PCTI-250-Technology Transfer Workshops on Enzyme Activity, Digestive Capacity, and In Vitro Digestibility

PCTI 250

HECTOR NOLASCO SORIA

Autor de Correspondencia

Dr. Héctor Nolasco Soria

Editor

31/10/2025

Fecha de Aprobación

Biotecnología y Ciencias Agropecuarias

Categoría

Autores

Héctor Nolasco Soria

PCTI

hnolascosoria@gmail.com

Most studies in digestive enzymes in aquatic organisms, either omit a clear definition of digestive capacity or conflate it with digestive organ morphology or isolated enzyme activities. We compared indicators and assay conditions (substrate type, pH, temperature, and expression of units), revealing significant inter-study variability.

Abstract

According to Nolasco et al. (2025). Most studies either omit a clear definition of digestive capacity or conflate it with digestive organ morphology or isolated enzyme activities. The comparison of indicators and assay conditions (substrate type, pH, temperature, and expression of units), revealing significant variability. There is a proposal of four operational definitions: (a) Extract Theoretical Volume (ETV), (b) digestive capacity (U)—the total catalytic activity present in the digestive tract at the moment of sampling, where 1 U is the amount of enzyme catalyzing the formation of 1 µmol of product per minute under species-specific physiological pH, ionic strength, and temperature, with the total activity expressed as U fish⁻¹, U organ⁻¹, or U g⁻¹ fish or U g⁻¹ organ. (c) Digestive Processing (DP)—the total number of bonds hydrolyzed during a given digestion time, whether instantaneous or over a defined period; and (d) Digestive Processing Index (DPI, U-min or U-h), which integrates digestive capacity over time. This framework provides a proposal for assay standardization and advances comparative studies in aquatic organism digestive physiology.

Problemática

There is no standardization of enzymatic methods for quantifying the activity, digestive capacity, and in vitro digestibility of ingredients and foods

Usuarios

Research groups, biochemical analysis laboratories, and food companies.

Proyecto

Technology Transfer Workshops on Enzyme Activity, Digestive Capacity, and In Vitro Digestibility

Instructor: Héctor Nolasco-Soria

General objective

To exchange experiences aimed at standardizing methods and forms of expression of enzymatic activity, digestive capacity, and digestibility in vitro in aquatic organisms.

Objectives

To develop technology transfer workshops, at the facilities of interested laboratories, on: I. Digestive enzymes and methods, II. Digestive capacity, and III. In vitro digestibility of dietary proteins, lipids, and carbohydrates in fish, crustaceans, and mollusks. The workshops are geared toward designing methods tailored to the species of interest, optimizing methods based on the laboratory’s capabilities, and standardizing methods and formulas for expressing units of enzyme activity, digestive capacity, and in vitro digestibility.

Transfer Format

On-site Theoretical-Practical Workshops.

Target Audience:

Research groups, biochemical analysis laboratories, and food companies.

MODULES

MODULE I. Digestive Enzymes and Methods

MODULE II. Digestive Capacity.

MODULE III. In vitro digestibility of dietary proteins, lipids, and carbohydrates.

TOPICS

MODULE I.

Digestive Enzymes and Methods

Digestive enzymes in the digestive tract
Enzyme extracts and preservation
Kinetic vs. endpoint methods
Spectrophotometric and fluorogenic methods
Catalytic reaction and hydrolysis products
Optimization of reaction volume, enzyme:substrate ratio (E:S), and incubation time
Determination of environmental conditions in the reaction mixture (temperature, pH, ionic strength, etc.).
Definition of enzyme units.
Standard curves and molar extinction coefficient (MEC).
Spreadsheets for determining the activity of acid protease (pepsin-like), alkaline protease, chymotrypsin, trypsin, leucine aminopeptidase, amylase, lipase, acid phosphatase, and alkaline phosphatase.

Duration: 3-5 days. Theory (2 days, 4.5 h/day), Practice (1-3 days, 4 h/day, 2-3 enzymes/day).

MODULE II. Digestive Capacity

Definition of digestive capacity
Digestive anatomy and function
Origin, feeding history, time of slaughter, weight, and size.
Dissection of digestive organs
Preparation of extracts
Enzyme análisis
Concepts of extract theoretical volume (ETV), digestive capacity (DC), digestive processing (DP), digestive processing index (DPI).
Spreadsheets for determining digestive capacity.

Duration: 2-3 days. Theory (1 day, 4.5 h/day), Practice (1-2 days, 4 h/day, 2-3 digestive capacity enzymes/day).

MODULE III. In vitro digestibility of dietary proteins, lipids, and carbohydrates.

Food polymers.
Types of in vitro digestibility methods (pH stat, closed reactor, semipermeable membrane digestion reactor), according to the food polymer.
Intake, digestion time.
Design of the in vitro digestibility method for the species.
- Type of food polymer.
- Origin, feeding history, time of slaughter, weight, size.
- Digestive organs for the enzymatic reagent.
- Determination of environmental conditions in the digestion mixture (temperature, pH, ionic strength, etc.).
- Determination of the E:S ratio in the digestion mixture.
- Digestion time.
- Method for quantifying in vitro
- Standard molecules (Reference) for relative digestibility.
- Spreadsheets for quantifying in vitro

Duration: 3-5 days. Theory (1 day, 4.5 h/day), Practice (1-4 days, 4 h/day, 1-2 digestibility methods/day)

Interested parties should contact:

Dr. Héctor Nolasco-Soria, hnolascosoria@gmail.com

National Researcher (SNII), Level 3

ORCID: https://orcid.org/0000-0003-1584-4647

RG: https://www.researchgate.net/profile/Hector-Nolasco

Scopus: https://www.scopus.com/authid/detail.uri?authorId=8968967900

Scholar: https://scholar.google.com/scholar?q=H%C3%A9ctor+G.+Nolasco-Soria

Editorial Board for FISHES, for the ‘Nutrition and Feeding’ section: https://www.mdpi.com/journal/fishes/sectioneditors/Nutrition_Feeding

PCTI: https://pcti.mx/wp-admin/post.php?post=67&action=edit

Impacto Socioeconómico

To facilitate comparative studies on enzymatic activity, digestive capacity and in vitro digestibility of dietary proteins, polysaccharides and lipids

References (Related Instructor published articles)

Nolasco, H., Kushner, D.J., and Ochoa, J.L. 2002. Purification and properties of an extracellular halophilic serine-protease from Haloferax mediterranei. Revista de la Sociedad Química de México. 46(3): 202-211. ISSN 0583-7693

Del monte, Alberto, Héctor Nolasco, Alina Forrellat, Carlos Aragón, Aliosha García, Joaquín Diaz y Olimpia Carrillo. 2002. Evidencias de la presencia de lipasas en el hepatopáncreas de Litopenaeus schmitti. Simposium Virtual de Acuacultura CIVA 2002. ISSN: 1578-4541

López-López, S., H. Nolasco and Vega-Villasante, F. Characterization of digestive gland esterase-lipase activity of Cherax quadricarinatus juveniles. 2003. Compartive Biochemistry and Physiology 135: 337-347. Doi: 10.1016/S 1096-4959(03)00087-3, ISSN: 1531-4332-CORRESPONDING AUTHOR

Del Monte A., A. Forrellat, H. Nolasco, T. Estevez, B. Boburg, J. Díaz, O. Carrillo. 2003. Aislamiento y caracterización de lipasas en el hepatopáncreas de Litopenaues vannamei. Revista Acuatic (electrónica) /CIVA 2003. P. 755-767. (http://www.revistaaquatic.com/civa2003). ISSN: 1578-4541

Forrellat-Barrios, A., A. del Monte.Martínez, T. Estevez-Lao, B. Boburg Castroconde, H. Nolasco-Soria, O. Carrillo-Farnes. 2004. Caracterización de lipasas en tres especies de camarones pendidos. Su importancia en la digestión. Revista Acuatic (electrónica) /CIVA 2004. p 767-776. (http://www.revistaaquatic.com/civa2004).

López-López, S., Nolasco, H., Villarreal-Colmenares, H., Civera-Cerecedo, R. 2005. Digestive enzyme response to suplemental ingredients in practical diets juvenile freshwater crayfish Cherax quadricarinatus. Aquaculture Nutrition 19: 87-95. ISSN: 1365-2095

Alvarez-González, C.A., M Cervantes-Trujano, D. Tovar-Ramirez, D.E. Conklin, H. Nolasco, E. Gisbert, and R Piedrahita. 2006. Development of digestive enzymes in California Halibut Paralichthys californicus larvae. Fish Physiol. Biochem. 31: 83-93. FI: 1.607, ISSN: 0920-1742

Casillas-Hernández, R., H. Nolasco-Soria, F. Lares-Villa, T. García-Galano, O. Carrillo-Farnes y F. Vega-Villasante. 2006. Ritmo circadiano de la actividad enzimática digestiva del camarón blanco Litopenaeus vannamei y su efecto en el horario de alimentación. Revista Latinoamericana de Recursos Naturales, (2) 53-62. ISSN 2594-0384

Del Monte Martinez, Alberto, Jorge González Bacerio, Lázara Romero del Sol, Carlos Aragón Abreu, Fernando Vega, Héctor Nolasco Soria, Joaquín Dáz Brito. 2008. Extraction Systems For Isolating esterases having interfacial adsorption. Rev Colomb. Biotecnol. Vol. XI, Julio, No. 1: 7:16. 1909-8758

Alvarez-González, C.A., F. J. Moyano-López, R. Civera-Cerecedo, V. Carrasco-Chávez, J. L. Ortíz-Galindo, H. Nolasco-Soria, D. Tovar-Ramírez, S. Dumas. 2009. Development of digestive enzyme activity in larvae of spotted sand bass Paralabrax maculatofasciatus. II: Electrophoretic análisis. Fish Physiol. Biochem. (DOI 10.1007/s10695-008-9276-4, F.I: 1.607). ISSN: 0920-1742

Nolasco, Héctor, Francisco Moyano-López and Fernando Vega-Villasante. 2011. Partial characterization of pyloric-duodenal lipase of gilthead seabream (Sparus aurata). (DOI: 10.1007/s10695-010-9414-7). Fish Physiology and Biochemistry, 2011, Volume 37, Number 1, Pages 43-52 FI: 1.607. ISSN: 0920-1742 SSN: 0920-1742

Cuenca-Soria, Carlos Alberto, Carlos Alfonso Alvarez-González, José Luis Ortíz-Galindo, Hector Nolasco-Soria, Dariel Tovar-Ramírez, Rocío Guerrero-Zárate, Alfonso Castillo-Domínguez, Martha Alicia Perera-García, Raul Hernández-Gómez, Enric Gisbert. 2013. Partial characterization of digestive proteases of the Mayan cichlid Cichlasoma urophthalmus. Fish Physiology and Biochemistry, DOI 10.1007/s10695-013-9876-5., ISSN 0920-1742

Cuenca Soria, Carlos Alberto, Carlos Alfonso Alvarez, Jose Luis Ortíz Galindo, Rocío Guerrero Zarate, Martha Alicia Pereira García, Raúl Enrique Hernández Gómez, Héctor Nolasco Soria. (2013). Digestibilidad in vitro de ingredientes proteínicos en la mojarra castarrica Chichlasoma urophthalmus.Universidad, Ciencia y Desarrollo Trópico Húmedo (Ecosistemas y Recursos Agropecuarios). 29(3): 263-275, ISSN 0186-2979.

Ayala-Borboa EG, Tovar-Ramírez D, Civera-Cerecedo R, Rojas-Contreras M, Cadena-Roa MA, Nolasco-Soria H, Goytortúa-Bores E. (2013). Effect of red crab meal (Pleuroncodes planipes) on growth and digestive enzyme expression in the intestine of white shrimp (Litopenaeus vannamel). Communications in agricultural and applied biological sciences. 78(4):19-22.

Moguel-Hernández, R. Peña, H. Nolasco-Soria, S. Dumas, I. Zavala-Leal. 2014. Development of digestive enzyme activity in spotted rose snapper, Lutjanus guttatus (Steindachner, 1869) larvae, Fish Physiol Biochem. DOI 10.1007/s10695-013-9890-7

Sacristan, Hernán, Hector Nolasco-Soria, Laura López Greco. (2014). Effect of attractant stimuli, starvation period and food availability on digestive enzymes in the redclaw crayfish Cherax quadricarinatus (Parastacidae). Aquatic Biology, Vol. 23: 87–99, 2014, doi: 10.3354/ab00611

Silva, B.C, H. Nolasco-Soria, F.Magallón-Barajas, R. Civera-Cerecedo, R. Casillas-Hernández, W. Seiffert. (2015). Improved digestion and initial performance of whiteleg shrimp using organic salt supplements. Aquaculture Nutrition, doi: 10.1111/anu.12315. ISSN: 1365-2095

Frías-Quintana, C. A., G. Márquez-Couturier, C. A. Alvarez-González, D. Tovar-Ramírez, H. Nolasco-Soria, M. A. Galaviz-Espinosa, S. Camarillo-Coop, R. Martınez-Garcıa, E. Gisbert, R. Martínez-Yanez. (2015). Development of digestive tract and enzyme activities during the early ontogeny of the tropical gar Atractosteus tropicus. Fish Physiol Biochem. DOI 10.1007/s10695-015-0070-9

Silva, B. C., G. F. A. Jesus, W. Q. Seiffert, F. N. Vieira, J. L. P. Mouriño, A.Jatobá & H. Nolasco-Soria (2016). The effects of dietary supplementation with butyrate and polyhydroxybutyrate on the digestive capacity and intestinal morphology of Pacific White Shrimp (Litopenaeus vannamei), Marine and Freshwater Behaviour and Physiology, DOI: 10.1080/10236244.2016.1257759. ISSN: 1023-6244 (Print) 1029-0362 (Online) Journal homepage: http://www.tandfonline.com/loi/gmfw20. To link to this article: http://dx.doi.org/10.1080/10236244.2016.1257759.

Montoya-Mejía, Magnolia, Hervey Rodríguez-González, and Héctor Nolasco-Soria. (2016). Circadian cycle of digestive enzyme production at fasting and feeding conditions in nile tilapia, Oreochromis niloticus (actinopterygii: perciformes: cichlidae). Acta Ichthyologica Et Piscatoria, 46 (3): 163–170. DOI: 10.3750/AIP2016.46.3.01, ISSN: 0137-1592 (printed), 1734-1515.

De los Santos-Romero, Rodolfo B., Fernando Vega-Villasante, Marcelo García-Guerrero, & Héctor Nolasco-Soria. (2017). Effect of dietary chitin on digestive enzyme activity, growth and survival of Macrobrachium tenellum juvenile prawns. Lat. Am. J. Aquat. Res., 45(1): 130-138, 2017, DOI: 10.3856/vol45-issue1-fulltext-12.

Álvarez-Sánchez, Ana, Héctor Nolasco-Soria*, Humberto Mejía-Ruíz. 2017. In vitro digestibility of specific dsRNA by enzymes of digestive tract of shrimp Litopenaeus vannamei. Digestibilidad in vitro del dsRNA especifico por enzimas del tracto digestivo del camarón Litopenaeus vannamei. MVZ Cordoba, 22(2):5867-5880, 2017. ISSN: 0122-0268. DOI:doi.org/10.21897/rmvz.1024.

Espinosa-Chaurand, Daniel, Fernando Vega-Villasante, Olimpia Carrillo-Farnés, Héctor Nolasco-Soria (2017). Eﬀect of circadian rhythm, photoperiod, and molt cycle on digestive enzymatic activity of Macrobrachium tenellum juveniles, Aquaculture 479 (2017) 225–232.

Montoya-Mejía, Magnolia, Hervey Rodríguez-González, and Héctor Nolasco-Soria. (2017). Digestibility, growth, blood chemistry and enzyme activity of juvenile Oreochromis niloticus fed isocaloric diets containing animal and plant by-products. Revista Brasileira de Zootecnia. 46(12):873-882, http://dx.doi.org/10.1590/S1806-92902016000800001, BRASIL, ISSN: 1806-9290

Jesús-Ramirez, Fidel, Carlos Alfonso Álvarez-González, Héctor Gerardo Nolasco-Soria, Emyr Peña, Rafael Martínez-García, Susana Camarillo-Coop, Kevin Conway y Camilo Pohlenz (2017). Caracterización parcial de proteasas digestivas del chucumite (Centropomus parallelus). Partial characterization of digestive proteases of fat snook (Centropomus paralellus). Hidrobiológica, 27 (3): 419-427.

Álvarez-Sánchez, Ana, Héctor Nolasco-Soria*, Alberto Peña-Rodríguez, Humberto Mejía-Ruíz (2018). In vitro Digestibility of Yarrowia lipolytica Yeast and Growth Performance in Whiteleg shrimp Litopenaeus vannamei. Turkish Journal of Fisheries and Aquatic Sciences, 18: 395-404. E-ISSN: 2149 – 181X, DOI: 10.4194/1303-2712-v18_3_05 CORRESPONDING AUTOR.

Hernández-Hurtado, Pablo S., Héctor Nolasco-Soria, Olimpia Carrillo-Farnés, Helios Hernández-Hurtado, Rafael García de Quevedo-Machain, Gustavo Casas-Andreu, Cynthia Montoya-Martínez y Fernando Vega-Villasante (2018). Contributions to the nutrition of the American crocodile Crocodylus acutus (Cuvier, 1807). Latin American Journal of Aquatic Research 46(1): 15-19, 2018. ISSN 0718-560X

Gisbert, Enric, Hector Nolasco, Mikhail Solovyev (2018).Towards the standardization of brush border puriﬁcation and intestinal alkaline phosphatase quantiﬁcation in ﬁsh with notes on other digestive enzymes, Aquaculture 487: 102–108. ISSN: 0044-8486, doi.org/10.1016/j.aquaculture.2018.01.004

Montoya-Martínez, Cynthia, Héctor Nolasco-Soria, Fernando Vega-Villasante, Olimpia Carrillo-Farnés, Alfonso Álvarez-González & Roberto Civera-Cerecedo. 2018. In vitro protein digestibility of animal, vegetal and microbial feed ingredients for Macrobrachium tenellum. Lat. Am. J. Aquat. Res., 46(3): 495.501. DOI: 10.3856/vol46-issue3-fulltext-1. ISSN 0718-560X.

Nolasco-Soria H, Moyano-López F, Vega-Villasante F, Del Monte A, Espinoza-Chaurand D, Gisbert E (2018). Lipase and Phospholipase Activity Methods for Marine Organisms. Chapter 7. Georgina Sandoval (ed.), Lipases and Phospholipases: Methods and Protocols, Methods in Molecular Biology, vol. 1835, https://doi.org/10.1007/978-1-4939-8672-9_7, © Springer Science+Business Media, LLC, part of Springer Nature 2018. Electronic ISSN 1940-6029

Espinosa-Chaurand D, Nolasco-Soria H (2019). In vitro digestibility of oils used in feed for Macrobrachium tenellum, Macrobrachium americanum and Litopenaeus vannamei adults. Aquaculture 512 (2019) 734303. https://doi.org/10.1016/j.aquaculture.2019.734303 ISSN 0044-8486.

Nolasco-Soria, H., Héctor-Rodrigo Nolasco-Alzaga, and Enric Gisbert (2020). The importance of pepsin-like acid protease quantification in aquaculture studies: a revision of available procedures and presentation of a new protocol for its assessment. Reviews in Aquaculture,12: 1928-1943. https://doi: 10.1111/raq.12417, Online ISSN:1753-5131

Nolasco-Soria, H. (2020) Improving and standardizing protocols for alkaline protease quantification in fish. Reviews in Aquaculture,13: 43-65. https://doi: 10.1111/raq.12463, Online ISSN:1753-5131

Nolasco-Soria, H. (2021) Amylase quantification in aquaculture fish studies: A revision of most used procedures and presentation of a new practical protocol for its assessment. Aquaculture, Volume 538, 30 May 2021, 736536, https://doi.org/10.1016/j.aquaculture.2021.736536, ISSN 0044-8486

Aréchiga-Palomera, Martín A., Héctor Nolasco-Soria, Olimpia Carrillo-Farnés, Karen N. Nieves-Rodríguez & Fernando Vega-Villasante (2023). Contribution to some aspects of the digestive tract anatomy and food evacuation of the Pacific fat sleeper Dormitator latrifrons. Latin american journal of aquatic research. ISSN 2594-0384. Latin American Journal of Aquatic Research, 51(4): 521-529, DOI: 10.3856/vol51-issue4-fulltext-3038

Nolasco-Soria, H. (2023). Nolasco-Soria H (2023) Fish digestive lipase quantiﬁcation methods used in aquaculture studies. Front. Aquac. 2:1225216. doi: 10.3389/faquc.2023.1225216

Perales-García, N., Tovar-Ramírez, D., Martínez-Morales, M.G., Ceballos-Vázquez, B.P., Corona-Rojas, D,A., Salcedo-Meza, M.A., Garrido-Mora, A., Vega-Villasante, F., Nolasco- Soria, H. (2023). pH evaluation in the digestive tract of the pygmy octopus, Paractopus digueti. Comparative Biochemistry and Physiology, ISSN: 1531-4332 https://doi.org/10.1016/j.cbpb.2023.110881.

Nolasco-Soria, H.; Alvarez-González, C.A.; Tovar-Ramírez, D.; González-Bacerio, J.; del Monte-Martínez, A.; Vega-Villasante, F. Optimization of Classical Lipase Activity Assays for Fish Digestive Tract Samples. Fishes 2024, 9, 261. https://doi.org/10.3390/fishes9070261. ISSN: 2410-3888.

Nolasco-Soria, H.; Yúfera, M.; Nolasco-Alzaga, H.R. Fish Digestive Capacity: Definition and Methods. Fishes 2025, 10, 546. https://doi.org/10.3390/fishes10110546. ISSN: 2410-3888.

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