Acid phosphatase activity in pre- and post-spawning hake (Merluccius hubbsi)

1. The specific activity of muscle acid phosphatase (E.C. 3.1.3.1.) in pre- and post-spawning hake was investigated. 2. Specific activity values, 0.54 in pre-spawning hakes and 1.64 in post-spawning fish, have been obtained. 3. Using a histochemical test, acid phosphatase has been localized mainly in the connective tissue. The strongest reaction has been observed in post-spawning hake. 4. The increase of the lysosomal activity in the connective tissue may be related to the changes found in the muscle texture associated with spawning.     

SSU rDNA analysis of Kudoa rosenbuschi (Myxosporea) from the Argentinean hake Merluccius hubbsi

The cloning and sequencing of the small subunit (SSU) ribosomal DNA gene from Kudoa rosenbuschi (myxosporean species associated with post-mortem myoliquefaction process in the Argentinean hake Merluccius hubbsi) is reported. The SSU rDNA was found to contain 1740 bp with a single polymorphic site with either a C or T at position 221. The sequence data obtained in this study and those known sequences of Kudoa species deposited in the GenBank were all analyzed to construct a phylogenetic tree. Nucleotide sequences showed the highest degree of identity with K. funduli, followed by K. miniauriculata, K. clupeidae and K. dianae. Phylogenetic analysis placed K. rosenbuschi in the same branch of K. clupeidae and K. funduli, and showed it to be closely related to K. dianae, K. paniformis and K. miniauriculata.     

Cytoskeletal ultrastructure and lipid composition of I-Z-I fraction in muscle from pre- and post-spawned female hake (Merluccius hubbsi)

Myofibrils from pre- and post-spawned female hake were depleted of thin and thick filaments by KI-treatment and the cytoskeletal ultrastructure of the preparations was investigated by electron microscopy. Irrespective of the gonadal stage of the fish, transmission (TEM) and scanning (SEM) electron micrographs showed an extensive network of filaments connecting Z-structures, which appeared as two circular rings held together. Z-structures of KI-treated myofibrils from post-spawned hake were more compact than those from pre-spawned fish. This ultrastructural difference was absent when KI-treated myofibrils from hake in both gonadal conditions were prepared in the presence of a proteinase inhibitor cocktail (1 mM EDTA+1 mM PMSF+1 mM iodoacetic acid). The total lipid (TL) content of the I-Z-I fraction from fish in pre-spawned condition was about 3.7 mg/g I-Z-I and non-polar lipids (NPL) represent 86.5% of TL. TL and NPL contents of the I-Z-I fraction from post-spawned hake were about 50% and 78.2% lower than those obtained from this fraction in pre-spawning fish. No significant changes (p>0.05) were observed in phospholipids (PL) and acidic phospholipids (AP). No significant differences (p<0.01) were observed among the corresponding lipid fractions of I-Z-I purified from pre- and post-spawning fish in the presence of a protease inhibitor cocktail.     

Fish protein hydrolysis by a psychrotrophic marine bacterium isolated from the gut of hake (Merluccius hubbsi)

The aims of this study were to identify a psychrotrophic bacterium, strain CR41, producing a cold adapted protease during growth at low temperatures and to evaluate the ability of the cells to hydrolyze hake fish protein. The strain was isolated from the intestinal tract of hake collected from the San Jorge Gulf (Patagonia, Argentina) and it was identified as Pseudoalteromonas. Growth and fish protein hydrolysis were determined using an aerated simple mineral medium plus 10% fish protein concentrate. Proteolytic activity was measured at 7 and 22 degrees C during culture in the concentrate. Protease production started in the exponential growth phase and reached a maximum during stationary phase. Protease activity at 7 degrees C was lower than at 22 degrees C. After 8 h of incubation, the percentage of hydrolyzed protein was 84% at 7 degrees C and 95% at 22 degrees C. Electrophoresis detection showed that degradation of muscle hake proteins was complete at both temperatures, and in gelatin zymograms extracellular activity showed two proteolytic bands with apparent molecular masses of approximately 31.6 and 62 kDa.     

Fatty acid composition of phospholipids of myelin and synaptosomal proteolipid complexes from vertebrate brain.

1. Fatty acid composition of five main phospholipids of vertebrate brain myelin and synaptosomal proteolipids and membranes was studied. 2. Higher content of monoenoic and lower content of saturated and polyenoic fatty acids was found to be characteristic of phospholipids from myelin and myelin proteolipids as compared to phospholipids from synaptosomal proteolipids and membranes of vertebrates (from fishes to mammalians). Fatty acid composition of phospholipids of proteolipid complexes and of the membranes, from which they were isolated, were found to be similar in various species studied. 3. Microviscosity was found to be higher in myelin as compared to synaptosomal membranes of frog Rana temporaria and in rabbit Lepus cuniculus. It appears to be due to the difference in proteolipid content and in lipid composition of myelin and synaptosomal membranes.     

Changes in lipids and biochemical properties of actomyosin from pre- and post-spawned hake (Merluccius hubbsi Marini)

The total lipid contents of muscle, the liver somatic index and the total lipid contents of actomyosin in hakes changed with the gonadal condition of the fish. Non-polar lipids in actomyosin from pre-spawned hake were 145% higher than in actomyosin from post-spawned hake; polar lipids were 30% higher. The relative percentage of phospholipids changed from 32% in pre-spawned hake to 48% in post-spawned hake. The Mg²⁺-ATPase activity in actomyosin increased from the pre-spawned to the post-spawned conditions, and this could be related to a higher phospholipids to neutral lipids ratio in post-spawned hake. Gradual decreases in both Ca²⁺-ATPase activity and myosin: actin ratio during the gonadal development of fish were found, suggesting a partial loss of myosin functionality of the actomyosin complex.     

Effect of triton X-100 treatment on the lipid composition,biochemical and functional properties of hake (Merluccius hubbsi) myofibrils purified from pre- and post-spawned fish

• 1.1. The effects of Triton X-100 treatment on the lipid contents and functional properties of hake myofibrils from pre- and post-spawned fish were investigated.
• 2.2. Differences in lipids, biochemical and functional properties of hake myofibrils related to the gonadal condition of fish were observed.
• 3.3. Triton X-100 treatment removed 65% of polar lipids in myofibrils from pre-spawned fish and only 10% in myofibrils from post-spawned fish.
• 4.4. Triton X-100 increased the Hill coefficient to 1.5 in an allosteric type of reaction for the myofibrillar Mg²⁺-ATPase from pre-spawned hake.
• 5.5. The detergent effect observed on the contraction response was greater in myofibrils from prespawned fish than in post-spawned fish.

     

Sexual differences in the increase of white muscle fibres in Argentine hake, Merluccius hubbsi, from the San Matias Gulf (Argentina)

Growth dynamics of white fibres from axial muscle has been investigated in Argentine hake, Merluccius hubbsi , discriminating between sexes for the first time. The frequency distributions of fibre diameters are remarkably different in both sexes at sizes between 42 and 63·9 cm total length (T.l.). Males have a much lower proportion of newly recruited fibres (0–10 μm) than females; at 52–53·9 cm T.l., females have 4% of fibres in that category and males 0·5%. It appears that, from this size interval onwards, the increase in muscle mass is due only to the increase in diameter of individual fibres, which may exceed 300 μm. The lower recruitment rate of new fibres in males, and its relationship to lower growth rates and smaller final sizes, are discussed, and possible effects of reproductive activity are considered.     

Feeding strategy of juvenile (age‐0+ year) Argentine hake Merluccius hubbsi in the Patagonian nursery ground

Age‐0+ year juvenile Argentine hake Merluccius hubbsi (60–150 mm total length, L_T) from San Jorge Gulf, north Patagonian shelf region of the Argentine Sea, had an almost exclusively pelagic diet dominated by the hyperiid amphipod Themisto gaudichaudii and the euphausiid Euphausia lucens. This suggested that final settlement and permanent demersal habitat utilization might not, as previously reported, occur at earlier sizes (c. 20 mm L_T). Their feeding strategy involves specialization at a population level towards both the main pelagic prey, indicating a narrow trophic niche. Novel data are provided which contribute to the growing body of information in relation to the age‐0+ year transitional stage in demersal fishes and particularly to M. hubbsi recruitment in the Argentine Sea.     

Morphological and biochemical approaches to assess the nutritional condition of the Argentine hake Merluccius hubbsi larvae from two different nursery areas

The Argentine hake, Merluccius hubbsi, is one of the most important fishing species in the Argentine Sea due to its great abundance and high-quality meat. The study of the nutritional condition is widely used to determine the physiological state of the fish larvae and to estimate their survival possibilities. The larval nutritional condition reflects the environmental conditions to which they have been exposed and represents a useful instrument to determine favourable nursery areas. It also provides tools for the comprehensive management of a population subjected to fishing exploitation. This study aimed to determine potential differences in the nutritional condition of M. hubbsi larvae from the two fishing stocks (Northern and Southern) of the species. The authors assessed the nutritional condition of larvae captured during the 2012 main reproductive peak in the nursery areas of each population. Two different methodologies were applied: a morphometrical approach, by recording five morphometric variables, and a biochemical technique, employing the RNA/DNA index (RD_s) and its derived index of growth performance. The morphometrical indexes did not show differences in the larval condition between both stocks. Nonetheless, the RD_s index did detect differences in the nutritional condition of larvae from different stocks. The RD_s index of larvae in pre-flexion and flexion stages showed significant differences between stocks, indicating that these stages are more susceptible to starvation. The results suggest that the biochemical indexes prove to be more sensitive than the morphometric indexes to detect slight differences in hake larvae nutritional condition. The scope and limitations of these techniques for the analysis of the nutritional condition of larvae are discussed.     

Occurrence of N-acylethanolamine phospholipids in fish brain and spinal cord

N-Acylethanolamine phospholipids were identified in the central nervous system of the fresh water fish, pike (Esox lucius) and carp (Cyprinus carpio), at levels ranging from 0.1 to 0.9% of total phospholipid. The N-acylethanolamine phospholipids of carp brain were isolated and characterized by chemical, biochemical and spectroscopic methods. Two major species, 1,2-diacyl-sn-glycero-3-phospho(N-acyl)ethanolamines (approx. 30%) and 1-O-(1'-alkenyl)-2-acyl-sn-glycero-3-phospho(N-acyl)ethanolamines (approx. 70%) were identified. The N-acyl groups of each species consisted primarily of 16:0 (approx. 60%) but also contained 16:1, 18:0 and 18:1 (approx. 10% each) and a number of trace constituents. The N-acylethanolamine phospholipids had O-acyl and O-alkenyl group compositions similar but not identical to those of the ethanolamine phospholipids of the same tissue. N-Acylethanolamine phospholipids were present in all subcellular fractions of carp brain, except mitochondria.     

Fatty acid composition of myelin lipids from developing rat forebrain and spinal cord: Influence of experimental hyperphenylalaninaemia

The fatty acid composition of individual myelin lipids from rat forebrain and spinal cord was analysed at 20 and 30 days p.p. During this phase of rapid myelination the proportions of C 160 and C 180 fatty acids decreased whereas the relative amounts of long chain and unsaturated fatty acids increased in most lipid classes. This developmental shift was more pronounced in the forebrain and was different with respect to both magnitude and direction for each myelin lipid. The experimental induction of chronic hyperphenylalaninaemia (hyper-Phe) from day 3 p.p. lead to alterations in the rate of myelination, which were most pronounced in the forebrain. At 20 days, especially in the forebrain, chain elongation and desaturation of fatty acids were delayed. This delay was maximal in sphingomyelin fatty acids, C 180 and C 241. In hydoxycerebrosides, the shift in the fatty acid composition from C 160 to C 220 was accelerated in hyper-Phe rats. No significant difference in the fatty acid composition of any myelin lipid was found in the more mature myelin (spinal cord, 30 days p.p.). No evidence was found of a primary effect of hyper-Phe on myelination.In memoriam of M. Klapproth who died by a tragic accident.     

Changes in the fatty acid composition of phospholipids in tissues of farmed sea bass (Dicentrarchus labrax) during an annual cycle. Roles of environmental temperature and salinity

We quantified seasonal effects on fatty acid composition of tissue phospholipids in farmed sea bass. Major changes in percent phosphatidylethanolamine and phosphatidylcholine were observed in all tissues between February and March, and the phosphatidylcholine/phosphatidylethanolamine ratio was drastically reduced at this time. Different changes in the fatty acid composition of total phospholipids were observed in all tissues examined. Fish fed all year on the same commercial diet showed a significant correlation between water salinity and percentage of 22:6n-3 in muscle, liver and gill phospholipids, but no correlation was found between percent 22:6n-3 of phospholipids and water temperature. In each tissue, we observed annual variation in the 20:5n-3/20:4n-6 ratio in phospholipids, but maximum and minimum values occurred at different times in each organ. From these results, we conclude that salinity can play a significant role in modulating the activities of enzymes acting on lipid metabolism during their natural circannual cycles.     

Effect of diets containing different oils on brain fatty acid composition in sea bass (Dicentrarchus labrax L.)

Four groups of adult sea bass were given diets containing about 8% of one of four different oils having a different fatty acid composition: linseed oil, grape-seed oil, containing high amounts of linolenic and linoleic acids respectively, hydrogenated coconut oil, mainly containing saturated fatty acids, and cod liver oil which was considered as reference. Total lipid, phospholipid and polar lipid contents of the brain of the different groups of sea bass were unaffected. The fatty acid composition of the brain agreed with the dietary history of sea bass: thus adult sea bass brain is capable of incorporating dietary fatty acids. Sea bass brain and structural lipids of the liver appeared to be similarly sensitive to the dietary input in contrast with mammalian brain which was reported to be more resistant than other tissues. The more striking dietary effect on liver total lipid fatty acid composition is ascribed to the incorporation of dietary fatty acids in depot fats.     

Fatty acid composition of different serum phospholipids in men and women

1. Eight young healthy persons, four men and four women, were maintained for a total of 2 months on a diet in which 40%, 16% and 44% of the total calories were present as fats, proteins and carbohydrates respectively. The ratio of complex to simple carbohydrates in the diet was 1:4. 2. The fatty acids of serum kephalins, lecithins, lysolecithins and sphingomyelins were determined by gas-liquid column chromatography. 3. Lysolecithins in both men and women had the highest content of saturated acids, followed by sphingomyelins, lecithins and kephalins in that order. The degrees of saturation and of polyunsaturation of the fatty acids in the different phospholipid fractions were significantly different, except for the differences in the polyunsaturation of the kephalins and lecithins. 4. No sex difference was found in the fatty acid composition of the different phospholipids.     

Larval anisakids and other helminths in the hake, Merluccius gayi (Guichenot) from Chile

One hundred and twenty hake Merluccius gayi caught off central Chile were examined for helminth parasites. Sixty hake were kept in ice from the catch of a trawler were examined upon arrival at port, while the other 60 hake were dissected on board immediately after collection. Ninety one per cent of hake were infected with one or more species of the following helminths: trypanorhynch plerocerci of Grillotia dollfusi Carvajal, 1971; adult worms of the genus Clestobothrium; larval nematodes of the genera Anisakis and Phocanema; 86% of the total were infected with Anisakis sp. and 42.5% with Phocanema sp. Infection with larval nematodes increased with length of the fish and there was a greater incidence of infection among the females. There was a greater frequency of occurrence of the parasites in the viscera than in the musculature.     

Fatty acid composition of galactolipids and phospholipids in neoplasmatic plant tissues (cecidia) and normal leaf tissue

Insect-elicited modifications of leaf cells that produce leaf galls on higher plants are concurrent with changes at the biochemical level. The neoplasmatic alterations affect the composition and concentration of membrane phospho and galactolipids. In vitro labeling of developing leaf galls from oak [induced by cympids (Hymenoptera) on L.] with [¹⁴C]-acetate and [³H]-oleate was used to study the label distribution in monogalactosyldiacylglyceride, digalactosyldiacylglyceride, phospha-tidylglycerol and phosphatidylinositol in these tissues and in the normal host organs (leaf laminae). The incorporation of both labels was higher into the leaf galactolipids than into gall galactolipids, whereas the incorporation of oleate was slightly increased in the 2 phospholipids in cynipid galls. The degree of acyl chain unsaturation of individual endogenous phosphoglycerides (phosphatidylcholine, phosphatidyletha-nolamine. phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol) was, in most cases, higher in the gall tissues. Differences in the endogenous fatty acid composition of plant polar lipids and the molar concentrations of labeled fatty-acid precursors in galacto- and phospholipids are discussed with regard to modifications of the leaf phenotype and cecidogenesis on Quercus palustris L.     

Amino acid composition and endocrine control of vitelline envelope proteins in European sea bass (Dicentrarchus labrax) and gilthead sea bream (Sparus aurata)

The vitelline envelopes of European sea bass and gilthead sea bream are both composed of mainly four proteins with the molecular masses of 90, 52, 48, 45 kDa and 75, 50, 48, 44 kDa, respectively. Each protein has an amino acid composition that is characterized by a high content of proline and glutamic acid and a low content of cysteine, similar to the whole vitelline envelope of both species. The amino acid composition suggests that each protein is distinct but related to the other vitelline envelope proteins. The use of homologous antisera shows that both species have vitelline envelope proteins that are induced by estradiol-17β. As males of both species synthesize these proteins after treatment with estradiol-17β, the origin is not restricted to the ovaries. Vitellogenin of both Eurpoean sea bass and gilthead sea bream has the apparent molecular mass of 170 kDa. © 1995 Wiley-Liss, Inc.