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Histopathological Finding of Liver and Kidney Tissues of the Yellow Mystus, 1 Hemibagrus Filamentus (Fang and Chaux, 1949)

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Histopathological Finding of Liver and Kidney Tissues of the Yellow Mystus, 1 Hemibagrus Filamentus (Fang and Chaux, 1949) 1 1 Histopathological finding of liver and kidney tissues of the yellow mystus, 2 Hemibagrus filamentus (Fang and Chaux, 1949), from the Tapee River, Thailand 3 4 Sinlapachai Senarat1, Jes Kettratad1, Pisit Poolprasert2, 5 Watiporn Yenchum3, Wannee Jiraungkoorskul4,* 6 7 1Department of Marine Science, Faculty of Science, Chulalongkorn University, 8 Bangkok, Thailand 9 2Faculty of Science and Technology, Pibulsongkram Rajabhat University, Phitsanulok, 10 Thailand 11 3 Bio-Analysis Laboratory, Department of Chemical Metrology and Biometry, National 12 Institute of Metrology (Thailand), Pathum Thani, Thailand 13 4Department of Pathobiology, Faculty of Science, Mahidol University, Bangkok, 14 Thailand 15 16 17 18 19 20 21 22 Corresponding author: Wannee Jiraungkoorskul, Tel: +6622015550, 23 e mail: [email protected] 2 1 Abstract: A histological finding of liver and kidney tissues of yellow mystus, 2 Hemibagrus filamentus (Fang and Chaux, 1949), collected from the Tapee River 3 (Chawang district, Nakhon Si Thammarat province, Thailand) was conducted to 4 examine the pathological lesions. Lesions including cellular swelling, hydropic 5 degeneration or vacuolation of hepatocytes were found. From this, coagulative necrosis 6 with pyknotic nuclei, infiltration of macrophage, constriction of sinusoidal capillaries 7 and damage of endothelial cells of blood vessels were observed in liver tissues. The 8 dilation of both Bowman’s space and the glomerulus were observed from kidney 9 lesions. Moreover, necrosis in renal tubular cells was also observed in some areas. 10 Although preliminary, the results from this study clearly show the need for further 11 investigation into stress factors and vectors affecting the health of H. filamentus and 12 other fish species within the Tapee River. 13 14 Key words: Hemibagrus filamentus, fish, histopathology, kidney, liver, Tapee River 15 16 17 18 19 20 21 22 23 24 3 1 1. Introduction 2 The Tapee River is the longest in southern Thailand, and it is recognized as the most 3 ecologically and economically important freshwater body of water in the region. Its 4 headwaters start from Khao Luang Mountain (Nakhon Si Thammarat Province), where 5 it supplies water for people who live along of this river, and it eventually flows to the 6 Gulf of Thailand. The Tapee River is a total length of 230 km, and drains 13,454 km2 7 that includes a forested area of 4,072 km2 and an agricultural area of 7,086 km2 8 covering parts of Surat Thani, Nakon Si Thammarat, and Krabi provinces (Laudee and 9 Prommi, 2011). The water quality and trophic status of the Tapee River has been 10 classified as having a clean-moderate and oligotrophic-mesotrophic status 11 (Leelahakriengkrai and Peerapornpisal, 2011). Despite its water quality and trophic 12 status, the Tapee River is vulnerable to pesticides pollution, which receives agricultural 13 runoff from paddy fields, Indian rubber plantations, and vegetable and fruit crops. 14 Common agricultural practices within Nakhon Si Thammarat Province involve the 15 application of herbicides such as glyphosate, paraquat, and atrazine to promote, and, 16 these agricultural chemicals have been extensively used over a long period of time. 17 Hence, it is likely that the Tapee River receives substantial inputs from herbicide 18 residue residing in surrounding soil and sediment as well as directly from surface water 19 runoff. Even though low to high concentration levels of herbicides remain in the river, 20 the chronic input and presence of these chemicals coupled with the potential for 21 biomagnification through the food web is of great concern. Similarity, the chronic 22 presence of agricultural chemicals can have a direct negative effect on both river 23 systems and on the health of specific species or groups, such as fishes that live in 24 proximity to such areas (McGlashan and Hughies, 2001). 4 1 One of the economically important freshwater fish in the Tapee River is the 2 yellow mystus, freshwater catfish or green catfish, Hemibagrus filamentus (Fang and 3 Chaux, 1949) belonging to the Order Siluriformes and Family Bagridae. This species is 4 found in the basin and also in a variety of lotic and lentic ecosystems with substrates of 5 sand and mud, feeding mainly on fish, crustaceans, larval aquatic insects and plant 6 matter (Hee and Rainboth, 1999). According to the International Union for 7 Conservation of Nature, the yellow mystus is also listed as a Threatened Species 8 (IUCN, 2013). Moreover, as the most widely consumed fish with a high economic value 9 for Thai people, the yellow mystus has been selected as a sentinel species for pesticides 10 contamination. As mentioned above, it is probable that the environmental contamination 11 in the Tapee River may negatively impact fish. Sublethal affects may include 12 histological alterations that can provide an early warning of stress and potential overall 13 impact to the population. Currently, the histopathological biomarker is one useful tool 14 that can indicate environmental pollutants, especially for sub-lethal and chronic effects 15 (Fatma, 2009; Nikalje et al., 2012). This histotechnique can also diagnose and predict 16 vital changes in histological structure of both tissues and organs (Hinton et al., 2001; 17 Adams, 2002; Dietrich and Krieger, 2009). 18 Although field surveys can provide presence-absence, histological methods are a 19 more effective precursor measure of fish health. Within the Tapee River, this may best 20 provide measures of impact to fish relative to agriculture zones. The histopathology 21 involving liver and kidney tissue of the yellow mystus is still not well known and 22 warrants further investigation. Therefore, the aim of this study was to investigate the 23 histopathological alteration in liver and kidney as a biomarker of the yellow mystus, 24 H. filamentus, obtained from the Tapee River, Thailand. 5 1 2. Materials and methods 2 2.1. Animal collection and study site 3 Mature fish samples (n = 20, 16 - 20 cm. standard length) were collected from the 4 Tapee River, Chawang district, Nakhon Si Thammarat province, Thailand (8°28'10" N, 5 99°29'45" E). The representative study site was selected during the rainy season (June- 6 July, 2013) and divided into three sampling sites. The length of each sampling site was 7 about six kilometers and each collecting site was two kilometers apart. All fish was 8 euthanized by the rapid cooling technique (Wilson et al., 2009). 9 2.2. Histological study 10 Upon euthanasia, each fish had both the liver and kidney immediately removed and 11 fixed in Davidson's fixative for 48 hours at room temperature. All specimens were 12 processed using standard histological techniques by being dehydrated through a graded 13 series of ethanol, cleared with xylene, embedded with melted paraffin (Bancroft and 14 Gamble 2002; Humason, 1979). Sections at 5 - 6 µm thick were cut by rotary 15 microtome and stained with Harris's hematoxylin and eosin (H&E). Finally, the 16 histological structure and histopathological lesion recorded as a mean prevalence in 17 liver and kidney was investigated by using a light microscope. 18 19 3. Results 20 3.1. Histopathology of liver 21 The liver was encapsulated with a thin layer of connective tissues. The polygonal 22 hepatocytes arranged along the sinusoid, arraying from the hepatic central vein. The 23 hepatocytes typically contained central nuclei with densely stained chromatin. 24 Cytoplasm of hepatocyte was slightly eosinophilic stained. Histopathological alterations 6 1 of the liver in H. filamentus from the Tapee River consisted of cellular swelling, 2 eosinophilic cytoplasm of hepatocytes, constriction of sinusoidal capillaries and damage 3 of endothelial cell of blood vessels. Some liver areas showed focal necrosis and 4 contained severe infiltration of macrophage, pyknotic nuclei and large lipid vacuoles in 5 the cytoplasm of hepatocytes (Figure A). The detected prevalence in liver tissues of this 6 fish is shown in Table 1. 7 8 3.2. Histopathology of kidney 9 Our research team has previously reported the histology of H. filamentus kidney 10 (Senarat et al., 2013). Histopathological alterations of kidney in H. filamentus from the 11 Tapee River showed kidney degeneration, glomerular alterations, Bowman’s capsule 12 rupture, hydropic swelling and cellular debris accumulation in lumen. In some cases, the 13 tubular cells showed the pyknosis and hyaline droplet accumulation, dilation of the 14 Bowman’s capsule and glomerulus. The dilation of the lumen in the tubules, 15 degeneration in the hemopoitic tissue, rupture in the collecting tubules and necrosis are 16 presented (Figures B - D). The prevalence of several lesions was observed in kidney 17 tissues of this fish. The details of its prevalence are given in Table 1. 18 19 4. Discussion 20 Freshwater fish are critical as a protein-rich food source but they are also susceptible to 21 the acute and chronic presence of pollutants (Thingran, 1974). The release of pollutants 22 to the aquatic environment is particularly toxic to aquatic organisms including fish. Fish 23 mortalities have been associated with the river receiving various pollutants (Prashanth et 24 al., 2005; Marina and Martinez, 2007), and the results of our study show that fish from 7 1 the Tapee River express histopathological alterations in liver and kidney during rainy 2 season. It is plausible that a lot of water during rainy reason could dilute pesticides and 3 other pollutants. In the same manner, the atrazine - herbicide group - has also been 4 reported from rainy season in both sediment and fish tissue (Puntioplites proctozyston) 5 from Nan river, northern Thailand (Senarat, 2011). However, our observation is not 6 unreasonable that the lesions on the target organs might have been a direct outcome of 7 pollutants including pesticides, metals and fertilizers, which enter the river with runoff. 8 Fish are susceptible to change in their surrounding environments as well as an increase 9 in pollution, and fish health provides a good precursory measure of the health status of a 10 specific aquatic ecosystem.
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