REPORT ON GUT ASSOCIATED LYMPHOID TISSUE (GALT) IN .

International Journal of Pure and Applied ZoologyVolume 2, Issue 2, pp: 95-99, 2014 2013 Rishan PublicationsISSN (Print) : 2320-9577ISSN (Online): 2320-9585http://www.ijpaz.comResearch ArticleREPORT ON GUT ASSOCIATED LYMPHOID TISSUE (GALT) INFRESHWATER FISH CHANNA PUNCTATUS (BLOCH)P. Venkatesh, S.P. Jeyapriya, N. Suresh* and T. VivekananthanDepartment of Zoology, Annamalai University, Annamalai Nagar-608002, Tamilnadu, India.Article History: Received 19th February 2014; Accepted 2nd April 2014; Published online 10th April 2014ABSTRACTThe present study describes the presence of intraepithelial leucocytes (IEL) and interlaminal leucocytes (ILL) inthe various gut regions of Channa punctatus. Routine histological techniques were adopted, section were stainedwith haematoxylin and eosin (H&E) and Giemsa. Apart from the IEL and ILL other cell types were observedalong the entire gut regions, such as mucus cells, goblet cells, enterocytes, mast cells, granular cells and plasmacells. The presence of lymphoid cells in dispersed manner indicates the presence of gut associated lymphoidtissues and common mucosal immune responses.Key Words: IEL, ILL, GALT, Channa.INTRODUCTIONThe mucosal immunology of vertebrates gainedmore attention in the past decades. The mucosalimmune system is beneficial to fish because itslives in pathogenic rich aquatic environment(Rombout et al., 2011). Still there is a lack ofdetails about this system due to its diversity. It iswell documented that teleost fish are able tomount humoral and cell mediated response(Fournier Betz et al., 2000).In fish, mucus associated lymphoid tissue(MALT) is subdivided into several anatomicallocation of lymphoid tissue- Gut associatedlymphoid tissue (GALT). The presence ofmucosal immunoglobulin was first reported inPlaice Pleuronectes platessa (Fletcher and Grant,1969). The mucosal immune system is morecomplex than its systemic counterpart both interms of effectors and anatomy (Brandtzaeg,2009). The MALT contains variety of leucocytes(T cells and B cells), plasma cells, macrophages,granulocytes and enterocytes (Salinas et al.,2011), but still considerable debate regardingwhich cells are the main antigen colleting cells inteleost gastro intestinal track. The adaptive*Corresponding author e-mail: cnsuresh2006@rediffmail.comimmune system of several teleost has beenreported by either histology or molecularanalysis (Lin et al., 2005).Fish IELs represent a potential model systemfor comparative studies of immunology, whichcould assist in unravelling the complexities of themammalian immune system (Frane Bozic, 1999).In view the above, the present study is aimed todeal with the GALT in relation to the frequencyof intraepithelial leucocytes (IELs) in differentregion of gut in Channa punctatus.MATERIALS AND METHODSC. punctatus weighing about 150.0 10.0 gmand length about 18-20 cm were collected fromthe local fish ponds situated in the vicinity ofAnnamalai Nagar. The fishes were brought to thelaboratory, healthy and without sign of anydisease were selected as a sample. The fisheswere anesthetized with Tricane MethaneSulphonate (TMS) and the gut was dissected out.The gut was washed thoroughly in the saline toremove the gut content without any damage tothe gut. The entire gut was cut into the differentpieces and fixed in Bouin’s fixatives or Neutralbuffered formalin for 24 hours or more. The

Venkatesh et al.Int. J. Pure Appl. Zool., 2(2): 95-99, 2014fixed tissue washed in tape water, dehydrated ingraded series of ethyl alcohol, embedded inparaffin wax and the sections were cut at 6µthickness in a Rotary microtome (Spencer). Thesections were stained in delafieled haematoxylinand counter stain in eosin. Few sections werealso stained with Giemsa stain.The entire length of gut in C. punctatus wasmorphologically divided into five regions;oesophagus, anterior intestine (AI), middleintestine (MI), posterior intestine (PI) andrectum. Histomorphologically the gut ofC.punctatus consisted of four strata as like inother teleost fish viz., serosa, muscularis,submucosa and mucosa. Serosa outermost layerconsisted by loose connective tissues. Muscularisconsisted of circular and longitudinal musclelayer and submucosa consisted of looseconnective tissue as a mesh network. In mucosaregion, the mucosal fold or villi made up ofcolumnar epithelial cells (Figure 1). The mucosalfolds were varied in their length and numbers ineach region of the gut. The distinct strata in thegut of C.punctatus consisted of several cells suchas, mucus cells, goblet cells, granular cells, mastcells, lymphoid cells-leucocytes and plasma cellswere found along the entire gut region.Accumulation or aggregation of lymphoid cellsor lymphocytes was not observed along the entiregut region of C. punctatus. The leucocytes werefound dispersed along the entire gut asintraepithelial leucocytes (IELs) located alongwith the epithelial cells of the mucosa andintralaminal leucocytes were found in the laminapropria of mucosal fold (Figure 2 and 3). Therelative percentage of IEL found entire gutregionweregiveninFigure4.Figure 1. Showing mucosal region of gut inC. punctatus. (H&E Ca 400) G- Goblet cells;IEL- Intera Epithelial Leucocytes; Lp- Laminapropria; Mc- Mucus cells.Figure 2. Showing the presence of ILL Intralaminal leucocytes in gut of C. punctatus.(H&E Ca 400). ILL- Intera leucocytes (IELs) were carried out by countingthe number of IEL per 100 epithelial cells(Doggett, 1989).RESULTS96

Venkatesh et al.Int. J. Pure Appl. Zool., 2(2): 95-99, 2014Figure 3. showing the presence of IEL andgranular cells in the submucosa of gut inC.punctatus gut. (H&E Ca 400). Mu- Mucosa;IEL- Inter Epithelial Leucocytes; M- Muscular;Gc- Granular cells.Figure 4. showing the relative proportation of the intraepithelial leucocytes (IELs)in the various region of gut in C. punctatus.of mucus for the protective role against themicroorganism (Diaz et al., 2003).DISCUSSIONIn the present report, the histological features ofthe gut of C.punctatus showed distinct strata viz.,serosa muscularis, submucosa and mucosa weresimilar with the other teleost fishes and also withmammalian species reported earlier (Suresh andRanganathan, 2003; Domeneghini et al., 2005;Raji and Norouzi, 2010; Lokka et al., 2013).In the present study, no visible accumulationof leucocytes or lymphocytes was found in theentire gut of C.punctatus. Suresh andRanganathan, (2005) reported that no suchaggregation of leucocytes in the gut of Catlacatla. In contradict to the above, accumulation ofleucocytes found in the gut of Oreochromismossambicus and Acanthopagrus latus (Doggett,and Harris, 1991; Salamat et al., 2011). Thepresence of IEL and intralaminal leucocytes inthe gut of C. punctatus were consistent with theother reports (Suresh and Ranganathan, 2005;Salamat et al., 2011).The different types of cells such as mucuscells, goblet cells, plasma cells, granular cellsand mast cells in the different strata of the gutalong the entire length were consistent withearlier studies on other fish species (Gargiulo etal., 1998; Cao et al., 2011; Lokka et al., 2013).The presence of mucus cells indicates secretion97

Venkatesh et al.Int. J. Pure Appl. Zool., 2(2): 95-99, 2014Doggett, T.A. and Harris, J.E., 1991.Morphology the gut associated lymphoidtissue of Oreochromis mossambicus and itsrole in antigen absorption. Fish and ShellfishImmunol., 1: 213-227.The numbers of teleost fishes have exists adiffused distribution of leucocytes, lymphocytes,plasma cells, granulocytes and macrophages(Petrie and Ellis, 2006). Similar observationswere made in A.latus (Salamat et al., 2011) andAtlantic halibut Hippoglossus hippoglossus(Grove et al., 2006).Domeneghini, C., Arrighia, S., Radaelli, G.,Bosia, G., and Veggetti, A., 2005.Histochemical analysis of glycoconjugatesecretion in the alimentary canal of Anguillaanguilla L. Acta. Histochem., 106: 477–487.Fish lack the organised GALT like organ likeother vertebrates such as Peyer’s patches ormesenteric lymph node. But in fishes lymphoidcells are present in scattered along the alimentarycanal (Salinas et al., 2011; Rombout et al.,2011). The presence of lymphocytes and IEL inthe gut especially in lamina propria indicates thepresence of GALT and involved in mucosalimmune response (Fournier-Betz et al., 2000).Fletcher, T.C. and Grant, P.T., 1969.Immunoglobulins in the serum and mucus ofthe plaice (Pleuronectes platessa). Biochem.J., 115: 65-69.Fournier-Betz, V., Quentel, C., Lamour, F. andLe Ven, A., 2000. Immunocytochemicaldetection of Ig-positive cells in blood,lymphoid organs and the gut associatedlymphoid tissue of the turbot (Scophthalmusmaximus). Fish Shellfish Immunol. 10, 187–202.CONCLUSIONIn present study the gut of C. punctatus consistedof four distinct layers were observed and thecells important for local immune response suchas IELs and ILL, goblet cells, mucus cellspresent.Frane Bozic, 1999. Isolation of carp ytes. Veterinarski Arhiv, 69 (2):97-103.CONFLICTS OF INTERESTThe authors declare that there are no conflicts ofinterest associated with this article.Gargiulo, A. M., Ceccarelli, P., Dall Aglio, C.,and Pedini, V., 1998. Histology andultrastructure of the gut of the tilapia (Tilapiaspp.), a hybrid teleost. Anat. Histol.Embryol., 27: 89-94.ACKNOWLEDGEMENTThe authors are indebted to University GrantCommission (UGC), New Delhi, for providingfinancial support under UGC Major ResearchProject.Grove, S., Johansen, R., Reitan, L.J. and Press,C.M., 2006. Immune- and enzymehistochemical characterisation of leucocytepopulations within lymphoid and mucosaltissues of Atlantic halibut (Hipoglossushipoglossus). Fish Shellfish Immunol. 20:693-708.REFERENCESBrandtzaeg, P., 2009. Mucosal immunity:induction, dissemination, and effectorsfunctions. Scand. J. Immunol., 70, 505–515.Cao, X.J., Wang, W.M., and Song, F., 2011.Anatomical and Histological characteristicsof the intestine of Topmouth culture (Culteralburnus). Anat. Histol.Embryl., 40:292-298.Lin, H.T., Lin, H.Y. and Yang, H.L., 2005.Histology and histochemical enzyme stainingpatterns of major immune organs inEpinephelus malabaricus. J. Fish. Biol., 66(3): 729-735.Diaz, A.O., Gracia, A.M., Devincenti, C.V., andGoldemberg, A.L., 2003. Morpholgical andhistochemical characterization of the mucousof the digestive tract in Engraulis anchoita.Aant. Histol. Embryol., 32 (6):341-346.Lokka, G., Austbo, L., Falk, K., Bjerkas, I., andKoppang, E.O., 2013. Intestinal morphologyof the wild Atlantic Salmos (Salmo salar). J.Morphol., 274: 859-876.Doggett T.A., 1989. The structure and function ofthe peripheral blood leucocytes and gutassociated lymphoid tissue in the cichilidOreochromis mossambicus. Ph.D thesis,Playmouth polytechnic University.Petrie,A.G. and Ellis, A.E., 2006. Evidence ofparticulate uptake by the gut of Atlanticsalmon (Salmo salar L.). Fish ShellfishImmuno., 20: 660-664.98

Venkatesh et al.Int. J. Pure Appl. Zool., 2(2): 95-99, 2014Raji, A. R. and Norouzi, E., 2010. Histologicaland histochemical study on the alimentarycanal in Walking catfish (Claris batrachus)and piranha (Serrasalmus nattereri). IranianJ. Vete. Res., Shiraz University, 11: 3-32,255.Salinas, I., Yong-An Zhang., Oriol Sunyer J.O.,2011. Mucosal immunoglobulins and B cellsof teleost fish. Dev. Comp. Immunol.35:1346-1365.Suresh, N. and Ranganathan, L.S., 2003. Studieson the histo architecture of the gastrointestinal tract of Catla catla (Hamilton).Aquaclt., 4(2): 209-213.Rombout, J.H.W.M., Abelli, L., Picchietti, S.,Scapigliati, G. and Kiron, V., 2011. Teleostintestinal immunology. Fish ShellfishImmunol., 31: 616-626.Suresh. N. and Ranganathan L.S., 2005. Lightmicroscopic examination of Gut associatedlymphoid tissues (GALT) in Indian majorcarp Catla catla (Hamilton) Aquacult., 6 (2)257-260.Salamat, N., Darbeh, R., Majd, N.M., andSafahieh, A.R., 2011. Histological Aspectsof Gut Associated Lymphoid Tissue inAcanthopagrus latus. Global Veterinaria., 7(4): 381-385.99

FRESHWATER FISH CHANNA PUNCTATUS (BLOCH) P. Venkatesh, S.P. Jeyapriya, N. Suresh* and T. Vivekananthan Department of Zoology, Annamalai University, Annamalai Nagar-608002, Tamilnadu, India. Article History: Received 19 th February 2014; Accepted 2 nd April 2014; Published online 10 th April 2014 ABSTRACT