几种固氮菌突变种固氮酶的特性及晶体生长

一，缺失nifZ的棕色固氮菌突变种钼铁蛋白的晶体生长的研究进展 在一定的结晶条件下，缺失nifZ的棕色固氮菌突变种钼铁蛋白将从溶液中结晶出深棕色的短斜四棱柱晶体。PEG 8000、MgCl2、NaCl、Tris的浓度及缓冲液的pH对该蛋白的结晶及晶体生长影响的系统研究表明，它们的浓度和pH较低时，不出晶体;当pH高于8.0，随着前三种化合物浓度的逐渐提高，便在一周内出现大量小品体;再提高浓度后，便延长结晶时问但出质好、数少而个大的晶体;然后晶体随浓度的提高而又变小、变多、甚至晶质变差，直至不再出晶体。影响晶体生长的各因子的最适浓度随其它条件的改变而有所不同。当缓冲液的pH为8.2而PEG 8000，MgCl’、NaCI、蛋白质和Tris的浓度分别为1.86%、300 mmol/L、400 mmoUL、4.64g/L、53 mmo/L时，首次发现在一滴悬滴结晶液中只有一颗较大的优质晶体（最大两边线度均为0.16mm）。 二，从分别含Mn和Cr的培养基中生长的固氮菌突变种UW3中纯化的固氮酶的特性和结晶 缺失nifH基因的棕色固氮菌突变种UW3，在有钼环境中不能固氮生长，但能在无钼而含MnS01或Na2Cr01的无氮培养基中固氮生长。分别经超声破碎、加热除去部分杂蛋白、离子交换柱层析和Sephacryl S-200或S-300柱层析的分离纯化，分别得到二种固氮酶组分l蛋白。金属元素测定表明，这两种蛋白除含铁元素外还分别含有锰和铬元素。它们的吸收光谱、CD和AR谱互不完全相同，并都与OP-MoFe蛋白存在较大差异。含Mn固氮酶也能还原C2H2、质子和N2，对它们的还原的比活性都分别约为MoFe蛋白活性的50%。初步结果表明，这一突变种在这两种培养条件下都可能已表达了不同于已发现的三种固氮酶的新固氮酶组分l蛋白-MoFe、VFe和FeFe蛋白，分别为含Mn或Cr的固氮酶组分1蛋白，分别被称为uw3，-MnFe蛋白和UW3-CrFe蛋白。通过对PEG 8000、MgCI2、NaCI和缓冲液的种类和浓度等结晶条件的大量优化组合实验，首次获得uw3-MnFe蛋白和UW3-CrFe蛋白的的晶体。最大晶体为21.4×14.3×2.1μm。

棕色固氮菌突变株DJ35 中钼铁蛋白和HBP59 蛋白的研究—纯化特性及晶体生长

我们从A. vinelandii突变株DJ35中纯化得到了ΔnifE Av1，并通过与OP Av1相比较对其特性进行了研究。虽然ΔnifE Av1以四聚体形式(α2β2)存在，但却表现出两种天然电泳行为。与OP Av1相比，ΔnifE Av1中的金属含量较低，每个蛋白分子仅含9.96个铁原子、0.36个钼原子。OP Av1和ΔnifE Av1的吸收光谱相似，均在280 nm附近出现蛋白吸收峰，在300-600 nm波长范围内光吸收普遍降低，并无特征峰出现。虽然ΔnifE Av1可见光区域CD谱的摩尔消光系数（Δε）总比OP Av1小，但在520nm区域附近摩尔消光系数减少的相对值明显大于在450nm附近摩尔消光系数减少的相对值。ΔnifE Av1的EPR信号明显与OP Av1不同，在g≈3.7处的EPR信号完全消失，在g≈4.3 和2.0处的EPR信号强度也分别降低了75%和50%以上。ΔnifE Av1不能与Fe蛋白组成活性单位，但被FeMoco激活后可与Fe蛋白组成活性单位。上述结果表明, ΔnifE Av1不含FeMoco，但具有与OP Av1相同的P-cluster。 我们在纯化ΔnifE Av1的过程中，发现一个类似OP Av1的β亚基的污染蛋白，经MALDI-TOF质谱鉴定这种蛋白是棕色固氮菌中一预测基因的产物。通过改进纯化方法, 我们首次得到80%电泳纯的蛋白，并将其命名为HBP59蛋白。HBP59为一单体蛋白，分子量约为59k Da。金属测定结果表明每个蛋白分子中含有0.42个铁原子。HBP59蛋白的可见光谱表现出典型的血红素蛋白光谱特征，还原态的HBP59蛋白在421nm处有最大吸收峰，同时在517nm、556nm处有两个伴随肩峰出现, A421/A280仅为0.146。HBP59蛋白氧化后，吸收峰发生蓝移，最大吸收峰从421nm移到413nm，517nm和556nm处的肩峰消失，A413/A280为0.168。HBP59蛋白的可见光区圆二色谱比较复杂，正峰出现在420nm, 406nm, 379nm 和364nm; 其摩尔消光系数分别为0.75, 0.94, 0.68 和 0.99;负峰出现在433 nm 和392nm，其摩尔消光系数分别为-1.13 和-0.78。血红素滴定实验结果说明每个HBP59蛋白分子结合了0.1个血红素，但每个蛋白分子具有最大结合1个血红素的能力。这低结合能力与这低比率的比率是非常一致的。该蛋白对温度相对敏感，高于40℃蛋白开始沉淀。上述结果说明HBP59是一个结合具有低自旋和不对称的血红素的蛋白，它在菌体内可能并未扮演贮藏血红素的角色，而是可能参与血红素的运输或附着。 此外，经过结晶条件的大量筛选后获得了ΔnifE Av1和ΔnifH Av1的优质小单晶,并对HBP59的晶体培养进行了初步探索。

含铬或锰的新型固氮酶的纯化, 特性和结晶—棕色固氮菌突变种UW3中CrFe蛋白和MnFe蛋白的研究

以含MnSO4或Na2CrO4的无钼无氨的修改Burk’s培养基， 培养不能合成含钼固氮酶体系的棕色固氮菌 (Azotobacter vinelandii Lipmann) 突变种UW3， 发现在一定浓度范围内， MnSO4或Na2CrO4的加入有助于促进其生长。 菌体生长和C2H2还原活性曲线测定结果表明， 这种促进作用很可能是通过Mn或Cr取代Mo， 参与组装固氮酶中心原子簇， 从而影响固氮活性而实现的。 利用阴离子交换 (DEAE-52和Q-Sepharose FF) 和凝胶过滤 (Sephacryl S-200) 柱层析从两种菌体中纯化得到固氮酶组分Ⅰ蛋白 (分别命名为MnFe蛋白和CrFe蛋白)， 并对其进行了特性研究。 厌氧天然聚丙烯酰胺凝胶电泳 (PAGE) 和SDS-PAGE结果显示， 两种蛋白均为两种亚基组成的四聚体。 亚基可以与OP MoFe蛋白抗体发生免疫反应， 分子量分别略小于野生种OP MoFe蛋白的α和β亚基。 CrFe蛋白的C2H2还原活性， Ar下放氢活性和固氮活性分别相当于OP MoFe蛋白的36%， 38%和43%， 而MnFe蛋白活性相当于OP MoFe蛋白的50%左右， 并且两种蛋白与OP MoFe蛋白具有相似的固氮电子利用率。 对两种蛋白金属含量的测定证实其中分别含有Mn和Cr， 但仍存在少量Mo污染。 与OP MoFe蛋白相比，这两种蛋白圆二色谱的摩尔椭圆率 ([θ]) 除在450nm较接近外，在可见光区的其它波长处均显著降低。 与DT还原OP MoFe蛋白相似， CrFe蛋白和MnFe蛋白具有g≈4。3、3。7和2。0的特征EPR信号， 但各处信号强度比例不同。 在对污染Mo可能引起的信号进行校正后，CrFe蛋白的三个信号强度分别相当于DT还原OP MoFe蛋白的20%， 0%和10%， 而MnFe蛋白则分别相当于112%， 49%和65%。 上述结果表明， CrFe蛋白和MnFe蛋白与OP MoFe蛋白金属原子簇的主要差异很可能在于FeMco (M=Cr， Mn或Mo)的M种类， 而P-cluster结构和组成均未见大的差异。 利用气相扩散悬滴法对MnFe蛋白和CrFe蛋白结晶条件进行了筛选和初步优化， 确定了以Tris/Hepes， NaCl， MgCl2和PEG 8000为主要变量的沉淀剂体系， 寻找各组分对于晶体生长的最适浓度。 以此为基础探讨了应用气相扩散坐滴法和液-液扩散法对两种蛋白结晶条件的优化。 在一定条件下， 两种蛋白分别通过液-液扩散法获得了优质大单晶。 对从CrFe蛋白和MnFe蛋白制备物中培养出的蛋白质晶体的SDS-PAGE鉴定显示， 晶体由与OP MoFe蛋白相似的两种亚基组成。 通过 “神舟三号” 飞船搭载实验探讨了空间微重力对于厌氧蛋白质结晶的影响， 结果表明， 微重力有助于避免孪晶形成， 并具有长期培养后获得适于进行X-射线衍射分析的优质大单晶的潜在前景。 结合空间科学使固氮酶结构与功能研究得以发展， 这项工作是有意义并且可行的。

缺失nifZ的棕色固氮菌突变株DJ194的钼铁蛋白研究—纯化、结晶及金属簇的组成与分布

从棕色固氮菌DJ194菌株得到的固氮酶粗提液经DEAE－52、Sephacryl S－200及Q－Sepharose等柱厌氧层析，分离纯化得到nifZ基因缺失的固氮酶MoFe蛋白（ΔnifZ Av1）制备物。通过天然电泳和SDS变性电泳发现早期纯化所得的ΔnifZ Av1制备物中残存相当比例的三个主要污染蛋白：属于热激蛋白60家族（Hsp 60 family）成员的分子伴侣GroEL、糖酵解过程的一个多功能酶——6-磷酸葡萄糖异构酶（6-Phosphate Glucose Isomerase，PGI）及棕色固氮菌细菌铁蛋白（Bacterioferritin，Bfr）。初步鉴定表明，它们分别为由约55kD亚基组成的14聚合体，62kD亚基组成的10聚体和20kD亚基组成的24聚体。首次发现PGI有如此高的聚合体。虽然GroEL和PGI在天然电泳中的迁移率小于ΔnifZ Av1蛋白，但它们的亚基在SDS变性电泳中与ΔnifZ Av1亚基具有相似的迁移率，互相重叠，从而使变性电泳比天然电泳显出更高的ΔnifZ Av1纯度;而细菌铁蛋白虽然不会在变性电泳中污染ΔnifZ Av1，但往往会在ΔnifZ Av1制备物的结晶中优先或较多地结晶出来,从而给它的晶体生长和解析研究带来干扰（Zhao等，2004）。 通过Sephacryl S－200柱洗脱峰收集精度的调整及Q－Sepharose柱的NaCl浓度梯度洗脱，得到了纯度大于90%的ΔnifZ Av1制备物。它的厌氧天然电泳及其免疫印渍（Western blotting），以及SDS-变性凝胶电泳显出，ΔnifZ Av1的电泳迁移率、分子量和亚基组成等均与野生种钼铁蛋白（OP Av1）相似，表明nifZ基因缺失并未改变ΔnifZ Av1的α2β2四聚体构成。ΔnifZ Av1的Mo含量、EPR信号（g≈4.3, 3.65和2.01）和520-660 nm附近的圆二色摩尔消光系数（Δε）也都与OP Av1较相似，从而表明ΔnifZ Av1含有与OP Av1数量相当的具有3/2自旋态的还原FeMoco。然而，ΔnifZ Av1的Fe含量和对底物（C2H2、H+和N2）的还原活性都较低, 分别约为OP Av1的74%和46-50%;而反映P-cluster状况的450nm附近的Δε也明显低于OP Av1。此外，与OP Av1相同的ΔnifZ Av1在g≈2.01的EPR信号却与推测含由双[4Fe-4S]簇组成的P-cluster前体的His-tagged ΔnifZ Av1（Hu等，2004）的信号明显不同。这就表明，ΔnifZ Av1与OP Av1的差别不在于FeMoco的结构、含量和氧还状态，也不在于P-cluster的结构和氧还状态，而仅在于ΔnifZ Av1中P-cluster数目的减少（约一半）。据此推测出与国外提出的His-tagged ΔnifZ Av1模型不同的ΔnifZ Av1(DJ194)的如下结构模型。一个αβ亚基对含有一个FeMoco和一个P-cluster，而另一个αβ亚基对只含FeMoco，其P-cluster区域则是空的。由于nifZ基因的缺失只造成了钼铁蛋白中两个P-cluster中的一个不能组装，因此推测P-cluster的组装可能不是受单一基因产物的影响。根据Lee等（1998）对nifZ产物（NifZ）和nifW产物（NifW）可以形成[NifWx-NifZy]多聚体，并可能是通过同一途径来影响固氮酶的合成的研究，提出NifZ的如下的可能作用机理。[NifWx-NifZy]多聚体影响与P-cluster合成相关的金属簇(如[4Fe-4S])的进入和P-cluster的最终合成，而其中的一个αβ对上的P-cluster的形成可能较多的受到NifZ的影响;nifZ的某些突变或缺失虽然不影响[NifWx-NifZy]多聚体的形成，但对于更依赖于NifZ的那个αβ对上的P-cluster的合成可能会产生不同的影响。 对这一高纯度的ΔnifZ Av1制备物结晶的研究表明，使用Tris缓冲系统的25%PEG 6K/MgCl2晶体培养液可以在较短的晶体培养时间内得到较大的晶体;在一定条件下，pH为7.5和8.3的培养液对出晶数和晶体大小的影响不明显;PEG 6K的浓度与MgCl2的浓度对出晶大小的影响有一定的相关性，即较低的PEG浓度在较低的MgCl2浓度下和较高的PEG浓度搭配较高的MgCl2浓度较易长出较大的晶体。虽然ΔnifZ Av1制备物的纯度达到90%以上，并且在染铁实验中表明其基本不含细菌铁蛋白，但我们在对得到的晶体进行电泳鉴定中仍然发现了一种与以前报导的砖红色晶体不同的深棕红色的细菌铁蛋白晶体，之所以颜色不同可能和所含的铁元素的氧化状态有关。不过在所鉴定的5支结晶管中只在一个管内发现了与固氮酶晶体同时存在的这种细菌铁蛋白晶体，说明通过蛋白纯度的提高减少细菌铁蛋白的含量以及晶体培养条件的优化，细菌铁蛋白晶体形成的几率就可大大降低。

Digestible protein and energy value of fish meal, dextrin, fish oil and soybean oil for Thai sharpunti (Puntius gonionotus Bleeker)

A laboratory trial was conducted to determine the digestible protein and energy value of fish meal, dextrin, fish oil and soybean oil for Thai sharpunti (Puntius gonionotus Bleeker). A reference diet containing 35% protein was formulated in which fish meal was the sole source of protein. Five test diets were formulated using reference diet and individual test ingredients (fish meal, dextrin, fish oil and soybean oil). Each treatment had three replicates with 15 fish per replicate. Fish were fed twice daily at the rate of 5% of their body weight. The result of the study indicated that the dietary protein in both reference and test diets were well digested and the apparent protein digestibility (APD) values of test diets ranged between 82.81 and 85.99%. The APD value of fish meal protein was 88.05%. The apparent digestible energy (ADE) value for the test ingredients ranged between 70.79 and 85.80% with soybean oil having the highest and fish meal the lowest value. The ADE values calculated in terms of Kcal/g of ingredients were 3.68, 3.22, 4.38 and 4.44 Kcal/g for fish meal, dextrin, fish oil and soybean oil respectively.

Studies on the optimum protein to energy ratio of African catfish (Clarias batrachus Burchell)

A laboratory trial was conducted to determine the optimum dietary protein to energy (P/E) ratio of African catfish, Clarias gariepinus. The experiment was carried out in a flow-through system for 6 weeks. There were 12 treatments each with two replicates having 10 fish each with a mean initial weight of 1.80 ± 0.02g. Twelve semi-purified diets were formulated with four digestible crude protein levels (23, 26.5, 30 and 33.5%) and three digestible energy levels (2.25, 2.75 and 3.25 Kcal/g). The fish were fed three times daily at satiation level. The results of the study showed that, diet containing 33.5% digestible protein and 2.75 kcal/g digestible energy with a protein to energy ratio of 121 .8 (mg protein/kcal) appeared to be best utilized for growth.

Feeding metabolism in an Indian major carp (Catla catla Lin.) fed on different protein diets

Feeding metabolism in an Indian major carp, Catla catla fingerlings of 10.8+0.56g was investigated in a flow-through water recirculating system. The metabolic energy loss in resting metabolism and feeding metabolism were determined by the indirect method of oxygen consumption followed by multiplication by suitable oxycalorific coefficient. This was done in four metabolic chambers of a respirometer system. Ten fish fingerlings of mean total weight of 109.5, 110.4 and 112.8g/chambers respectively each in two experimental runs of three treatments a, b and c were used. The mean resting metabolic rate during unfed condition showed no significant variation in different treatments. The fish in three treatments a, b and c fed on diets containing 28, 33 and 38% crude protein had significantly different (p<0.05) post-fed SDA magnitude of 497.7, 638.7 and 735.5 mgO2/chamber/day having an equivalent energy loss of 12.68, 14.68 and 15.86 KJ respectively. The SDA co-efficient in three treatments a, b and c were 14.95, 19.00 and 22.36% respectively whereas, respiratory energy - 'R' as % of mean total ingested energy in three treatments were 26.93, 31.17 and 34.74% respectively showing a significant increase (p<0.05) with increase of protein. Feeding metabolism in an Indian major carp (Catla catla Lin.) fed on different protein diets.

Effects of animal and plant protein diets on growth of Indian major carp (Labeo rohita Ham.) fingerlings

An experiment was conducted with Labeo rohita fingerlings in an indoor static fish rearing water system of glass made aquaria. Five experimental diets A, B, C, D and E were formulated containing 33% dietary protein level in five treatments each having two replicates containing 12 fingerlings of mean total initial weight of 13.00±0.2g. Sixty days of feeding trial in this experiment showed that fish fed on diet 'A' containing fish meal and diet 'E' containing mixed plant sources protein had significantly highest and lowest growth respectively. However, no significant difference of growth was found in fish fed on diets C and D containing meat and bone meal, and mix of animal protein source diets respectively. The result showed that the apparent protein digestibility (APD) of diets 'A' and 'E' had significantly best and least values respectively. Food conversion ratio (FCR) and protein efficiency ratio (PER) ranged between 1.37 to 2.17 and 1.38 to 2.18 respectively. On the basis of observed FCR and PER diets 'A' and 'E' produced significantly highest and lowest growth respectively.

Cloning and characterization of a blood coagulation factor IX-binding protein from the venom of Trimeresurus stejnegeri

A blood coagulation factor IX-binding protein (TSV-FIX-BP) was isolated from the snake venom of Trimeresurus stejnegeri. On SDS-polyacrylamide gel electrophoresis, TSV-FIX-BP showed a single band with an apparent molecular weight of 23,000 under non-reducing conditions. and two distinct bands with apparent molecular weights of 14,800 and 14,000 under reducing conditions. cDNA clones containing the coding sequences of TSV-FIX-BP were isolated and sequenced to determine the structure of the precusors of TSV-FIX-BP subunits. The deduced amino acid sequences of two subunits of TSV-FIX-BP were confirmed by N-terminal protein sequencing and trypsin-digested peptide mass fingerprinting. TSV-FIX-BP was a nonenzymatic C-type lectin-like anti-coagulant. The anti-coagulant activity of TSV-FIX-BP was mainly caused by its dose dependent interaction with blood coagulation factor IX but not with blood coagulation factor X. (C) 2003 Elsevier Science Ltd. All rights reserved.

Molecular cloning and characterization of a platelet glycoprotein Ib-binding protein from the venom of Trimeresurus stejnegeri

A platelet glycoprotein Ib-binding protein, termed TSV-GPIb-BP, was isolated from the venom of Trimeresurus stejnegeri. On SDS-polyacrylamide gel electrophoresis, TSV-GPIb-BP showed a single band with an apparent molecular weight of 28,000 and two distinct bands with apparent molecular weights of 16,000 and 15,000 under non-reducing and reducing conditions, respectively. cDNA clones containing the coding sequences for both TSV-GPIb-BP subunits were isolated and sequenced. The deduced amino acid sequences of TSV-GPIb-BP subunits were confirmed by N-terminal protein sequencing and trypsin-digested peptide mass fingerprinting. Interestingly, the a subunit of TSV-GPIb-BP is identical to that of alboaggregin-B, and the sequence identity of their beta subunits is 94.3%. TSV-GPIb-BP inhibited ristocetin-induced human platelet agglutination in platelet-rich plasma under lower dosages (<5 mug/ml). On the other hand, it directly aggregated washed human platelets in the absence of additional Ca2+ or any other cofactors under higher dosages (>5 mug/ml). This platelet aggregation activity was dose-dependently inhibited by specific GPIbalpha antibodies, but not by those antibodies against platelet GPIa, GPIIa, GPIIb and GPIIIa. (C) 2003 Elsevier Science Ltd. All rights reserved.

Maximin 9, a novel free thiol containing antimicrobial peptide with antimycoplasma activity from frog Bombina maxima

Amphibian skin is a rich resource of antimicrobial peptides, like maximins and maximin Hs from frog Bombina maxima. Novel cDNA clones encoding a precursor protein, which comprises a novel maximin peptide (maximin 9) and reported maximin H3, were isolated from two constructed skin cDNA libraries of B. maxima. The predicted primary structure of maximin 9 is GIGRKFLGGVKTTFRCGVKDFASKHLY-NH2. A surprising substitution is at position 16, with a free cysteine in maximin 9 rather than usual conserved glycine in other reported maximins. Maximin 9, the homodimer form and its Cys(16) to Gly(16) mutant were synthesized and their antimicrobial activities were evaluated. Unlike previously reported maximin 3, the tested bacterial and fungal strains were resistant to maximin 9, its homodimer and the Cys(16) to Gly(16) mutant (with MICs > 100 mu M). On the other hand, interestingly, while eight clinical Mollicutes strains were generally resistant to maximin 9 homodimer and its Cys(16) to Gly(16) mutant, most of them are sensitive to maximin 9 at a peptide concentration of 30 mu M, especially in the presence of dithiothreitol. These results indicate that the presence of a reactive Cys residue in maximin 9 is important for its antimycoplasma activity. The diversity of antimicrobial peptide cDNA structures encountered in B. maxima skin cDNA libraries and the antimicrobial specificity differences of the peptides may reflect well the species' adaptation to the unique microbial environments. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

An ARGONAUTE4-containing nuclear processing center colocalized with Cajal bodies in Arabidopsis thaliana.

ARGONAUTE4 (AGO4) and RNA polymerase IV (Pol IV) are required for DNA methylation guided by 24 nucleotide small interfering RNAs (siRNAs) in Arabidopsis thaliana. Here we show that AGO4 localizes to nucleolus-associated bodies along with the Pol IV subunit NRPD1b; the small nuclear RNA (snRNA) binding protein SmD3; and two markers of Cajal bodies, trimethylguanosine-capped snRNAs and the U2 snRNA binding protein U2B''. AGO4 interacts with the C-terminal domain of NRPD1b, and AGO4 protein stability depends on upstream factors that synthesize siRNAs. AGO4 is also found, along with the DNA methyltransferase DRM2, throughout the nucleus at presumed DNA methylation target sites. Cajal bodies are conserved sites for the maturation of ribonucleoprotein complexes. Our results suggest a function for Cajal bodies as a center for the assembly of an AGO4/NRPD1b/siRNA complex, facilitating its function in RNA-directed gene silencing at target loci.

Comparative evaluation of fish protein concentrate and functional fish protein concentrate from cat fish

Fish protein concentrate and functional fish protein concentrate samples were prepared from eviscerated meat of cat fish (Tachysurus jella Day). Functional fish protein concentrate is found to be lighter, less gritty and rehydrates more rapidly than fish protein concentrate. Functional FPC is seen to have higher PER and biscuits containing it at levels of 5 and 7 percent are less hard compared to FPC.

Formulation, processing and water stability of pelleted feeds with varying levels of protein used in carp nutrition studies

Four dry pelleted feeds containing 20%, 30%, 40% and 45% protein were formulated incorporating casein as the main source of protein for use in carp nutrition studies. The caloric content in all the feeds was maintained constant. The method of processing is described. The formulated diets were tested for water stability. This test has revealed that the diet containing 20%, 30% and 40% protein had better stability than that containing 45% protein. This was due to the relatively higher fat content in the former three diets. However, all the feeds were sufficiently stable at the end of one hour in which time carps are known to utilise supplementary diets.

A preliminary study on the protein requirement of Chanos chanos (Forskal) fry in a controlled environment

Growth rate of fish appeared to be related to the levels of the protein in the diet up to 40%. Fish fed diets containing 50 and 60% grew slower than those fed 40%, and the optimum level appears to be 40% when fed to fry at a rate of 10% of body weight. Best feed conversion of 1.96 was also obtained from the 40% protein diet. Mean survival rates were low in all treatments, but highest for the 40% protein diet. The competition of 5 isocaloric experimental diets containing various levels of protein are tabulated, as are weight gains, diet conversions and survival rates for milkfish fry fed various dietary levels of protein. Growth curves for milkfish fry are shown, and the relationship between weight gains of milkfish fry and the dietary levels of protein are illustrated.