Manual for starch gel electrophoresis: A method for the detection of genetic variation

The procedure to conduct horizontal starch gel electrophoresis on enzymes is described in detail. Areas covered are (I) collection and storage of specimens, (2) preparation of tissues, (3) preparation of a starch gel, (4) application of enzyme extracts to a gel, (5) setting up a gel for electrophoresis, (6) slicing a gel, and (7) staining a gel. Recipes are also included for 47 enzyme stains and 3 selected gel buffers. (PDF file contains 26 pages.)

Effect of gibbing on protein hydrolysis during the ripening of salted herring at 4°C in polypropylene barrels

Effect of gibbing process on the protein hydrolysis in terms of free alpha amino nitrogen (FAN) content during the ripening of barrel salted herring at low temperature (4°C) was investigated. For this purpose North Sea herring (Clupea harengus) from north-east British coast was salted in polypropylene barrels and allowed to ripen at 4°C. This process of barrel salting was carried out for whole fish in one batch and gibbed fish in another batch. The investigation was performed by using new salt and used salt in separate barrels for each batch of experimental fish. Results of the present study show that protein hydrolysis was significantly higher in the ripened salt-herring produced from whole fish which was found to have more characteristic sensory properties than those produced from gibbed fish. Similar result (proteolysis) was obtained when the investigation was repeated for the spent herring although the spent herring fails to produce a ripened product with the desired characteristic sensory attributes, compared to those of pre-spawning herring.

Effect of modified starch on the shelf life of frozen fish Kamaboko with prepared spinach

Fish kamaboko with spinach was prepared by mixing fish kamaboko with spinach prepared with different combinations of ordinary starch (OS) and modified starch (MS), viz., 40:60, 50:50, 60:40, 100%) MS and 100% OS. The products were frozen at -40°C and stored at -20°C, and subjected to biochemical (peroxide value, total titratable acidity, pH and moisture) organoleptic (appearance, taste and colour), microbiological (total plate count) and physical (folding and expressible water) tests at monthly intervals. Among the different combinations tried, it was observed that fish kamaboko with spinach prepared with 50:50 and 60:40 combinations of OS and MS could be stored for six months at-20°C.

Lipid hydrolysis in mackerel (Rastrelliger kanagurta) during frozen storage

The hydrolytic changes in the lipids of mackerel (Rastrelliger kanagurta) during storage at -l8°C were studied with a view to understand the factors involved in the formation of free fatty acids. Only the phosphorylated fraction did undergo hydrolysis at an appreciable rate. It was found that the free fatty acid production was mainly associated with the phospholipid hydrolysis. As regards the triglycerides and unsaponifiable matter, there was no significant change in levels during frozen storage.

Selective release of fatty acids during lipid hydrolysis in frozen-stored milk fish (Chanos chanos)

Lipid hydrolysis and the nature of fatty acids lost as a result of lipid hydrolysis in milk fish (Chanos chanos) during frozen storage at -20°C is discussed in this paper. There was a preferential loss of saturated acids during the first three weeks of storage. This was followed by loss of polyunsaturated acids during the next seven weeks. Sharp decrease in the levels of monounsaturated acids was observed from the 10th week of frozen storage. These observations are due to the preferential hydrolysis of phospholipids with relatively high proportion of saturated acids during the first three weeks, followed by the hydrolysis of phospholipids with high proportions of polyunsaturated fatty acids from the 3rd to the 10th week, and finally, predominant hydrolysis of neutral lipids from the 10th week onwards. Storage of fish in the ice prior to freezing was found to accelerate lipid hydrolysis, especially that of neutral lipids, during frozen storage.

Effects of crude, semipurified and purified starch of sago (Metroxylon sagu Rottb.) on the water stability of pelleted shrimp diets

A study was conducted to determine the comparative effectiveness of purified, semipurified and crude starch of sago as binders for pelleted shrimp diets. The diet containing semipurified sago starch had the highest water stability (79.1%). The values were nearly the same for the pellets bound with purified and crude sago starch. Reasons for the low binding capacity of purified and crude sago could be that the gel of purified sago is weakened due to purification, and that of the crude sago is due to the spongy material present in the product. Thus, semipurified sago starch is a better source of binder and purified crude sago. From the economic viewpoint, the cost of purified sago is prohibitive for use as binder. Both semipurified and crude sago palm starch are acceptable. Composition of shrimp diets containing various sources of sago palm starch, and binder cost and water stability of shrimp diets containing various sources of sago palm starch tested at 3, 6 and 12 hours, are tabulated.

Gum arabic, carrageenan of various types and sago palm starch as binders in prawn diets

An experiment was undertaken in order to investigate the use of sago palm starch, gum arabic and carrageenans as binders in prawn diets. Water stability data are presented; EPT-2 carrageenan was found to be the best binder for both steamed and unsteamed pellets.

Preliminary investigation into implication of a single cell organism in fish feed bouyancy and flotation

Two Isocaloric Isoproteic 30% crude protein diets were formulated for Clariid catfish and Tilapia with wheat grain starch (WGS) and cassava tuber starch (CTS) incorporated at 10 percent as binding agents. Saccharomyces cerevisiae was included at 2% as floating additive. The water stability, nutrient retention and flotation of pelleted feeds were observed for 60 minutes. There were generally decreasing trends in stability and retention at increasing time of immersion in water. The lipid retention was higher (P>0.05) than proteins in both diets. WGS diet was better (P>0.05) than CTS diet in flotation, which has attributed to the presence of gluten protein in wheat products. It is envisaged that a break through in floating feed development in Nigeria aquaculture would save the Nigeria economy from extruded (floating) feed importation

Genetic variation of rougheye rockfish (Sebastes aleutianus) and shortraker rockfish (S. borealis) inferred from allozymes

Rougheye rockfish (Sebastes aleutianus) and shortraker rockfish (Sebastes borealis) were collected from the Washington coast, the Gulf of Alaska, the southern Bering Sea, and the eastern Kamchatka coast of Russia (areas encompassing most of their geographic distribution) for population genetic analyses. Using starch gel electrophoresis, we analyzed 1027 rougheye rockfish and 615 shortraker rockfish for variation at 29 proteincoding loci. No genetic heterogeneity was found among shortraker rockfish throughout the sampled regions, although shortraker in the Aleutian Islands region, captured at deeper depths, were found to be significantly smaller in size than the shortraker caught in shallower waters from Southeast Alaska. Genetic analysis of the rougheye rockfish revealed two evolutionary lineages that exist in sympatry with little or no gene f low between them. When analyzed as two distinct species, neither lineage exhibited heterogeneity among regions. Sebastes aleutianus seems to inhabit waters throughout the Gulf of Alaska and more southern waters, whereas S. sp. cf. aleutianus inhabits waters throughout the Gulf of Alaska, Aleutian Islands, and Asia. The distribution of the two rougheye rockfish lineages may be related to depth where they are sympatric. The paler color morph, S. aleutianus, is found more abundantly in shallower waters and the darker color morph, Sebastes sp. cf. aleutianus, inhabits deeper waters. Sebastes sp. cf. aleutianus, also exhibited a significantly higher prevalence of two parasites, N. robusta and T. trituba, than did Sebastes aleutianus, in the 2001 samples, indicating a possible difference in habitat and (or) resource use between the two lineages.

Genetic analysis of juvenile coho salmon (Oncorhynchus kisutch) off Oregon and Washington reveals few Columbia River wild fish

Little is known about the ocean distributions of wild juvenile coho salmon off the Oregon-Washington coast. In this study we report tag recoveries and genetic mixed-stock estimates of juvenile fish caught in coastal waters near the Columbia River plume. To support the genetic estimates, we report an allozyme-frequency baseline for 89 wild and hatchery-reared coho salmon spawning populations, extending from northern California to southern British Columbia. The products of 59 allozyme-encoding loci were examined with starch-gel electrophoresis. Of these, 56 loci were polymorphic, and 29 loci had P0.95 levels of polymorphism. Average heterozygosities within populations ranged from 0.021 to 0.046 and averaged 0.033. Multidimensional scaling of chord genetic distances between samples resolved nine regional groups that were sufficiently distinct for genetic mixed-stock analysis. About 2.9% of the total gene diversity was due to differences among populations within these regions, and 2.6% was due to differences among the nine regions. This allele-frequency data base was used to estimate the stock proportions of 730 juvenile coho salmon in offshore samples collected from central Oregon to northern Washington in June and September-October 1998−2000. Genetic mixed-stock analysis, together with recoveries of tagged or fin-clipped fish, indicates that about one half of the juveniles came from Columbia River hatcheries. Only 22% of the ocean-caught juveniles were wild fish, originating largely from coastal Oregon and Washington rivers (about 20%). Unlike previous studies of tagged juveniles, both tag recoveries and genetic estimates indicate the presence of fish from British Columbia and Puget Sound in southern waters. The most salient feature of genetic mixed stock estimates was the paucity of wild juveniles from natural populations in the Columbia River Basin. This result reflects the large decrease in the abundances of these populations in the last few decades.

Functional properties of enzymatically modified protein from fish waste

Fish flour from dried waste consisting of head, tail, fins and entrails was enzimatically hydrolysed using various proteases and the hydrolysate was spray dried. The functional properties such as water-fat absorption ratio, foaming and solubility index of the hydrolysates and fish flour revealed that some of the products might find significant uses in the food and/or cosmetics industry. Electrophoretic separation of the proteins from the fish flour and of the hydrolysates indicated that almost all the flour proteins are susceptible to proteolytic cleavage with the exception of one or two. The extent of degree of hydrolysis from 43-70.3% with a simultaneous decrease in unpleasant smell suggest an economical tool for minimizing odour pollution due to fish industry waste deterioration.

Utilisation of trash fish. Pt. 1. Preparation of fish flake

The paper deals with the investigations carried out on the preparation of odorless fish-starch flakes using partially deodorized trash fish meat and different sources of starch like corn, tapioca, maida and black gram. It has been found that the products using corn and tapioca are better compared to those prepared using other two starches, the product from corn being the best. The product has a protein content of about 20% and has been found to have a storage life of 4 months at 37°c.

Surimi based dried fish cake

Minced fish prepared from the fillets of the sciaenid fish (Lutjanus sp.) was washed with cold water (<10 °C) three times. The washed muscle was pressed through a piece of fine cloth to a moisture content around 80%. The pressed cake (Surimi) was ground with 2.5% sodium chloride and 3% tapioca starch. The mixed material was shaped in the form of a cake and left for one hour for the gel to set. The cakes were then steamed. The cooled cakes were cut into pieces of 1 cm length x 1 cm width x 0.5 cm thick. The pieces were either dried in an electrical oven at 50°C or dried in sun to a moisture content of 11-12%. Biochemical, bacteriological and organoleptic evaluation revealed that the cakes were in very good acceptable form for 8 months. The cakes could be rehydrated by soaking in water at ambient temperature for half an hour and boiling in water for 10 minutes.

Dried fish cakes with improved texture

To overcome the problem of underutilization of marine by-catches, Basu et al. (1985) developed fish cube from minced fish meat. Although the product was acceptable, it had little rubbery texture. An attempt was made to improve the texture of the cake by several methods. It was found that 5% tapioca starch along with 3% texturised soybean protein improved the texture and juiciness of the rehydrated product. Preheating of the minced meat at 70°C for 30 minutes also improved the texture appreciably. It was also found that mixing of the ingredients at low speed (less than 100 rpm) in a dough mixer gave the best texture, higher speed and sharp blades leading to rubbery texture. The dehydrated product (moisture 19-20%) thus prepared had a shelf life of six months at ambient temperature.