PHYSALOPTERA SPP. CHANGE THE MORPHOLOGY OF STOMACH TISSUES OF RATTUS NORVEGICUS (BERKENHOUT, 1769)

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1 PHYSALOPTERA SPP. CHANGE THE MORPHOLOGY OF STOMACH TISSUES OF RATTUS NORVEGICUS (BERKENHOUT, 1769) Erum Khadijah* and M. Arshad Azmi Department of Zoology, University of Karachi, Karachi *Corresponding author: Abstract: In the present investigation the histophathology of stomach cells of the Rattus norvegicus (Berk.) was studied infected with nematodes (Physaloptera spp. Histopathological section of infected stomach were prepared by fixing small pieces of stomach in buffer formalin for 24 h. The fixed material was dehydrated and prepared for block making by usual procedure. Eosin and Hematoxylin stained serial section were mounted permanently in DPX. Sections were examined and photomicrographs of selected section were taken by Nikon (Aptiphot-2) photomicroscope using Fuji color flim. These nematodes were found attach to the mucosa of the stomach and caused serious damage in the mucous layers. Nematodes infection caused severe damaged in stomach. Nematodes damaged the four layers of the stomach wall. During getting meal from the host and through migration from one organ to another organ in the host body, so completely washed villous architecture and changed the morphology of gland cells was also observed. Nematodes caused inflammation, focal hemorrhagic, atrophy, hypertrophy and necrosis in stomach cells. High power view of mucosal surface of stomach with thread like worms were seen attached to the luminal surface of mucosa. Scattered non specific inflammatory cells were also seen in the stroma and can be observed in photomicrogphs. Keywords: Rattus norvegicus, stomach, nematode, histopathology. INTRODUCTION Family Muridae includes rats and mice; the most common and most important murid rodent is Rattus norvegicus (Berkenhout, 1769), the Brown rat, Norway rat, and Sewer rat. Brown rats are inherent form northern china. In the early 18th century, brown rat was found only in the Eastern Europe and at the end of this century brown rat has spread in all European countries. Now the brown rats are found throughout the world except "Antarctica". Brown rats are also known as Sewer rat (Fragasgy and Perry, 2003). In Asia sewer rat disappeared from forest and bushy areas. Sewer rats are mostly found in every part of city in all over the world (Southern, 1964). Their association with humans has been so close and lasting that this species depends upon man for food and shelter. Sewer rats live at different places, such as basements, wood lands, open fields, garbage, dumps, and also make burrows. Most of the carnivores, mammals, birds and reptiles preyed on the brown rats such as cats, owls, and snakes etc. Because of their foraging habits, brown rats are major pest of stored products. Often they spoil more through pollution and contamination with their droppings and hairs than what they actually consume. 29

2 Brown rats are hazardous to humans and animal health (Webster et al., 1995). Brown rats play most important role in spreading host to a plethora of diseases, in which plague and typhus are important. Bacteria are present in the saliva, when they bite cause rat bite fever. They also destroy fires and electrical wires in buildings by crushing of teeth. Brown rats are pathogens which are responsible for many serious diseases, such as Hantavirus, pulmonary syndrome, rat bite fever, Q fever, Weil's disease and cryptosporidiosis. In America brown rats are responsible for Q fever and Toxoplasmosis. Q fever is caused by Coxiella burnetii and Toxoplasmosis is caused by Toxoplasma gondii, the parasite that cause Toxoplasmosis. Toxoplasmosis is transferred from rats to humans when cats prey on infected sewer rat. Trichinella pseudospiralis a parasite has been found to be pathogenic in humans and carried by the brown rats. This rat is some times a major reservoir of bubonic plague, Yersinia pestis found in few endemic species of rodents such as ground squirrels, black rats and wood rats. Yersinia pestis is mostly transmitted zoonotically by rat fleas. Brown rat may be suffered by plague through non infected rodent species and also including dog, cats and humans. Natale et al., (2001) reported the histomorphometry of whole rat stomach, an absolutely and relevant procedure for quantitative analysis of mucosal injury. Spencer et al., (2001) reported interlukin-4 receptor - stage 6 signaling in murine disease spread by parasites with a tissue-dwelling nematode. Similarly MacDonald et al., (2002) studied immunology of parasitic helminth disease. Silveria et al., (2002) reported the disease spread with Strongyloides venezuelensis caused transient airway eosinophilic inflammation, higher in immunoglobulin E, and hyper responsiveness in rats. Suzuki et al., (2008) reported the effectors reaction for the gastrointestinal nematode parasites, Trichinella spiralis expulsion in rats. Takeda et al., (2010) studied directly effects of IL-4/IL-13 and the nematode Nippostrongylus brasiliensis on intestinal epithelial cells in vitro. Garcia et al., (2014) studied the metabolic and histopathological profile of Rattus norvegicus (Wistar) experimentally infected by Angiostrongylus cantonensis. Rats are widely distributed all over the world and are associated with disease organisms. These include Hemoparasites, protozoans, cestodes and nematodes and trematodes born diseases. Present investigation deals with helminth parasites (nematodes) present in Rattus norvegicus in Karachi, and study the histophatholgy of infected organs i.e., stomach. MATERIAL AND METHODS Rats (Rattus norvegicus, Berkenhout) were trapped from different locations of Karachi during Two rats stomach were found infected with Physaloptera spp. Pieces of infected stomach were fixed in 5% buffer formalin for 24h, then washed several time with 70% alcohol to remove as much as possible the fixative. The material was dehydrated in graded series of ethyl alcohol (70%, 90% and 100%). Small pieces of stomach were transferred to cedar wood oil for hours. Cedar wood oil was slowly removed by placing there is a mixture of equal amounts of cedar wood oil and xylol and then into pure xylol for 2 h. Wax block of specimens was made in L-blocks by usual procedure. Section were gently cut a thickness of 6-8 microns and stained with hematoxylin and eosin of infected permanently in Canada balsam by usual procedure. For preparing photomicrograph of selected portion of infected stomach 400 colour film was used on Nikon (Optiphot-2) photomicrograph. 30

3 RESULTS (1) A small section of stomach of Rattus norvegicus shows the full thickness of wall of stomach with intact mucosa, submucosa, muscular layer and outer most serosa, focal hemorrhagic area are identifies in the luminal surface of mucosa. Scattered non specific inflammatory cell are seen in the stroma. (2) A small section of stomach of Rattus norvegicus of high power view shows, thread like worms is seen to the luminal surface of mucos. Scattered non specific inflammatory cell and necrotic cellular debris present of stomach of Rattus norvegicus of high power view shows the walls of stomach with thread like worms are also observed. Stomach with thread like worms is seen penetrating the muscular layer of stomach. In photomicrograph-3: A small section of stomach of Rattus norvegicus of high power view shows the mucosal surface of stomach with thread like worms are attached to the luminal surface of mucosa. Scattered non specific inflammatory cell are seen in the stroma. In photomicrograph-4: A small section of stomach of Rattus norvegicus of high power view shows the mucosal surface of stomach with thread like worms are seen and attached to the luminal surface of mucosa. Scattered non specific inflammatory cell are also seen in the stroma. Photomicrograph-1: A small section of stomach of Rattus norvegicus of intermediate power view of full thickness of wall of stomach with intact mucosa, sub mucosa, focal hemorrhagic area (a) identifies in the luminal surface of mucosa, (b) scattered non specific inflammatory cell seen in the stroma. Photomicrograph-2: A small section seen penetrating the muscular layer of stomach. Scattered non specific inflammatory cell and necrotic cellular debris present in stomach of Rattus norvegicus of high power view wall of stomach with thread like worm (arrow). Inflammatory cell seen in the stroma. (A) Physaloptera spp. 31

4 Photomicrograph-3: A small section of stomach of Rattus norvegicus of high power view of mucosal surface of stomach with thread like worms seen attached to the luminal surface of mucosa (arrow). Scattered non-specific inflammatory cell are seen in the stroma. Photomicrograph-4: A small section of stomach of Rattus norvegisus of high power view of mucosal surface of stomach with thread like worms (arrow) seen attached to the luminal surface of mucosa. Scattered non-specific inflammatory cell are seen in the stroma. DISCUSSION In the present investigation study the histophathology of stomach cells of the Rattus norvegicus. Sample 1 and 2 of stomach infected with nematodes (Physaloptera spp.) Photograph A. These nematodes attached to the mucosa of the stomach and caused serious damage in the mucous layers. Nematodes infection caused severe damaged in stomach. Nematodes damaged the four layers of the stomach wall. During getting meal from the host and through migration from one organ to another organ in the host body, so completely washed villous architecture and changed the morphology of gland cells was also observed. Besides in early study Lafferly et al., (2010) reported nematodes encysted in the stomach lining of native geckos and suspect the geckos could be serving host for Mastophorus muris in rats (Rattus rattus). Rothenburger et al., (2014) recorded hyperkeratosis, mucosal hyperplasia and keratin pustules and submucosal inflammation in the non-glandular stomach of rat (Rattus rattus) with Eucoleus spp. In present study nematodes caused inflammation, focal hemorrhagic, atrophy, hypertrophy and necrosis in stomach cells. High power view of mucosal surface of stomach with thread like worms seen attached to the luminal surface of mucosa. Scattered non specific inflammatory cell are seen in the stroma was showed in photomicrogphs. 32

5 LITERATURE CITED Fragaszy, D.M. and Perry, S The Biology of Traditions: Model and Evidence. Cambridge University Press. p Garcia, J.S., Lucio, C.S., Bonfim, T.C., Junior, A.M., Tunholi, V.M., Tunholi-Alves, V.M., Mota, E.M., Simoes, R.O., Suntana, A.C., Pinherio, J. and Bioa, M.N Metabolic and histopathological profile of Rattus norvegicus (Wistar) experimentally infected by Angiostrongylus cantonensis (Chen. 1935). Exp. Parasitol., 137: Laffertry, K.A., Hathaway, S.A., Wegmann, A.S., Shipley, F.S., Backlin, A.R., Helm, J. and Fisher, R.N Stomach nematodes (Mastophorus muris) in rats (Rattus rattus) are associated with coconut (Cocos nucifera) habitat at Palmyra Atoll. J. Parasitol., 96: MacDonald, A.S., Aranjo, M.I. and Pearce, E.J Immunology of parasitic helminth infection. Infect. Immune., 70 (2): Natale, G., Lazzeri, G., Blandizzi, C., Gheradi, G., Lenzi, P., Pellegrini, A. and Tacca, M.D Histomorphometry of whole rat stomach an accurate and reliable method for quantitative analysis of mucosal damage. Tox. Appl. Pharmacol., 174 (1): Rothenburger, J.L., Himsworth, C.G., Lejeune, M., Treuting, P.M. and Leighton, F.A Lesions associated with Eucoleus sp. in the non-glandular stomach of wild brown rats (Rattus norvegicus). Int. J. Parasitol. Parasites Wildl., 3: Silveira, M.R., Nunes, K.P., Cara, D.C., Souza, D.G., Correa, A., Teixeira, M.M. and Correa, D.N Infection with Strongyloides venezuelensis induced transient airway eosinophilic inflammation, an increase in immunologlobulin E and hyperresponsiveness in rats. Infect immune., 70 (11): Spencer, L., Shultz, L. and Rajan, T.V Interlukin-4 receptor-stage-6 signaling in murine infection with a tissue-dwelling nematode parasite. Infect. Immun., 69 (12): Suzuki, T., Sasaki, T., Takagi, H., Sato, K. and Ueda, K The effectors responsible for gastrointestinal nematode parasites, Trichinella spiralis, expulsion in rats. Parasitol. Res., 103 (6): Takeda, K., Hashimoto, K., Uchikawa, R., Tegoshi, T., Yamada, M. and Arizono, N Direct effects of IL-4/IL-13 and the nematoda Nippostrongylus brasiliensis on intestinal epithelial cells in vitro. Parasite Immunology., 32 (6): Southern, N.H The Handbook of British Mammals. Oxford: Blackwell Scientific. Webster, J.P., Lloyd, G. and Macdonald, D.W Q fever (Coxiella burnetii) reservoir in wild brown rat (Rattus norvegicus) populations in UK. Parasitol., 110: (Received in July 2017 and accepted for publication in November 2017) 33