Fish viscera are produced in large quantities in the fishing industry and represent a waste disposal and environmental pollution problem. However, this material is a rich source of trypsins that may have some unique properties, such as high molecular activity at low processing temperature, low thermostability, and high pH optimum/pH stability, for both basic research and industrial applications. The main objectives of this project were to extract, purify and characterize trypsin from the pyloric ceca of hoki (Macruronus novaezealandiae ), which is by far the most important commercial fish in New Zealand. / Trypsin was purified from the pyloric ceca of hoki by ammonium sulfate fractionation, followed by acetone fractionation and affinity chromatography on SBTI-Sepharose 4B. The purified extract was simultaneously desalted and concentrated by ultrafiltration, and then characterized using N-alpha-benzoyl-DL-arginine-p-nitroanilide (BAPNA) as substrate. The affinity fraction migrated as a signal band in SDS-PAGE gels as well as in isoelectric focusing gels. The molecular weight of the isolated trypsin was determined by SDS-PAGE to be approximately 26,000 Da, whereas the MALDI-TOF MS method of analysis indicated a molecular weight of 23,791 Da. The isoelectric point was determined as 6.5. / The kinetic properties, temperature, pH and inhibition effects on the activity of the purified trypsin were verified. On the basis of the kinetic properties, hoki trypsin showed better amidase activity than bovine trypsin. The hoki trypsin had alkaline pH optimum (pH 9.0) and was stable at a high pH. Hoki trypsin had a higher optimum temperature (60°C) and still had relative higher activity at lower temperature. On the other hand, hoki trypsin was unstable at higher temperature. The enzyme was inhibited by well known trypsin inhibitors (SBTI, aprotinin, benzamidine and PMSF). The N-terminal residues of hoki trypsin, IVGGQECVPNSQPFMASLNY, displayed considerable homology with other fish trypsins. Based on the above characteristics, it is suggested that the hoki enzyme is authentic trypsin with potential for use in food industry and related applications.

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Three New Zealand marine species, hoki (Macruronus novaezealandiae), kahawai (Arripis trutta) and trevally (Pseudocaranx dentex) were used to develop model fermented fish sausage. The formulation comprised fish mince, carbohydrate, minced garlic and salt in a mass ratio of 1 (fish): 0.15: 0.05: 0.03, respectively. The carbohydrate source was cooked rice or glucose. (Endogenous lactic acid bacteria (LAB) failed to ferment rice). Folate was also added to the mixture as a factor. The mixtures were extruded into 50 mL plastic syringes, where the needle end of the barrel had been excised by lathe. The lubricated barrel was overfilled to 60 mL, capped with a layer of ParafilmTM and aluminium foil, sealed tightly by rubber band and incubated at 30°C. Over time the piston was progressively advanced to yield samples for microbiological, physical, and chemical analysis. Over 96 hours an increase in the LAB count was observed with a concomitant decrease in pH. After fermentation was complete, the samples contained around 8.77 log cfu LAB g-1 with the pH range from 4.38 to 5.08. The microbiological and pH behaviour of each species varied between preparations. Hardness, adhesiveness, springiness and cohesiveness of the treatments increased with fermentation, except for hoki. The treatments showed different colour characteristics with fermentation. The light reflectance (L* values) of the trevally and kahawai treatments increased, while the a* (redness) and b* (yellowness) values decreased. Hoki exhibited smaller colour changes except for yellowness, which increased markedly. Proteolysis, measured colorimetrically by soluble peptide bonds, was greatest for trevally. Lipid oxidation, measured by the thiobarbituric acid method, was least for hoki, notably the species with the lowest fat content. Biogenic amines, which are a general quality indicator of fermented products, increased during fermentation. The trevally treatment generated the highest concentration of amines, but these values were lower than those reported for fermented fish sausage in Southeast Asia. Notably there were no important difference between folate treatments and those without folate. The results point to commercial opportunities and further research with New Zealand marine species, especially trevally. To improve the product quality and to show geographical exclusivity, further research could be done by using starter culture, and a New Zealand staple carbohydrate source such as kumara and potato, and spices and herbs which are commonly used in New Zealand, such as rosemary, thyme and sage or specific to New Zealand, such as horopito. In addition, sensory studies should also be performed before the products could be tested in the market.

Three New Zealand marine species, hoki (Macruronus novaezealandiae), kahawai (Arripis trutta) and trevally (Pseudocaranx dentex) were used to develop model fermented fish sausage. The formulation comprised fish mince, carbohydrate, minced garlic and salt in a mass ratio of 1 (fish): 0.15: 0.05: 0.03, respectively. The carbohydrate source was cooked rice or glucose. (Endogenous lactic acid bacteria (LAB) failed to ferment rice). Folate was also added to the mixture as a factor. The mixtures were extruded into 50 mL plastic syringes, where the needle end of the barrel had been excised by lathe. The lubricated barrel was overfilled to 60 mL, capped with a layer of ParafilmTM and aluminium foil, sealed tightly by rubber band and incubated at 30°C. Over time the piston was progressively advanced to yield samples for microbiological, physical, and chemical analysis. Over 96 hours an increase in the LAB count was observed with a concomitant decrease in pH. After fermentation was complete, the samples contained around 8.77 log cfu LAB g-1 with the pH range from 4.38 to 5.08. The microbiological and pH behaviour of each species varied between preparations. Hardness, adhesiveness, springiness and cohesiveness of the treatments increased with fermentation, except for hoki. The treatments showed different colour characteristics with fermentation. The light reflectance (L* values) of the trevally and kahawai treatments increased, while the a* (redness) and b* (yellowness) values decreased. Hoki exhibited smaller colour changes except for yellowness, which increased markedly. Proteolysis, measured colorimetrically by soluble peptide bonds, was greatest for trevally. Lipid oxidation, measured by the thiobarbituric acid method, was least for hoki, notably the species with the lowest fat content. Biogenic amines, which are a general quality indicator of fermented products, increased during fermentation. The trevally treatment generated the highest concentration of amines, but these values were lower than those reported for fermented fish sausage in Southeast Asia. Notably there were no important difference between folate treatments and those without folate. The results point to commercial opportunities and further research with New Zealand marine species, especially trevally. To improve the product quality and to show geographical exclusivity, further research could be done by using starter culture, and a New Zealand staple carbohydrate source such as kumara and potato, and spices and herbs which are commonly used in New Zealand, such as rosemary, thyme and sage or specific to New Zealand, such as horopito. In addition, sensory studies should also be performed before the products could be tested in the market.

Westland petrels Procellaria westlandica breed only near Punakaiki on the West Coast of New Zealand. About 80 km offshore from their breeding colony, New Zealand's largest commercial fishery (for hoki Macruronus novaezelandiae) operates from mid June to early September, coinciding with the Westland petrel's breeding season. It has been assumed that Westland petrels feed extensively on fisheries waste and that this habit has been at least partly responsible for the increase in the Westland petrel population. Some seabird biologists have expressed concern that if a species comes to depend on scavenging at fishing vessels, such a species could experience a food crisis if fishing operations changed in a way that reduced the quantity of waste discharged. The aim of this research was to assess how dependent Westland petrels have become on fisheries waste for food. Diet studies showed that during the hoki fishing season, waste accounts for more than half by weight of the solid food Westland petrels bring back to the colony to feed their chicks. After the hoki season, waste contributes only about a quarter of their diet as birds switch to more natural prey and scavenge a wider variety of fish species presumably from smaller, inshore fishing vessels. Much of the fisheries waste eaten by Westland petrels was flesh which could not be identified using traditional techniques. The electrophoretic technique iso-electric focusing increased the number of fish samples that could be identified and consequently the diet was interpreted differently than it would have been had only traditional diet analysis been used. The survey of Westland petrel distribution off the west coast of the South Island, found that although hoki fishing vessels influence the distribution of Westland petrels, only a small proportion of the Westland petrel population appears to utilise this food resource at any one time. Westland petrels were tracked at sea by VHF radio telemetry and then by satellite tracking. Satellite tracking showed that there is considerable variation in the amount of time Westland petrels spend in the vicinity of fishing vessels. On average, satellite tracked birds spent one third of their time near vessels, but they foraged over much larger areas than that occupied by the West Coast South Island hoki fishing fleet. Although fisheries waste is an important component of the Westland petrel diet, it appears that the situation is one of opportunistic use of a readily available resource, rather than one of dependence. Several features of the Westland petrel's breeding biology and foraging ecology suggest that Westland petrels could compensate for a reduction in waste from the hoki fishery by switching to other sources of waste and increasing their consumption of natural prey. Nevertheless, much remains unanswered concerning the role of fisheries waste in the Westland petrel's diet. In particular, quantifying the waste available to seabirds, and the success of Westland petrels in acquiring that waste compared to other scavenging species, is needed in order to better predict the effect of a reduction in fisheries waste on Westland petrel population size.

Westland petrels Procellaria westlandica breed only near Punakaiki on the West Coast of New Zealand. About 80 km offshore from their breeding colony, New Zealand's largest commercial fishery (for hoki Macruronus novaezelandiae) operates from mid June to early September, coinciding with the Westland petrel's breeding season. It has been assumed that Westland petrels feed extensively on fisheries waste and that this habit has been at least partly responsible for the increase in the Westland petrel population. Some seabird biologists have expressed concern that if a species comes to depend on scavenging at fishing vessels, such a species could experience a food crisis if fishing operations changed in a way that reduced the quantity of waste discharged. The aim of this research was to assess how dependent Westland petrels have become on fisheries waste for food. Diet studies showed that during the hoki fishing season, waste accounts for more than half by weight of the solid food Westland petrels bring back to the colony to feed their chicks. After the hoki season, waste contributes only about a quarter of their diet as birds switch to more natural prey and scavenge a wider variety of fish species presumably from smaller, inshore fishing vessels. Much of the fisheries waste eaten by Westland petrels was flesh which could not be identified using traditional techniques. The electrophoretic technique iso-electric focusing increased the number of fish samples that could be identified and consequently the diet was interpreted differently than it would have been had only traditional diet analysis been used. The survey of Westland petrel distribution off the west coast of the South Island, found that although hoki fishing vessels influence the distribution of Westland petrels, only a small proportion of the Westland petrel population appears to utilise this food resource at any one time. Westland petrels were tracked at sea by VHF radio telemetry and then by satellite tracking. Satellite tracking showed that there is considerable variation in the amount of time Westland petrels spend in the vicinity of fishing vessels. On average, satellite tracked birds spent one third of their time near vessels, but they foraged over much larger areas than that occupied by the West Coast South Island hoki fishing fleet. Although fisheries waste is an important component of the Westland petrel diet, it appears that the situation is one of opportunistic use of a readily available resource, rather than one of dependence. Several features of the Westland petrel's breeding biology and foraging ecology suggest that Westland petrels could compensate for a reduction in waste from the hoki fishery by switching to other sources of waste and increasing their consumption of natural prey. Nevertheless, much remains unanswered concerning the role of fisheries waste in the Westland petrel's diet. In particular, quantifying the waste available to seabirds, and the success of Westland petrels in acquiring that waste compared to other scavenging species, is needed in order to better predict the effect of a reduction in fisheries waste on Westland petrel population size.