Incorporation and metabolism of (n-3) and (n-6) polyunsaturated fatty acids in phospholipid classes in cultured turbot (Scophthalmus maximus) cells

The incorporation and metabolism of various (n-3) and (n-6) polyunsaturated fatty acids (PUFA) supplemented to the culture medium was investigated in a turbot cell line (TF). The distribution, and the occurrence and extent of further metabolism of incorporated PUFA via desaturation/elongation mechanisms in specific phospholipid classes was determined from the different fatty acid compositions. The cells contained Δ6 and Δ4 desaturase activities but were generally deficient in C18–20 elongase activity. Δ5 Desaturase activity was generally masked by this deficiency but was present. The compositional data indicated that there was a high degree of specificity between individual phospholipid classes and particular fatty acids probably driven by the specificities of the acylating enzymes. The highest percentages of the supplemented acids were generally observed in the phosphatidic acid/cardiolipin fraction (PA/CL), suggesting a role for PA in the incorporation of the supplemented acids into the phospholipid pool. PI had a characteristic composition consistent with a putative role as a pool of precursor fatty acid for eicosanoid synthesis. Mechanisms were evident for generating and/or maintaining this composition.     

Incorporation and metabolism of (n-3) and (n-6) polyunsaturated fatty acids in phospholipid classes in cultured rainbow trout (Salmo gairdneri) cells

The incorporation and metabolism of various (n-3) and (n-6) polyunsaturated fatty acids supplemented to the culture medium was investigated in the rainbow trout cell line, RTG-2. The distribution, and the occurrence and relative extent of further desaturation and elongation of the incorporated acids was determined in individual phospholipid classes by analysis of the fatty acid compositions. RTG-2 cells exhibited Δ6 and Δ5 desaturase activities whereas Δ4 desaturase activity was almost totally absent. The percentage of precursor acids was greatest in the phosphatidic acid/cardiolipin fraction (PA/CL), suggesting a role for possibly PA in the initial incorporation of these acids into the phospholipid pool. The compositional data indicated that individual intermediates and products of the desaturation pathways were associated with specific phospholipid classes probably via mechanisms depending upon the specificities of the acylating enzymes. The composition of phosphatidylinositol (PI) and the tightly controlled mechanisms for generating/maintaining it are consistent with a role for this phospholipid in providing precursor fatty acid for eicosanoid synthesis.     

The incorporation and metabolism of polyunsaturated fatty acids in phospholipids of cultured cells from chum salmon (Oncorhynchus keta)

The incorporation and metabolism of (n-3) and (n-6) polyunsaturated fatty acids were studied in a cell line derived from chum salmon heart (CHH-1). Supplementing media with 25 M fatty acid considerably altered the cellular fatty acid composition but did not affect the lipid class composition or cause the appearance of cytoplasmic lipid droplets. CHH-1 cells exhibited considerable -6-desaturase activity but showed no preference between (n-3) and (n-6)PUFA substrates. CHH-1 cells also possess -5-desaturase activity which showed preference towards (n-3)PUFA, but -4-desaturase activity was totally absent. Elongation of 20-carbon PUFA was especially active in CHH-1 cells with 22-carbon PUFA being specifically incorporated into PE and PS lipid classes. The fatty acid composition of PI indicated specific incorporation of 20-carbon PUFA into this lipid class. Supplementation with 22:6(n-3) generated fatty acid compositions more closely resembling those of intact salmonid hearts. Substantial chain shortening of 22:6(n-3) to 20:5(n-3) occurred.Abbreviations BHT butylated hydroxytoluene - BSA bovine serum albumin - CL cardiolipin - FCS fetal calf serum - PA phosphatidic acid - PC phosphatidylcholine - PE phosphatidylethanolamine - PI phosphatidylinositol - PS phosphatidylserine - PUFA polyunsaturated fatty acid - SM sphingomyelin     

The conversion of linoleic acid and linolenic acid to longer chain polyunsaturated fatty acids by Tilapia (Oreochromis) nilotica in vivo

Tilapia (Oreochromis) nilotica were fed either a commercial diet containing 2.2% (n-3) and 0.5% (n-6) polyunsaturated fatty acids (PUFA), or a diet containing 1.0% methyl linoleate as the only PUFA. The fatty acid composition of tissue lipids generally reflected that of the diet. Fish from both dietary groups were injected intraperitoneally with ¹⁴C-labelled linoleic acid, 18:2 (n-6), or linolenic acid, 18:3 (n-3), and the distribution of radioactivity in tissue lipids examined. The conversion of both 18:2 (n-6) and 18:3 (n-3) to longer chain PUFA was lower in fish fed the commercial diet than in those fed the diet containing only 18:2 (n-6). Half of the radioactivity from both substrates recovered in liver polar lipids was present in C20 and C22 PUFA with fish maintained on the experimental diet. It is concluded that T. nilotica is capable of elongating and desaturating both 18:2 (n-6) and 18:3 (n-3), but that this conversion is suppressed by dietary longer chain PUFA. NERC Unit of Aquatic Biochemistry     

The desaturation and elongation of 14C-labelled polyunsaturated fatty acids by pike (Esox lucius L.) in vivo

To examine the ability of pike (Esox lucius L.) to modify exogenous PUFA by desaturation and elongation, ¹⁴C-labelled 18:2(n-6), 18:3(n-3), 20:4(n-6) and 20:5(n-3) were injected intraperitoneally and the distribution of radioactivity in tissue lipid classes and liver PUFA measured. In all tissues examined, radioactivity from all ¹⁴C-PUFA was recovered in many classes of acyl lipids and the level of recovery generally reflected the relative abundance of the lipid classes. Triacylglycerols, CGP and EGP usually contained high levels of all incorporated ¹⁴C-PUFA. PI contained higher levels of radioactivity from ¹⁴C-20:4(n-6) than from other injected substrates. In liver lipid, the 6 desaturation products of ¹⁴C-18:2(n-6) and ¹⁴C-18:3(n-3) contained no measurable radioactivity although the elongation products of the 6 desaturation products were labelled, as were the direct elongation products of these injected substrates. No radioactivity from ¹⁴C-18:2(n-6) or ¹⁴C-18:3(n-3) was detected in C₂₀ or C₂₂ products of 5 and 4 desaturation. Almost all radioactivity from injected ¹⁴C-20:4(n-6) was recovered in this PUFA. Of the total radioactivity from ¹⁴C-20:5(n-3) incorporated into liver lipid, 7% was present as 24:5 and 16.4% was recovered in hexaenoic fatty acids. In liver, 24:5(n-3) and 24:6(n-3) each accounted for 1% of the mass of total fatty acids and were located almost exclusively in triacylglycerols. The presence of radioactivity in these C₂₄ PUFA suggests that in pike the synthesis of 22:6(n-3) from 20:5(n-3) may proceed without 4 desaturase via the pathway which involves chain shortening of 24:6(n-3). It is concluded that under the circumstances employed in this study pike, do not exhibit 5 desaturase activity and are unable to synthesize 20:4(n-6) and 20:5(n-3) from 18:2(n-6) and 18:3(n-3), respectively. This suggests that pike may require 20:4(n-6) and 20:5(n-3) preformed in the diet.Abbreviations CGP choline glycerophospholipids - CL cardiolipin - EGP ethanolamine glycerophospholipids - PG phosphatidylglycerol - PI phosphatidylinositol - PS phosphatidylserine - PUFA polyunsaturated fatty acids - SM sphingomyelin - TLC thin-layer chromatography     

The effect of culture on morphology, lipid and fatty acid composition, and polyunsaturated fatty acid metabolism of rainbow trout (Oncorhynchus mykiss) skin cells

Rainbow trout (Oncorhynchus mykiss) skin cell cultures were obtained by trypsinization of the tissue and grown in Leibovitz L-15 medium. Lipid class compositions, and fatty acid profiles of total lipids and individual phospholipid classes were determined at different times of culture. The metabolism of polyunsaturated fatty acids (PUFA) was investigated by incubating primary cultures after 7 and 14 days with [1-¹⁴C]18:2n-6 and [1-¹⁴C-]18:3n-3. The change in morphology between epithelial-like primary cultures and fibroblastic-like secondary subcultures was accompanied by alterations in the lipid composition. Polar lipids became predominant by 14 days in culture. The relative proportions of phosphatidylcholine (PC), the most abundant phospholipid, phosphatidylinositol and cholesterol increased significantly, while sphingomyelin decreased. Saturated fatty acids, 18:1n-9, n-6 and n-9PUFA were more abundant in total lipid in cultures at 14 days and 4 months than in cells initially isolated which contained higher percentages of longer chain monoenes and n-3PUFA. The changes in fatty acid composition with time in culture were observed in all the major phospholipid classes. Rainbow trout skin cells in culture desaturated and elongated both 18:2n-6 and 18:3n-3, with 20:4n-6 and 20:5n-3 being the most abundant products, respectively. PC presented the highest incorporation of radioactivity, especially following incubation with 18:3n-3. Lipid metabolism in general increased with the age of primary cultures, with both the amount of C18 PUFA incorporated and metabolized by desaturation/elongation significantly increased in 14 day cultures compared to 7 day cultures. Product/precursor ratios calculated for both n-6 and n-3 fatty acids showed that, while 6 desaturase activity was increased significantly with cell age, 5 desaturase activity was more affected by the fatty acid series, with 18:3n-3 being more readily transformed to 20:5n-3 than 18:2n-6 to 20:4n-6. Further desaturation of 20:5n-3 to hexaenes was low. Overall, the data suggested that the trout skin cell cultures were more similar to mammalian skin fibroblasts than mammalian epidermal/keratinocyte cultures.     

Organ and phospholipid class fatty acid specificity in response to dietary depletion of essential n‐3 fatty acids in Atlantic salmon (Salmo salar L.)

The cell membrane phospholipid composition is of major importance for normal cell functions. However, it is not known how complete depletion of both shorter and longer chain omega‐3 fatty acids in salmon diets influences fatty acid composition of phospholipid subclasses in different organs of Atlantic salmon. We describe here the fatty acid composition in phospholipid subclasses of liver, muscle, heart and intestine in Atlantic salmon after 18 months of dietary n‐3 essential fatty acids deprivation. The percentage of 22:6n‐3 was markedly reduced in almost all phospholipid subclasses, and except for muscle phosphatidylcholine, phosphatidylethanolamine (PE) and phosphatidylinositol (PI), phospholipids in deficient fish were totally devoid of 20:5n‐3. As compensation, we observed significant increases in 20:4n‐6, and especially in 20:3n‐9 (Mead acid) and 22:5n‐6, varying among phospholipids and organs. High amounts of 20:3n‐9 were found in liver and intestinal PE, little in PE from heart and muscle. For 22:5n‐6, we saw a small incorporation in PI in liver and intestine compared to heart and muscle. Generally in PI, the preference for 20:4n‐6 to 20:5n‐3 differed significantly between organs. Overall, changes upon n‐3 deprivation seemed to be strongest in liver and intestine, the lipid‐secreting organs, and less in muscle and heart.     

Muscle fatty acid composition and oxidative stress indices of Arctic charr, Salvelinus alpinus (L.), in relation to dietary polyunsaturated fatty acid levels and temperature

The influence of feeding high levels of polyunsaturated fatty acids (PUFA) on muscle fatty acid composition and indices of oxidative damage was examined in Arctic charr, Salvelinus alpinus (L.). All diets contained 100 g kg^?1 lipid of dry weight. Two diets contained marine fish oils giving a PUFA level of 250 g kg^?1 and 500 g kg^?1 of lipid. The remaining two diets contained vegetable oils high in either 18:2n-6 or 18:3n-3, giving a PUFA level of more than 500 g kg^?1 of dietary lipid. The charr were maintained at 8°C until their weight doubled, and were then transferred to 0.8°C for 30 days. Growth was similar in all groups. The fatty acid compositions of muscle were influenced by dietary PUFA but were less diverse than those of the diets. The overall pattern of fatty acid compositions indicated preferential desaturation and elongation of n-3 PUFA coupled with selective oxidation of 18:2n-6. Total n-3 PUFA content in TAG was always lowered compared with the diet, suggesting a specific mechanism for the removal of these fatty acids. Subjecting the fish to low temperature increased PUFA content in muscle of charr fed the 250 g kg^?1 marine n-3 PUFA diet, but had no effect on the other treatments. For fish at 8°C, no significant differences were found between groups in terms of haematocrit, plasma alanine aminotransferase (ALAT), and plasma and muscle thiobarbituric acid reactive substances (TBARS), although there was a tendency towards increased levels of TBARS in the group receiving 500 g kg^?1 marine n-3 PUFA of lipid. Subjecting the muscle to forced oxidative conditions resulted in increases in TBARS in all groups, particularly those fed 500 g kg^?1 marine n-3 PUFA. Lowering the environmental temperature corresponded with a further increase in the plasma ALAT and muscle TBARS in this group. It is concluded that feeding diets containing high levels of long-chain n-3 PUFA may be detrimental to the fish's health and flesh quality, particularly at low environmental temperatures.     

Incorporation and metabolism of14C-labelled polyunsaturated fatty acids in wild-caught juveniles of golden grey mullet,Liza aurata,in vivo

The incorporation, and the capacity for desaturation and elongation in vivo, of intraperitoneally-injected,¹⁴C-labelled n-3 and n-6 C₁₈ and C₂₀ polyunsaturated fatty acids (PUFA) were investigated in juvenile golden grey mullet,Liza aurata. The results indicate that juvenile mullet have only limited ability to convert C₁₈ polyunsaturated fatty acids to C₂₀ and C₂₂ highly unsaturated fatty acids (HUFA)in vivo. This suggests that juvenile golden grey mullet require the provision of preformed C_20/22 HUFA, such as eicosapentaenoic and docosahexaenoic acids, in the diet. The impairment in the desaturase/elongase pathway was similar to that found in turbot,Scophthalmus maximus, and gilthead sea bream,Sparus aurata, being primarily at the level of Δ5-desaturase. The data from the largely herbivorous golden grey mullet juveniles are consistent with the hypothesis that marine fish in general, irrespective of dietary habits, have limited capacity for the desaturation and elongation of C₁₈ PUFA. The defect in Δ5-desaturase activity combined with the consistent finding that arachidonic acid is selectively incorporated and retained in membrane phosphatidylinositol suggests that, like turbot and gilthead sea bream, golden grey mullet may also have a requirement for preformed arachidonic acid in the diet.     

Effects of dietary (n-3) and (n-6) polyunsaturated fatty acid ratio on the immune response of Atlantic salmon, Salmo salar L.

To examine the influence of the dietary ratio of (n-3) to (n-6) polyunsaturated fatty acids (PUFAs) on the immune system of Atlantic salmon, Salmo salar L., two dietary trials were carried out in which parr were maintained on diets containing either fish oil [(n-3)/(n-6) PUFA = 5.2] or sunflower oil [(n-3)/(n-6) PUFA = 0.3] and assessed for differences in immunological parameters. There were no significant differences in blood cell counts, differential leucocyte counts or haematocrit values between dietary groups, and while no apparent differences were observed in the non-specific immune parameters measured, there was a significantly higher number of B cells responding to Aeromonas salmonicida, in the kidney and spleen of vaccinated fish maintained on high (n-3)/(n-6) PUFAs diets. There was also a significant difference (P≤ 0.01) between the dietary groups in trial 1 and trial 2 when non-vaccinated fish were challenged with Aeromonas salmonicida and Vibrio anguillarum, respectively, with the (n-6) group succumbing to the bacterium before the (n-3) group. The results suggest that Atlantic salmon fed diets with a low ratio of (n-3)/(n-6) PUFA may be less resistant to infection than those fed diets containing lipid with a high (n-3)/(n-6) PUFA ratio.     

Effect of polyunsaturated fatty acids on the fecundity of the Amazon river prawn Macrobrachium amazonicum (Heller, 1862)

Effects of the ratio of dietary fatty acids, namely n‐3 (mainly long chain polyunsaturated – LC‐PUFA) to n‐6 PUFA on the fecundity of Macrobrachium amazonicum were evaluated. In T1, the diet had equal and low levels of dietary n‐3 and n‐6 fatty acids (6 mg g^?1). In T2, the concentration of n‐3 (6 mg g^?1) was a half of the concentration of the n‐6 (12 mg g^?1), and in T3, the diet had equal and high concentrations of n‐3 and n‐6 (12 mg g^?1). Females with ovaries in stages I, III and V were collected. Higher gonadosomatic index (GSI) (6.89%) was observed in females in ovarian stage V than at other ovarian stages; however, the hepatosomatic index (HIS) showed high values in all females with ovaries in the stage III. A ratio of 1:2 n‐3:n‐6 fatty acids increased the GSI of mature females and the number of eggs spawned. Raising the level of both n‐3 and n‐6 fatty acids from ~0.6% to ~1.2% of the diet did not produce any effect on the GSI or on fecundity, suggesting that the ratio is more important than the absolute value of these two families of fatty acids.     

Polyunsaturated fatty acid metabolism in a cell culture model of essential fatty acid deficiency in a freshwater fish, carp (Cyprinus carpio)

Proliferation of an essential fatty acid deficient cell line from carp (EPC-EFAD; epithelioma papillosum carp-essential fatty acid deficient) is stimulated by supplementing the cells with C₂₀, but not C₁₈ polyunsaturated fatty acids (PUFA). It is hypothesized that the differential ability of the PUFA to stimulate proliferation of the EPC-EFAD cells may be related to the extent of the cells' ability to desaturate and elongate C₁₈ PUFA. In the present study, the metabolism of ¹⁴C-labeled C₁₈ and C₂₀ PUFA was investigated in EPC-EFAD cells in comparison with normal EPC cells. The incorporation of all the PUFA was significantly greater in EPC-EFAD cells but the rank order, 20:5n-3 > 18:3n-3 = 18:2n-6 >20:4n-6 was the same in both cell lines. The proportion of radioactivity from all labeled PUFA recovered in phosphatidylethanolamine and total polar lipids was significantly lower in EPC-EFAD cells compared to EPC cells, whereas the proportion of radioactivity recovered in all the other phospholipid classes and total neutral lipid was greater in EPC-EFAD cells. Both cell lines desaturated[1-¹⁴C]18:3n-3 and [1-¹⁴C]20:5n-3 to a greater extent than the corresponding (n-6) substrates but the desaturation of all the ¹⁴ C-labeled PUFA was significantly greater in EPC-EFAD cells compared to EPC cells. The results showed that, although essential fatty acid deficiency had several significant effects on PUFA metabolism in EPC cells, the fatty acid desaturation/elongation pathway was not impaired in EPC-EFAD cells and so they can desaturate 18:3n-3 to 20:5n-3 and 22:6n-3, and 18:2n-6 to 20:4n-6. However, 20:4n-3 and 20:3n-6, and not 20:4n-6 and 20:5n-3, were the predominant C₂₀ PUFA produced by the elongation and desaturation of [1-¹⁴C]18:3n-3 and [1-¹⁴C]18:2n-6, respectively. Therefore, the previously reported inability of 18:3n-3 and 18:2n-6, compared to 20:5n-3 and 20:4n-6, to stimulate proliferation of the cells is apparently not due to a general deficiency in the fatty acid desaturation pathway in EPC-EFAD cells but may be related to potential differences in eicosanoid profiles in cells supplemented with C₁₈ PUFA compared to C₂₀ PUFA.     

Influence of dietary palm oil on growth, tissue fatty acid compositions, and fatty acid metabolism in liver and intestine in rainbow trout (Oncorhynchus mykiss)

This study aimed to investigate the effects of dietary crude palm oil (CPO) on fatty acid metabolism in liver and intestine of rainbow trout. Triplicate groups of rainbow trout for 10 weeks at 13 °C were fed on diets in which CPO replaced fish oil (FO) in a graded manner (0–100%). At the end of the trial, fatty acid compositions of flesh, liver and pyloric caeca were determined and highly unsaturated fatty acid (HUFA) synthesis and fatty acid oxidation were estimated in isolated hepatocytes and caecal enterocytes using [1‐¹⁴C]18:3n‐3 as substrate. Growth performance and feed efficiency were unaffected by dietary CPO. Fatty acid compositions of selected tissues reflected the dietary fatty acid composition with increasing CPO resulting in increased proportions of 18:1n‐9 and 18:2n‐6 and decreased proportions of n‐3HUFA, 20:5n‐3 and 22:6n‐3. Palmitic acid, 16:0, was also increased in flesh and pyloric caeca, but not in liver. The capacity of HUFA synthesis from 18:3n‐3 increased by up to threefold in both hepatocytes and enterocytes in response to graded increases in dietary CPO. In contrast, oxidation of 18:3n‐3 was unaffected by dietary CPO in hepatocytes and reduced by high levels of dietary CPO in enterocytes. The results of this study suggest that CPO can be used at least to partially replace FO in diets for rainbow trout in terms of permitting similar growth and feed conversion, and having no major detrimental effects on lipid and fatty acid metabolism, although flesh fatty acid compositions are significantly affected at an inclusion level above 50%, with n‐3HUFA reduced by up to 40%.     

银鲳卵黄发生期间组织中抗氧化水平的变化及饲料n-3 LC-PUFA对其的影响

研究了银鲳(Papus argenteus)肝及卵巢组织中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性、总抗氧化能力(T-AOC)及丙二醛(MDA)含量在整个卵黄发生过程中的变化情况,并分析了饲料n-3 LC-PUFA对卵黄发生期间组织中抗氧化水平的影响。分别以100%鱼油(FO组)、70%鱼油和30%大豆油(FSO组)、30%鱼油和70%大豆油(SFO组)、100%大豆油(SO组)为脂肪源,配制了4组等氮、等能及等脂的试验饲料。以1年龄雌性银鲳为试验对象,每组饲料设3重复,试验周期185 d。研究结果表明,肝与卵巢组织SOD、CAT活性、T-AOC水平及MDA含量在卵黄发生过程中均呈现逐渐升高的趋势,且卵黄发生后期各指标水平均显著高于卵黄发生前期(P0.05)。肝SOD(除SO饲料组外)、CAT活性、T-AOC水平及MDA含量在卵黄发生中期与前期之间未表现出显著性差异(P0.05),而卵黄发生中期FO与FSO饲料组卵巢SOD、CAT活性及T-AOC水平则均显著高于卵黄发生前期(P0.05)。FSO饲料组肝与卵巢SOD、CAT活性在卵黄发生过程中均为最高值,且在卵黄发生中、后期均显著高于SO饲料组(P0.05),但与FO饲料组无显著性差异(P0.05)。各饲料组间肝与卵巢T-AOC水平在卵黄发生前期均未表现出显著性差异(P0.05),而在卵黄发生后期,FO与FSO饲料组肝与卵巢T-AOC水平均显著高于SO饲料组(P0.05),但FO与FSO饲料组间无显著性差异(P0.05)。肝与卵巢中MDA含量随着饲料n-3 LC-PUFA含量的升高而呈现出升高趋势,且这种升高趋势在肝组织中表现更为明显,卵巢组织MDA含量在卵黄发生中、后期仅FO饲料组表现出显著性升高趋势(P0.05),其他各饲料组在卵黄发生各期均未表现出显著性差异(P0.05)。统计分析表明,银鲳卵黄发生过程中组织中抗氧化水平逐渐升高,适宜的饲料n-3 LC-PUFA含量(4.01%,FSO饲料组)可明显改善银鲳卵黄发生中期与后期组织中的抗氧化水平。双因素方差分析结果表明,试验饲料与卵黄发生时期对银鲳组织抗氧化水平均具有极显著性影响(P0.01),且两者对肝T-AOC水平与MDA含量存在显著性的交互作用(P0.05)。     

First isolation of Vibrio tapetis and an atypical strain of Aeromonas salmonicida from skin ulcerations in common dab (Limanda limanda) in the North Sea

Skin ulcerations rank amongst the most prevalent lesions affecting wild common dab (Limanda limanda) with an increase in prevalence of up to 3.5% in the Belgian part of the North Sea. A complex aetiology of these ulcerations is suspected, and many questions remain on the exact factors contributing to these lesions. To construct the aetiological spectrum of skin ulcerations in flatfish, a one‐day monitoring campaign was undertaken in the North Sea. Fifteen fish presented with one or more ulcerations on the pigmented and/or non‐pigmented side. Pathological features revealed various stages of ulcerations with loss of epidermal and dermal tissue, inflammatory infiltrates and degeneration of the myofibers bordering the ulceration, albeit in varying degrees. Upon bacteriological examination, pure cultures of Vibrio tapetis were retrieved in high numbers from five fish and of Aeromonas salmonicida in one fish. The V. tapetis isolates showed cross‐reactivity with the sera against the representative strain of serotype O2 originating form a carpet‐shell clam (Ruditapes descussatus). Moreover, the A. salmonicida isolates displayed a previously undescribed vapA gene sequence (A‐layer type) with possible specificity towards common dab. Further research is necessary to pinpoint the exact role of these agents in the development of skin ulcerations in common dab.     

Effects of olive pomace on growth performance,digestibility, body composition and fatty acid profile in yearling Siberian sturgeon,Acipenser baerii (Brandt 1896)

Replacement of olive pomace (OP) with wheat flour in diet was studied in diet of yearling Siberian sturgeon (Acipenser baerii). Fish (165 ± 6.8 g) were randomly allocated to 15 fibreglass tanks (15 fish/tank, in triplicate). Fish were assigned to one of five dietary treatments at 19.1 ± 1.5°C for 8 weeks: control diet (crude protein and crude lipid at 514.6 and 188 g/kg_, respectively) and four experimental diets with 20, 50, 75 and 100 g/kg OP to replace wheat flour in the experimental diets. No significant difference was observed in the final weight, feed conversion ratio, specific growth rate, hepatosomatic index, viscera‐somatic index and survival rate among the treatments (p > 0.05). No change was seen in digestibility of protein and lipid in diets containing 20 g/kg and 50 g/kg OP compared to control group, while these values decreased with increasing in OP above 50 g/kg. Digestibility of dry matter and gross energy among the treatments demonstrated no significant difference (p > 0.05). Polyunsaturated fatty acids, especially C22:6n3 (DHA), in the muscle of the fish fed 75 g/kg and 100 g/kg OP in diets were significantly higher than control group (p < 0.05). These data show that OP could be recommended as a substitute for wheat flour in diet of S. sturgeon but its utilization for other species warranted future works.     

A comparative study of Edwardsiella ictaluri parent (EILO) and E. ictaluri rifampicin-mutant (RE-33) isolates using lipopolysaccharides,outer membrane proteins,fatty acids,Biolog, API 20E and genomic analyses

The biological properties of Edwardsiella ictaluri RE-33 rifampicin-mutant and its parent strain EILO were analysed. RE-33 is an avirulent isolate used as a modified live vaccine against enteric septicaemia of catfish. Electrophoretic analysis of lipopolysaccharide (LPS) patterns showed high homology between both isolates. Further characterization of LPS by immunoblotting revealed the main differences in LPS composition. The RE-33 isolate lacks the high molecular weight bands of LPS (HMW-LPS). Outer membrane protein analysis also showed some immunological differences between RE-33 and the EILO parent strain. Only two fingerprinting techniques, fatty acid composition analysis and Biolog phenotypic profiles, were able to discriminate between both isolates.     

The effect of time‐dependent protein restriction on growth factors,nonspecific immunity,body composition,fatty acids and amino acids in the Siberian sturgeon (Acipenser baerii)

The effect of time‐dependent protein restriction (PR) and refeeding were investigated on growth, body composition, fatty acids (FA), amino acids (AA) and nonspecific immune functions in the juvenile Siberian sturgeon (Acipenser baeri; 75 ± 5 g). Test diets included: T1 (control) and T2 fish fed diets containing 400 and 300 g/kg protein respectively during the whole experimental period, T3 (every other day) and T4 (every other week) fish were fed diets containing 300 g/kg protein (PR) and refeeding with diet containing 400 g/kg protein respectively, and T5 fish fed a diet containing 300 g/kg protein for 3 weeks and a diet containing 400 g/kg protein for 5 weeks, were fed to the fish for 56 days by visual satiation. Unlike ration T2 and T5, feeding treatments of T3 and T4 showed an increase in fish growth and body composition (p < 0.05) and they were very close to the control group (p > 0.05). Regarding the fatty acid profile, although the percentage of highly unsaturated fatty acids (HUFA) and polyunsaturated fatty acids (PUFA) declined significantly in the T2 and T5 groups, the saturated fatty acid (SFA) and monounsaturated fatty acids (MUFA) indicated no change among the treatments. In addition, the T2 and T5 groups demonstrated a reduction in essential amino acids (EAA) with a simultaneous increase in nonessential amino acids (NEAAs), which was significant from that of controls and other groups (p < 0.05). In terms of nonspecific immune parameters (serum lysozyme activity and alternate complement activity [ACH50]), treatment 3 has an appropriate result that did not have a significant difference with the control group (p > 0.05), but there was significant difference with other groups (p < 0.05). As a result, the T3 treatment can be used in sturgeon aquaculture practically without any negative impacts on biological or physiological indices.     

Complete repertoire of long‐chain polyunsaturated fatty acids biosynthesis enzymes in a cyprinid,silver barb (Barbonymus gonionotus): Cloning,functional characterization and dietary regulation of Elovl2 and Elovl4

The silver barb (Barbonymus gonionotus) is a valuable cyprinid with good deposition of long‐chain polyunsaturated fatty acids (LC‐PUFA) such as docosahexaenoic acid (DHA). We have previously isolated a fatty acyl desaturase (Fads2) and elongase (Elovl5), which fulfils the production of eicosapentaenoic acid (EPA) and arachidonic acid (ARA) from C18 polyunsaturated fatty acid (PUFA) substrates. The isolation, functional characterization and dietary regulation of two elongases, Elovl2 and Elovl4, are reported here. The Elovl2 displayed the capacity to elongate C20 and C22 PUFA substrates, while the showed low activities towards saturated fatty acids and C22 PUFA substrates. This discovery validates the existence of a complete set of enzymes of LC‐PUFA in silver barb. The elovl2 showed higher expression in liver, in limited dietary LC‐PUFA intake conditions. As for Elovl4, tissue expression showed prominence in eye tissue. We also showed that the expression of both genes was upregulated when fish was fed with diets devoid of LC‐PUFA. Fatty acid composition analysis indicates the utilization of C18 polyunsaturated fatty acids for LC‐PUFA biosynthesis.