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Parasitic and Fungal Identification in Bamboo Lobster Panulirus Versicolour and Ornate Lobster P

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Parasitic and Fungal Identification in Bamboo Lobster Panulirus Versicolour and Ornate Lobster P Parasitic and Fungal Identification in Bamboo Lobster Panulirus versicolour and Ornate Lobster P. ornatus Cultures Indriyani Nur1)* and Yusnaini2) 1) Department of Aquaculture, Faculty of Fisheries and Marine Science, University of Halu Oleo, Kendari 93232, Indonesia Corresponding author: [email protected] (Indriyani Nur) Abstract—Lobster cultures have failed because of mortalities transmissible [5] including Vibrio, Aeromonas, Pseudomonas, associated with parasitic and fungal infections. Monitoring of spawned Cytophaga and Pseudoalteromonas [6,7]. Futher examination eggs and larva of bamboo lobsters, Panulirus versicolour, and ornate of these lobsters revealed four parasite species: Porospora lobsters, P. ornatus, in a hatchery was conducted in order to gigantea (Apicomplexa: Sporozoea), Polymorphus botulus characterize fungal and parasitic diseases of eggs and larva. One (Acanthocephala: Palaeacanthocephala), Hysterothylacium sp. species of protozoan parasite (Vorticella sp.) was identified from eggs while two species of fungi (Lagenidium sp. and Haliphthoros sp.) were (Nematoda: Secernentea), and Stichocotyle nephropis found on the lobster larvae. Furthermore, adult lobsters cultured in (Platyhelminthes: Trematoda) [8]. Therefore, the aim of this floating net cages had burning-like diseases on their pleopod, uropod, study was to examine and to characterize the infestation of and telson. Histopathological samples were collected for parasite parasites and fungus of potentially cultured lobsters (larval identification and tissue changes. There were two parasites found to stages and adult stages of the spiny lobsters), and also to look infect spiny lobsters on their external body and gill, they are Octolasmis sp. and Oodinium sp..Histopathology showed tissue at the histopathological changes of their tissues. This is changes, necrosis on the kidneys/liver/pancreas, necrosis in the gills regarded as a major cause of low production in aquaculture and around the uropods and telson. systems. Characterization and identification of pathogens is important in disease control and effective treatment. Keywords—Fungal, histopathology, lobster, parasite, infection. II. MATERIALS AND METHODS I. INTRODUCTION A. Broodstock and Larva HE bamboo lobster Panulirus versicolour and ornate Female lobsters that carried eggs under their abdomen were lobster P. ornatus, are common in the ocean. However, the T obtained from South East Sulawesi waters and Banda Sea, lobster has been captured and exploited in over 90 countries [1] Indonesia. They were caught at night and transported to the land which has lead to a loss of spawning stock and overfishing. using air circulation systems. Now, in many area, the population of lobsters has declined After the egg bearing females arrived at the hatchery, the dramatically due to: pollution, loss of habitat and susceptibility lobster were kept in the concrete tank for varying periods of to infection [2]; consequently, the lobsters are now considered time depending on the development stage of their eggs. The rare species. larvae were sampled at 1 – 6 days post hatching (dph). The lobsters remain a significant economic value source for many communities and they are a preferred seafood in the B. Lobster Sampling world. To address the increasing demand for food, lobsters have Adult lobsters were cultured in floating net cages. During the become a major sector of fisheries production through culture period the lobster were fed fresh fish and frozen squid aquaculture. Active research and development programs daily. The live lobsters were kept in aerated containers and worldwide have attempted to grow this sector, but so far none transferred to the laboratory where they were kept in aquariums has been successful. During the culture period, the lobsters for a maximum of two days until they were euthanatized to look often experience many serious problems, particularly those for parasites. Prior to dissection, all lobsters were weighed. caused by parasites and fungus infection. The challenges faced with raising lobsters is great. The most difficult stage to grow C. Examination for Parasites on Larvae and Adult Lobster these species is their larval stage [3]. To date, seed stock cannot yet be produced on demand. Larvae were investigated on outer layers attached to the It has been reported that black gills caused by fungus body. While adult lobsters were killed and then examined Fusarium sp. appeares to have caused mortality in 69.5% of externally and internally for clinical signs of disease and other lobster in cages in Vietnam [4]. Transmission of PaV1 virus abnormalities. Each lobster was necropsied and the following among lobsters also has been a detected [2]. Additionally, organs were checked: external body surface (carapace, gills, several types of bacteria have been isolated from lesions of and fins) as well as the kidneys, liver, and pancreas. Lobsters were affected lobsters of shell diseases and were known to be dissected using general methods [9]. Briefly, each lobster was systematically searched for parasites. First the carapace, gills and uropod/telson were examined using a combination of eye Vorticella sp. was observed in outer layers of larvae. and microscope. The outer layer of the carapace was scraped Vorticella sp. are small unicellular ciliates often found on with scalpel and then the scrapings were examined using a carapace and skin of many fish species. Vorticella sp. have low microscope. The gills were removed and examined individually host specificity and therefore are widely distributed. Most under microscope, and then scraped into a petri dish filled with families of freshwater fish harbor Vorticella sp. [12, 18]. They clean water to dislodge the parasites and examined under a have also been reported on amphibians, crustaceans, mollusks stereo microscope. The kidneys, liver, and pancreas were dissected and coelenterates. Vorticella sp. inhabit both fresh and salt and examined for parasites using a microscope. Squash water habitats. According to [18], some Vorticella sp. species preparations were prepared and examined under a microscope. are pathogenic. Classification of Vorticella sp. in detail were: All detected parasites were removed and treated with saline to Phylum Ciliophora; Order Peritrichida; Sub Order Sessilina; retain morphological definition. Where possible, photographs Family Vorticellidae; Genus Vorticella [10]. were taken of live specimens and then they were killed, fixed There are various species of Protozoan parasites, which are and preserved in 70% ethanol. Parasites were classed as distinguished by their cilia. Phylum Ciliophora was ectoparasites or endoparasites. characterized with short cilia. Genus Vorticella also consisted of numerous species, all of them are soliter, attach to the D. Identification and Determination of substrate (host) with stalked-like, namely slender, cylindrical, Prevalence and Intensity or contractile stalk [10;19]. External and internal parasites were identified using the Vorticella is bell shaped with big mouth (peristome). Cell following relevant works (e.g. [10-12]). Prevalence and mean color varies, but is usually yellow or green. They live in intensity for selected parasites were determined according to freshwater and sea water, and the adult stage is attach to an [13]. All parasites were identified to at least genus or species object, animal, or aquatic plant. Peritrich and suctorian ciliates and enumerated. are often found as commensalisms on the external surfaces of E. Fungal Disease Identification on Eggs crustacean embryos, such Corthunia sp, Epistylis sp. and Eggs with fungal infection showed signs of disease. The Vorticella sp. usually infect lobster cultures [20]. They attack symptoms were a cloudy color and cotton-like filaments the body including the: eyestalk, antenna, uropod, and eggs. In attached to the outer layers. Fungi of the lobster eggs were this research, we found that the stalked of parasite Vorticella isolated using PYGS agar (1.25 g peptone; 1.25 g yeast extract; penetrated the inner part of the larvae body. 3.00 g glucose; 12–15 g agar; 1,000 mL sea water). In order to B. Fungus on Lobster Eggs inhibit most bacterial growth antibiotics were added to the Fungus found on the agar media and in the stains was medium. After fungal colonies develop on the agar plates, each identified as two genera of fungi. They are derived from the was isolated onto fresh PYGS agar to obtain a pure culture. For order Lagenidiales, genus Haliphthoros and genus Lagenidium. morphological observations, the isolates were inoculated into Characterization of these fungi can be seen in Figure 1-4. PYGS broth and incubated at 25 ˚C for 3–5 days [14]. Fungal diseases that attack eggs are a serious problem in the Determination and characterization of pure culture colony hatchery because they reduce the success of the eggs. This study isolate based on macroscopic investigation including shape, found several eggs undergoing pathological disorders, size of the colony, and character of the hyphae. While, characterized by a white cloudy egg surface. microscopic morphology included the number of branches, This fungus group was also found infecting eggs and larvae septa, size, and diameter of hyphae. After this the colony was of mangrove crab at Coastal Fisheries Research Center in stained with cotton blue with the guidance of references Gondol, Indonesia [21]. Fungi produce zoospores in the water, [15,16]. and then swim freely to find and attach to the host
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