Species specific in vitro protein digestion (pH-stat) for fish: method development and application for juvenile rainbow trout (Oncorhynchus mykiss), cobia (Rachycentron canadum), and Nile tilapia (Oreochromis niloticus)

Yasumaru, Fanny; Lemos, Daniel

doi:10.1016/j.aquaculture.2014.01.012

Cited by 51 publications

(44 citation statements)

References 31 publications

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“…However, the structures and inner pH values of gastrointestinal tracts of these fish species are largely different. For example, common carp has no stomach and the intestinal pH value is 6.8–7.3 (Ji ), whereas the fish species having stomach, such as channel catfish, rainbow trout and tilapia, have lower pH values in stomach (Robinson & Wilson ; Yasumaru & Lemos ). Moreover, the properties and chemical composition of diets, optimum living temperature and digestive efficiency are also different among fish species.…”

Section: Discussionmentioning

confidence: 99%

The optimum active site of acidic phytase in the gastrointestinal tract of channel catfish (Ictalurus punctatus):in vitroandin vivostudies

et al. 2015

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This study was conducted to determine the optimum active site of acidic phytase in the gastrointestinal tract of channel catfish. The pH value, acidic phytase activity, resistance of acidic phytase against gastrointestinal protease, substrate availability, and enzymatic reaction products were measured in different gastrointestinal sections of channel catfish, respectively. The results indicated that in the whole gastrointestinal tract, only the pH value of stomach (4.2-4.7) lied in the optimum active pH range of acidic phytase (1.5-5.5). The resistant ability of acidic phytase against proteases in different sections of the gastrointestinal tract was comparable, and the highest activity of phytase was observed in the stomach compared with other gastrointestinal sections. Moreover, the highest concentration of soluble phytic acid as the substrate of phytase was also seen in the stomach. After feeding fish with the diet containing acidic phytase, the residual phytic acid in the stomach was significantly reduced. Correspondingly, the dietary addition of acidic phytase significantly improved the phosphorus absorption in the intestine. In short, this study verified that stomach is the optimum active site for acidic phytase in channel catfish, because it allows higher phytase activity and provides more soluble substrates of phytase than other gastrointestinal sections.

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Section: Discussionmentioning

confidence: 99%

The optimum active site of acidic phytase in the gastrointestinal tract of channel catfish (Ictalurus punctatus):in vitroandin vivostudies

et al. 2015

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“…It provides multiple advantages such as specific response using standardized enzymes, stable conditions, rapid, precise, test of small amounts of different ingredients and suitability for different sources of ingredients, including marine‐based animal and plant ingredients (Lemos, Lawrence, & Siccardi, ; Yasumaru & Lemos, ). There is currently great interest in efforts to standardize the pH‐Stat method in fish species (Dimes & Haard, ; El‐Mowafi, Dorrell, & Bureau, ; Tibbetts, Milley, Ross, Verreth, & Lall, ; Tibbetts, Verreth, & Lall, ; Yasumaru & Lemos, ) and crustaceans (Ezquerra, Garcia‐Carreño, Civera, & Haard, ; Lemos, Ezquerra, & García‐Carreño, ; Lemos et al., ). This is because the main limitation in pH‐Stat assays seems to be an incomplete understanding of assay results in relation to the activities and origin of various enzymes, given that variation in species, fish size/age and phenotype could generate poor reproducibility of the in vitro digestion assays (Tibbetts, Milley et al., ).…”

Section: Introductionmentioning

confidence: 99%

In vitro protein digestibility of different grow-out stages of spotted rose snapper (Lutjanus guttatus, Steindachner, 1869)

et al. 2017

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The aim of this study was to compare in vitro protein digestibility between two groups of fish, at early (21 g) and late stages (400 g) of spotted rose snapper Lutjanus guttatus, to evaluate the degree of hydrolysis (DH) and total amino acid release (TAAR) using crude extracts from stomach, pyloric caeca and intestine of 13 protein ingredients including marine, animal and plant meals. Degree of hydrolysis and TAAR were measured by a pH-Stat method, and the PAGE-Zymogram was also used as complementary technique. Differences in DH were found between both grow-out stages mainly in the alkaline hydrolysis phase. Fish and squid meals (marine sources) had the highest DH and TAAR, followed by porcine meat and poultry meal byproducts from recycling sources, and soybean and canola meals (plant sources), which represent better protein sources for use in practical diets. Stomach zymograms showed two pepsin isoforms in both grow-out stages. Pyloric caeca and intestine zymograms showed five bands with proteolytic activity in the early grow-out stage, whereas four additional bands were found in late grow-out stage. Alkaline proteases were identified as serine and metalloproteases. Thus, L. guttatus presents an ontogenetically differentiated digestive enzyme pattern that modifies the DH and TAAR of different protein sources. K E Y W O R D Salkaline proteases, amino acids, electrophoresis, Lutjanus guttatus, pepsin, pH-Stat

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“…In addition, the use of enzymatic cocktails from fish makes the methods more expensive and less reproducible (Gomes et al., ), since enzyme activity can change with fish age, sexual maturation, specie and procedures using these cocktails need standardization (Dimes & Haard, ). Although, Yasumaru and Lemos () standardized the hydrolytic capacity of the species‐specific enzyme extracts on purified protein substrates, most of the studies using fish enzymes did not. Another point with in vitro methods used for assess protein digestibility as pH‐stat or drop methods is the use of alkaline pH (>8.0), without gastric phase.…”

Section: Introductionmentioning

confidence: 99%

In vitro assays predicts mineral retention and apparent protein digestibility of different fish feed measured using a juvenileP. mesopotamicusmodel

et al. 2018

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Feed chemical composition only provides information about nutrient content of a diet, but not its bioavailability. The aims were to determine mineral bioaccessibility and protein digestibility of three different fish feed, using in vitro methods and to correlate them with feed mineral retention and apparent protein digestibility measured by juvenile Piaractus mesopotamicus model, considering elliptical joint confidence region (EJCR) of weighted least-squares analysis. For in vivo assays, juvenile P. mesopotamicus were randomly stocked into conical tanks and fed during 38 days with a commercial diet, an experimental diet, or the same experimental diet added with phytase (0.2 g/kg). Total body mineral retention (mineral bioavailability indicator) and apparent protein digestibility of different diets were measured. For in vitro methods, mineral dialysability (mineral bioaccessibility indicator) and protein digestibility of the same diets were determined. No significant difference in morphometric and haematological markers were detected between diets. For iron, zinc and phosphorous bioavailability, and protein digestibility, weighted least-squares analysis showed the optimal point (1, 0) was included in EJCR, indicating in vivo and in vitro methods were comparable and correlated satisfactorily. In vitro methods were able to reproduce accurately the results obtained from traditional in vivo assays using juvenile P. mesopotamicus model. K E Y W O R D Sfish feed, mineral dialysability, mineral retention, protein digestibility

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Species specific in vitro protein digestion (pH-stat) for fish: method development and application for juvenile rainbow trout (Oncorhynchus mykiss), cobia (Rachycentron canadum), and Nile tilapia (Oreochromis niloticus)

Cited by 51 publications

References 31 publications

The optimum active site of acidic phytase in the gastrointestinal tract of channel catfish (Ictalurus punctatus):in vitroandin vivostudies

The optimum active site of acidic phytase in the gastrointestinal tract of channel catfish (Ictalurus punctatus):in vitroandin vivostudies

In vitro protein digestibility of different grow-out stages of spotted rose snapper (Lutjanus guttatus, Steindachner, 1869)

In vitro assays predicts mineral retention and apparent protein digestibility of different fish feed measured using a juvenileP. mesopotamicusmodel

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