Int. J. Aquat. Biol. (2018) 6(3): 114-121 ISSN: 2322-5270; P-ISSN: 2383-0956 Journal homepage: www.ij-aquaticbiology.com © 2018 Iranian Society of Ichthyology Original Article Circulatory system of red tail catfish (Phractocephalus hemioliopterus Bloch & Schneider, 1801): a corrosion cast study Ali Taheri Mirghaed*1, Hooman Rahmati-Holasoo1, Mehrdad Ardeshiri1, Hoseinali Ebrahimzadeh Mousavi1, Javad Sadeghinezhad2 1Department of Aquatic Animal Health, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran. 2Department of Basic Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran. Article history: Received 21 February 2018 Accepted 24 June 2018 Available online 2 5 June 2018 Keywords: Circulatory system Red tail catfish Corrosion cast Physiology Abstract: Red tail catfish, Phractocephalus hemioliopterus, in one of the popular ornamental fish. The present study is aimed to describe and visualizes the cardiovascular system of this species with corrosion cast study method. For this purpose, 10 red tail catfish with 580 gr average weight were obtained and were filled their blood vessels and heart with fluid artificial resin made on the basis of methylmetacrylate after anaesthetizing and euthanizing. For complete polymerization and hardening of the methylmetacrylate, the fish were further submersed for 12-24 hrs in water bath following by 24-48 hrs submersion in a 25% solution of KOH to full maceration. Based on the results we describe the cardiovascular system i.e. the afferent and efferent vessels of gill, different parts of the heart, ventral aorta, dorsal aorta, intestinal and gastric vessels, liver, anterior and posterior parts of the kidneys, spleen, portal and hepatic vein. Introduction The circulatory system plays an important role in many modeling approaches, therefore understanding its anatomy can be useful for many basic and diagnostic studies (Nematollahi et al., 2003, 2005; Noestelthaller et al., 2005; Ghadam et al., 2016). Vascular corrosion casting consists of injecting a liquid polymer into the circulatory system of a whole animal or an organ, allowing it to harden and then corroding away the overlying tissue (typically with potassium hydroxide solution) to produce a 3D cast of the internal structure of the vessels (Hossler et al., 1986, 1991; Zeindler et al., 1989). It will be possible to study the vascular structures in 3D by corrosion cast studies. This would give more information about the complexity of the vascular system in various species (Schmidt et al., 1980). This method has been used in many studies on human and animals. Few studies have been performed on the cardiovascular system of fishes (Seyed Ali et al., 1987; Passantino et al., 2000; Basten et al., 2009) such as light and scanning electron microscopic observations on methyl-methacrylate corrosion casts *Corresponding author: Ali Taheri Mirghaed DOI: https://doi.org/10.22034/ijab.v6i3.445 E-mail address: mirghaed@ut.ac.ir of the blood vessels in the gills of channel catfish (Ictalurus punctatus) (Boland and Olson, 1979), corrosion cast of the entire blood vascular system by a single injection of resin from the heart of a Japanese catfish (Silurus asotus) (Iwamizu and Itazawa, 1986), gill vasculature of yellow stingray (Urolophus jamaicensis) using resin casts under SEM (Sherman and Spieler, 1998), spatial organization of the microcirculation in gills of striped mullet (Mugil cephalus) by scanning electron microscopic analysis of corrosion cast prepared by intravascular injection of methylmethacrylate (Passantino et al., 2000), structural variation in gill vasculature among some elasmobranchs using corrosion casting and SEM (Sherman et al., 2005), vascular pattern of the recirculating system of rainbow trout (Oncorhynchus mykiss) by corrosion cast methodology (Nematollahi et al., 2011), the efferent branchial arteries and splanchnic arteries in the yellow stingray (Urobatis jamaicensis) using SEM to study the vascular corrosion casts (Basten et al., 2009), and comparative study of the circulatory system in common carp (Cyprinus carpio) and beluga (Huso huso) with 115 Int. J. Aquat. Biol. (2018) 6(3): 114-121 emphasis on the heart and main blood vessels, employing the corrosion cast methodology (Ghadam et al., 2016). Red tail catfish, Phractocephalus hemioliopterus is a large Neotropical catfishes of the family Pimelodidae, inhabiting the Amazon and Araguaia- Tocantins River basins. This fish is characterized by having a strong skull ossification, a huge and bony predorsal plate, and red-orange tail and dorsal fins (Goulding, 1980). It has a great economic importance as an ornamental fish, sport fishing or fish production, providing food and a valuable source of income for coastal populations, and also in some regions of the Amazon, its fat and leather extracted are used as folk medicine for treating respiratory problems (Souza et al., 2012). The present study describes and visualizes a 3D view of the circulation system of this fish using the corrosion cast methodology as an important Figure 1. Injection of artificial resin fluid into the heart (a), branchial vessels (b), liver (c), spleen (d), anterior kidney (e) and posterior kidney (f). (L) Liver, (SB) Swim bladder, (ST) Stomach. 116 Taheri Mirghaed et al/ Circulatory system of Phractocephalus hemioliopterus using corrosion cast study ornamental fish. Materials and Methods Ten red tail catfish with average weight of 580 g were obtained from ornamental fish supply stores in Tehran, Iran. Fish were kept for 2 weeks prior to the experiment in 1000-L tanks supplied with filtered, recirculated tap water (23±1°C). Dissolved oxygen and pH were 6.5±0.5 mg/L and 7±0.3, respectively. Wet smears from skin, fins and gills was prepared for parasitological examination under light microscope (Nikon, E600). No ectoparasite were observed. The fish were anesthetized using PI222 (Pars Imen Daru, Iran) in dosage of 100 ppm and transferred to the operating table. To prevent the formation of blood clots in the vessels, the fish were injected intraperitoneally with heparin (5000 Iu/kg) and euthanized with an overdose of the aforementioned anesthetic solution. Then the heart and blood vessels were filled with artificial resin fluid made on the basis of methylmetacrylate. This solution was injected in the heart, branchial vessels, liver, spleen, anterior, and posterior part of the kidney (Fig. 1a-f). Fish were left for 12-24 hrs in a 20-24°C tap water bath to complete polymerization of the injected resin. Then dipped in 25% potassium hydroxide (KOH) for 24-48 hrs to maceration of the tissues (Ghadam et al., 2016). The resin did not dissolve in potassium hydroxide, thus the heart and vascular casts remained in their natural positions. These structures were studied by gross examination and stereomicroscope. Results The results are illustrated in Figures 1-12 to provide information on the cardiovascular system. Different parts of the heart and related vessels, including Atrium (A), Ventricle (V), Bulbus arteriosus (BA) and Ventral aorta (VA) were shown in Figure 2a, b. The VA continues from the heart to the afferent branchial arteries (ABAs) (Fig. 2c). The length of VA is short. In a short distance from the heart, a common trunk (CT) is separated from VA. The common trunk is divided into two branches (ABA 1 and 2) and the rest of VA separately made ABA 3 and 4 (Fig. 2c). The DA continues along the vertebral column and supplied different parts of the body. In a short distance from the beginning of DA, a branch viz. Subclavian artery (SCA) was separated to supply the cranial parts of body such as pectoral fins (Fig. 5). In the middle of the body, it passed through a Longitudinal groove (LG) in WB (Fig. 6a). It supplies this part of the body through some pores (P) in the bones (Fig. 6a, b, c). In caudal area, the DA passed through haemal arches (HA). DA and Caudal vein (CV) which carried the Figure 2. A, B and C: the main parts of the heart and related vessels. A (Atrium), BA (Bulbus arteriosus), CT (Common trunk), V (Ventricle), VA (Ventral aorta) and ABAs (1-4). Each ABAs itself constituted afferent filament arteries (AFAs) (secondary lamella), that provide more surface for oxygen exchange (Fig. 3a and b). 117 Int. J. Aquat. Biol. (2018) 6(3): 114-121 deoxygenate blood, are passed together in HA (Fig. 7). The venous blood from the caudal area such as caudal fin, ribs, trunk muscles and bones is collected by CV. Many small vessels collect the venous blood, forming CV that goes to the anterior parts of the body. CV in its path is unified with the venous vessels from the spleen, stomach, intestine, gall bladder, and mesenteric vein and then carries as Portal vein (PV) to the liver (Fig. 8). The collected blood in the liver is entered to the heart via Hepatic vein (HV) (Fig. 8). The blood from the posterior kidneys is collected by the Renal portal vein (RPV) and then is carried by the right post cardinal vein (RPCV) and left post cardinal vein (LPCV) to the Common cardinal vein (CCV) and then to the heart. RPCV and LPCV are passed inside the bone plates on both right and left sides of the body. In total, RPCV is thicker than LPCV (Fig. 8).On the Figure 3. ABA (Afferent branchial arteries) and AFA (Afferent filament arteries) are seen. After oxygen exchange, ABAs joined together and made Efferent branchial arteries (EBAs). EBAs joined together and formed dorsal aorta (DA) at the beginning of Weber bone (WB) (Fig. 4). Some branches such as Orbital artery (OA) and Encephalic artery (EA) separated from EBAs and supplied blood to different anterior parts of the body. (Fig. 4). Figure 4. Formation of dorsal aorta by joining of EBAs. DA (Dorsal aorta), EBA (Efferent branchial arteries), EA (Encephalic artery) and WB (Weber bone). ABAs and AFAs were removed. Figure 5. DA (Dorsal aorta), EBAs (Efferent branchial arteries), SCA (Subclavian artery) and WB (weber bone) are seen. 118 Taheri Mirghaed et al/ Circulatory system of Phractocephalus hemioliopterus using corrosion cast study other hand, the blood from anterior part of the kidneys (AK) and subclavian vein (SCV) are joined to the RPCV and LPCV entering CCV (Fig. 9). The pecardinal vein (PrCV) transports the major parts of the blood from the head and brain to the heart through Figure 6. LG (longitudinal groove) and WB (Weber bone) and P (pores of the weber bone and other vertebrae). Figure 7. Vessels passing through the haemal arches. CV (Caudal vein), DA (Dorsal aorta), HA (Haemal arch) and S (Spleen). Figure 8. The path of the blood vessels from caudal area to the heart. A (Atrium), ABAs (Afferent branchial arteries), BA (Bulbus arteriosus), CCV (Common cardinal vein), CV (Caudal vein), DA (Dorsal aorta), GV (Gastric vein), HV (Hepatic vein), IV (Intestinal vein), LPCV (Left post cardinal vein), PV (Portal vein), RPCV (Right post cardinal vein), RPV (Renal portal vein), SCV (Subclavian vein) and V (Ventricle). Figure 9. AK (Anterior kidney), CCV (Common cardinal vein, PV (portal vein), PrCV (Precardinal vein), RPCV (Right post cardinal vein), RPV (Renal portal vein) and SCV (Subclavian vein). Figure 10. AK: Anterior kidney, CCV: Common cardinal vein, HV: Hepatic vein, JV: Jugular vein, PrCV: Precardinal vein, RPCV: Right post cardinal vein and PV: portal vein. 119 Int. J. Aquat. Biol. (2018) 6(3): 114-121 CCV. The right PrCV and RPCV are joined as a common part constituting CCV (Figs. 9, 10). Jugular vein (JV) transports the rest part of the blood from the head to CCV separately (Fig. 10). Finally, the collected blood in CCV is transported to the heart on right and left sides. The casts from liver showed wide vessels plexus (Fig. 11a, b, c). Bone plates had a dorso-ventral appearance extending from the head to the anterior part of the tail (Fig. 12). Discussion The corrosion cast study method can be used in pathological malformations and comparative anatomy (Zeindler et al., 1989; Hossler and Douglas, 2001; Nematollahi et al., 2011). In present study, the circulatory system of P. hemioliopterus including the different parts of heart and its vessels such as branchial arteries, dorsal aorta, common cardinal vein and hepatic vein were studied by corrosion cast methodology. We obtained a full casts of blood vessels perfusing various organs such as liver, stomach, spleen, posterior kidney and anterior kidney. According to observations, the length of the ventral aorta was short and after a short distance from the heart, a common trunk is separated from the ventral aorta. This branch is divided into two branches forming the branchial artery 1 and 2. In the following, the branchial artery 3 and 4 divided separately. These observations is correspond with those vascular pattern of common carp, but different with those of beluga and yellow stingray, showing similar pattern in bony fishes. However, there are some structural differences with common carp. The results also showed that the liver has two lobes of the same size locating behind of the heart. The caudal vein joins to the portal vein at the left side. As regards one of the most important problem in this fish is swallowing foreign bodies and subsequently it needs to surgical or non-surgical procedures (Chansue and Tangtrongpiros, 2005; Ebrahimzadeh et al., 2006; Wildgoose, 1998), therefore the results suggests that during surgical procedures it would be better that the incision must be done at right side of midline of this catfish. In most studies, the remained casts from liver show a wide vessel plexus because of high metabolic activity of this organ. According to the result, the right post cardinal vein (RPCV) was thicker than left post cardinal vein (LPCV) in P. hemioliopterus showing that major part of the blood of the renal portal vein is carried by (RPCV) that has not been reported in Figure 11. Vessels plexus in the heart and the liver. (a and b): liver and (c) heart. Figure 12. Dorsal view of the predorsal plate of red tail catfish. 120 Taheri Mirghaed et al/ Circulatory system of Phractocephalus hemioliopterus using corrosion cast study previous studies. However, presentation of a complete blood vessel system using a single corrosion cast can be led artifacts due to the extremely fine capillaries. Furthermore researches are suggested to be done on other catfishes for comparison purpose. Acknowledgments Authors are thank S. Ezhari for assisting with the figures. References Basten B.L., Sherman R.L., Lametschwandtner A., Spieler R.E. (2009). A unique vascular conFiguration among the efferent branchial arteries and splanchnic arteries in the yellow stingray (Urobatis jamaicensis). Microscopy and Microanalysis, 15: 194-196. Boland E.J., Olson K.R. (1979). Vascular organization of the catfish gill filament. Cell and Tissue Research, 198: 487-500. Chansue N., Tangtrongpiros J. (1998). Case report: Surgical removal of plastic plant and clay pot from red tail catfish (Phractocephalus hemioliopterus) stomach. In 24. Thai Veterinary Medical Association and 4. Thailand Veterinary Practitioner Association Meeting, Bangkok (Thailand), 5-7 Aug 1998. Ebrahimzadeh M.H., Vajhi A.R., Hosseini F., Sharifpour I., Tajik P. (2006). Non-surgical removal of some stones from a red tailed catfish (Phractocephalus hemioliopterus) stomach as gastric foreign bodies. Iranian Journal of Fisheries Sciences, 6: 35-42. Ghadam M., Ebrahimzadeh M.H.A., Rahmati‐Holasoo H., Shafiei S. (2016). Comparative study of the circulatory system of common carp (Cyprinus carpio Linnaeus, 1758) and beluga (Huso huso Linnaeus, 1758): a corrosion cast study. Journal of Applied Ichthyology, 32: 11-17. Gouldin M. (1980). The fishes and forest: explorations in Amazonian natural history. University of California Press, Berkeley. 280 p. Hossler F.E., Douglas J.E., Douglas L.E. (1986). Anatomy and morphometry of myocardial capillaries studied with vascular corrosion casting and scanning electron microscopy: a method for rat heart. Scanning Electron Microscopy, 4: 1469-1475. Iwamizu M., Itazawa Y. (1986). Corrosion casting method for the gross anatomy of the blood vascular system of fish. Japanese Journal of Ichthyology, 32: 431-436. Lloyd R., Lloyd C. (2011). Surgical removal of a gastric foreign body in a sand tiger shark, Carcharias taurus Rafinesque. Journal of Fish Disease, 34: 951–953. Nematollahi A., Decostere A., Pasmans F., Ducatelle R., Haesebrouck F. (2003). Adhesion of high and low virulence Flavobacterium psychrophilum strains to isolated gill arches of rainbow trout Oncorhynchus mykiss. Diseases of Aquatic Organisms, 55: 101-107. Nematollahi A., Decostere A., Ducatelle R., Haesebrouck F., Pasmans F. (2005). Development of a gut perfusion model as an alternative to the use of live fish. Laboratory Animals, 39: 194-199. Nematollahi A., Shadkhast M., Shafeie S., Majidian F. (2011). Circulatory system of rainbow trout Oncorhynchus mykiss (Walbaum): a corrosion cast study. Journal of Applied Ichthyology, 27: 916-919. Passantino L., Abbate F., Cianciotta A., Germana G.P., Patruno R., Passantino G.F. (2000). Corrosion cast of the vascularization of Mugil cephalus gills. Italian Journal of Anatomy and Embryology, 105: 121-129. Noestelthaller A., Probst A., Koenig H.E.S. (2005). Use of corrosion casting techniques to evaluate coronary collateral vessels and anastomoses in hearts of canine cadavers. American Journal of Veterinary Research, 66: 1724-1728. Schmidt S., Lohse C.L., Suter P.F. (1980). Branching patterns of the hepatic artery in the dog: arteriographic and anatomic study. American Journal of Veterinary Research, 41: 1090-1097. Sherman R.L., Lametschwandtner A., Spieler R.E. (2005). Structural variation in gill vasculature among some batoid elasmobranchs examined using corrosion casting and SEM. Microscopy and Microanalysis, 11: 1218- 1219. Sherman R.L., Spieler R.E. (1998). Examination of gill vasculature of yellow stingray, Urolophus jamaicensis (Urolophidae), by SEM observations of resin casts. Italian Journal of Zoology, 65: 431-434. Souza C.A., Hashimoto D.T., Pereira L.H.G., Oliveira C., Foresti F., Porto-Foresti F. (2012). Development and characterization of microsatellite loci in Phractocephalus hemioliopterus (Siluriformes: Pimelodidae) and their cross-species amplification in six related species. Conservation Genetics Resources, 4: 499-501. Syed Ali S., Korf H.W., Oksche A. (1987). Microvasculature of the pineal organ of the rainbow trout (Salmo gairdneri). Cell and Tissue Research, 250: 121 Int. J. Aquat. Biol. (2018) 6(3): 114-121 425-429. Wildgoose W.H. (1998). Successful removal of a gastric foreign body from a red tailed catfish Phractocephalus hemioliopterus. Fish Veterinary Journal, 3: 20-26. Yoon H.Y., Jeong S.W., Choi Y.M., Jeong M.k., Kim J.Y., Han H.J., Hwang M., No B.K., Park S.H. (2004). Surgical removal of stones in the stomach of a tiger shovelnose catfish. Journal of Veterinary Science, 5: 275-277 Zeindler C.M., Kratky R.G., Roach M.R. (1989). Quantitative measurements of early atherosclerotic lesions on rabbit aortae from vascular casts. Atherosclerosis, 76: 245-255. Int. J. Aquat. Biol. (2018) 6(3): 114-121 E-ISSN: 2322-5270; P-ISSN: 2383-0956 Journal homepage: www.ij-aquaticbiology.com © 2018 Iranian Society of Ichthyology چکیده فارسی مطالعه(: Phractocephalus hemioliopterus Bloch & Schneider, 1801) ردتیل ماهی گربه خون گردش سیستم عروق تحلیلی گیریقالب 2نژادصادقی جواد ،1موسوی زادهابراهیم حسینعلی ،1اردشیری مهرداد ،1هوالسو رحمتی هومن، 1*میرقائد طاهری علی .ایران تهران، تهران، دانشگاه ،دامپزشکی دانشکده ،آبزیان هایبیماری و بهداشت گروه1 .ایران تهران، تهران، دانشگاه دامپزشکی، دانشکده پایه، علوم گروه2 چکیده: هدف از این تحقیق مشاهده و توصیف سیستم . استعروف ( یکی از ماهیان زینتی مPhractocephalus hemioliopterus) گربه ماهی ردتیل گرم استفاده 580عدد ماهی ردتیل با متوسط وزن 10. برای این منظور ازباشدمی گیری تحلیلی عروققالب گردش خون این گونه با استفاده از روش برای تکمیل پلیمریزه و سخت شدن متیل متاکریالت پر شد. عروق خونی ماهیان با رزین مصنوعی ساخته شده از متیلشد. پس از بیهوشی، قلب و غوطه ور شدند. در نهایت KOH %25ساعت در محلول 48-24ساعت در حمام آب و سپس به مدت 12-24مدت متاکریالت، ماهیان در ادامه به ورت پشتی، عروق روده، معده و کبد، عروق ئسیستم گردش خون به شکل عروق آوران و وابران آبشش، بخش های مختلف قلب، آئورت شکمی، آ ه، طحال و نیز سیاهرگ پورتال و کبدی مشخص و نشان داده شد.خلفی و قدامی کلی .فیزیولوژی عروق، تحلیلی گیریقالب ردتیل، ماهی گربه خون، گردش متسیس :کلمات کلیدی