Ectoparasite and endoparasite between black and brown water in Batang Kerang, Balai Ringin, Serian, Sarawak

Ectoparasite and endoparasite between black and brown water in Batang Kerang, Balai Ringin, Serian, Sarawak Annia Awing Merang SH 175 A615 loll Bachelor of Science with Honours (Aquatic Resource Science and Management Programme) 2011

Pusat Khidmlt M.klumat Akademik,. UNlVEwsm MALAYSIA SARAWAK Ectoparasite and endoparasite between black and brown water in Batang Kerang, Balai Ringin, Serian, Sarawak Annia Awing Merang P.KHIDMAT MAKLUMAT AKADEMIK 1111110 I 111111111 1000235726 A report submitted in partial fulfillment of the Final Year Project 2 (STF 3015)..., Supervisor: Mr Khairul Adha A. Rahim Aquatic Resource Science and Management Programme Department of Aquatic Science Faculty of Resource Science and Technology Universiti Malaysia Sarawak 2011

Acknowledgement I feel grateful to God for giving me strength during the Final Year Project process. Thank you to my family support during this an time. Here, I would like to express my gratitude and appreciation to those who bring successful comp]etion to my final year project. I would like to thank my supervisor, En. Khairul Adha A. Rahim for his help, advices and encouragement during the whole project. Thank you also to Dr. Siti Akmar for guideline to make this report. I would like to thank the boatmans; En. Jeman Mardzuki, En. Nakhoda Jeman and local people of Balai Ringin for helping us in collecting fish samples during sampling at Batang Kerang floodplain. I would also like to thank to laboratory assistants, En. Zulkifli Ahmad, En. Nazri Latip and En. Richard Toh for the guidance in laboratory works and field works. Not forget also expressing my gratitude to my course mate and master students for guidance me in order to complete my Final Year Project. Lastly, I would like to express my gratitude to UNIMAS who support, educates me while I am in UNIMAS. Thank you all.

DECLARATION No portion of the work referred to this dissertation has been submitted in support of an application for another degree of qualification of this or any other university or institutions of higher learning. Annia Awing Merang Programme of Aquatic Resource Science and Management Department of Aquatic Science Faculty of Resource Science and Technology Uninversiti Malaysia Sarawak II

Pusat Khidmat Maklumlt Akademik UNTVERSm MALAYSIA SARAWAK Table of Content Acknowledgements... Declaration... Table ofcontents...,... "......... List ofabbreviations......... List oftables... I II III V VI List offigures... VII Abstract... '"......... 1 1.0 Introduction...................... 2 1.1 Background... 2 1.2 Problem statement................................ 3 1.4 Objective...... 3 2.0 Literature Review...'" 4 2.1 Fish in Malaysia.............................. 4 2.2 Distribution and habitat............. 5 2.3 Parasites in fish................... 5 2.4 Previous parasites research in Malaysia.................... 7 --. "\ 2.5 Parasites of freshwater fish...:'........... ~ 7 2.5.1 Monogeneans...... 8 2.5.2 Nematode...... 9 2.6 Parasites in Aquaculture...... 10 3.0 Material and Method... 11 3.1 Study Site...... 11 3.2 Sampling...... 12 3.3 Laboratory Work... '" 13 3.4 Data Analysis....... 13 5.0 Result and Discussion............... 14 5.1 Sample number for parasites study......................... 14 III

5.2 Ectoparasites and Endoparasites offish samples... 15 5.3 Parasites in Specimen........... 16 5.4 Important ofstudy fish parasite in Balai Ringin... 28 6.0 Conclusion.................................... 29 7.0 Recommendation...... 29 8.0 Reference...................................................... 30.. "" " IV

List of Abbreviations cm centimetre g gram SL Standard Length SE Standard Error C degree Celsius SD Standard Deviation TL Total Length --. v

List of Tables Table 2 3 4 5 6 7 Description Fish sampl for parasites examination were collected from black and brown water habitat at Batang Kerang, Balai Ringin, Serian, Sarawak Number of parasites infection on fish sample at Brown water Balai Ringin, Serian Number of parasites infection on fish sample at Black water Balai Ringin, Serian Number of fish examined, number of infected fish, site of infection length (centimetre), total weight (gram) and number of parasites in fish samples from Brown water Balai Ringin, Serian Number of fish examined, number of infected fish, site of infection length (centimetre), total weight (gram) and number of parasites in fish samples from Black water Balai Ringin, Serian Species parasites and the host (fish species) Water quality of brown and black water at Batang Kerang, Balai Ringin, Serian. (Mean ± SD) Page 14 16 16 19 20 25 26 t VI

List of Figure Figure Description Page Monogenea 8 2 Nematoda 9 3 Maps of Batang Kerang floodplain, Balai Ringin, Sarawak 11 4 Monogenea (Dactylogyrus) (Gill) with 100 magnificant 15 5 Nematoda (intestine) with 40 magnificant 15 6 Fish species and total number of parasites in fish species. 18 Appendixes Description List of Appendices Fish species with total length, standard length, weight and parasites in Brown and Black water Balai Ringin, Serian. 2 Water quality parameter Brown water Balai Ringin, Serian. 3 Water quality parameter Black water Balai Ringin, Serian. 4 Unidentified monogenean with 100 magnificant 5 Dactylogyrus monogenean (anterior part) with 400 magnificant 6 Dactylogynls monogenean (posterior part) with 400 magnificant -. t 7 Unindentified monogenean with 100 magnificanto VII

Ectoparasite and endoparasite between brown and black water in Batang Kerang, Balai Ringin, Serian, Sarawak Annia Awing Merang Department of Aquatic Science Faculty of Resource Science and Technology Universiti Malaysia Sarawak (UNIMAS) ABSTRACT Fish samples were collected in 6 to 7 August 2010 and 12 March 2011 from Batang Kerang Floodplain, Balai Ringin, Serian, Sarawak for the parasites analysis. A total of 46 samples were examined for the study and only 27 fish samples were infected. Two species (monogenean and nematode) of parasites were found observed in this study. The result showed monogenean species is the abundance parasites from brown water and black water. Hampala bimaculata and Cyclocheilichthys apogon fish were the most infected by parasites. Key words: Ectoparasites, endoparasites ABSTRAK lkan ditangkap pada 6 hingga 7 Ogos 2010 dan 12 Mac 2011 dari Batang Kerang, Balai Ringin, Serian, Sarawak untuk tujuan mengkaji jangkitan..parasit. Sejumlah 46 sampel telah digunakan untuk kajian ini dan hanya 27 sampel telali dijangkiti parasit. Dua specid (monogeneandan nematode) parasit ditemui dalam kajian ini. Keputusan menunjukkn monogenean species adalah parasit yang banyak ditemui di air keruh dan air hitam. lkan Hampala bimakulata dan Cyclocheilichthys apogon adalah ikan yang paling dijangkiti parasit. Kata kunci: Ektoparasit, endoparasit

1.0 Introduction 1.1 Background Parasites are group of pathogen that can cause diseases to fishes. Parasites classify as ectoparasites are monogenean, protozoa, copepod, and hirudinae commonly found on external body part of the fish. The most abundance ectoparasites were monogenea. The monogean parasites mainly attack gill and body surface of fish and cause damage to fish skin (Chandra, 2006). Endoparasites can be found in the host body within the intestine, stomach and inner skin fish. Parasites classify as endoparasites are cestoda, digenea, acanthocephalan, nematode and myxozoa. Ectoparasites generally found more than endopasites (Pazooki et at., 2007). The outbreak parasites can cause a lot of problem to human especially in aquaculture sector by killed most of the pond fish (Leong and Wong, 1988). But in wild environment, the infection of parasites to fi.sh is much lower compare to pond fishes. In wild environment, the population density and space of movement is not limited. The study of parasites can provide information about the fish condition in river. The parasites infection can relate to water quality of water (Galli et at., 2001; Martin et at., 2002) Batang Kerang is located at Balai Ringin, Serian, Sarawak. Balai.. Ringin River can divide into two: brown water and black.~.water. The black water is tributary to the brown water. The common fish in Balai Ringin water is from Family Cyprinid (Khairul Adha, 2009). 2

1.2 Problem Statement There no study about parasites fauna in Batang Kerang, Balai Ringin floodplain. The aims of this study are to provide information about the parasites fauna at Balai Ringin. 1.3 Objective The objectives ofthis study are: 1) To study the composition of parasites in fishes obtained from black water and brown water of Batang Kerang. 2) To analysis the occurrence ofparasites species at Batang Kerang.... 3

2.0 Literature Review 2.1 Fish in Malaysia The information about fish species in natural habitat is still lacking in Malaysia. Moshin and Ambak (1983) listed that Peninsular Malaysia has 382 species of freshwater fish and representing 56 families. However, Lee et ai, (1993) listed there are 292 species of freshwater fish representing 43 families included 13 introduced species found in wild habitat in Peninsular Malaysia. In Sabah, Chin and Inger (1990) listed a total of 155 species of freshwater fish included three sub-species and 13 introduced species. In Sarawak, however a total of 249 species of freshwater fish has been listed including fish species from Brunei (Kottlet et al., 1995). Ahmad and Khairul Adha (2007) listed about 280 species of freshwater fishes in Peninsular Malaysia, with more than 100 species reported in Sabah and 200 species reported from Sarawak. The latest compilation about Malaysia fishes was done by Chong et ai, (2010) listed 470 species of freshwater with 81 endemic species. The freshwater fishes of Malaysia are threatened by a variety of anthropogenic -. process. The water quality of most rivers in Malaysia is declining as a result of land clearing and eutrophication from agricultural and urban area (Chong et al., 2010). This condition has change the nature origin condition of river ecosystem. The fishes will have to change to adapt in the new environment or become extinct. 4

2.2 Distribution and habitat Freshwater fish in Malaysia has great variety of adaptation in different environment such as torrential stream, paddy field, peat swamp and human made pool. In Malaysia, the most dominant fish families are Cyprinids and followed by Channidae and Anabantidae (Salam et ai., 2006). Some areas in Sarawak are black water peat swamp area with amazing biomass in tropical rainforest. The fishes in black water are stenotopic to it environment (Chong et ai., 2010). This black water peat swamp characterised by its unique environment such substrate consist of detritus plant which release organic acids into poor buffer water system that contribute to the lower ph and low dissolved oxygen (William et al., 2003), resulting in poor species abundance and diversity. Fishes communities in black water of Malaysia are very different even with similar habitat and physico-chemical environment (Chong et ai., 2010). The endemic fishes in North Selangor, South-East Pahang and South- Western Johor peat swamp have different species even they have similar environment (Chong et ai., 2010). The North Selangor peat swamp forest has recorded more than 100 fish species (Ng et ai., 1992). According to William et al. (2003) the most abundance fish in peat swamp areas are Puntius johorensis followed by Belrmtia hasseilii, Silurichthys -. indragiriensis and Puntius hexazona. 2.3 Parasites in fish The parasites fauna of freshwater fishes in Sarawak is very poorly known. There have been limited number parasites fauna studied in Sarawak. Parasite is a group of pathogen that causes infection, diseases to freshwater fish (Chandra, 2006) and affect fish health, growth and survival (Mayo et al., 2009). There are 1211 species of different parasites from 5 phyla and 11 classes of invertebrates (Ahmed et al., 2007). The main 5

parasites member found in freshwater fishes are trematodes, cestodes, acanthocephalans and nematodes (Ahmed et at., 2007). The amount of parasites that found in intestines could be associated with the digestions of food intake of fish and resulting on the release of parasites cysts in food particles (Onyedinake et al., 2010). The parasite that was found in gill however is few due to continue movement of water current through the gill which may not encourage the attachment and survival of the parasites on gill (Onyedinake et ai, 2010). The size and weight of fish also has influences the number of parasites in fish. Increase in weight and size also increase fish susceptibility to parasitization (Onyedinake et al., 2010). Edema et al. (2008) stated that all fish species are susceptible to various infection of the parasites but depend on fish species, type of stream inhabited and environment. It is important to study the parasites in wild and culture environment. Freshwater fish culture in South Asia is one of the fast growing economic. Freshwater fish that are cultured in tropical countries are Oreochromis spp., Clarias spp., Pangasius spp., Tor tambroides, Trichogaster trichopterus, Barbonymus gonionatus and Leptobarbus hoevenii.. (Szekely et al., 2009a). Outbreaks of disease in cultures can :,cause financial loss to fish t farmers in Malaysia. The outbreaks of parasites in culture are due to low dissolved oxygen of water, high loading of organic matter, poor environment condition and anthropogenic changes in water environment (Edema et al., 2008; Lymbery et al., 2010). In Asian aquaculture, the dominant parasite species is from genera Myxobolus and Henneguya (Molnar, 2006). Research about monogenean in Peninsular Malaysia has been done by Lim (1990) and first monogenean described in Peninsular is Gyrodactylus fernandoi (Hanek and Furtado, 1973). 6

2.4 Previous parasites research in Malaysia Lim (1990) studied on the freshwater monogenean parasites at Peninsular Malaysia. Rahman and Bakri (2008) have examined the parasites on Clarias batrachus, Trichogaster pectoralis, Channa striata, Anabas testudineus, Notopterus notopterus (Asian Knifefish) and Puntius gonionotus of paddy-field fish from Kedah. Si et al. (1997) found that freshwater fish community of a reservoir along Perak River such as Puntius schwanenfeldii, Hampala marcoepidota, Puntioplites bulu, Osteochilus hassetlii, Cyclocheilichthys apogon and Hemibargus nemurus were infected by parasites. Szekelyet al. (2009a) found that fishes cultured in pond and net-cage in Malaysia such as Pangasianodon hypophthalmus, Leptobarbus hoevenii, Barbonymus gonionotus, Monopterus albus, Trichogaster trichopterus and Tilapia sp. has also been infected by parasites. Studies of myxozoan infection were examined by Szekely (2009b) on Osteochilus vittatus, Oxyeleotris marmoratus and Hemibargus nemurus. 2.S Parasites of freshwater fish The studies of parasites fauna in Sarawak are poorly known compare to Peninsular.. Malaysia. According to Rahman and Bakri (2008), parasites were very species specific because there were no fish have the same species of parasites and the parasites infections occur due to the fish feeding habit. Bu and Leong (1997) found that fish species is dominated by specific parasites. The studied shown that Puntius schwanenfeldii were infected by Dactylogyrus lampan and Dactylogyrus megavesicularis, Puntioplites bulu were infected by Dactylogyrus pahengenensis, Dactylogynls fascicule and Dactylogyrus perakensis, Osteochilus hasseltii were infected by Dactylogyrus torquegenitalis, and Hemibargus nemurus were infected by Silurodiscoides sp. and Cornudisco ides sundanensis. A study by Bu and Leong (1997) and Lerssutthichawal (2008) found that ~... 7

monogenean species is the most common and abundance parasites on freshwater fish in tropical reservoir along the Perak River, Malaysia and at Nakhonsithammarat River, Southern Thailand. Dactylogyridae is the most diverse monogenean family that infected fishes in Malaysia and Thailand (Lim, 1990 and Lerssutthichawal, 2008). Lim (2006) studied shown that the archerfish Toxotes jaeufatrix has infected by diplectanids monogenean (Heteropfeetanum flabelliforme, Dipfeetanum sumpit, Dipfeetanum jaeufator and Dipfeetanum toxotes). Rahman and Bakri (2008) found that Cfarias batraehus were infected by trematoda (Orientocreadium spp.) and nematode (Procamallanus clarias), Channa striata were infected by acanthocephalan (Pfallisentis sp.) and nematode (Camallanus yehi), Anabas testudineus and Notopterus notopterus were infected by nematode Zeyfanema anabantis and Spinitectus sp. In Edema et af. (2008) studied, nematodes (Procamallanus sp. followed by Cucullanus barbi and Spinitectus sp.) were most abundance parasites found to infected most fish species in Okhou River, Benin City, Nigeria. 2.S.1 Monogenean ~ Kingdom: Animalia. Phylum: Platyhelminthes Class: Monogenea Figure 1: Monogenea 8

Most of monogeneans are ectoparasites and commonly known as flukes. Monogenean parasites usually infected at the gills, skin and fin of fishes. The basis morphology character of monogeans is a posterior apparatus called an opisthaptor. There are two most common genera of monogenean on freshwater fishes: Dactylogyrus and Gyrodactylus (Klinger and Floyd, 1994; Marina, 2008). Study done by Lim (1990) listed 18 genera ofmono gene an exist in Peninsular Malaysia. 2.5.2 Nematode Kingdom: Animalia Phylum: Nematoda Intestine '-malo r.olal oland Figure 2: Nematode Nematode are endoparasites and known as roundworm. Nematodes usually observed in intestine of the host. Nematode has high concern from human because nematode can transfer from fish to human by consume infected fish (Marina, 2008). 9

2.6 Parasites in Aquaculture The examples of introduced fish species for aquaculture are common carp (Cyprinus carpio), Nile til apia (Oreachromis niloticus) and African shafttoothed catfish (Clarias gariepinus) (Lerssutthichawal, 2008). Infections of parasites in aquaculture fish are due to poor management water, overstocking fish in pond and nutritional deficiency (Martin et al., 2002 and Ghiraldelli et ai., 2006). A study by Leong et al. (1988) revealed that Grouper (Epinephelus malabaricus) in culture habitat has show high number of infection compare to grouper from the wild habitat. Nile tilapia, Oreochromis niloticus (Ghiraldelli, 2006) culture in Brazil were most infected by trichodinids Trichodina magna and Trichodlna truncate follow by monogenean Cichlydogyrus sclerosus and crustacea Lamproglena sp. and Arglt/us spinulosus. Szekely et al. (2009a) has discover four new species of myxozoan pathogen (Myxobolus omari sp., Myxobolus leptobarbi sp., Thelohanellus zahrahae sp. and Henneguya daoudi sp.) in freshwater fish culture in Malaysia. Ahmed et al (2007) reported nematode parasites, Procamallanus heteropneustus were infected Heteropneustes fossilis in pond near area of Kasur. Kabata (1991) discover the infection by Gorgorhynchus ophiocephali on intestine of Channa micropeltes from.. Tasik Bera and infection by Pallisentis gaboes on Cha;zn~ striatus from Melaka and' Seremban. 10

3.0 Materials and Method 3.1 Study site I, I, J. I,,.. I' I " I \, \.... ' J a.skm oj Sg. Ensebang Bare -! I '.. 0. f,~..., I \ \ I,.,..... ;, -- I, I I I I \.. \,.., t Figure 3: Maps of Batang Kerang floodplain, Balai Ringin, Sarawak (Khairul Adha et ai, 2009) 11

The studies of parasite were conducted at Batang Kerang, Balai Ringin, Serian, Sarawak. Balai Ringin water system can divide into two different river systems; these are black water and brown water. The tidal in Balai Ringin River were influence by tide season called low water and high water season (Khairul Adha et al., 2009). Characteristic of brown water river at Batang Kerang is muddy due to high sedimentation, while the black water river is acidic with low ph and low dissolved oxygen. The black color of black water is due to high concentration of humid acid (Khairul Adha et al., 2009). Along the brown water Batang Kerang floodplain, there are floating vegetation such as Hanguana malayana and Eichhomia crassipes (Khairul Adha et ai., 2009). The local village uses Balai Ringin as transportation and source of family income by selling freshwater fish. 3.2 Sampling The water quality parameter reading such as dissolved oxygen, ph, transparency, temperature, turbidity, total suspended solid and current were taken during the sampling. Various species of freshwater fishes at Batang Kerang were collected from three stations each in brown water and black water habitat during high tide season and low tide season. 3 layers of monofilament gill nets (4 em and 25.5cm) and gi.ll net (25.5 cm) were used and the nets were placed overnight at the selected section. The total length and standard length of fish were measures to the nearest centimeter. The weights of fish were measures to the nearest gram. The species of fish were identified according to Mohsin and Ambak (1983). The fish were examined for the ectoparasites and endoparasites. The fish were dissected to take the gills, body mucus, swim bladder and intestine then preserve in 10% formalin. Fish that do not dissect in-situ were brought back to laboratory for further identification by preserve in cool box.. '"'. ~ 12

3.3 Laboratory Work Parasite examinations were followed according to Kabata (1991) and Williams et al. (1994). The body surface, mouth, nostrils and fins of the fish samples were observed by naked eyes and stereo microscope (Leica ZOOM 2000 Model no. Z45V). The mucus from the body surface were collected and preserved in 10% formalin. The eyes, gill and fins from fish samples were dissected and preserved in 10% formalin. The preservation of mucus body, eyes, gills and fins samples were put in different bottle for each fish samples and organs. The bottles contain gill were shaken to let the parasites retain in formalin. The formalin of the body mucus and gill were put on slide with cover slip and observed under \ compound microscope. Then, the gill was scraped with scalpel and put on slide with cover slip then observed using compound microscope. The magnifications of compound microscope were use between 40 to 400 magnifications. The present or absent, number of parasites and type of parasites in fish sample were recorded. For the endoparasites examination, the fish samples were opened on ventral part and then the swim bladder and intestine were preserved in 10% formalin. The swim bladder and intestine were dissected, both were tore apart in Petri dish!mq examined under a stereo microscope (Leica ZOOM 2000 Model no. Z45V) and compound microscope. The magnifications of compound microscope were use between 40 to 400 magnifications. The parasites samples were identified according to Kabata (1991) and Williams et al. (1994). The present or absent, numbers of parasites and type of parasites in fish sample were recorded. 3.4 Data analysis The t-test statistic (p>0.05) were used to compare the number of parasites in brown and black water in Balai Ringin floodplain by using the SPSS 17. 13

5.0 Result and Discussion 5.1 Sample number for parasites study The fish samples were taken in 6 to 7 August 2010 (high water season) and 12 March 2011(low water season). A total of 46 fish samples belonging to seven species were examined for the parasites infection. Thirty-three fishes were collected from brown water and 13 fishes were collected from black water. The most abundance fish in Balai Ringin floodplain was from Family Cyprinidae (56.52%) similar with the finding by Khairul Adha el ai, (2009). The habitat and number of fish samples are shown in Table 1. Table 1: Fish sample for parasites examination were collected from black water and brown water habitat at Batang Kerang, Balai Ringin, Serian, Sarawak. Family Species Brown Black Anabantidae 8agridae Anabas testudineus 3 2 Hemibagrus baramensis 5 0 Cyprinidae Belostomatidae Parachela oxygastroides ~.. ", 5 0 t Cyc/ocheilichthys apogon 15 3 Hampala bimaculata 0 3 Helostoma temminckii 0 5 Durldae Krytopterus schilbeides 5 0 Total 33 13 14

5.1 Ectoparasites and Endoparasites of fish samples A total of three types of parasite were found on 27 out of 46 fish samples collected from Balai Ringin floodplain. Two type of class monogenea parasites (ectoparasites) and one phylum nematode (endoparasites) were observed. Parasites were found on this studies are showned below, where Figure 4 was an ectoparasites and the Figure 5 were an endoparasites. Figure 4: Monogenea (Dacty/ogyrus) (Gill) with 100 magnificant Figure 5: Nematoda (intestine) with 40 magnificant 15