Next Article in Journal
Research on the Behavior Recognition of Beef Cattle Based on the Improved Lightweight CBR-YOLO Model Based on YOLOv8 in Multi-Scene Weather
Previous Article in Journal
Captive Breeding and Early Developmental Dynamics of Cirrhinus mrigala: Implications for Sustainable Seed Production
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Animals
Volume 14
Issue 19
10.3390/ani14192798
animals-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Histochemical Analysis and Distribution of Digestive Enzymes in the Gastrointestinal System of the European Barracuda Sphyraena sphyraena (Linnaeus, 1758)

by
Ivana Tlak Gajger
1,*,
Srebrenka Nejedli
2,
Zvonimir Kozarić
2,† and
Josipa Vlainić
3
1
Department for Biology and Pathology of Fish and Bees, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova 55, 10 000 Zagreb, Croatia
2
Department of Anatomy, Histology and Embryology, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova 55, 10 000 Zagreb, Croatia
3
Division of Molecular Medicine, Ruđer Bošković Institute, Bijenička Cesta 54, 10 000 Zagreb, Croatia
*
Author to whom correspondence should be addressed.
†
Retired.
Animals 2024, 14(19), 2798; https://doi.org/10.3390/ani14192798
Submission received: 31 July 2024 / Revised: 8 September 2024 / Accepted: 25 September 2024 / Published: 27 September 2024
(This article belongs to the Section Aquatic Animals)
Browse Figures
Versions Notes

Abstract

:

Simple Summary

The European barracuda (Sphyraena sphyraena) is a wild fish species that is both commercially and ecologically valuable and is found in the Adriatic Sea. To better understand its distribution and physiology, this study examined the digestive enzymes in various parts of the barracuda’s gastrointestinal system using histochemical analyses. This is the first known study to evaluate the activity of aminopeptidase, alkaline phosphatase, acid phosphatase, and non-specific esterase in the pyloric caeca and the anterior, middle, and posterior segments of the European barracuda’s gastrointestinal tract.

Abstract

In this study, we examined the gastrointestinal tract of the European barracuda (Sphyraena sphyraena) living in the Adriatic Sea near Dubrovnik, Croatia. The study aimed to identify the presence and distribution of four digestive enzymes: alkaline phosphatase, aminopeptidase, acid phosphatase, and non-specific esterase. We found that alkaline phosphatase activity was present in the brush border of the enterocytes in all the investigated intestinal segments. The activity of the alkaline phosphatase was the strongest in the pyloric caeca but strong only in the basal part of the intestinal villi in the anterior and middle intestinal segments. In the posterior intestinal segment, alkaline phosphatase had strong activity along the entire villi. The activity of acid phosphatase was weak in all the investigated parts of the intestine, except in the posterior part, where it was moderate. Aminopeptidase was detected in the brush border of the intestinal epithelium, with stronger activity in the pyloric caeca and anterior part of the intestine and weaker activity posteriorly. The activity of the non-specific esterase was moderate in the pyloric caeca and anterior part of the intestine, while it was weak in the posterior segment and the lamina propria in all parts of the digestive tract. Weak acid phosphatase activity was observed only in the lamina propria of the posterior part of the intestine. This study is the first to evaluate the activity of digestive enzymes in the European barracuda.

1. Introduction

Within the family Barracudas (Sphyraenidae), there are 28 different species [1] varying in size from less than 50 cm to 1.8 m in body length and weighing eight kilograms [2]. The European barracuda or Mediterranean barracuda (Sphyraena sphyraena) is an elongated pelagic and predatory fish with two separated dorsal fins, a pointed head, a large mouth, and sharp protruding teeth. This species is globally distributed but mainly inhabits the Mediterranean basin [3,4,5]. Barracudas are popular as food for humans and have gained huge fishery importance. Kalogirou et al. (2012) investigated the feeding habits of the European barracuda originating from the eastern Mediterranean Sea. They concluded that the diet of this fish species is based on the fishes (99%), mainly on pelagic and supra-benthic species, and rarely on cephalopods [6].
The fish gastrointestinal tract’s main functions are digestion, absorption, and transportation of the digested nutrients [7]. Changes in the dietary components with relatively broad diets can modulate fish digestive enzyme activities [8]. The utilization of nutrients depends on the digestive enzyme activity in the proper micro locations along the lumen of the gastrointestinal tract [9,10,11,12,13,14], and it could vary with feeding habits [15], taxonomy [16], pH values [17], and aging [18]. The reproduction success, survival, and growth rates depend on the incoming energy, which is represented by the feeding activity and nutrient quantity and quality. The degree of nutrition digestibility could be explained by understanding the types and functions of digestive enzymes [19,20]. Also, digestive enzymes may be used as an indicator of food acceptance, digestive capacity, and development rate, as well as for survival [21,22]. The digestive enzymes physiology indicates the correlation between their activity and feeding ecology [14].
Previously published study results have described this fish species’ morphology, bite mechanism, predator–prey interactions, length–weight relationships in different microenvironments, behavior and adaptation patterns, and mitogenomic architecture, as well as parasitological examinations and consumers’ risk health assessments with heavy metals linked with fish meat consummation [6,23,24,25,26,27,28,29,30].
To understand fish digestive physiology, histochemical analyses of the gastrointestinal tract are necessary, and currently, for European barracuda, this knowledge gap exists. The present study aimed to determine the histochemical localization and the possible role of four digestive enzymes, namely aminopeptidase (AP), alkaline phosphatase (ALP), acid phosphatase (ACP), and non-specific esterase (NSE), on the intracellular digestion and transport of digested nutrients in the pyloric caeca and anterior, middle, and posterior segments of the intestine in the European barracuda. This study for the first time presents novel information about the nutritional biology of these commercially and ecologically very interesting species, important for their management and conservation in the Adriatic Sea.

2. Materials and Methods

This study was carried out on the free-living adult European barracuda (S. sphyraena Linnaeus, 1758), caught with fish for commercial purposes during morning hours, near the town of Dubrovnik, in Croatia (Figure 1). The total length (TL) of the investigated fish (n = 12) was from 20.5 to 30.5 cm, and body weight (BW) was from 34 to 75 g.
Immediately after catching, fish were held at a lower temperature on ice, and the dissection was accomplished. Samples of the three segments of the fish specimen intestine (anterior, middle, and posterior) and pyloric caeca were removed (Figure 2) and fixed in cold (4 °C) formol calcium for 24 h and then transferred to an ice-cold sucrose solution. The mentioned tissues were sectioned with a cryocut (Leica Cryocut 1800 Cryostat, Nussloch, Germany) into 10 μm thick slides and used for histochemical detection of enzymatic activity. Histochemical techniques for detecting enzymatic activities were taken from Pearse (1968) [31] and for the AP supstrate-L-Leu-4-MNA (pH 6.5). For the enzymes ALP (E.C.3.1.3.1)-substrate, sodium β-glycerolphosphate (pH 9.4); ACP (E.C.3.1.3.2.)-substrate, sodium β-glycerolphosphate (pH 5.5) and NSE (E.C.3.1.1.)-substrate 1-naphthyl-acetate (pH 6.5), the method according to the Loyda et al. (1979) was used [32]. AP was incubated for 60 min at the temperature of 37 °C. ACP was incubated for 60 min and ALP and NSE for 30 min, at room temperature. After incubation, the sections were rinsed and mounted in glycerin jelly (Canada balsam).
According to the intensity of the color reaction, the enzymatic activity was analyzed visually, using a light microscope (Olympus BX41 phase contrast microscope; Olympus Corporation, Tokyo, Japan) and photographed with an Olympus DP12 U-TVO camera (Olympus Corporation, Tokyo, Japan). All stained sections were examined, with at least ten visual microscopic fields per slide and ten different slides of the same anatomical area, under 10× to 40× magnification. Observations of enzymatic activity were evaluated according to its staining intensity through the adoption of the score ranging from 1 to 4 plus signs and described as very strong reaction (++++), strong reaction (+++), moderate reaction (++), and weak (barely detectable reaction) (+). A score of ++++ was given to those with the highest enzyme activity demand of crucial relevance following the experience of the researcher. Two researchers performed all microscopic observations. To avoid bias, all scores were provided without allowing the view of the scores given by another researcher. The results were calculated from scores received and sorted according to the relevant criteria.

3. Results

3.1. Morphology and Enzyme Histochemistry

The pyloric caeca and intestine exhibited a uniform histological structure throughout its entire length in all the investigated fish specimens. The mucosa was thrown into many thin folds and contained only two types of cells, columnar epithelial and goblet cells. The numerous goblet cells were distributed across the whole length of the intestine with the lowest number in the pyloric caeca. The major part of the mucosa was formed of columnar epithelial cells. The intestinal layer called tunica muscularis includes the internal circular muscle layer and the external longitudinal muscle bundles. Enzymatic activity in the epithelium of pyloric caeca and all segments of the intestine were related to the columnar epithelial cells.

3.2. Aminopeptidase Histochemistry and Distribution

Aminopeptidase activity was observed as present in all investigated parts of the gastrointestinal tract. It was visible along the intestinal epithelium brush border. The intensity of AP activity was strong at the top of the absorptive cells in the pyloric caeca (Figure 3a) and in the anterior part of the intestine (Figure 3b). The enzymatic activity of AP was moderately intense in the middle intestine (Figure 3c) and barely detectable in the posterior part of the intestine (Figure 3d).

3.3. Alkaline Phosphatase Histochemistry and Distribution

Intestinal ALP was detected in the brush border of the enterocytes in all the investigated intestinal segments. This activity was very strong in the pyloric caeca (Figure 4a). In the anterior and middle segments of the intestine, activity was strong just on the basal part of the intestinal villi (Figure 4b,c). In the posterior intestinal segment, ALP activity was strong along the whole villi (Figure 4d) and weak in the goblet cells (Figure 4d).

3.4. Acid Phosphatase Histochemistry and Distribution

The activity of ACP was detected as a fine-granular reaction product in the supranuclear region of enterocytes (Figure 5a–c). The activity was weak in all the investigated parts of the digestive tract, except in the posterior part of the intestine, where it was moderate with weak enzyme activity in the lamina propria (Figure 5c).

3.5. Non-Specific Esterase Histochemistry and Distribution

The activity of NSE was found in the cytoplasm of enterocytes in the pyloric caeca and all the examined intestinal segments. The activity was moderate in the pyloric caeca and the anterior segment of the intestine (Figure 6a,b) with weak activity in the posterior segment (Figure 6c). Weak enzymatic activity was seen in the lamina propria of all the examined parts of the digestive tract (Figure 6a–c).

4. Discussion

The utilization of nutrients in the gastrointestinal tract of the European barracuda depends on the fish’s age [18]. The growth rate is highest during the first year of life but decreases later [33]. According to measurements of the body weight and length of the S. sphyraena in the investigations of Allam et al. (2004) [34], we could estimate that our investigated fish were between one and two years old.
The pyloric caeca were in the form of multiple finger pouches, which are often present in fish. Through increasing the intestine surface [35], pyloric caeca has an important role in digestion and absorption. Many fish do not have a pyloric caecum. The pyloric caeca and intestine of the S. sphyraena showed a uniform histological structure throughout the entire length. The tunica muscularis in the S. sphyraena includes the internal circular muscle layer and the external longitudinal muscle bundles.
In our study, goblet cells were distributed along the whole length of the pyloric caeca, and it was previously realized that they secrete mucous for the protection of gut epithelium and enhancement of nutrition substances transportation into the next parts of a gastrointestinal tube [36]. Cho et al. (2023) suggested that a huge number of goblet cells have the function of lubrication in the esophagus of marbled flounder (Pseudopleuronectes yokohamae) [37]. Many goblet cells were found throughout all segments of the intestine, like predator fishes such as S. dumerili [38]. At the posterior part of the intestine, most herbivorous fishes have a small number of goblet cells [35,39]. In general, mucus substances that are produced by the goblet cells have the function of moistening for facilitating the excretion and protecting the tunica mucosa against parasites, chemicals, and acidity [40,41,42].
Alkaline digestion and nutrient absorption occur in the intestine [43]. From the anterior intestine, the remnants of food particles that were not absorbed migrate into the intermediate and posterior intestines, where the absorption process continues [44]. The possibility to predict the ability of a fish to use different nutrients from the profile of digestive enzymes is suggested by Hofer and Köck (1989) [45]. The intensity and distribution of digestive enzymes vary with intestinal morphology and feeding habits [46]. For digestion, the food must be in direct contact with the digestive enzymes for a certain period. Lower enzyme activity can be compensated with prolonged food retention in the digestive tract [10].
The brush border structure of the marine teleost is linked with the presence of peptidase enzymes, which are important for maximizing the digestion and absorption processes [47]. According to Bates et al. (2007) [48], intestinal AP in zebrafish prevents inflammation caused by the gut microbiota by detoxifying the lipopolysaccharides. Gylfason et al. (2010) found that the intestinal enterocytes of the Gadus morhua AP were enriched with lipid raft fractions [49]. The activity of AP in the European barracuda was visible along the intestinal epithelium brush border with strong intensity in the pyloric caeca and the anterior part of the intestine. Moderately intense AP activity was detected in the middle intestine, and it was low in the posterior part of the intestine, suggesting that these segments may also be responsible for peptide digestion. The European barracuda is a predatory fish, and its food is largely made of proteins [6]. The protein content in its diet is probably responsible for the higher activity of AP, an enzyme important for the digestion and absorption of proteins. This enzyme catalyzes the hydrolysis of phosphoric acid monoesters from carbohydrates, fats, and proteins in an alkaline medium and plays a role in the metabolism of calcium, phosphorus, and fatty acids. Also, it participates in regulating pH values in the lumen of the intestine, detoxifying inflammatory components, and modulating the composition of the intestinal microflora [50].
Intestinal ALP is a primary brush border enzyme, and it is important in the absorption of nutrients such as glucose, lipids, inorganic phosphate, and calcium [51]. It also plays many roles in the intestine such as pH regulation, fat acquisition, and anti-inflammatory response, as well as the regulation of the gut microbiome and homeostatic enzymes [52,53,54,55]. ALP contains hydrolase, which is most effective in an alkaline environment [56]. More than half of the reviewed publications reported the highest ALP activity in the anterior part, moderate in the middle, and lowest in the posterior segment of the intestine in both juvenile and older fish [50]. The activity of ALP in European barracuda was strong along all parts of the intestine and very strong in the pyloric caeca. In the anterior and middle segments of the intestine, the activity was strong just on the base of the intestine villi. In these segments, the food probably stands for a longer time than in the posterior intestinal segment in which ALP activity was strong along the whole villi. These results indicate that intracellular protein decomposition occurs along the entire intestine. Higher ALP activity across the intestine has also been reported in farmed Anguilla anguilla [12]. In the European barracuda weak activity of ALP was visible in the goblet cells of the posterior part of the intestine which was in accordance with the study which also mentioned ALP activity in the goblet cells of fish [50].
ACP is a lysosomal enzyme that hydrolyzes organic phosphates in an acidic pH [56]. The ACP is one of the marker enzymes for lysosomes, and the activity of this enzyme is related to intracellular digestion [12,13]. Also, it can be linked with pinocytic activity, which could be an alternative pathway for protein digestion [57]. In our study, ACP was detected as a fine granular reaction product in the supranuclear region of enterocytes, which agrees with the previous studies on Merluccius merluccius [9], A. anguilla [12], Diplodus vulgaris [13], and Cynoglossus semilaevis [56]. ACP in the European barracuda was weak in all the examined segments of the intestine and moderate in the macrophages of the lamina propria in the posterior segment. In the posterior part of the intestine, ACP activity was also visible in the lamina propria of Hippoglossus hippoglossus [58]. Wang et al. (2019) found a similar condition in Cynoglossus semilaevis, which coincided with the presence of pinocytotic processes [56]. The presence of proteins within vesicles of the enterocyte’s cytoplasm in the posterior parts of the fish intestine may be an explanation for the stronger enzymatic activity of ACP in these segments [40,59].
NSE is involved in the digestion of glycerol esters of fatty acids [60]. These enzymes may be of particular importance for fish, which can utilize lipids [13]. The intestinal activity of NSE in fishes may influence different feeding styles and environmental conditions [56]. The activity of NSE in the European barracuda was detected in the cytoplasm of enterocytes in the pyloric caeca and in all intestinal segments, which comply with the studies on Lepidotrigla Cavillone [11], Diplodus vulgaris [13], Piaractus mesopotamicus [61], and Cyprinus carpio var. koi [62]. The activity of NSE in the European barracuda was moderate in the pyloric caeca and the anterior part of the intestine. In the posterior segment, the activity of the NSE was weak. Similar findings were observed in the wild A. anguilla [12]. The weak NSE activity in the posterior part of the intestine leads us to the conclusion that the hydrolysis of carboxylic esters (by non-specific esterase) occurs to a small degree in these parts of the intestine in the European barracuda. The weak activity of the NSE was found in the lamina propria of all the investigated parts of the digestive tract.
In the European barracuda, the final stage of digestion occurs in the enterocytes and depends on the activity of digestive enzymes, which showed segmental variation in the activity. In the pyloric caeca, the activity of ALP was very strong, AP activity was strong, that of NSE was moderate, and ACP activity was weak. The strong activity of the AP and ALP was found in the anterior part of the intestine with weak activity of the ACP and NSE. In the middle segment, the activity of the ALP was strong, AP activity was moderate, and those of NSE and ACP were weak. The activity of the ALP in the posterior segment was strong, the ACP activity was moderate, and those of NSE and AP were weak. Based on the obtained results, we can conclude that the pyloric caeca and all parts of the intestine of the European barracuda are very convenient for the absorption of nutrients such as glucose, lipids, and inorganic phosphate. The digestion and absorption of proteins decrease toward the posterior intestine. In the posterior part of the intestine, the digestion of glycerol esters of fatty acids is a little more intensive than in the other parts of the intestine.
As a recommendation to ensure consistency and reproducibility in the scoring of microscopic slide staining intensity, computer software can be used for quantitative analysis. To the best of our knowledge, this is the first study evaluating the activity of the main digestive enzymes in the described fish species.

Author Contributions

Conceptualization, Z.K.; methodology, Z.K.; investigation, I.T.G., Z.K., S.N. and J.V.; resources, Z.K.; S.N. and J.V.; writing—original draft preparation, S.N. and I.T.G.; writing—review and editing, I.T.G., Z.K., S.N. and J.V.; visualization, I.T.G. and Z.K.; funding acquisition, Z.K.; J.V. and S.N. All authors have read and agreed to the published version of the manuscript.

Funding

This study was supported by the Ministry of Science, Education and Sport of Croatia (project No. 053-0010501-2107).

Institutional Review Board Statement

The study protocol was approved by the Ethics Committee of the Faculty of Veterinary Medicine University of Zagreb (no. 640-01/21-02/09; 1 October 2021).

Informed Consent Statement

Not applicable.

Data Availability Statement

Data are presented in this article.

Acknowledgments

The authors thank Nada Crnogaj and Gordana Husinec for their technical support.

Conflicts of Interest

The authors declare no conflicts of interest.

References

  1. Fricke, R.; Eschmeyer, W.; Fong, J.D. Species by Family/Subfamily in Eschmeyer’s Catalog of Fishes. Available online: http://researcharchive.calacademy.org/research/ichthyology/catalog/SpeciesByFamily.asp (accessed on 7 May 2020).
  2. Zonn, I.S.; Kostianoy, A.G.; Semenov, A.V.; Joksimović, A.; Đurović, M. European Barracuda or Mediterranean Barracuda (Sphyraena sphyraena). In The Adriatic Sea Encyclopedia. Encyclopedia of Seas; Springer: Cham, Switzerland, 2021. [Google Scholar] [CrossRef]
  3. Osman, H.M.; El Ganainy, A.A.; Amin, A.M. A study on Diet Composition and Feeding Habits of Barracuda fish (Sphyraena chrysotaenia and S. flavicauda) in the Gulf of Suez. Egypt. J. Aquat. Biol. Fish 2019, 23, 223–232. [Google Scholar] [CrossRef]
  4. Aggrey-Fynn, J.; Fynn-Korsah, S.; Appiah, N. Length-Weight Relationships and Food Preference of Two Coastal Marine Fishes, Galeoides decadactylus (Polynemidae) and Sphyraena sphyraena (Sphyraenidae) off Cape Coast, Ghana, West African. J. Appl. Ecol. 2013, 21, 87–96. [Google Scholar]
  5. Mohammadizadeh, F.; Valinassab, T.; Jamili, S.; Matinfar, A.; Bahri-Shabanipour, A.H.; Mohammadizadeh, M. A Study on Diet Composition and Feeding Habitats of Sawtooth Barracuda (Sphyraena putnamae) in Bandar-Abbas (North of Persian Gulf). J. Fish Aquat. Sci. 2010, 5, 179–190. [Google Scholar]
  6. Kalogirou, S.; Mittermayer, F.; Pihl, L.; Wennhage, H. Feeding ecology of indigenous and non-indigenous fish species within the family Sphyraenidae. J. Fish Biol. 2012, 80, 2528–2548. [Google Scholar] [CrossRef]
  7. Wilson, J.M.; Castro, L.F.C. Morphological diversity of the gastrointestinal tract in fishes. In The Multifunctional Gut of Fish, 1st ed.; Grosell, M., Farrell, A.P., Brauner, C.J., Eds.; Academic Press: Boston, MA, USA, 2011; Volume 30, pp. 2–44. ISBN 9780080961361. [Google Scholar]
  8. German, D.P.; Horn, M.H.; Gawlicka, A. Digestive enzyme activities in herbivorous and carnivorous prickleback fishes (Teleostei: Stichaeidae): Ontogenetic, dietary, and phylogenetic effects. Physio. Biochem. Zool. 2004, 77, 789–804. [Google Scholar] [CrossRef] [PubMed]
  9. Kozarić, Z.; Kužir, S.; Nejedli, S.; Petrinec, Z.; Srebočan, E. Histochemical distribution of digestive enzymes in hake, Merluccius merluccius L. 1758. Vet. Arhiv 2004, 74, 299–308. [Google Scholar]
  10. Furné, M.; Hidalgo, M.C.; Lopez, A.; Garcia-Gallego, M.; Morales, A.E.; Domezain, A.; Domezaine, J.; Sanz, A. Digestive enzyme activities in Adriatic sturgeon Acipenser naccarii and rainbow trout Oncorhynchus mykiss, a comparative study. Aquaculture 2005, 250, 391–398. [Google Scholar] [CrossRef]
  11. Kozarić, Z.; Petrinec, Z.; Kužir, S.; Gjurčević, E.; Baždarić, B. Histochemical Analyses of Digestive Enzymes in the Intestine of Adult Large-Scaled Gurnard (Lepidotrigla Cavillone, Lacepède, 1801). Anat. Histol. Embryol. 2011, 40, 314–320. [Google Scholar] [CrossRef]
  12. Kužir, S.; Gjurčević, E.; Nejedli, S.; Baždarić, B.; Kozarić, Z. Morphological and histochemical study of intestine in wild and reared European eel (Anguilla anguilla L.). Fish. Physiol. Biochem. 2012, 38, 625–633. [Google Scholar] [CrossRef]
  13. Tlak Gajger, I.; Nejedli, S.; Kozarić, Z. Histochemical Distribution of Digestive Enzymes in the Intestine of the Common Two-Banded Sea Bream, Diplodus vulgaris, Geoffroy St-Hilaire 1817. Anat. Histol. Embryol. 2013, 42, 161–167. [Google Scholar] [CrossRef]
  14. Weinrauch, A.M.; Schaefer, C.M.; Goss, G.G. Activity and post-prandial regulation of digestive enzyme activity along the Pacific hagfish (Eptatretus stoutii) alimentary canal. PLoS ONE 2019, 14, e0215027. [Google Scholar] [CrossRef] [PubMed]
  15. Ray, A.K.; Ringø, E. Aquaculture Nutrition: Gut Health, Probiotics and Prebiotics. In The Gastrointestinal Tract of Fish, 1st ed.; Merrifield, D., Ringø, E., Eds.; Wiley Blackwell: Oxford, UK, 2014; pp. 1–13. ISBN 978-0-470-67271-6. [Google Scholar]
  16. Al Abdulhadi, H.A. Some comparative histological studies on alimentary tract of tilapia fish (Tilapia spilurus) and sea bream (Mylio cuvieri). Egypt. J. Aquat. Res. 2005, 31, 387–397. [Google Scholar]
  17. Solovyeva, M.M.; Kashinskayaa, E.N.; Izvekovab, G.I.; Glupova, V.V. pH Values and Activity of Digestive Enzymes in the Gastrointestinal Tract of Fish in Lake Chany (West Siberia). J. Ichthyol. 2015, 55, 251–258. [Google Scholar] [CrossRef]
  18. Kuz’mina, V.V. Influence of age on digestive enzyme activity in some freshwater teleosts. Aquaculture 1996, 148, 25–37. [Google Scholar] [CrossRef]
  19. Kolkovski, S. Digestive enzymes in fish larvae and juveniles—Implications and applications to formulated diets. Aquaculture 2001, 200, 181–201. [Google Scholar] [CrossRef]
  20. Solovyev, M.; Gisbert, E. Influence of time, storage temperature and freeze/thaw cycles on the activity of digestive enzymes from gilthead sea bream (Sparus aurata). Fish Physiol. Biochem. 2016, 42, 1383–1394. [Google Scholar] [CrossRef]
  21. Wei, L.; Xiu-Mei, Z.; Li-Bo, W. Digestive enzyme and alkaline phosphatase activities during the early stages of Silurus soldatovi development. Zool. Res. 2010, 1, 627–632. [Google Scholar]
  22. Gisbert, E.; Morais, S.; Moyano, F.J. Feeding and Digestion. In Larval Fish Aquaculture; Qin, J.G., Ed.; Nova Publishers: New York, NY, USA, 2013; pp. 73–124. ISBN 978-1-62417-899-3. [Google Scholar]
  23. Wadie, W.F.; Dowidar, N.M.; Rizkalla, S.I. Morphometric variations of the family Sphyraenidae from the South-eastern part of the Mediterranean Sea. Folia Morphol. 1987, 35, 124–132. [Google Scholar]
  24. Grubich, J.R.; Rice, A.N.; Westneat, M.W. Functional morphology of bite mechanics in the great barracuda (Sphyraena barracuda). Zoology 2008, 111, 16–29. [Google Scholar] [CrossRef]
  25. Fanti, F.; Minelli, D.; Conte, G.L.; Miyashita, T. An exceptionally preserved Eocene shark and the rise of modern predator-prey interactions in the coral reef food web. Zool. Lett. 2016, 2, 9. [Google Scholar] [CrossRef]
  26. Hosseini, A.; Kochanian, P.; Marammazi, J.; Yavari, V.; Savari, A.; Salari-Aliabadi, M.A. Length-weight relationship and spawning season of Sphyraena jello C., from Persian Gulf. Pak. J. Biol. Sci. 2009, 12, 296–300. [Google Scholar] [CrossRef] [PubMed]
  27. Hanafy, B.G. Gross morphological and surface ultrastructural investigation on the gills of the European barracuda Sphyraena sphyraena. Microsc. Res. Tech. 2022, 85, 917–926. [Google Scholar] [CrossRef] [PubMed]
  28. Kundu, S.; Kim, H.-W.; Lee, J.; Chung, S.; Lee, S.R.; Gietbong, F.Z.; Wibowo, A.; Kang, K. Mitogenomic Architecture and Phylogenetic Relationship of European Barracuda, Sphyraena sphyraena (Teleostei: Sphyraenidae) from the Atlantic Ocean. Fishes 2023, 8, 573. [Google Scholar] [CrossRef]
  29. Corner, R.D.; Cribb, T.H.; Cutmore, S.C. A new genus of Bucephalidae Poche, 1907 (Trematoda: Digenea) for three new species infecting the yellowtail pike, Sphyraena obtusata Cuvier (Sphyraenidae), from Moreton Bay, Queensland, Australia. Syst. Parasitol. 2020, 97, 455–476. [Google Scholar] [CrossRef] [PubMed]
  30. Nyarko, E.; Boateng, C.M.; Asamoah, O.; Edusei, M.O.; Mahu, E. Potential human health risks associated with ingestion of heavy metals through fish consumption in the Gulf of Guinea. Toxicol. Rep. 2023, 10, 117–123. [Google Scholar] [CrossRef]
  31. Pearse, A.G.E. Histochemistry, Theoretical and Applied, 3rd ed.; J. and A. Churchill Ltd.: London, UK, 1968; ISBN 0700013261. [Google Scholar]
  32. Loyda, Z.; Gossrau, R.; Schiebler, T.H. Enzyme Histochemistry. A Laboratory Manual, 1st ed.; Springer: New York, NY, USA, 1979; pp. 53–243. ISBN 978-3-540-09269-8. [Google Scholar]
  33. Wadie, W.F.; Reskalla, S.I.; Dowidar, N.M. Age and growth studies of the Sphyraenidae family in the southeastern Mediterranean based on otolith measurements. Folia Morphol. 1989, 37, 38–56. [Google Scholar]
  34. Allam, S.M.; Faltas, S.N.; Ragheb, E. Age and growth of barracudas in the Egyptian Mediterranean waters. Egypt. J. Aquat. Res. 2004, 30, 281–289. [Google Scholar]
  35. Purushothaman, K.; Lau, D.; Saju, J.M.; Musthaq, S.K.S.; Lunny, D.P.; Vij, S.; Orbán, L. Morpho-histological characterization of the alimentary canal of an important food fish, Asian seabass (Lates calcarifer). PeerJ 2016, 24, e2377. [Google Scholar] [CrossRef]
  36. Løkka, G.; Austbø, L.; Falk, K.; Bjerkås, I.; Koppang, E.O. Intestinal morphology of the wild Atlantic salmon (Salmo salar). J. Morphol. 2013, 274, 859–876. [Google Scholar] [CrossRef]
  37. Cho, J.-H.; Park, J.W.; Ryu, Y.-W.; Kim, K.-W.; Hur, S.-W. Morphology, Histology, and Histochemistry of the Digestive Tract of the Marbled Flounder Pseudopleuronectes yokohamae. Animals 2023, 13, 936. [Google Scholar] [CrossRef]
  38. Grau, A.; Crespo, S.; Sarasquete, M.C.; De Canales, M.L.G. The digestive tract of the amberjack Seriola dumerili, Risso: A light and scanning electron microscope study. J. Fish Biol. 1992, 41, 287–303. [Google Scholar] [CrossRef]
  39. Amer, F.I.; Naguib, S.A.A.; El Ghafar, F.A.A. Comparative study on the intestine of Schilbe mystus and Labeo niloticus in correlation with their feeding habits. J. Am. Sci. 2011, 7, 465–484. [Google Scholar] [CrossRef]
  40. Murray, H.M.; Wright, G.M.; Goff, G.P. A comparative histological and histochemical study of the post-gastric alimentary canal from three species of pleuronectid, the Atlantic halibut, the yellowtail flounder and the winter flounder. J. Fish Biol. 1996, 48, 187–206. [Google Scholar]
  41. Petrinec, Z.; Nejedli, S.; Kužir, S.; Opačak, A. Mucosubstances of the digestive tract mucosa in northern pike (Esox lucius L.) and European catfish (Silurus glanis L.). Vet. Arhiv. 2005, 75, 317–327. [Google Scholar]
  42. Cardoso, N.D.N.; Firmiano, E.M.D.S.; Gomesna, I.D.; Scimento, A.A.D.; Sales, A.; Araujo, F.G. Histochemical and immunohistochemical study on endocrine cells (5HT, GAS and SST) of the gastrointestinal tract of a teleost, the characin Astyanax bimaculatus. Acta Histochem. 2015, 117, 595–604. [Google Scholar] [CrossRef]
  43. Canan, B.; Da Nascimento, W.S.; Da Silva, N.B.; Chellappa, S. Morphohistology of the digestive tract of the damsel fish Stegastes fuscus (Osteichthyes: Pomacentridae). Sci. World J. 2012, 3, 787316. [Google Scholar] [CrossRef]
  44. Nasruddin, N.S.; Azmai, M.N.A.; Ismail, A.; Saad, M.Z.; Daud, H.M.; Zulkifli, S.Z. Histological Features of the Gastrointestinal Tract of Wild Indonesian Shortfin Eel, Anguilla bicolor bicolor (McClelland, 1844). Sci. World J. 2014, 2014, 312670. [Google Scholar] [CrossRef]
  45. Hofer, R.; Köck, G. Method for quantitative determination of digestive enzymes in fish larvae. Pol. Arch. Hydrobiol. 1989, 36, 439–441. [Google Scholar]
  46. Gawlicka, A.; Herold, M.A.; Barrows, F.T.; De La Noṻe, J.; Hung, S.S.O. Effects of dietary lipids on growth, fatty acid composition, intestinal absorption and hepatic storage in white sturgeon (Acipenser transmontanus R.) larvae. J. Appl. Ichthyol. 2002, 18, 673–681. [Google Scholar] [CrossRef]
  47. Infante, J.L.Z.; Cahu, C.L. Ontogeny of the gastrointestinal tract of marine fish larvae. Comp. Biochem. Phys. Part C Tox. Pharmacol. 2001, 130, 477–487. [Google Scholar]
  48. Bates, J.M.; Akerlund, J.; Mittge, E.; Guillemin, K. Intestinal alkaline phosphatase detoxifies lipopolysaccharide and prevents inflammation in zebra fish in response to the gut microbiota. Cell Host Microbe 2007, 2, 371–382. [Google Scholar] [CrossRef] [PubMed]
  49. Gylfason, G.A.; Knútsdóttir, E.; Ásgeirsson, B. Isolation and biochemical characterization of lipid rafts from Atlantic cod (Gadus morhua) intestinal enterocytes. Comp. Biochem. Physiol. B BMB 2010, 155, 86–95. [Google Scholar] [CrossRef]
  50. Lallès, J.P. Intestinal alkaline phosphatase in the gastrointestinal tract of fish: Biology, ontogeny, and environmental and nutritional modulation. Rev. Aquacult. 2019, 2, 555–581. [Google Scholar] [CrossRef]
  51. Dupuis, Y.; Tardivel, S.; Porembska, Z.; Fournier, P. Effect of some phosphatase inhibitors on intestinal calcium transfer. Int. J. Biochem. 1991, 23, 175–180. [Google Scholar] [CrossRef] [PubMed]
  52. Yang, Y.Y.; Wandler, A.M.; Postlethwait, J.H.; Guillemin, K. Dynamic Evolution of the LPS-Detoxifying Enzyme Intestinal Alkaline Phosphatase in Zebrafish and Other Vertebrates. Front. Immunol. 2012, 3, 314. [Google Scholar] [CrossRef] [PubMed]
  53. Lallès, J.P. Intestinal alkaline phosphatase: Novel functions and protective effects. Nutr. Rev. 2014, 72, 82–94. [Google Scholar] [CrossRef] [PubMed]
  54. Rader, B.A. Alkaline phosphatase, an unconventional immune protein. Front. Immunol. 2017, 8, 897. [Google Scholar] [CrossRef]
  55. Lallès, J.P. Intestinal alkaline phosphatase, inflammation and nutrition: Recent advances. Nutr. Rev. 2019, 77, 710–724. [Google Scholar] [CrossRef]
  56. Wang, Y.Z.; Sun, J.F.; Lv, A.J.; Zhang, S.L.; Sung, Y.Y.; Shi, H.Y.; Hu, X.C.; Chen, S.J.; Xing, K.Z. Histochemical distribution of four types of enzymes and mucous cells in the gastrointestinal tract of reared half-smooth tongue sole Cynoglossus semilaevis. J. Fish Biol. 2017, 92, 3–16. [Google Scholar] [CrossRef]
  57. Ribeiro, L.; Sarasquete, C.; Dinis, M.T. Histological and histochemical development of the digestive system of Solea senegalensis (Kaup 1858) larvae. Aquaculture 1999, 171, 293–308. [Google Scholar] [CrossRef]
  58. Grove, S.; Johansen, R.; Reitan, L.J.; Press, C.M. Immune-and enzyme histochemical characterization of leukocyte populations within lymphoid and mucosal tissues of Atlantic halibut (Hippoglossus hippoglossus). Fish Shellfish Immunol. 2006, 20, 693–708. [Google Scholar] [CrossRef] [PubMed]
  59. Georgopoulou, U.; Dabrowski, K.; Sire, M.F.; Vernier, J.M. Absorption of intact proteins by the intestinal epithelium of trout, Salmo gairdnera. A luminescence enzyme immunoassay and cytochemical study. Cell Tissue Res. 1986, 251, 145–152. [Google Scholar] [CrossRef] [PubMed]
  60. Chakrabarti, I.; Gani, M.A.; Chaki, K.K.; Sur, A.; Misra, K.K. Digestive enzymes in 11 freshwater teleost fish species in relation to food habit and niche segregation. Comp. Biochem. Physiol. Part A 1995, 112, 167–177. [Google Scholar] [CrossRef]
  61. Moretti, D.; Nordi, B.; Cruz, T.M.P.; Cyrino, J.E.P.; Machado-Neto, R. Histochemical distribution of intestinal enzymes of juvenile pacu (Piaractus mesopotamicus) fed lyophilized bovine colostrum. Fish Physiol. Biochem. 2014, 40, 1487–1493. [Google Scholar] [CrossRef]
  62. Sun, J.; Wang, Y.; Lv, A.; Xian, J.A.; Wang, Q.; Zhang, S.; Guo, Y.; Xing, K. Histochemical distribution of four types of enzymes and mucous cells in the intestine of koi carp (Cyprinus carpio var. koi). Fish Physiol. Biochem. 2019, 45, 367–1376. [Google Scholar] [CrossRef]
Animals 14 02798 g001
Figure 1. Specimen of examined European barracuda caught with fish for commercial use, intended for human consumption.
Figure 1. Specimen of examined European barracuda caught with fish for commercial use, intended for human consumption.
Animals 14 02798 g001
Animals 14 02798 g002
Figure 2. Anatomical macro morphology of investigated features of the gastrointestinal tract of European barracuda.
Figure 2. Anatomical macro morphology of investigated features of the gastrointestinal tract of European barracuda.
Animals 14 02798 g002
Animals 14 02798 g003
Figure 3. Activity of the aminopeptidase in the gastrointestinal tract of European barracuda: (a) pyloric caeca (+++); (b) anterior part of the intestine (+++); (c) middle part of the intestine (++); (d) posterior part of the intestine (+).
Figure 3. Activity of the aminopeptidase in the gastrointestinal tract of European barracuda: (a) pyloric caeca (+++); (b) anterior part of the intestine (+++); (c) middle part of the intestine (++); (d) posterior part of the intestine (+).
Animals 14 02798 g003
Animals 14 02798 g004
Figure 4. Activity of alkaline phosphatase in the gastrointestinal tract of European barracuda: (a) pyloric caeca (++++); (b) anterior part of the intestine, base part (1) of the intestine villi (+++); (c) middle part of the intestine base part (1) of the intestine villi (+++); (d) posterior part of the intestine (+++), goblet cell (+) (2).
Figure 4. Activity of alkaline phosphatase in the gastrointestinal tract of European barracuda: (a) pyloric caeca (++++); (b) anterior part of the intestine, base part (1) of the intestine villi (+++); (c) middle part of the intestine base part (1) of the intestine villi (+++); (d) posterior part of the intestine (+++), goblet cell (+) (2).
Animals 14 02798 g004
Animals 14 02798 g005
Figure 5. Activity of acid phosphatase in the intestine of European barracuda: (a) anterior part (+); (b) middle part (+); (c) posterior part (++) and in the lamina propria (+).
Figure 5. Activity of acid phosphatase in the intestine of European barracuda: (a) anterior part (+); (b) middle part (+); (c) posterior part (++) and in the lamina propria (+).
Animals 14 02798 g005
Animals 14 02798 g006
Figure 6. Activity of non-specific esterases in the gastrointestinal tract of European barracuda: (a) in pyloric caeca (++), enzyme activity is observed in the lamina propria (+); (b) in anterior part of the intestine (++), enzyme activity is observed in the lamina propria (+); (c) in posterior part of the intestine (+), enzyme activity is observed in the lamina propria (+).
Figure 6. Activity of non-specific esterases in the gastrointestinal tract of European barracuda: (a) in pyloric caeca (++), enzyme activity is observed in the lamina propria (+); (b) in anterior part of the intestine (++), enzyme activity is observed in the lamina propria (+); (c) in posterior part of the intestine (+), enzyme activity is observed in the lamina propria (+).
Animals 14 02798 g006
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Tlak Gajger, I.; Nejedli, S.; Kozarić, Z.; Vlainić, J. Histochemical Analysis and Distribution of Digestive Enzymes in the Gastrointestinal System of the European Barracuda Sphyraena sphyraena (Linnaeus, 1758). Animals 2024, 14, 2798. https://doi.org/10.3390/ani14192798

AMA Style

Tlak Gajger I, Nejedli S, Kozarić Z, Vlainić J. Histochemical Analysis and Distribution of Digestive Enzymes in the Gastrointestinal System of the European Barracuda Sphyraena sphyraena (Linnaeus, 1758). Animals. 2024; 14(19):2798. https://doi.org/10.3390/ani14192798

Chicago/Turabian Style

Tlak Gajger, Ivana, Srebrenka Nejedli, Zvonimir Kozarić, and Josipa Vlainić. 2024. "Histochemical Analysis and Distribution of Digestive Enzymes in the Gastrointestinal System of the European Barracuda Sphyraena sphyraena (Linnaeus, 1758)" Animals 14, no. 19: 2798. https://doi.org/10.3390/ani14192798

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop