Marine Protein Research Articles

Synthesis of Sequential Polypeptides Containing O‐Phospho‐L‐Serine

AbstractThe chemistry of phosphoserine [Ser(P)] containing peptides and polypeptides was extensively investigated to explore a new biomineralization material science. The selective cleavage of the O,O′‐diphenyl phospho‐protecting groups of Ser(PO3Ph2) was examined using hydrogenolysis catalysts. Among the catalysts examined, only PtO2 in 50% trifluoroacetic acid (TFA)/AcOH successfully cleaved the protecting group of Ser(PO3Ph2) to give Ser(P). Based on these characteristic new findings, Ser(P)‐containing dipeptides such as Gly‐Ser(P), Ala‐Ser(P), Ser‐Ser(P), Asp‐Ser(P), Glu‐Ser(P), and Lys‐Ser(P), and tetrapeptide [Asp‐Ser(P)]2 were synthesized by a facile method. When we used the Ser(PO3Ph2) residues at the C terminals, the amino functional groups of amino acids and peptides can be coupled by the unsymmetric mixed anhydride using isobutyl chloroformate but cannot be by the symmetric anhydride method using dicyclohexylcarbodiimide. Neither unsymmetric mixed anhydride nor symmetric anhydride can be coupled with p‐nitrophenol at their C terminals. High‐molecular‐weight sequential polypeptides containing Ser(P) such as poly[Ser(P)‐Xaa] (Xaa: Gly, Ala, Ser, Lys, Asp, Glu) and poly[Gly‐Ser(P)‐Gly] were first synthesized by the polycondensation of the di‐ and tripeptide p‐nitrophenyl active esters, followed by the quantitative elimination of the diphenyl protecting groups by PtO2 in TFA/AcOH. The new strategy to synthesize Ser(P)‐containing peptides and model proteins may help the development of hybrid formulations of marine and biomimetic protein minerals.magnified image

Potential of meat meal to replace fish meal in extruded dry diets for barramundi,Lates calcarifer(Bloch). II. Organoleptic characteristics and fatty acid composition

The organoleptic quality of barramundi fed for 66 days on pelleted diets containing varying amounts of fish meal and meat meal was determined in two experiments (E1 and E2). Each compared four diets: a 430 g kg1 crude protein (CP) control diet (containing 35% Chilean anchovy fish meal); two diets containing high inclusions (40% or more) of meat meal; and a proprietary barramundi diet. In E1, the two meat meal diets contained 10% Chilean fish meal whereas the two meat meal diets in E2 had no marine protein ingredients. Panellists identified and rated the colour of flesh, and scored odour, flavour and texture characteristics and overall liking on structured graphic line scales (0-100). Fish fed the high-meat meal diets were sweeter and firmer than those fed the high-fish meal control diet in E1 (P 60) for overall liking and low (< 10) for undesirable odours and tastes. Exclusion of all sources of marine protein from the diet in E2 did not detract from the sensory value of the fish. The influence of diet on the fatty acid profile of the fish was examined in E2. Compared with fish fed the control diet, the neutral lipid fraction of those fed the meat meal diets had higher proportions of saturated and short-chain monounsaturated fatty acids at the expense of longer chain fatty acids, especially 22:6n-3. Polar lipids showed only subtle dietary effects, which were confined to the long-chain unsaturated fatty acids.

Potential of meat meal to replace fish meal in extruded dry diets for barramundi, Lates calcarifer (Bloch). I. Growth performance

Juvenile barramundi (220-280 g start weight) were fed extruded dry-pelleted diets containing varying amounts of fish meal and meat meal in three experiments (E). E1 and E2 were each 66-day farm studies utilizing 16 floating cages (400 fish per cage) in an aerated freshwater pond. E3 examined the same diets as fed in E2 but under controlled water temperature (28 ± 0.7 °C) and photoperiod (12:12) laboratory conditions in a 42-day study involving 24 aquaria (eight fish per aquarium). In all studies, the same 430 g kg1 crude protein (CP), 15 kJ g1 digestible energy (DE) control (Ctl) diet (containing 35% Chilean anchovy fish meal) was compared with two high-inclusion meat meal diets and a proprietary diet. The meat meal diets evaluated in E1 were a high-ash (260 g kg1) meat meal that contained 520 g kg1 CP and a low-ash (140 g kg1) meat meal that contained 600 g kg1 CP when included at either 450 or 400 g kg1, respectively, in combination with 100 g kg1 Chilean fish meal in diets that were isonitrogenous and isoenergetic with the Ctl diet. Growth rates and feed conversions were similar (P > 0.05) for all diets. In E2 and E3, the 520 g kg1 CP meat meal was included at 500 g kg1 without any marine protein source in diets formulated to provide either 15 or 16.2 kJ g1 DE and the same CP/DE ratio (29 mg kJ1) as the Ctl diet. Fish performance ranking of diets was similar in both experiments, with the 16.2 kJ g1 DE diet supporting better (P < 0.05) growth rates than the Ctl diet and feed conversion ratios equivalent to the Ctl diet but better (P < 0.05) than all other diets.

Dogs, Ducks, Deer and Diet: New Stable Isotope Evidence on Early Mesolithic Dogs from the Vale of Pickering, North-east England

We present new carbon and nitrogen isotope values for two domestic dogs (Canis familiaris) and an aquatic bird (Grus grus) from the Mesolithic sites of Star and Seamer Carr, Yorkshire, England. Previous measurements of the carbon isotopes of the Seamer Carr dog were undertaken by Clutton-Brock & Noe-Nygaard (1990). They interpreted the δ13C values as indicating a partly marine diet, and suggested that this was evidence for a seasonal movement of people and their domestic dogs between the coast and the inland Seamer Carr site. Day (1996) questioned this interpretation, arguing that the dog's δ13C values resulted from the consumption of aquatic foods from the ancient lake in the Vale of Pickering, thereby undermining any suggestion of seasonal movements to the coast. Day's argument is based on the assumption that the freshwater lake near Seamer/Star Carr contained δ13C-enriched dissolved CO2 from limestone, resulting in plants and aquatic birds with values mimicking those of a marine system. Here, we re-measure the carbon values of the Seamer Carr dog and provide new data for a dog and an aquatic bird from Star Carr. These new carbon and nitrogen isotope values indicate that the Seamer Carr dog did indeed consume significant amounts of marine protein, while the Star Carr dog did not. The aquatic bird did not have enriched δ13C values as Day predicted, and therefore we found no evidence to support Day's assumptions. Our new data supports the position that there could have been seasonal movements between the coast and inland during the Early Mesolithic at the site of Seamer Carr.

Dating Women and Becoming Farmers: New Palaeodietary and AMS Dating Evidence from the Breton Mesolithic Cemeteries of Téviec and Hoëdic

This paper presents and discusses the results of a palaeodietary and AMS dating study of burials from the Mesolithic sites of Téviec and Hoëdic, Brittany, France. In common with other Mesolithic coastal populations in Europe, isotopic analysis demonstrates the significant use of marine resources by the sites' inhabitants. Greater interest, however, is provided by the inter- and intrasite details of the analysis. There is an unexpected difference between the two sites, with the inhabitants of Hoëdic deriving 70 to 80% of their protein from the sea, while the inhabitants of Téviec appear to show a more balanced use of marine and terrestrial protein. At the intrasite level, women, and particularly young women, were found to exhibit less use of marine foods. It is suggested that this could indicate an exogamous, patrilocal marriage pattern, with some women marrying in from more inland communities. The AMS dating program shows that the sites were roughly contemporaneous but were used for burial over a longer period of time than originally anticipated. Two cases could suggest the reuse of graves after the passage of centuries, a practice more typically associated with Neolithic passage graves. Unresolved issues remain surrounding the calibration of the dates, complicated by the inclusion of marine protein in the diet, but even before correction for this effect a number of dates overlap with the earliest Neolithic of the region. This raises a number of possible scenarios for the Mesolithic-Neolithic transition in Brittany.

Neolithic Diet at the Brochtorff Circle, Malta

From Neolithic Malta, there is evidence of increasing population size accompanied by increasingly elaborate material culture, including the famous megalithic architecture. Stoddart et al. (1993) argued that social tensions and controls increased as food resources diminished. One important requirement of this argument is that the Neolithic inhabitants of Malta depended entirely on domesticated plants and animals for subsistence and therefore, with increased population sizes, the poor agricultural potential of these islands was stretched. However, it is possible that the consumption of wild foods, particularly marine resources, in the Neolithic would make up any shortfall in the agricultural foods. A direct way of measuring the amounts of marine protein in human diets is through chemical analysis of human bone. Stable isotope analyses undertaken on seven Accelerator Mass Spectrometer (AMS) radiocarbon dated humans from the Neolithic at the Brochtorff Circle indicated that there is no evidence for the significant use of marine foods by these Neolithic individuals. These new data indicate that agricultural foods were the dietary staple for this sample of the Maltese Neolithic population and therefore support the argument that increasing population during the Neolithic could have resulted in increasing resource stress.

Neolithic diet at the Brochtorff Circle, Malta

From Neolithic Malta, there is evidence of increasing population size accompanied by increasingly elaborate material culture, including the famous megalithic architecture. Stoddart et al. (1993) argued that social tensions and controls increased as food resources diminished. One important requirement of this argument is that the Neolithic inhabitants of Malta depended entirely on domesticated plants and animals for subsistence and therefore, with increased population sizes, the poor agricultural potential of these islands was stretched. However, it is possible that the consumption of wild foods, particularly marine resources, in the Neolithic would make up any shortfall in the agricultural foods. A direct way of measuring the amounts of marine protein in human diets is through chemical analysis of human bone. Stable isotope analyses undertaken on seven Accelerator Mass Spectrometer (AMS) radiocarbon dated humans from the Neolithic at the Brochtorff Circle indicated that there is no evidence for the significant use of marine foods by these Neolithic individuals. These new data indicate that agricultural foods were the dietary staple for this sample of the Maltese Neolithic population and therefore support the argument that increasing population during the Neolithic could have resulted in increasing resource stress.

Diet, Body Size, and Landscape Use among Holocene People in the Southern Cape, South Africa

Diet, Body Size, and Landscape Use among Holocene People in the Southern Cape, South Africa

Diet, Body Size, and Landscape Use among Holocene People in the Southern Cape, South Africa

Diet, Body Size, and Landscape Use among Holocene People in the Southern Cape, South Africa

Assessment of anadromous salmon resources in the diet of the Alexander Archipelago wolf using stable isotope analysis.

The Alexander Archipelago wolf (Canis lupus ligoni) is unique to southeast Alaska, occurring on islands south of Frederick Sound and along the mainland between Dixon Entrance and Yakutat Bay. Sitka black-tailed deer (Odocoileus hemionus sitkensis) are an important prey species for wolves across the southern part of the region. Spawning salmon (Onchorynchus sp.) are seasonally available but their presence in wolf diets has not previously been quantified. We examined the range of bone collagen δ13C and δ15N values for wolves throughout southeast (n = 163) and interior (n = 50) Alaska and used a dual-isotope mixing model to determine the relative contribution of salmon-derived marine protein in the diet. Southeast Alaska wolves consumed significantly more salmon (mean ± SE: 18.3 ± 1.2%) than did wolves from interior Alaska (9.1 ± 0.6%, P<0.001). Wolves on the southeast Alaska mainland appeared to have higher marine isotopic signatures than island wolves, although this difference was not significant. Variation among individual wolf diets was higher for southeast than for interior Alaska wolves, and variation was highest in coastal mainland wolf diets (P<0.001). Marine resources may augment the diet of southeast Alaska wolves during seasonal or annual fluctuations in the availability of deer, particularly in those areas on the mainland where densities of terrestrial ungulates are relatively low.

Nouvelles dates AMS à Téviec et Hoëdic (Quiberon, Morbihan)

Abstract A new series of 14 accelerator dates have been obtained on human bone from the Breton Late Mesolithic sites of Téviec and Hoëdic. The dates from Téviec range between 6740 and 5680 B.P., while those from Hoëdic range between 7165 and 5080 B.P.; most dates from both sites fall within the second half of the 7th millennium B.P. This presents a longer period of use for these spatially well- defined burial grounds than might have been expected. Moreover, at least three dates appear to overlap with what is traditionally defined as the Middle Neolithic I. However, a full assessment of the place of Téviec and Hoëdic in the Breton sequence must await calibration of the new dates, a procedure complicated by the considerable use of marine protein in the human diet.

Stable isotopes and the seasonality of the Oronsay middens

Research on six late Mesolithic shell middens on the small Hebridean island of Oronsay address questions of permanent and seasonal occupation. Stable isotope analysis of human bones shows marine resources providing the majority of protein, supporting year-round occupation of Oronsay. One individual, however, demonstrated a mixed diet of marine and terrestrial protein, suggesting seasonal visits and different patterns of site occupation.

Stable isotopic analysis of human diet in the Marianas Archipelago, Western Pacific

Proportions of marine vs. terrestrial resources in prehistoric human diets in the southern Mariana Islands (Guam, Rota, Saipan), Micronesia, have been estimated by analysis of stable isotope ratios of carbon and nitrogen in bone collagen and of carbon in apatite. The isotopic composition of marine and terrestrial food resources from the Marianas have also been determined. Experimental evidence shows that collagen carbon isotopes mainly reflect those of dietary protein sources and thus overestimate the contribution of marine animal foods. Marine protein consumption apparently ranges from approximately 20% to approximately 50% on these islands. Experiments also demonstrate the carbon isotope ratio of bone apatite carbonate accurately reflects that of the whole diet. Carbonate carbon isotope data suggest some individuals consumed significant amounts of 13C-enriched (C4) plants or seaweeds. Sugar cane is an indigenous C4 crop and seaweeds are eaten throughout the Pacific, but they have not been considered by archaeologists to have been prehistoric dietary staples. Apatite carbon isotope analysis has apparently identified previously unrecognized prehistoric dietary adaptations in the Mariana Islands, but this must be confirmed by archaeobotanical evidence.

Paleodiet of Hunter-Gatherers in Japan Estimated by 13C-15N and Lipid Analyses.

Reconstruction of paleodiets for hunter-gatherers in Japan is carried out using the major nutritive elements. (1) Carbohydrate sources are examined using carbon isotope data from charcoal in burnt “cookies” and pottery residues. Measurements of charcoal in burnt cookies indicated no evidence of C4 plant use during the prehistoric age in Japan. (2) Protein sources are identified using carbon and nitrogen isotope ratios measured on collagen extracted from human bones. Among hunting-fishing-gathering people in Japan, marine protein intake was about 70-90% for people in Hokkaido including the Ainu people, while Jomon people in Honshu and Kyushu had a wider variation of 20-60% between coastal and inland people. (3) Lipid sources are identified from fatty acids and sterols extracted from pottery residues and coprolites by GC and GC-MS. Coprolites and pottery sherds from shellmidden sites indicated that animal foods were dominant lipid sources in the diet, and marine food resources were not constantly, but occaisionally used.

Synthesis and adhesive studies of barnacle model proteins

The marine adhesive barnacle model protein has been synthesized by polycondensation. The O-benzyl, O- and N-carbobenzoxy, and toluenesulphonyl groups were used to protect the sidechains of serine and threonine, tyrosine and lysine, and arginine and histidine, respectively. Eighteen kinds of N-carboxyanhydrides (NCA) were synthesized according to two different procedures, using phosgene gas and phosphorus pentachloride respectively, followed by polycondensation. The protecting groups of the resulting polypeptide with a degree of polymerization of 45 were simultaneously cleaved by trifluoromethanesulphonic acid. Bonding strenghts of the model polypetide were measured on test pieces and exhibited a tensile strength of 15–19 kg cm −2 on iron, 5–7 kg cm −2 on alumina, and 3–7 kg cm −2 on glass and shell, whereas the model protein was found to have small tensile strength on woods and plastics. The bonding strength is compared with those of natural marine adhesive proteins and the factors important for effective adhesion are discussed.

An assessment of processing effects on the nutritive value of marine protein sources for juvenile chinook salmon ( Oncorhynchus tshawytscha)

This study was undertaken to assess the nutritional value of marine fish protein sources processed in different ways for juvenile chinook salmon reared in fresh water. The protein sources compared were a freeze-dried pollock muscle and euphausid mix (9:1) (FPE), and three whole herring meals processed differently from a common lot of raw herring. A casein-gelatin mix supplemented with arginine and DL-methionine (CS) served as a reference protein source. The five protein sources were each tested at three protein concentrations in isocaloric diets. FPE was also assessed at two additional protein levels. Protein was replaced by dextrin and glucose on an estimated metabolizable energy basis. Each diet was fed to duplicate groups of fish held at 10.5°C on a natural photoperiod for 42 days. All fish were fed daily to satiation. Two groups received Oregon moist pellets (OMP-2), a practical hatchery diet. Each of the various methods employed to evaluated protein quality yielded different relative values in comparison to FPE, which was judged to be the best protein source on the basis of growth rate response and assays based on body protein gain. Low temperature (75°C) dried herring meal had slightly reduced protein quality compared to freeze-dried herring meal, whereas the high temperature (150°) dried meal had dramatically reduced protein quality. High estimates of protein quality were obtained for CS. However, this protein source was not as palatable as the low temperature dried marine protein sources and consequently growth was reduced.

The utilization of blood meal as a protein ingredient in the diet of the marine shrimp Penaeus vannamei

Four blood meal products were compared and tested as protein substitutes for a marine protein mix in grow-out rations for the marine shrimp Penaeus vannamei. The blood meal products tested were a ring-dried blood meal (RD), an acidulated, sun-dried blood meal (AS), an acidulated, sundried blood meal with crystalline methionine added (ASAM), and an acidulated, sun-dried blood meal with covalently linked methionine (ASCM). There were no significant differences in weight gain (0.62–0.75 g per week), survival (96.8–100%), or feed conversion ratios (1.6–2.0) during the 42-day trial. The AS and ASAM blood meal diets produced significantly ( P < 0.05) lower normalized biomasses (total biomass gained by surviving shrimp divided by the initial number of animals stocked) than the ring-dried blood meal and the control diets. The ASCM and RD blood meal diets were statistically indistinguishable ( P > 0.05) from the control diet. The results suggest that blood meal products can replace marine proteins in grow-out rations for medium sized (3–4 g) Penaeus vannamei. Results also indicate that the blood meal product containing covalently attached methionine may be a potentially useful way to supplement limiting amino acids in shrimp diets.

The Importance of Marine Protein in the Diet of Coastal Australian Aborigines

The Importance of Marine Protein in the Diet of Coastal Australian Aborigines

Use of stable-carbon isotope analysis to estimate marine and terrestrial protein content in gull diets

Stable-carbon isotope analyses of bone collagen of gulls (Larus spp.) were used to estimate the relative proportion of marine and terrestrial protein in the diets of gulls that are known to use both types of food sources. Mean δ13C values for Glaucous-winged Gulls (n = 23) collected at a Vancouver dump and for Western Gulls (n = 18) from Southeast Farallon Island were −15.0 ± 1.3 and −15.1 ± 0.5‰, respectively. No significant difference in δ13C values was found between adult male and female Western Gulls. Archaeological gull bones (n = 3) show a mean δ13C value of −13.6 ± 1.0‰ and support the assumed marine end point of −13.0‰. The range of terrestrial protein in the diets of coastal gulls was 0–61%. The narrow distribution of δ13C values for Western Gulls from the Southeast Farallon Island colony suggests that gull colonies may be calibrated so that changes in dependence on terrestrial protein can be monitored.

Use of stable-carbon isotope analysis to estimate marine and terrestrial protein content in gull diets : Hobson, K.A., 1987. Can. J. Zool., 65(5):1210–1213

Use of stable-carbon isotope analysis to estimate marine and terrestrial protein content in gull diets : Hobson, K.A., 1987. Can. J. Zool., 65(5):1210–1213