Microsc. Microanal. Res. 2019, 32(1) 26-29 Microscopy and Microanalysis Research Microsc

Microsc. Microanal. Res. 2019, 32(1) 26-29 Microscopy and Microanalysis Research Microsc. - The Journal of The Microscopy Society of Thailand Microanal. Journal Home Page: https://www.tci-thaijo.org/index.php/mmres/ Res.

Histology study of Siamese freshwater batfishOreoglanis ( siamensis)

Wasin Srirattanasart 1, Worawit Maneepitaksanti 1*, Rung Hiranwong 2 and Sakuldech Nunta 3

1 Animal and Aquatic Sciences Department, Agriculture Faculty, Chiang Mai University, Chiang Mai, 50200, Thailand. 2 Doi Inthanon National Park, Ban Luang, Chom Thong District, Chiang Mai, 50160, Thailand. 3 Mae Aum Watershed Management Unit, Protected Areas Regional Office 16 Chiang Mai, Department of National Park, Wildlife and Plant Conservation, Chiang Mai, 50120, Thailand *Corresponding author’s e-mail address: [email protected] (W. Maneepitaksanti)

ABSTRACT ARTICLE INFO Article history Submitted: 9 April 2019 Siamese freshwater batfish (Oreoglanis siamensis) is considered as endangered species due to the Revised: 6 August 2019 limited habitat such as highland running water sources of Inthanon mountain, Chiang Mai province, Accepted: 6 August 2019 Thailand. Moreover, this fish is threatened by the environmental changes and human hunting for food Available online: 26 September 2019 source. Therefore, this fish is listed in the wildlife protection of Thailand. Based on those reason this fish must be enhanced for conservation. Unfortunately, the information of this fish is little such as the feature of habitat, food item, reproduction, and diseases. Basically, histological study makes us Keywords: know the information of animal cell and tissue for baseline of further study. Hence, this study aims Mekong giant catfish ; histopathology ; to provide the information of Siamese freshwater batfish histology. Thirty fish samples were collected aquaculture ; columnaris bacteria ; disease; from Huay Khang Chang, Inthanon mountain using scoop net in October 2017. The fish organ samples Chiang Mai province were fixed in 10% neutral buffer formalin, dehydrated throughout of alcohol series by automatic tissue processing machine, embedded in paraffin block, 5 micrometer thickness, and stained by Hematoxylin & Eosin. The sections were observed under the microscope (Drawell, Smart series) and recorded by the camera (Zeiss, Scope.A1 and Zeiss, Axiocam 305 color). The histology of nasal barbel, gills, skin, brain, heart, liver, kidney, intestine, and stomach of Siamese freshwater fish is documented for the first time in Thailand.

INTRODUCTION

Siamese freshwater batfish Oreoglanis( siamensis) is sisorid catfish 98๐29’56”E at 1,470 m above sea levels) in October 2017 by scoop from Inthanon mountain and considered as endangered species in nets [3]. The fish samples were measured and weighed. After that, the Thailand. According to the study of this fish species biology, it lives in fish organs, including barbel, gills, skin, brain, heart, liver, kidney, lotic water habitat with water temperature during 18-20๐C at 1600 stomach and intestine were fixed in 10% neutral buffered formalin for mater above sea level and contain a sand, gravel and rocks that effect 24 hours. The organ tissues were dehydrated with ethanol series, cleared on natural selection to be alive [1] Based on these conditions, this with xylene, embedded in paraffin, cut at 5 µm thickness by microtome species is limited in some upper streams of the mountain. The characteristics and stained with hematoxylin and eosin (H&E) [4]. All stained slides of this species are strongly depressed head and body, inferior mouth, were observed under the light compound microscope (Drawell, Smart greatly enlarged paired fins, which modified to an adhesive apparatus, series) and recorded by camera (Zeiss, Scope.A1 and Zeiss, Axiocam notch on posterior margin of lower lip are presence, nasal barbel reaching 305 color). The experiment was performed according to the protocol midway between its base and anterior orbital margin, and emarginate of Laboratory animal center Chiang Mai University (2561/AQ-0002). shaped caudal fin [2]. However, the biological data of this species is scanty, especially the histology study which is an important information RESULTS for further histopathological study. Therefore, this study aims to demonstrate the histology information of Siamese freshwater batfish The section of fish nasal barbel was composed of taste bud barbel collected in Thailand for the first time. enclose to dermis layer, several goblet cells, melanophore pigments, numerous alarm substance cells. In addition, the connective tissue and MATERIALS AND METHODS a cartilage rod were central area of barbel were found (Figure 1). Gills were flaps of tissue packed full of capillaries that located Thirty adult fish samples were collected from Huay Khang Chang, beside the skull, enclose to nostril and mouth cavity and under the Inthanon mountain, Chiang Mai province, Thailand (18๐30’20”N, operculum. Gill filament were covered by squamous epithelium with Microscopy and Microanalysis Research Microsc. Microanal. Res., 2019, Vol. xx, 1 – 6. Microscopy and Microanalysis Research -The Journal of Microscopy Society of Thailand Microsc. Microanal. Res., 2019, Vol. xx, 1 – 6. -The Journal of Microscopy Society of Thailand

and contains the pillar cells inside. The efferent arteriole, which by outer layer or epicardium and the inner layer, myocardium, Histology study of Siamese freshwater batfish (Oreoglanis siamensis) take the blood away from gills, was found at the apex of gill which comprised of loose cardiac muscle (Figure 5). 1 1 2 3 filament (Figure 2). Wasin Srirattanasart , Worawit Maneepitaksanti *, Rung Hiranwong , Sakuldech Nunta 1 Animal and Aquatic Sciences Department, Agriculture Faculty, Chiang Mai University, Chiang Mai, 50200, Thailand. 2 Doi Inthanon National Park, Ban Luang, Chom Thong District, Chiang Mai, 50160, Thailand. 3 Mae Aum Watershed Management Unit, Protected Areas Regional Office 16 Chiang Mai, Department of National Park, E

Wildlife and Plant Conservation, Chiang Mai, 50120, Thailand P *Corresponding author’s e-mail address: [email protected]

ABSTRACT

GF Siamese freshwater batfish (Oreoglanis siamensis) is considered as endangered species due to the limited habitat such as highland running water sources of Inthanon mountain, Chiang

Mai province, Thailand. Moreover, this fish is threatened by the environmental changes and human GL hunting for food source. Therefore, this fish is listed in the wildlife protection of Thailand. Based

on those reason this fish must be enhanced for conservation. Unfortunately, the information of this fish is little such as the feature of habitat, food item, reproduction, and diseases. Basically, histological study makes us know the information of animal cell and tissue for baseline of further study. Hence, this study aims to provide the information of Siamese freshwater batfish histology. Thirty fish samples were collected from Huay Khang Chang, W.Inthanon Srirattanasart mountain using et al. scoop / Microsc. net Microanal. Res. 2019, 32(1) 26-29 27 Figure 4 The telencephalon or cerebrum of mature Siamese in October 2017. The fish organ samples were fixed in 10% neutral buffer formalin, dehydrated freshwater batfish showed numerous nerve cells. 20x. throughout of alcohol series by automatic tissue processing machine, embedded in paraffin block, Keywords: 5 micrometer thickness, gobletand stained or mucous by Hematoxylin cells presented & Eosin especially. The sections at the were apex observed of the filament.under epidermisFigure 2 andGill muscle. filament In of addition, mature muscleSiamese fibre freshwater bundles batfish. with numerous The Siamese freshwater batfish; Histology; the microscope (Drawell,Gill Smart lamellae series) includesand recorded an envelopeby the camera of squamous (Zeiss, Scope.A1 epithelium and Zeiss, and few peripheralmajor histological nuclei were components observed. Theare hypodermisefferent arteriole contained (E); thepillar loose Inthanon mountain; Chiang Mai province. Axiocam 305 color). Thegoblet histology cells, of nasalusually barbel, thickness gills, skin, layer brain, and containsheart, liver, the kidney, pillar intestine,cells inside. connectivecells (P); gill tissue filament (Figure (GF); 3). gill lamellae (GL). 40x. and stomach of Siamese Thefreshwater efferent fish arteriole, is documented which for take the thefirst bloodtime inaway Thailand. from gills, was found CentralFish nervousskin was system composed (CNS) containedof three neurons layers and including non-neurons at the apex of gill filament (Figure 2). suchepidermis, as neuroglia, dermis endothelia, and hypodermis ependymal. The middlecells and layer periocytes of epidermis of blood Fish skin was composed of three layers including epidermis, vesselwas composedwalls. The oftelencephalon large alarm (cerebrum) substance includedcells, which numerous usually nerve

INTRODUCTION dermis and hypodermis. The middle layer of epidermis was composed cellsround that or located oval shape, in olfactory having lobes the (Figuredistinct 4). nuclei in the central ofRESULTS large alarm substance cells, which usually round or oval shape, locationHeart. ofAlthough, this fish thiswas locatedlayer inw theas chambercovered immediatelyby squamous behind Siamese freshwater batfish (Oreoglanis siamensis) is having the distinct nuclei in the central location. Although, this layer gillepithelium arch and, betweeninvisible operculum. goblet cell The, usually heart wasmelanin covered pigment by outer also layer B The section of fish nasal barbel was composed of taste bud sisorid catfish from Inthanon mountain and considered as was covered by squamous epithelium, invisible goblet cell, usually orfound epicardium in this layer.and the Dermis inner layer,were locatedmyocardium, between which epidermis comprised and of barbel enclose to dermis layer, several goblet cells, melanophore endangered species in Thailand. According to the study of this melanin pigment also found in this layer. Dermis were located between loosemuscle. cardiac In muscleaddition, (Figure muscle 5). fibre bundles with numerous pigments, numerous alarm substance cells. In addition, the fish species biology, it lives in lotic water habitat with water peripheral nuclei were observed. The hypodermis contained the temperature during 18-20๐C at 1600 mater above sea level and connective tissue and a cartilage rod were central area of barbel loose connective tissue (Figure 3). contain a sand, gravel and rocks that effect on natural selection to were found (Figure 1). be alive [1] Based on these conditions, this species is limited in CM some upper streams of the mountain. The characteristics of this species are strongly depressed head and body, inferior mouth, Me greatly enlarged paired fins, which modified to an adhesive E TB S apparatus, notch on posterior margin of lower lip are presence, CT Figure 5 The heart of mature Siamese freshwater batfish. The De major histological components are epicardium (E); cardiac nasal barbel reaching midway between its base and anterior ASC orbital margin, and emarginate shaped caudal fin [2]. However, ASC CR muscle (CM); blood cells (B). 20x the biological data of this species is scanty, especially the histology study which is an important information for further Me De histopathological study. Therefore, this study aims to demonstrate the histology information of Siamese freshwater batfish collected GC in Thailand for the first time. H

MATERIALS AND METHODS Ep N Thirty adult fish samples were collected from Huay Khang N Chang, Inthanon mountain, Chiang Mai province, Thailand (18๐

30’20”N, 98๐29’56”E at 1,470 m above sea levels) in October V

2017 by scoop nets [3]. The fish samples were measured and Figure 3 Muscular system of mature Siamese freshwater batfish. Figure 1 The nasal barbel of mature Siamese fresh water batfish. weighed. After that, the fish organs, including barbel, gills, skin, Figure 1. The nasal barbel of mature Siamese fresh water batfish. FigureThe major 3. Muscularhistological system components of mature are Siamese squamous freshwater epithelium batfish. (S); The The major histological components visible are epidermis (EP); brain, heart, liver, kidney, stomach and intestine were fixed in MicroscopyThe majorand Microanalysishistological components Research visible are epidermis alarm substancemajorMicrosc. cells histological (ASC); Microanal. melanincomponents Res pigment., 2019, are squamous (Me); Vol. dermisxx ,epithelium 1 – (De); 6. dermis-The Journal (De); of Microscopytaste bud Society(TB); ofmelanophore Thailand (Me); alarm 10% neutral buffered formalin for 24 hours. The organ tissues Microscopy(EP); dermis and (De); Microanalysis taste bud (TB); Research melanophore (Me); peripheral (S);nuclei alarm Microsc.(N). substance 40x. Microanal. cells (ASC); Res ., melanin2019, Vol.pigment xx, (Me);1 – 6. substance cell (ASC); goblet cell (GC); connective tissue (CT); -The Journal of Microscopy Society of Thailand dermis (De); peripheral nuclei (N). 40x. were dehydrated with ethanol series, cleared with xylene, cartilage rodalarm (CR) substance 20x. cell (ASC); goblet cell (GC); connective Central nervous system (CNS) contained neurons and non- embedded in paraffin, cut at 5 µm thickness by microtome and and containstissue the (CT);pillar cartilagecells inside. rod (CR)The efferent 20x. arteriole, which by outer layer or epicardium and the inner layer, myocardium, stained with hematoxylin and eosin (H&E) [4]. All stained slides Gills were flaps of tissue packed full of capillaries that neurons such as neuroglia, endothelia, ependymal cells and take andthe containsblood away the pillarfrom cellsgills, inside. was found The efferentat the apex arteriole of gill, which which by comprised outer layer of orloose epicardium cardiac muscleand the (Figure inner layer,5). myocardium, located beside the skull, enclose to nostril and mouth cavity and periocytes of blood vessel walls. The telencephalon (cerebrum) were observed under the light compound microscope (Drawell, filamenttake (Figurethe blood 2). away from gills, was found at the apex of gill which comprised of loose cardiac muscle (Figure 5). under the operculum. Gill filament were covered by squamous included numerous nerve cells that located in olfactory lobes Figure 6 The liver of mature Siamese freshwater batfish. The Smart series) and recorded by camera (Zeiss, Scope.A1 and Zeiss, filament (Figure 2). Axiocam 305 color). The experiment was performed according to epithelium with goblet or mucous cells presented especially at the (Figure 4). major histological components are hepatocytes (H); hepatic apex of the filament. Gill lamellae includes an envelope of the protocol of Laboratory animal center Chiang Mai University portal vein (V); peripheral nuclei (N). 20x. squamous epithelium and few goblet cells, usually thickness layer Heart of this fish was located in the chamber immediately (2561/AQ-0002). E behind gill arch and between operculum. The heart was covered E P

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GL GL

Figure 4 The telencephalon or cerebrum of mature Siamese freshwaterFigure batfish 4 The showed telencephalon numerous or nerve cerebrum cells. 20x.of mature Siamese Figure 2 Gill filament of mature Siamese freshwater batfish. The Figure freshwater 4. The batfish telencephalon showed numerous or cerebrum nerve cells. of mature 20x. Siamese freshwater batfish showed numerous nerve cells. 20x. FiguremajorFigure histological2. Gill 2 Gill filament filament components of ofmature mature are Siamese efferent Siamese freshwater arteriole freshwater (E);batfish. batfish. pillar The The major histological components are efferent arteriole (E); cellsmajor (P); gill histological filament (GF); components gill lamellae are efferent(GL). 40x. arteriole (E); pillar pillar cells (P); gill filament (GF); gill lamellae (GL). 40x. cellsFish (P); skin gill filamentwas composed (GF); gill oflamellae three (GL).layers 40x. including epidermis, F dermisish skin and hypodermiswas composed. The middleof three layer layersof epidermis including was epidermis,composed dermisof large and alarm hypodermis substance. The cells,middle which layer ofusually epidermis roundwas or composedoval shape, of havinglarge alarmthe distinct substance nuclei cells, in thewhich central usually location. Although, this layer was covered by squamous round or oval shape, having the distinct nuclei in the central epitheliumlocation, invisible. Although, goblet this cell layer, usually was melanin covered pigment by s quamousalso B B foundepithelium in this layer., invisible Dermis goblet were celllocated, usually between melanin epidermis pigment and also muscle. In addition, muscle fibre bundles with numerous found in this layer. Dermis were located between epidermis and peripheralmuscle. nuclei In addition,were observed muscle. The fibre hypodermis bundles contained with numerous the looseperipheral connective nuclei tissue were (Figure observed 3). . The hypodermis contained the loose connective tissue (Figure 3).

E S FigureE 5 The heart of mature Siamese freshwater batfish. The S major histological components are epicardium (E); cardiac ASC Figure 5 The heart of mature Siamese freshwater batfish. The musclemajor (CM); histological blood cells components (B). 20x are epicardium (E); cardiac ASC muscle (CM); blood cells (B). 20x

Me De Me De

H H N N N N V

Figure 3 Muscular system of mature Siamese freshwater batfish. V

The Figuremajor histological 3 Muscular components system of mature are squamous Siamese epithelium freshwater (S);batfish. alarm substance cells (ASC); melanin pigment (Me); dermis (De); The major histological components are squamous epithelium (S); peripheralalarm substancenuclei (N). cells 40x. (ASC); melanin pigment (Me); dermis (De); peripheral nuclei (N). 40x. Central nervous system (CNS) contained neurons and non-

neurons suchCentral as nervousneuroglia, system endothelia, (CNS) containedependymal neurons cells andand non - periocytes of blood vessel walls. The telencephalon (cerebrum) neurons such as neuroglia, endothelia, ependymal cells and includeperiocytesd numerous of blood nerve vessel cells walls.that locatedThe telencephalon in olfactory ( cerebrumlobes ) Figure 6 The liver of mature Siamese freshwater batfish. The (Figureinclude 4). d numerous nerve cells that located in olfactory lobes major Figure histological 6 The livercomponents of mature are Siamese hepatocytes freshwater (H); batfish.hepatic The (Figure 4). portalmajor vein (V);histological peripheral components nuclei (N) .are 20x. hepatocytes (H); hepatic Heart of this fish was located in the chamber immediately portal vein (V); peripheral nuclei (N). 20x. behind gillHeart arch andof this between fish was operculum. located in The the heart chamber was immediatelycovered behind gill arch and between operculum. The heart was covered

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Microscopy and Microanalysis Research Microsc. Microanal. Res., 2019, Vol. xx, 1 – 6. -The Journal of Microscopy Society of Thailand

and contains the pillar cells inside. The efferent arteriole, which by outer layer or epicardium and the inner layer, myocardium, take the blood away from gills, was found at the apex of gill which comprised of loose cardiac muscle (Figure 5). filament (Figure 2).

Microscopy and Microanalysis Research Microsc. Microanal. Res., 2019, Vol. xx, 1 – 6. -The Journal of Microscopy Society of Thailand

Figure 4 The telencephalon or cerebrum of mature Siamese freshwater batfish showed numerous nerve cells. 20x. and contains the pillar cells inside. The efferent arteriole, which by outer layer or epicardium and the inner layer, myocardium, take the blood away from gills, was found at the apex of gill whichFigure comprised 2 Gill filament of loose ofcardiac mature muscle Siamese (Figure freshwater 5). batfish. The filament (Figure 2). major histological components are efferent arteriole (E); pillar cells (P); gill filament (GF); gill lamellae (GL). 40x. Fish skin was composed of three layers including epidermis, dermis and hypodermis. The middle layer of epidermis E was composed of large alarm substance cells, which usually

P round or oval shape, having the distinct nuclei in the central location. Although, this layer was covered by squamous B epithelium, invisible goblet cell, usually melanin pigment also found in this layer. Dermis were located between epidermis and muscle. In addition, muscle fibre bundles with numerous peripheral nuclei were observed. The hypodermis contained the GF loose connective tissue (Figure 3).

GL CM

E S Figure 5 The heart of mature Siamese freshwater batfish. The 28Figure 4 The telencephalonW. or Srirattanasart cerebrum of mature et al. / SiameseMicrosc. Microanal.major histologicalRes. 2019, 32(1)components 26-29 are epicardium (E); cardiac ASC freshwater batfish showed numerous nerve cells. 20x. muscle (CM); blood cells (B). 20x

Figure 2 Gill filament of mature Siamese freshwater batfish. The major histological components are efferent arteriole (E); pillar De cells (P); gill filament (GF); gill lamellae (GL). 40x. Me

Fish skin was composed of three layers including epidermis, dermis and hypodermis. The middle layer of epidermis H was composed of large alarm substance cells, which usually round or oval shape, having the distinct nuclei in the central N location. Although, this layer was covered by squamous epithelium, invisible goblet cell, usually melanin pigment also B N found in this layer. Dermis were located between epidermis and muscle. In addition, muscle fibre bundles with numerous V peripheral nuclei were observed. The hypodermis contained the Figure 3 Muscular system of mature Siamese freshwater batfish. Microscopy and Microanalysis Research loose connective tissue (Figure 3). The major histological components are squamous epithelium (S); Microsc. Microanal. Res., 2019, Vol. xx, 1 – 6. -The Journal of Microscopy Society of Thailand alarm substance cells (ASC); melanin pigment (Me); dermis (De); peripheral nuclei (N). 40x. CM

Fish liver was located on dorsal side of the body cavity with comparison, the lamina propria of this fish also larger than some Central nervous system (CNS) contained neurons and non- gastrointestinal tract. The hepatocytes showed peripheral nuclei carnivorous fishes such as Channa striata and C. batrachus, but

Eneurons such as neuroglia, endothelia, ependymal cells and and hepatic portal vein was present. All blood from the hepatic the muscularis layer of this fish was thicker than submucosa. The S periocytes of blood vessel walls. The telencephalon (cerebrum) Figure 5 The heart of mature Siamese freshwater batfish. The portal vein through the blood sinusoid and drains into the central small vessels were observed between muscularis and serosa. included numerous nerve cells that located in olfactory lobes Figuremajor 5.histological The heart ofcomponents mature Siamese are epicardium freshwater (E);batfish. cardiac The FigureFigurevein (Figure 6. 6 The 6) liver.liver of of mature mature Siamese Siamese freshwater freshwater batfish. batfish. The Themajor Blood vessels were larger than other catfish such as Clarias ASC (Figure 4). muscle (CM);major blood histological cells (B). components 20x are epicardium (E); cardiac major histologicalhistological components components are arehepatocytes hepatocytes (H); (H); hepatic hepatic batrachus [7]. The kidney of this fish was located close to the vertebral muscle (CM); blood cells (B). 20x portal veinportal (V); peripheralvein (V); peripheral nuclei (N) nuclei. 20x. (N). 20x. Heart of this fish was located in the chamber immediately column. The renal corpuscle contains a glomerulus that behind gill arch and between operculum. The heart was covered surrounded by Bowman’s capsule. The renal tubule connected to Me De the renal corpuscle (Figure 7).

The Microscopy Society of Thailand 2 www.mmres.org Fish liver wasH located on dorsal side of the body cavity with M

gastrointestinal tract. The hepatocytes Nshowed peripheral nuclei and hepatic portal vein was present. All blood from the hepatic portal vein through the blood sinusoid and drains into the central vein (Figure 6). RC N The kidney of this fish was located close to the vertebral column. H The renal corpuscle contains a glomerulusV that surrounded by Bowman’s

capsule. The renal tubule connected to the renal corpuscle (Figure 7). Figure 3 Muscular system of mature Siamese freshwater batfish. SB The stomach of this fish was Y-shaped and consisted of mucosa The major histological components are squamous epithelium (S); layer with the stratified columnar epithelial. The mucus cell was covered MR alarm substance cells (ASC); melanin pigment (Me); dermis (De); peripheral nuclei (N). 40x. with cilia. The numerous gastric glands were inserted in lamina propria, deep pink stained, and some connective tissues were found (Figure 8).

Central nervous system (CNS) contained neurons and non- The mucosa layer of intestine was composed of high squamous RT columnar epithelial whereas the laminar propria consisted of connective neurons such as neuroglia, endothelia, ependymal cells and periocytes of blood vessel walls. The telencephalon (cerebrum) tissue. The submucosa had some lymphocytes and erythrocytes (Figure 9). Figure 9 The intestine of mature Siamese freshwater batfish. included numerous nerve cells that located in olfactory lobes Figure 6 The liver of mature Siamese freshwater batfish. The The major histological components are mucosa layer (M); DISCUSSIONS (Figure 4). major histological components are hepatocytes (H); hepatic submucosa (SB); muscularis (MR). 20x. portal vein (V); peripheral nuclei (N). 20x. Heart of this fish was located in the chamber immediately The histology of Siamese freshwater batfish showed the same Figure 7 The kidney of mature Siamese freshwater batfish. The Pillar cells was observed inside the gill lamellae which Figure 7. The kidney of mature Siamese freshwater batfish. The behind gill arch and between operculum. The heart was covered characteristics as carnivorous scaleless fishes. The stomach of this fish major histological components are renal tubule (RT); renal function to sustain the gill branch and make the small pore to major histological components are renal tubule (RT); was abundant of gastric glands similar Clarias gariepinus in fundic corpuscle (RC); haemopoietic tissue (H). 20x. transfer red blood cells in respiratory systems [7]. It was the same renal corpuscle (RC); haemopoietic tissue (H). 20x. regions [5], Claris batrachus [6]. By comparison, the lamina propria in general fishes in Silurid fish, such as C. lazera presented the The stomach of this fish was Y-shaped and consisted of secondary lamellae and interlamellar epithelium with pillar cells The Microscopy Society of Thailand 2of this fish also larger than some carnivorous www. fishesmmres.org such as Channa mucosa layer with the stratified columnar epithelial. The mucus striata and C. batrachus, but the muscularis layer of this fish was thicker in normal tissue [8], whereas the armoured catfish (Hypostomus cell was covered with cilia. The numerous gastric glands were plecostomus) demonstrated the pillar cells in second gill lamellae than submucosa. The small vessels were observed between muscularis of this fish showed taste bud on tip of barbel, this organ was found in inserted in lamina propria, deep pink stained, and some [9]. and serosa. Blood vessels were larger than other catfish such asClarias theconnective same of other tissue catfishs were suchfound as (Figurelctalurus 8). punctatus [12]. batrachus [7]. Liver of this fish including hepatocytes that control metabolism Numerous alarm substrance cells in dermis layer were Pillar cells was observed inside the gill lamellae which function and macromolecules analysis for living activities. Vein was inserted found in the skin of this fish indicate that this fish was susceptible to sustain the gill branch and make the small pore to transfer red blood between hepatocytes that receive red blood cell from heart and transferred for the stimulator for example the water factor changes [10]. Also, G cells in respiratory systems [7]. It was the same in general fishes in to blood sinusoids by branchingMu vein [7]. Kidney contained the renal Plotosus lineatus was showed a lot of alarm substance cells in Silurid fish, such as C. lazera presented the secondary lamellae and corpuscle included the numerous of glomerulus for ammonia and waste dermis [11]. The nasal barbel of this fish showed taste bud on tip of barbel, this organ was found in the same of other catfish such interlamellar epithelium with pillar cells in normal tissue [8], whereas elimination by the circulatory systems. For the kidney GGstructure was the armoured catfish Hypostomus( plecostomus) demonstrated the pillar also same in others fish [11,12] as lctalurus punctatus [12]. cells in second gill lamellae [9]. Liver of this fish including hepatocytes that control Numerous alarm substrance cells in dermis layer were found in ACKNOWLEDGEMENTS metabolism and macromolecules analysis for living activities. the skin of this fish indicate that this fish was susceptible for the stimulator Vein was inserted between hepatocytes that receive red blood cell for example the water factor changes [10]. Also, Plotosus lineatus was This research was supported by National research council of from heart and transferred to blood sinusoids by branching vein showed a lot of alarm substance cells in dermis [11]. The nasal barbel Thailand, NRT [347909] who providedSB insight and expertise that greatly [7]. Kidney contained the renal corpuscle included the numerous of glomerulus for ammonia and waste elimination by the

circulatory systems. For the kidney structure was also same in others fish [11,12] Figure 8 The Stomach of mature Siamese freshwater batfish. The major histological components are mucus cell (Mu); goblet cell ACKNOWLEDGEMENTS (G); gastric glands (GG); submucosa (SB). 40x. This research was supported by National research council of Thailand, NRT [347909] who provided insight and expertise The mucosa layer of intestine was composed of high that greatly assisted the research. We are thankful to Comparisons squamous columnar epithelial whereas the laminar propria of Egg Production and Quality Among Four Popular Commercial consisted of connective tissue. The submucosa had some Layer breeds in Thailand Project for our work suppurations. We lymphocytes and erythrocytes (Figure 9). thank Entomology and Plant Disease Laboratory and Animal Disease Diagnostic and Histology Preparation Laboratory, DISCUSSIONS Department of Animal and Aquatic Sciences, Faculty of Agriculture, Chiang Mai University for scientific instruments The histology of Siamese freshwater batfish showed the supporting in this study, We would also like to show our gratitude same characteristics as carnivorous scaleless fishes. The stomach to Doi Inthanon National Park and Mae Aum Watershed of this fish was abundant of gastric glands similar Clarias Management Unit, Protected Areas Regional Office 16 Chiang gariepinus in fundic regions [5], Claris batrachus [6]. By Mai for sample collection in this study.

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Fish liver was located on dorsal side of the body cavity with comparison, the lamina propria of this fish also larger than some gastrointestinal tract. The hepatocytes showed peripheral nuclei carnivorous fishes such as Channa striata and C. batrachus, but and hepatic portal vein was present. All blood from the hepatic the muscularis layer of this fish was thicker than submucosa. The portal vein through the blood sinusoid and drains into the central small vessels were observed between muscularis and serosa. vein (Figure 6). Blood vessels were larger than other catfish such as Clarias batrachus [7]. The kidney of this fish was located close to the vertebral column. The renal corpuscle contains a glomerulus that surrounded by Bowman’s capsule. The renal tubule connected to the renal corpuscle (Figure 7).

Figure 9 The intestine of mature Siamese freshwater batfish.

The major histological components are mucosa layer (M); submucosa (SB); muscularis (MR). 20x.

Figure 7 The kidney of mature Siamese freshwater batfish. The Pillar cells was observed inside the gill lamellae which major histological components are renal tubule (RT); renal function to sustain the gill branch and make the small pore to corpuscle (RC); haemopoietic tissue (H). 20x. transfer red blood cells in respiratory systems [7]. It was the same in general fishes in Silurid fish, such as C. lazera presented the The stomach of this fish was Y-shaped and consisted of secondary lamellae and interlamellar epithelium with pillar cells mucosa layer with the stratified columnar epithelial. The mucus in normal tissue [8], whereas the armoured catfish (Hypostomus cell was covered with cilia. TheW. numerousSrirattanasart gastric et glands al. / Microsc.were plecostomus Microanal.) demonstratedRes. 2019, 32(1) the pillar 26-29 cells in second gill lamellae 29 inserted in lamina propria, deep pink stained, and some [9]. connective tissues were found (Figure 8). REFERENCESNumerous alarm substrance cells in dermis layer were found in the skin of this fish indicate that this fish was susceptible for the[1] stimulator W. Srirattanasart, for example R. theJodchaiyaphum, water factor changes A. Sanarsa, [10]. S.Also, Arungamol, G Mu Plotosus R.lineatus Hiranwong, was showed S. Nunta, a lot W. of Maneepitaksanti, alarm substance Somecells in biological characteristics of Siamese freshwater batfish (Oreoglanis siamensis). dermis [11]. The nasal barbel of this fish showed taste bud on tip of barbel,Journal this organ of Agri. was foundResearch in the and same Extension of other, 2018, catfish 35(2) such (suppl. 2), GG as lctalurus1003-1014. punctatus [12].

[2]Liver C. Vidthayanon,of this fish P.including Saenjundaeng, hepatocytes H. H. Ng,that Eight control new species metabolismof theand torrent macromolecules catfish genus analysisOreoglanis for (Teleostei:living activities Sisoridae). from Vein was Thailand.inserted between Ichthyol. hepatocytes Explor. Fres that., 2009,receive 20(2), red blood 127. cell from heart and transferred to blood sinusoids by branching vein SB [7]. [3]Kidney J. Chervinski,contained the Environmental renal corpuscle physiology included the of numeroustilapias. In: Pullin, of glomerulusR S V andfor Lowe-McConnell ammonia and wasteR H (eds.), elimination Proceedings, by Internationalthe Microscopy and Microanalysis Research Microsc. Microanal. Res., 2019, Vol. xx, 1 – 6. -The Journal of Microscopy Society of Thailand circulatoryCenter systems. for LivingFor the Aquatic kidney Resourcesstructure was Management also same (ICLARM),in Manila, Philippines, The Biology and Culture of Tilapias. 1982, others fish [11,12] (7), 119-128. Figure 8 The Stomach of mature Siamese freshwater batfish. The Figure 8. The Stomach of mature Siamese freshwater batfish. The ACKNOWLEDGEMENTS Fish liver was located on dorsal side of the body cavity with comparison,major histological the lamina components propria of are this mucus fish also cell larger (Mu); than goblet some cell [4] G. L. Humason, Animal Tissue Techniques, 4th ed. W. H. Freeman gastrointestinal tract. The hepatocytes showed peripheral nuclei carnivorousmajor fishes histological such as Channa components striata and are C . batrachusmucus cell, but(Mu); (G); gastric glands (GG); submucosa (SB). 40x. and Company, San Francisco. 1979. and hepatic portal vein was present. All blood from the hepatic the muscularisgoblet layer cell of (G);this fishgastric was thickglandser (GG);than submucosa. submucosa The (SB). This research was supported by National research council of Thailand, NRT [347909] who provided insight and expertise portal vein through the blood sinusoid and drains into the central small vessels40x. were observed between muscularis and serosa. [5] D. M. Mokhtar, E. A. Abd-Elhafez. A. H. S. Hassan, A histological, The mucosa layer of intestine was composed of high that greatly assisted the research. We are thankful to Comparisons vein (Figure 6). Blood vessels were larger than other catfish such as Clarias histochemical and ultrastructural study on the fundic region of squamous columnar epithelial whereas the laminar propria of Egg Production and Quality Among Four Popular Commercial batrachus [7]. the stomach of Nile catfishClarias ( gariepinus). J. Cytol. Histol., The kidney of this fish was located close to the vertebral consisted of connective tissue. The submucosa had some Layer breeds in Thailand Project for our work suppurations. We 2015, 6(4), 1. column. The renal corpuscle contains a glomerulus that lymphocytes and erythrocytes (Figure 9). thank Entomology and Plant Disease Laboratory and Animal surrounded by Bowman’s capsule. The renal tubule connected to Disease Diagnostic and Histology Preparation Laboratory, DISCUSSIONS [6] A. R. Raji, E. Norouzi, Histological and histochemical study on the renal corpuscle (Figure 7). Departmentthe alimentaryof Animal canal and inAquatic Walking Sciences, catfish (Claris Faculty batrachus of ) and Agriculture, Chiang Mai University for scientific instruments The histology of Siamese freshwater batfish showed the piranha (Serrasalmus nattereri). Iran. J. Vet. Res., 2010. 11(3), supporting in this study, We would also like to show our gratitude same characteristics as carnivorous scaleless fishes. The stomach 255-261. M to Doi Inthanon National Park and Mae Aum Watershed of this fish was abundant of gastric glands similar Clarias Management[7] S. Chinabut, Unit, Protected C. Limsuwan, Areas P.Regional Kitsawat, Office Histology 16 Chiang of the walking gariepinus in fundic regions [5], Claris batrachus [6]. By Mai for sample collection in this study. catfish,Clarias batrachus, Department of Fisheries, 1991. RC [8] A. M. Abdel-Moneim, N. M. Abou Shabana, S. E. M. Khadre M.,

H The Microscopy Society of Thailand 3 H. H. Abdel-Kader, Physiological www. and histopathologicalmmres.org effects in catfish Clarias( lazera) exposed to dyestuff and chemical SB wastewater. Int. J. Zool. Res, 2008, 4(4), 189-202.

[9] M. N. Fernandes, S. A. Perna-Martins, Epithelial gill cells in the MR armored catfish, Hypostomus cf. plecostomus (Loricariidae). Revista Brasileira de Biologia, 2001, 61(1), 69-78.

RT [10] R. J. F. Smith, Alarm signals in fishes,Fish. Biol. Fisher., 1992, 2(1), 33-63. Figure 9 The The intestine intestine of ofmature mature Siamese Siamese freshwater freshwater batfish. batfish. The Figure 9. [11] G. Zaccone, P. L. Cascio, Studies on the structure and histochemistry The major histologicalmajor histological components components are mucosa are layermucosa (M); layer (M); of the epidermis in the marine catfishPlotosus lineatus (Thunberg, submucosasubmucosa (SB); muscularis (SB); muscularis(MR). 20x. (MR). 20x. 1791) (Plotosidae, Pisces). Acta. Histochem., 1981, 69(1), 106- Figure 7 The kidney of mature Siamese freshwater batfish. The Pillar cells was observed inside the gill lamellae which 118. major histological components are renal tubule (RT); renal function to sustain the gill branch and make the small pore to [12] E. C. Joyce, G. B. Chapman, Fine structure of the nasal barbel of corpuscle (RC); haemopoietic tissue (H). 20x. transfer red blood cells in respiratory systems [7]. It was the same assistedin general the fishresearch.es in SiluriWe ared thankfulfish, such to asComparisons C. lazera presentedof Egg Production the the channel catfish,Ictalurus punctatus. J. Morphol., 1978, 158(2), The stomach of this fish was Y-shaped and consisted of andsecondary Quality Amonglamellae Four and Popular interlamellar Commercial epithelium Layer with breeds pillar in Thailandcells 109-153. mucosa layer with the stratified columnar epithelial. The mucus Projectin normal for ourtissue work [8] , suppurations.whereas the armoured We thank catfish Entomology (Hypostomus and Plant cell was covered with cilia. The numerous gastric glands were Diseaseplecostomus Laboratory) demonstrated and Animal the pillar Disease cells Diagnosticin second gill and lamellae Histology inserted in lamina propria, deep pink stained, and some Preparation[9]. Laboratory, Department of Animal and Aquatic Sciences, connective tissues were found (Figure 8). Faculty of Agriculture, Chiang Mai University for scientific instruments supporting Numerous in this study,alarm substranceWe would alsocells like in todermis show layerour gratitude were to Doifound Inthanon in the sNationalkin of this Park fish indicateand Mae that Aum this Watershed fish was susceptib Managementle for the stimulator for example the water factor changes [10]. Also, G Unit, Protected Areas Regional Office 16 Chiang Mai for sample Mu collectionPlotosus inlineatus this study. was showed a lot of alarm substance cells in dermis [11]. The nasal barbel of this fish showed taste bud on tip of barbel, this organ was found in the same of other catfish such GG as lctalurus punctatus [12].

Liver of this fish including hepatocytes that control metabolism and macromolecules analysis for living activities. Vein was inserted between hepatocytes that receive red blood cell from heart and transferred to blood sinusoids by branching vein SB [7]. Kidney contained the renal corpuscle included the numerous of glomerulus for ammonia and waste elimination by the

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