Homeopathy 2020; 109(01): 003-013
DOI: 10.1055/s-0039-1692998
Original Research Article
The Faculty of Homeopathy

Evaluation of Homeopathic Phosphoric Acid, Silica and Pathogenic Vibrio on Digestive Enzyme Activity of Longfin Yellowtail Fish (Seriola rivoliana)

José Manuel Mazón-Suástegui
1   Centro de Investigaciones Biológicas del Noroeste S.C. (CIBNOR), La Paz, Baja California Sur, Mexico
Joan Salas-Leiva
1   Centro de Investigaciones Biológicas del Noroeste S.C. (CIBNOR), La Paz, Baja California Sur, Mexico
Andressa Teles
1   Centro de Investigaciones Biológicas del Noroeste S.C. (CIBNOR), La Paz, Baja California Sur, Mexico
Dariel Tovar-Ramírez
1   Centro de Investigaciones Biológicas del Noroeste S.C. (CIBNOR), La Paz, Baja California Sur, Mexico
› Author Affiliations
Funding The study was funded by Sectoral Fund for Science and Education, Basic Science Project No. 258282 “Experimental evaluation of homeopathy and new probiotics in the cultivation of molluscs, crustaceans and fish of commercial interest”, and PROINNOVA-CONACyT/PEASA No. 241777 “Innovation and continuous improvement of hatchery production technology”, under the academic responsibility of author J.M.M.S. J.S.Ll is a recipient of a Postdoctoral Fellowship (AMEXCID 811-06-9616; CIBNOR 241777) and A.T. is a Doctoral Fellow (CONACYT 335728).
Further Information

Publication History

06 March 2019

22 May 2019

Publication Date:
27 August 2019 (online)


Background This research aimed to observe the effect of homeopathically prepared Vibrio parahaemolyticus (ViP) and V. alginolyticus (ViA) and the commercial homeopathic compound Similia (Phosphoricum acidum and Silicea terra) on the digestive enzyme activities of Seriola rivoliana juveniles under usual culture conditions.

Materials and Methods Biochemical analysis was used to study the effect of highly diluted substances (7C potency) prepared from ViP and ViA (Treatment 1: T1) and the homeopathic compound Phosphoricum acidum and Silicea terra (Treatment 2: T2) on changes in the main digestive enzymes on weaning-state fish (WS; 30 days post-hatching [DPH]) and early juveniles (EJ; 62 DPH) versus a reference control group that received no homeopathic medicines.

Results Treatment T2 significantly increased the activity of trypsin and lipase and decreased the activity of amylase, whereas treatment T1 increased the activity of chymotrypsin and reduced the activity of aminopeptidase-N in WS fish. Except for alkaline phosphatase, which was significantly reduced in the intestine, no significant differences in enzymatic activity were found between treated EJ fish and controls. The fish of the WS group had a higher growth rate with the T2 treatment.

Conclusions T1 treatment stimulated chymotrypsin in EJ fish and T2 promoted intestinal maturation of WS fish. Higher growth rate with the T2 treatment may be associated with the stimulation of trypsin activity. Thus, T2 may be applied, under hatchery conditions, during larval stages with an aim to enhance digestion and assimilation of inert food.


• Effects of two homeopathic treatments on the digestive enzyme activity in Seriola rivoliana were evaluated.

• Homeopathically prepared Vibrio pathogenic strains caused stimulation of chymotrypsin in Seriola rivoliana.

Phosphoricum acidum and Silicea terra significantly increased the activity of trypsin and lipase.

• Homeopathy may be an ecological alternative to antibiotic use in aquaculture.

Supplementary Material

Supplementary Material

  • References

  • 1 Roo J, Fernández-Palacios H, Hernández-Cruz CM. , et al. First results of the spawning and larval rearing of the longfin yellowtail Seriola rivoliana as a fast-growing candidate for European marine finfish aquaculture diversifications. Aquacult Res 2014; 45: 689-700
  • 2 Teles A, Salas-Leiva J, Alvarez-González CA. , et al. Histological study of the gastrointestinal tract in longfin yellowtail (Seriola rivoliana) larvae. Fish Physiol Biochem 2017; 43: 1613-1628
  • 3 Mesa-Rodríguez A, Hernández-Cruz CM, Socorro JA. , et al. Skeletal development and mineralization pattern of the vertebral column, dorsal, anal and caudal fin complex in Seriola rivoliana (Valenciennes, 1833) larvae. J Aquac Res Dev 2014; 5: 6
  • 4 Mesa-Rodríguez A, Hernández-Cruz CM, Betancor MB, Fernández-Palacios H, Izquierdo MS, Roo J. Bone development of the skull, pectoral and pelvic fins in Seriola rivoliana (Valenciennes, 1833) larvae. Fish Physiol Biochem 2016; 42: 1777-1789
  • 5 Quiñones-Arreola MF, Arcos-Ortega GF, Gracia-López V. , et al. Reproductive broodstock performance and egg quality of wild-caught and first-generation domesticated Seriola rivoliana reared under same culture conditions. Lat Am J Aquat 2015; 43: 953-962
  • 6 Kissinger KR, García-Ortega A, Trushenski JT. Partial fish meal replacement by soy protein concentrate, squid and algal meals in low fish-oil diets containing Schizochytrium limacinum for longfin yellowtail Seriola rivoliana . Aquaculture 2016; 452: 37-44
  • 7 Silva FCP, Nicoli JR, Zambonino-Infante JL. , et al. Influence of partial substitution of dietary fish meal on the activity of digestive enzymes in the intestinal brush border membrane of gilthead sea bream, Sparus aurata and goldfish, Carassius auratus . Aquaculture 2010; 306: 233-237
  • 8 Bowyer JN, Qin JG, Smullen RP. , et al. The use of a soy product in juvenile yellowtail kingfish (Seriola lalandi) feeds at different water temperatures: 1. Solvent extracted soybean meal. Aquaculture 2013; 384: 35-45
  • 9 Novelli B, Otero-Ferrer F, Díaz M. , et al. Digestive biochemistry as indicator of the nutritional status during early development of the long snouted seahorse (Hippocampus reidi). Aquaculture 2016; 464: 196-204
  • 10 Tovar D, Zambonino J, Cahu C. , et al. Effect of live yeast incorporation in compound diet on digestive enzyme activity in sea bass (Dicentrarchus labrax) larvae. Aquaculture 2002; 204: 113-123
  • 11 Suzer C, Çoban D, Kamaci HO. , et al. Lactobacillus spp. bacteria as probiotics in gilthead sea bream (Sparus aurata, L.) larvae: effects on growth performance and digestive enzyme activities. Aquaculture 2008; 280: 140-145
  • 12 Mohapatra S, Chakraborty T, Prusty AK. , et al. Use of different microbial probiotics in the diet of rohu, Labeo rohita fingerlings: effects on growth, nutrient digestibility and retention, digestive enzyme activities and intestinal microflora. Aquacult Nutr 2012; 18: 1-11
  • 13 Soleimani N, Hoseinifar SH, Merrifield DL, Barati M, Abadi ZH. Dietary supplementation of fructooligosaccharide (FOS) improves the innate immune response, stress resistance, digestive enzyme activities and growth performance of Caspian roach (Rutilus rutilus) fry. Fish Shellfish Immunol 2012; 32: 316-321
  • 14 Castillo S, Rosales M, Pohlenz C, Gatlin III DM. Effects of organic acids on growth performance and digestive enzyme activities of juvenile red drum Sciaenops ocellatus . Aquaculture 2014; 433: 6-12
  • 15 Teles A, Salas-Leiva J, Alvarez-González CA, Tovar-Ramírez D. Changes in digestive enzyme activities during early ontogeny of Seriola rivoliana . Fish Physiol Biochem 2019; 45: 733-742
  • 16 Valentim-Zabott M, Vargas L, Ribeiro RPR. , et al. Effects of a homeopathic complex in Nile tilapia (Oreochromis niloticus L.) on performance, sexual proportion and histology. Homeopathy 2008; 97: 190-195
  • 17 Júnior RP, Vargas L, Valentim-Zabott M, Ribeiro RP, da Silva AV, Otutumi LK. Morphometry of white muscle fibers and performance of Nile tilapia (Oreochromis niloticus) fingerlings treated with methyltestosterone or a homeopathic complex. Homeopathy 2012; 101: 154-158
  • 18 Braccini GL, Natali MRM, Ribeiro RP. , et al. Morpho-functional response of Nile tilapia (Oreochromis niloticus) to a homeopathic complex. Homeopathy 2013; 102: 233-241
  • 19 Feitosa KC, Povh JA, de Abreu JS. Physiological responses of pacu (Piaractus mesopotamicus) treated with homeopathic product and submitted to transport stress. Homeopathy 2013; 102: 268-273
  • 20 Andretto AP, Fuzinatto MM, Bonafe EG. , et al. Effect of an homeopathic complex on fatty acids in muscle and performance of the Nile tilapia (Oreochromis niloticus). Homeopathy 2014; 103: 178-185
  • 21 Mazón-Suástegui JM, Bernal MG, Abasolo-Pacheco F. , et al. Homeopathy for shrimp aquaculture: Increased survival and superoxide dismutase activity in juvenile white shrimp Litopenaeus vannamei during a bacterial pathogen-challenge. Homeopathy 2016; 105: 33
  • 22 Mazón-Suástegui JM, Rosero-García A, Avilés-Quevedo A. , et al. Homeopathy for marine fish aquaculture: increased growth and survival of juvenile spotted rose snapper Lutjanus guttatus . Homeopathy 2016; 105: 32-33
  • 23 Mazón-Suástegui JM, Salas-Leiva J, Teles A, Tovar-Ramírez D. Immune and antioxidant enzyme response of the Longfin yellowtail (Seriola rivoliana) juveniles to ultra-diluted substances derived from phosphorus, silica and pathogenic Vibrio . Homeopathy 2019; 108: 43-53
  • 24 Ortíz-Cornejo NL, Tovar-Ramírez D, Abasolo-Pacheco F, Mazón-Suástegui JM. Homeopatía, una alternativa para la acuicultura. Rev Médica Homeopat 2017; 10: 18-24
  • 25 Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72: 248-254
  • 26 Anson ML. The estimation of pepsin, trypsin, papain, and cathepsin with hemoglobin. J Gen Physiol 1938; 22: 79-89
  • 27 Vega-Villasante F, Nolasco H, Civera R. The digestive enzymes of the Pacific brown shrimp Penaeus californiensis: I—Properties of amylase activity in the digestive tract. Comp Biochem Physiol Part B Comp Biochem 1993; 106: 547-550
  • 28 Maroux S, Louvard D, Baratti J. The aminopeptidase from hog intestinal brush border. Biochim Biophys Acta 1973; 321: 282-295
  • 29 Toledo Cuevas EM, Moyano López FJ, Tovar-Ramírez D. Development of digestive biochemistry in the initial stages of three cultured Atherinopsids . Aquacult Res 2011; 42: 776-786
  • 30 Benítez-Hernández A, Jimenez-Bárcenas L, Sánchez-Gutiérrez EY. , et al. Use of marine by-product meals in diets for juvenile longfin yellowtail Seriola rivoliana . Aquacult Nutr 2017; 24: 562-570
  • 31 Bowyer JN, Qin JG, Smullen RP. , et al. The use of a soy product in juvenile yellowtail kingfish (Seriola lalandi) feeds at different water temperatures: 2. Soy protein concentrate. Aquaculture 2013; 410: 1-10
  • 32 Abbink W, Garcia AB, Roques JAC. , et al. The effect of temperature and pH on the growth and physiological response of juvenile yellowtail kingfish Seriola lalandi in recirculating aquaculture systems. Aquaculture 2012; 330: 130-135
  • 33 Frías-Quintana CA, Domínguez-Lorenzo J, Álvarez-González CA, Tovar-Ramírez D, Martínez-García R. Using cornstarch in microparticulate diets for larvicultured tropical gar (Atractosteus tropicus). Fish Physiol Biochem 2016; 42: 517-528
  • 34 Ma Z, Qin JG, Hutchinson W. , et al. Responses of digestive enzymes and body lipids to weaning times in yellowtail kingfish Seriola lalandi (Valenciennes, 1833) larvae. Aquacult Res 2014; 45: 973-982
  • 35 Galaviz MA, López LM, García Gasca A, Álvarez González CA, True CD, Gisbert E. Digestive system development and study of acid and alkaline protease digestive capacities using biochemical and molecular approaches in totoaba (Totoaba macdonaldi) larvae. Fish Physiol Biochem 2015; 41: 1117-1130
  • 36 Kolkovski S. Digestive enzymes in fish larvae and juveniles—implications and applications to formulated diets. Aquaculture 2001; 200: 181-201
  • 37 Falk-Petersen IB. Comparative organ differentiation during early life stages of marine fish. Fish Shellfish Immunol 2005; 19: 397-412
  • 38 Ogiwara K, Takahashi T. Specificity of the medaka enteropeptidase serine protease and its usefulness as a biotechnological tool for fusion-protein cleavage. Proc Natl Acad Sci U S A 2007; 104: 7021-7026
  • 39 Cara B, Moyano FJ, Zambonino JL, Fauvel C. Trypsin and chymotrypsin as indicators of nutritional status of post-weaned sea bass larvae. J Fish Biol 2007; 70: 1789-1808
  • 40 Zambonino Infante JL, Cahu CL. Ontogeny of the gastrointestinal tract of marine fish larvae. Comp Biochem Physiol C Toxicol Pharmacol 2001; 130: 477-487
  • 41 Rahimi-Yadkoori N, Zanguee N, Mousavi SM, Zakeri M. Effects of ginger (Zingiber officinale) extract on digestive enzymes and liver activity of Mesopotamichthys sharpeyi fingerlings. J Persian Gulf 2015; 6: 1-10
  • 42 Mazón-Suástegui JM, García-Bernal M, Saucedo PE, Campa-Córdova Á, Abasolo-Pacheco F. Homeopathy outperforms antibiotics treatment in juvenile scallop Argopecten ventricosus: effects on growth, survival, and immune response. Homeopathy 2017; 106: 18-26
  • 43 Kil DY, Piao LG, Long HF. , et al. Effects of organic or inorganic acid supplementation on growth performance, nutrient digestibility and white blood cell counts in weanling pigs. Asian-Australas J Anim Sci 2006; 19: 252-261
  • 44 Lemieux H, Blier P, Dutil JD. Do digestive enzymes set a physiological limit on growth rate and food conversion efficiency in the Atlantic cod (Gadus morhua)?. Fish Physiol Biochem 1999; 20: 293-303
  • 45 Tovar-Ramırez D, Infante JZ, Cahu C. , et al. Influence of dietary live yeast on European sea bass (Dicentrarchus labrax) larval development. Aquaculture 2004; 234: 415-427
  • 46 Cahu CL, Infante JLZ, Peres A. , et al. Algal addition in sea bass (Dicentrarchus labrax) larvae rearing: effect on digestive enzymes. Aquaculture 1998; 161: 479-489
  • 47 Langeland M, Lindberg JE, Lundh T. Digestive enzyme activity in Eurasian perch (Perca fluviatilis) and Arctic charr (Salvelinus alpinus). J Aquac Res Dev 2013; 5: 208
  • 48 Bowyer JN, Booth MA, Qin JG. , et al. Temperature and dissolved oxygen influence growth and digestive enzyme activities of yellowtail kingfish Seriola lalandi (Valenciennes, 1833). Aquacult Res 2014; 45: 2010-2020
  • 49 Sunde J, Taranger GL, Rungruangsak-Torrissen K. Digestive protease activities and free amino acids in white muscle as indicators for feed conversion efficiency and growth rate in Atlantic salmon (Salmo salar L.). Fish Physiol Biochem 2001; 25: 335-345
  • 50 Ribeiro L, Zambonino-Infante JL, Cahu C, Dinis MT. Digestive enzymes profile of Solea senegalensis post larvae fed Artemia and a compound diet. Fish Physiol Biochem 2002; 27: 61-69
  • 51 Kuz'mina VV. Influence of age on digestive enzyme activity in some freshwater teleosts. Aquaculture 1996; 148: 25-37
  • 52 Cuvier-Péres A, Kestemont P. Development of some digestive enzymes in Eurasian perch larvae Perca fluviatilis . Fish Physiol Biochem 2001; 24: 279-285
  • 53 Liu W, Zhang XM, Wang LB. Digestive enzyme and alkaline phosphatase activities during the early stages of Silurus soldatovi development. Dongwuxue Yanjiu 2010; 31: 627-632
  • 54 Chaudhuri A, Mukherjee S, Homechaudhuri S. Diet composition and digestive enzymes activity in carnivorous fishes inhabiting mudflats of Indian Sundarban estuaries. Turk J Fish Aquat Sci 2012; 12: 265-275
  • 55 Chen BN, Qin JG, Kumar MS. , et al. Ontogenetic development of digestive enzymes in yellowtail kingfish Seriola lalandi larvae. Aquaculture 2006; 260: 264-271
  • 56 Namulawa VT, Kato CD, Rutaisire J. , et al. Enzyme activity in the Nile perch gut: implications to Nile perch culture. Int J Fish Aquac 2013; 5: 221-228
  • 57 Bowyer JN, Qin JG, Smullen RP, Stone DAJ. Replacement of fish oil by poultry oil and canola oil in yellowtail kingfish (Seriola lalandi) at optimal and suboptimal temperatures. Aquaculture 2012; 356: 211-222
  • 58 Kofuji PYM, Murashita K, Hosokawa H, Masumoto T. Effects of exogenous cholecystokinin and gastrin on the secretion of trypsin and chymotrypsin from yellowtail (Seriola quinqueradiata) isolated pyloric caeca. Comp Biochem Physiol A Mol Integr Physiol 2007; 146: 124-130
  • 59 Murashita K, Fukada H, Rønnestad I, Kurokawa T, Masumoto T. Nutrient control of release of pancreatic enzymes in yellowtail (Seriola quinqueradiata): involvement of CCK and PY in the regulatory loop. Comp Biochem Physiol A Mol Integr Physiol 2008; 150: 438-443
  • 60 Murashita K, Fukada H, Takahashi N. , et al. Effect of feed ingredients on digestive enzyme secretion in fish. Bull Fish Res Agenda 2015; 40: 69-74
  • 61 Furutani T, Masumoto T, Fukada H. Molecular cloning and tissue distribution of cholecystokinin-1 receptor (CCK-1R) in yellowtail Seriola quinqueradiata and its response to feeding and in vitro CCK treatment. Gen Comp Endocrinol 2013; 186: 1-8
  • 62 Alvarez-González CA, Moyano-López FJ, Civera-Cerecedo R, Carrasco-Chávez V, Ortiz-Galindo JL, Dumas S. Development of digestive enzyme activity in larvae of spotted sand bass Paralabrax maculatofasciatus. 1. Biochemical analysis. Fish Physiol Biochem 2008; 34: 373-384
  • 63 Midhun SJ, Arun D, Edatt L. , et al. Modulation of digestive enzymes, GH, IGF-1 and IGF-2 genes in the teleost, Tilapia (Oreochromis mossambicus) by dietary curcumin. Aquacult Int 2016; 24: 1277-1286
  • 64 Hoseinifar SH, Dadar M, Ringø E. Modulation of nutrient digestibility and digestive enzyme activities in aquatic animals: the functional feed additives scenario. Aquacult Res 2017; 48: 3987-4000
  • 65 Comabella Y, Mendoza R, Aguilera C. , et al. Digestive enzyme activity during early larval development of the Cuban gar Atractosteus tristoechus . Fish Physiol Biochem 2006; 32: 147
  • 66 Cui K, Cheng D, Ma Z. , et al. Ontogenetic development of digestive enzymes in larval and juvenile crimson snapper Lutjanus erythopterus (Bloch 1790). Aquacult Res 2017; 48: 4533-4544
  • 67 Bates JM, Akerlund J, Mittge E, Guillemin K. Intestinal alkaline phosphatase detoxifies lipopolysaccharide and prevents inflammation in zebrafish in response to the gut microbiota. Cell Host Microbe 2007; 2: 371-382
  • 68 Merlini LS, Vargas L, Piau Jr R, Ribeiro RP, Merlini NB. Effects of a homeopathic complex on the performance and cortisol levels in Nile tilapia (Oreochromis niloticus). Homeopathy 2014; 103: 139-142
  • 69 Auperin B, Baroiller JF, Ricordel MJ, Fostier A, Prunet P. Effect of confinement stress on circulating levels of growth hormone and two prolactins in freshwater-adapted tilapia (Oreochromis niloticus). Gen Comp Endocrinol 1997; 108: 35-44
  • 70 Barton BA. Stress in fishes: a diversity of responses with particular reference to changes in circulating corticosteroids. Integr Comp Biol 2002; 42: 517-525
  • 71 Firla B, Arndt M, Frank K. , et al. Extracellular cysteines define ectopeptidase (APN, CD13) expression and function. Free Radic Biol Med 2002; 32: 584-595
  • 72 Tang J, Qu F, Tang X. , et al. Molecular characterization and dietary regulation of aminopeptidase N (APN) in the grass carp (Ctenopharyngodon idella). Gene 2016; 582: 77-84
  • 73 Farhoudi A, Abedian Kenari AM, Nazari RM, Makhdoomi CH. Changes of digestive enzymes activity in common carp (Cyprinus carpio) during larval ontogeny. Iran J Fish Sci 2013; 12: 320-334
  • 74 Frías-Quintana CA, Márquez-Couturier G, Alvarez-González CA. , et al. Development of digestive tract and enzyme activities during the early ontogeny of the tropical gar Atractosteus tropicus . Fish Physiol Biochem 2015; 41: 1075-1091
  • 75 Bell IR, Koithan M. A model for homeopathic remedy effects: low dose nanoparticles, allostatic cross-adaptation, and time-dependent sensitization in a complex adaptive system. BMC Complement Altern Med 2012; 12: 191
  • 76 Bell IR, Schwartz GE. Adaptive network nanomedicine: an integrated model for homeopathic medicine. Front Biosci (Schol Ed) 2013; 5: 685-708
  • 77 Bell IR, Sarter B, Koithan M. , et al. Nonlinear response amplification mechanisms for low doses of natural product nanomedicines: dynamical interactions with the recipient complex adaptive system. J Nanomed Nanotechnol 2013; 4: 179
  • 78 Bell IR, Schwartz GE. Enhancement of adaptive biological effects by nanotechnology preparation methods in homeopathic medicines. Homeopathy 2015; 104: 123-138
  • 79 Ambrosone A, Scotto di Vettimo MR, Malvindi MA. , et al. Impact of amorphous SiO2 nanoparticles on a living organism: morphological, behavioral, and molecular biology implications. Front Bioeng Biotechnol 2014; 2: 37