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Histological and Histochemical Study on the Alimentary Canal in Walking Catfish (Claris Batrachus) and Piranha (Serrasalmus Nattereri)

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Histological and Histochemical Study on the Alimentary Canal in Walking Catfish (Claris Batrachus) and Piranha (Serrasalmus Nattereri) Iranian Journal of Veterinary Research, Shiraz University, Vol. 11, No. 3, Ser. No. 32, 2010 Histological and histochemical study on the alimentary canal in Walking catfish (Claris batrachus) and piranha (Serrasalmus nattereri) Raji, A. R.1* and Norouzi, E.2 1Department of Basic Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran; 2Graduated from Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran *Correspondence: A. R. Raji, Department of Basic Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran. E-mail: [email protected] (Received 23 Sept 2009; revised version 26 Jan 2010; accepted 17 Apr 2010) Summary The purpose of this study is to describe the histological and histochemical features of digestive tract in two aquarium fishes with different types of feeding habits which are omnivorous (Walking catfish) and carnivorous (piranha), respectively. Five adult Walking catfish (L: 26.4 ± 3.4 cm, W: 265 ± 55 g) and five adult piranha (L: 23.6 ± 4.5 cm, W: 247 ± 50 g) were used for this study. The fishes were killed by decapitation method and small pieces (0.5 cm × 0.5 cm) from the esophagus, cardiac, fundus and pyloric portions of the stomach, proximal, distal parts of intestine and pyloric caeca were fixed by immersion in Bouin’s solution. The specimens were processed through routine paraffin embedding technique and cut at 5-7 µm and stained with hematoxylin and eosin, Van Gieson, Verhoof, Alcian blue, PAS, and AB+PAS technique for different microstructural and microfibrilar structures. The morphological and histochemical structures of all specimens were studied with a light microscope and micrographs were prepared. The esophagus of Walking catfish and piranha are found to have numerous deep longitudinal folds, stratified squamous epithelium with numerous mucous cells which react positively to PAS and AB stains. Taste buds and muscularis mucosa were not seen in the esophagus of both species. The muscularis mucosa was organized in longitudinal and circular layers of striated muscular fibers. The mucosa of the stomach was formed by simple columnar epithelium with folds and gastric pits formed by the invagination of the mucosal layer into the lamina propria. The surface epithelium of the stomach reacted positively to PAS but negatively to AB staining. The mucosal surface of the intestine in Walking catfish and piranha has numerous folds lined by simple tall columnar cells, along with goblet cells which reacted positive to PAS and AB staining, so that goblet cells in the intestine of Walking catfish were significantly more than in piranha. The Caeca pyloric was found only in piranha and it showed an elongated fold lined with columnar cells and numerous mucous cells which reacted positively to PAS. Key words: Histology, Digestive tract, Walking catfish, Piranha Introduction in fish species (Hale, 1965; Caceci, 1984; Caceci and Hrubrec, 1990; Mojazi et al., Fish digestive tracts show remarkable 2005; Khojasteh et al., 2009). The feeding differences in morphology and function. activities of fish are classified most Differences observed at specific levels are commonly according to the type of food related to food, feeding habits, body weight, consumed. In simplest form, fish are either shape and sex (Grosh and Das, 1987; herbivore if they eat plant material, Boglione et al., 1992; Murray et al., 1996). carnivores if they consume animal material Furthermore, the presence of a relation or omnivores if they eat a combination of between feeding behavior and the basic plant and animal materials (Evans, 1998). histology view or the improbability of this The Walking catfish (Clarias batrachus) (Martin and Blaber, 1984) has been argued. is a species of freshwater air breathing The histology of the digestive tract has been catfish found primarily in Southeast Asia, studied with light and electron microscopes although known in English as “Walking 255 Iranian Journal of Veterinary Research, Shiraz University, Vol. 11, No. 3, Ser. No. 32, 2010 catfish”, its “Walk” is often just a sort of chemicals were purchased from sigma wriggling motion with a snakelike chemical company. movement, using its pectoral fins to keep it upright. In the wild, the natural diet habit of Results this creature is omnivorous (Mills, 1993). Piranha (Serrasalmus nattereri), which In the present study the digestive tract of include not only the carnivorous but also Walking catfish and piranha were divided large, peaceful herbivores, live in South into three parts: esophagus, stomach and American rivers (Bailey and Sandford, intestine (Figs. 1 and 2). Although each 1998). portion of the digestive tract has a special Walking catfish has an elongated body structure, the wall of the digestive tract as a shape. This catfish has long-based dorsal whole is composed of the mucosa, tela and anal fins as well as several pairs of submucosa, tunica muscularis and tunica sensory barbells, and the pectoral fins serosa. contain poisonous spines which are The esophagus of Walking catfish was especially stout on the male. Piranha is found to have numerous deep longitudinal snub-nosed and the lower jaw is longer than folds; it was lined by a few layers of the top jaw, and both jaws contain very stratified squamous epithelium with sharp teeth. numerous superficial mucous cells, so that it The main purpose of this study is to changed to simple columnar epithelium at describe the histological and histochemical the end of the esophagus. Taste buds were features of the digestive tract in two absent in the epithelium of the esophagus of aquarium fish with different types of feeding Walking catfish. The lamina propria was habits which are omnivorous (Walking formed by loose connective tissue without catfish) and carnivorous (piranha), elastic fibers. The lamina muscularis mucosa respectively. was not present. The tunica muscularis with striated muscular fibers had two layers; a Materials and Methods thick inner circular and thinner outer longitudinal layer. Outer serosa was a thin Five adult Walking catfish (L: 32 cm, layer of connective tissue surrounded by W: 240 g) and five adult piranha (L: 22 cm, mesothelium. The mucous cells were stained W: 190 g) were used for this study. The with AB and PAS in the lower part of the fishes were killed by decapitation and the mucosal fold, but stained only with AB in entire digestive tract was exposed through a the upper part of the mucosal folds, mid-ventral incision and removed from the indicating the presence of neutral and acidic body. Small pieces from the esophagus, (carboxyl and sulphate groups) glycoprotein cardiac, fundus and pyloric portion of the (Fig. 3). stomach, proximal and distal parts of the The stomach of Walking catfish intestine and pyloric caeca after washing in included the cardiac, fundus and pyloric phosphate buffer were fixed by immersion region. The mucosa of the cardiac and in Bouin’s solution. After fixation, the fundus were formed by a single layer of specimens were rinsed in water, dehydrated columnar epithelium with folds (Fig. 4). in graded ethanol solutions (50% to There were mucosal folds in three portions absolute), cleared in xylene and embedded that were shallow in the pyloric region. in paraffin. Transverse paraffin sections (5-7 There were numerous gastric pits of the µm) of each portion of the digestive tract stomach formed by the invaginations of the were prepared and processed for staining mucosal layer into the lamina propria, which through hematoxylin and eosin (H&E), Van were lined to the opening of tubular Gieson (VG), Verhof (V), Alcian blue (AB), branched glands formed by the columnar Periodic acid Schiff (PAS), AB+PAS (Lee cells (Fig. 4). The columnar epithelial cells and Luna, 1968), and technique micrographs contain a layer of cells with pepsinogen were taken with a video camera connected to granules. In the pyloric region the gastric a microscope (Olympus, Tokyo, Japan). All glands were scarce, but did not disappear 256 Iranian Journal of Veterinary Research, Shiraz University, Vol. 11, No. 3, Ser. No. 32, 2010 completely. Although the lamina muscularis The intestine in Walking catfish had no mucosa was inconspicuous, a few muscular macroscopically differentiable regions. fibers were present and in the pylorus, the However, there were microscopic muscularis mucosa was complete. The differences that characterize the intestine submucosa is formed by loose connective tissue. The surface epithelium of the stomach reacted positively to PAS but was negative to AB. Fig. 1: Walking (a) catfish and piranha (b) Fig. 3: Transverse section of the esophagus in Walking catfish with alcian blue (AB) and periodic acid schiff + AB (PAS+AB) staining. Epithelium (EP), lamina propria (LP), mucosal cells (MC), muscular layer (ML), serosa (S), circle (PAS positive), triangle (AB positive), and rectangle (AB and PAS positive, ×64) Fig. 2: Gross photograph of digestive tract in Fig. 4: Transverse section of the cardiac Walking catfish (a) and piranha (b). region in piranha showing epithelium (EP), Esophagus (E), stomach (S), caeca pyloric (c), gastric pit (GP), lamina propria (LP), mucosal liver (L), proximal intestine (PI), distal layer (ML), sub mucosa (SM), muscularis intestine (DI), and swim bladder (SB) layer (ML) and serosa (S), (PAS+VG, ×320) 257 Iranian Journal of Veterinary Research, Shiraz University, Vol. 11, No. 3, Ser. No. 32, 2010 Fig. 7: Transverse sections of caeca pyloric in piranha showing lumen (L), mucosal fold (MF), epithelium (EP), and columnar cell (CC), lamina propria (LP), muscularis layer (ML), serosa (S), (H&E, ×320) the mucosal surface were stained by AB and PAS (Fig. 5). The absence of muscularis mucosa prevented the distinct separation of lamina propria from the submucosa. The tunica muscularis was organized in two Fig. 5: Transverse section of the proximal distinct layers of smooth muscle; inner intestine in Walking catfish with PAS and AB circular and outer longitudinal layers.
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