle amount of activity in the hindgut. The pH ranges for the maximum activity of tilapia protease in the stomach and intestine were 1.6–3.3 and 9.5–10.5 respectively and the optimal temperature in these organs was 55°C. The pH ranges for the maximum amylase activity in the intestine and hepatopancreas were 6–7 and 7.5 respectively, and high amylase activity in these organs was found at 25–35°C. The activity of lipase in the intestine remained unchanged at pH 6.0–9.0 and with temperatures between 25 and 35°C. These results suggest that the ability of tilapia to utilize feed varies at different growth stages and remains relatively stable with changes in environmental pH and temperature, which should be taken into account when designing feed formulas for tilapia.