Evaluation of growth performance and nutritional quality of diets using digestive enzyme markers and in vitro digestibility in Siamese fighting fish (Betta splendens Regan, 1910)

Digestive enzymes and their effects on in vitro digestibility of feeds and feedstuffs, as well as on growth performance quality were studied in Siamese fighting fish (Betta splendens Regan, 1910). The specific activities of total protease, amylase, trypsin and chymotrypsin increased during development and were higher in females than in males at maturation (P < 0.05). The activity ratio of trypsin to chymotrypsin corresponded to fish growth, and showed lower values in females than in males. White muscle levels of RNA decreased during development, with females having higher levels than males, while the protein levels increased with no difference between sexes. In the oocytes, trypsin-like and chymotrypsin-like specific activities were very low, and the concentrations of RNA, protein and protein/lipid ratio were higher than in the muscle. For in vitro digestibility, wheat gluten, soybean meal and fish meal were among good protein sources while the meals from peanut, fish and soybean were good carbohydrate sources. Golden apple snail meat was a good source for both protein and carbohydrate. The crude enzyme extracts from different growth stages and sexes had different abilities to digest the same feeds and feedstuffs. This will make it possible to preliminarily study the authenticated nutritional quality of raw materials for future feed formulations for B. splendens.Keywords: Betta splendens, digestive enzymes, feedstuff, in vitro digestibility, muscle quality, oocyte quality, Siamese fighting fishAfrican Journal of Biotechnology Vol. 12(14), pp. 1689-170

Proteomic and Characterization of α α α α α-amylase Inhibitor from Mungbean (Vigna radiate)

ABSTRACT Callosobruchus maculatus, the insect pest, causes damaging to stored mungbean seeds leading to economical losses. For this reason the use of α-amylase inhibitors to interfere the pest digestion process has been an interesting alternative as a biocontrolling agent. The studies of gamma irradiated mungbean mutant, M5-16, demonstrated that α-amylase inhibitor from the mutant line was more inhibitory effective on C. maculatus α-amylase than its wild type, KPS1. To investigate this mechanism, both proteomics and biochemical characterization were employed. The α-amylase inhibitor from mungbean seed, M5-16, was purified in a two-step procedure involving ammonium sulfate precipitation and gel filtration chromatography using a sephadex G-100 column. This α-amylase inhibitor existed as a monomer and had a molecular weight of 27 kDa. Moreover, It was very interesting that α-amylase inhibitor from the mungbean seed inhibited C. maculatus α-amylase but did not inhibit human salivary α-amylase. 2D gel electrophoresis suggested that the gamma-induced mutant line has decreased expression of α-amylase inhibitor and other proteins comparing with its wild type

Characterization of Digestive Enzymes from Adult Freshwater Pearl Mussel Chamberlainia hainesiana (Lea, 1856)

Freshwater mussel Chamberlainia hainesiana is a vulnerable species in Thailand. The species has a lustrous nacreous shell that can be utilized for various purposes, and has a potential industrial use for producing freshwater pearl. Characterization of digestive enzymes would provide critical information prerequisite for studying the development of digestive enzymes during on–growing as well as for in vitro digestibility study important for the development of suitable artificial feed formulations for the commercial culture. The aim of this preliminary study was to determine optimum pH and temperature for three digestive enzymes: amylase, protease and lipase, from digestive gland including stomach of adult C. hainesiana under various pH’s (2–11) and temperatures (25–80°C). The results revealed at least three isoforms of amylase and protease, and at least 4–5 isoforms of lipase. No alkaline protease activity was detected. For future enzyme expression study, the suitable optimal conditions are 40°C pH 6 for amylase activity, 50°C pH 5 for protease activity, and 50°C pH 8 for lipase activity. At natural habitat of around 30°C, these enzymes are most active at pH 6, 6, and 8 (or 10), respectively, which are suitable conditions for future study on in vitro digestibility

Temperature and pH Characteristics of Amylase and Lipase at different developmental stages of Siamese fighting fish (Betta splendens Regan, 1910)

Temperature (20-80°C) and pH (2-12) profiles of amylase and lipase at three stages (10-dayold, 1.5-month-old and 3-month-old) in Siamese fighting fish Betta splendens Regan, 1910) were studied. At least nine amylase activities were observed during development: pH 7 at 40°C, pH 9 at 30°C and 50° C, pH 8 and pH 11 at 50°C in females and pH 8 and pH 11 at 40°C and 60°C in males. At the younger stage (≤ 1.5 month old), activity at pH 7 and 40°C, and at pH 9 and 30°C and 50°C was observed. At the older stage (≥ 1.5 month old), amylase activity at pH 8 and pH 11 in the temperature range of 40-60°C was found. Lipase had at least five levels of activity: pH 7 at 20°C and 40°C, pH 8 at 20-40°C and 60°C, and at pH 11, where the activity peak disappeared in the maturing stage (3 month old) in both sexes. Amylase had very low specific activity at the 10-day-old stage, while lipase had a high specific activity, similar to older stages. The most suitable pH to determine amylase and lipase activity in Siamese fighting fish was pH 8, whereas a temperature of 50°C was appropriate for amylase and 40°C was suitable for lipase, regardless of sex and age. This information is a prerequisite for future studies of the in vitro digestibility evaluation of nutrient utilization in Siamese fighting fish

Characterization and expression levels of protease enzymes at different developmental stages of Siamese fighting fish (Betta splendens Regan, 1910)

Characterization of total proteases, trypsin and chymotrypsin were performed at pH 2 12 and temperatures 20-80°C in three life stages of Siamese fighting fish (Betta splendens Regan, 1910). At least fourteen protease activities were detected, including seven acidic (pH levels 2, 4 and 5-6), one neutral (pH 7), and six alkaline (pH levels 8-12) activities in the optimal temperature range of 30-60°C. The neutral and alkaline pH profiles were different between sexes during the on-growing stage (1.5 month-old). The acidic proteases played a major role in digestion in the early life stage (10-day-old), while the alkaline proteases became more important toward older stages. Trypsin and chymotrypsin showed similar characteristics with at least five activities observed. For trypsin, activity was detected with pH 7 and 8 at 50°C, and pH 10 at 30, 50 and 60°C. For chymotrypsin, activity was found at pH levels 7 and 8 at 50°C, and pH 10 at 30, 40 and 50°C. Regardless of sex and age, the most suitable conditions for studying digestive enzyme activities in general were at pH 8 and 50°C for total proteases, pH 7-8 and 50°C or pH 10 and 30-35°C for trypsin and pH 7-8 and 50°C or pH 10 and 40°C for chymotrypsin. Trypsin and chymotrypsin are important proteases showing similar general optimal conditions to that of total proteases. This information provides elementary knowledge for studying protein digestibility by in vitro techniques and for determining growth performance quality using digestive enzyme markers in Siamese fighting fish

Characterization and Activity of Digestive Enzymes in Different Sizes of Nile Tilapia (Oreochromis niloticus L.)

ABSTRACT The activity of protease, amylase and lipase from the pseudostomach, upper intestine, lower intestine and liver of 5.7, 35.8 and 92.1 g male Nile tilapia (Oreochromis niloticus L.) was determined. Protease activity had an optimal pH of 12, 9 and 9 for the 5.7, 35.8 and 92.1 g fish respectively and the highest specific activity was found in the 35.8 g fish. Amylase activity had an optimal pH of 6, 7 and 2, respectively and the highest specific activity was found in the 92.1 g fish. Lipase activity had an optimal pH of 8, 7 and 8, respectively and the highest specific activity was found in the 35.8 g fish. In all sizes of tilapia, amylase activity was significantly higher in the liver than other organs (P<0.01). Protease and lipase activity in the lower and upper intestine respectively of the 5.7 and 35.8 g fish were significantly higher than in other organs (P<0.01). This study indicated that different sizes of fish had different levels of enzymatic activities. Middle-sized fish appeared to have the highest specific activity of protease and lipase, while the large fish had the highest specific activity of amylase. These results will be used as a basis for suitable feed formulation for different sizes of Nile tilapia so that optimum nutritional values and cost-effectiveness can be achieved

Changing in TSS, TA and Sugar Contents and Sucrose Synthase Activity in Ethephon-Treated 'Pattavia' Pineapple Fruit

ABSTRACT Exogenous ethylene increases endogenous ethylene which plays a crucial role on ripening in climacteric fruits. Although pineapple is a non-climacteric fruit, ethylene released from ethephon is effectively used to hasten the harvesting period. Effects from the use of a high concentration of ethephon on eating quality, fruit size and the reduction in harvesting period have been reported. In this paper, the effect of a low concentration of ethephon on pineapple fruit quality and sucrose synthase (SuSy) activity was investigated. Field experiment was arranged in split plot design. In the main plot, two levels of ethephon concentrations, i.e. 0 and 500 mg/l, were used by spraying at 110 days after forcing (DAF) fruits. The sub plot was harvesting time, i.e. 5 times of one-week intervals from 124 to 152 DAF. We found that the total soluble solid (TSS) was significantly increased in most of harvesting-treated fruits while the titratable acid (TA) was significantly increased at 131 DAF of harvesting-treated fruits. Only at 131 DAF harvesting time, the glucose content and SuSy activity of ethephon-treated fruits were significantly reduced and return to the control level afterward. However, ethephon had no effect on the fructose and sucrose contents at all harvesting times. In conclusion, fruit quality with shortening of harvesting time could be improved by applying 500 mg/l ethephon at 110 DAF since TSS content which is one of the parameter predicting eating quality of pineapple was increased without decreasing fruit quality