Occurrence and Molecular Identification of Anisakis Dujardin, 1845 from Marine Fish in Southern Makassar Strait, Indonesia

Anisakis spp. (Nematoda: Anisakidae) parasitize a wide range of marine animals, mammals serving as the definitive\ud host and different fish species as intermediate or paratenic hosts. In this study, 18 fish species were investigated for Anisakis infection. Katsuwonus pelamis, Euthynnus affinis, Caranx sp., and Auxis thazard were infected with high prevalence\ud of Anisakis type I, while Cephalopholis cyanostigma and Rastrelliger kanagurta revealed low prevalence. The mean intensity of Anisakis larvae in K. pelamis and A. thazard was 49.7 and 5.6, respectively. A total of 73 Anisakis type I larvae collected from K. pelamis and A. thazard were all identified as Anisakis typica by PCR-RFLP analysis. Five specimens of Anisakis from K. pelamis and 15 specimens from A. thazard were sequenced using ITS1-5.8S-ITS2 region and 6 specimens\ud from A. thazard and 4 specimens from K. pelamis were sequenced in mtDNA cox2 region. Alignments of the samples\ud in the ITS region showed 2 patterns of nucleotides. The first pattern (genotype) of Anisakis from A. thazard had 100% similarity with adult A. typica from dolphins from USA, whereas the second genotype from A. thazard and K. pelamis had 4 base pairs different in ITS1 region with adult A. typica from USA. In the mtDNA cox2 regions, Anisakis type I specimens from A. thazard and K. pelamis showed similarity range from 94% to 99% with A. typica AB517571/DQ116427. The difference\ud of 4 bp nucleotides in ITS1 regions and divergence into 2 subgroups in mtDNA cox2 indicating the existence of A. typica sibling species in the Makassar Strait

Multiple gene analyses of caligid copepods indicate that the reduction of a thoracic appendage in Pseudocaligus represents convergent evolution

Background: The Caligidae is a family of parasitic copepods containing over 30 recognised genera. They are\ud commercially important parasites as they cause disease in numerous finfish aquaculture facilities globally.\ud Morphological features are used to distinguish between the genera and Pseudocaligus has traditionally been\ud differentiated from Caligus solely by the presence of a much reduced form of the fourth thoracic leg. Currently\ud there are numerous DNA sequences available for Caligus spp. but only the type species, Pseudocaligus brevipedis,\ud has molecular data available, so systematic studies using molecular phylogenetic analyses have been limited.\ud Methods: Three gene regions, SSU rDNA, 16S and CO1, for Pseudocaligus fugu from puffer fish from Japan and\ud Pseudocaligus uniartus from rabbit fish from Indonesia are sequenced and molecular phylogenetic analyses\ud performed in order to infer phylogenetic relationships between Pseudocaligus and other caligid copepods.\ud Results: The analysis revealed that there was no discrete grouping of Pseudocaligus spp. and that they had a\ud polyphyletic distribution within Caligus taxa. Pseudocaligus fugu grouped with Caligus elongatus and contained a\ud unique synapomorphy in the SSU rDNA region only seen in members of that clade. Pseudocaligus uniartus\ud formed a well-supported group, in the SSU rDNA analyses, with a Caligus sp. that also infects rabbit fish, but was\ud unresolved in the other analyses. Pseudocaligus brevipedis consistently and robustly grouped with Caligus curtus\ud and C. centrodonti in all analyses. The majority of Lepeophtheirus spp. form a monophyletic sister group to the\ud Caligus clade; however, L. natalensis is unresolved in all analyses and does not form part of the main\ud Lepeophtheirus clade.\ud Conclusions: These findings do not support the morphological-based distinction between Pseudocaligus and Caligus,\ud suggesting that the reduced fourth leg is a feature that has evolved on multiple occasions throughout caligid evolution.\ud Congruent molecular phylogenetic data support groupings based on the presence of morphological features, such as\ud lunules, geography and host fish type rather than appendage morphology. Therefore, we support the synonymy of\ud Pseudocaligus with Caligus

IDENTIFIKASI MOLEKULER DENGAN TEKNIK PCR-RFLP LARVA PARASIT Anisakis spp (Nematoda: Anisakidae) PADA IKAN TONGKOL (Auxis thazard) DAN KEMBUNG (Rastrelliger kanagurta) DARI PERAIRAN MAKASSAR

Beragam spesies ikan laut dikenal sebagai inang perantara dari berbagai spesies parasit, termasuk nematoda Anisakis yang merupakan agen penyebab Anisakiasis pada manusia. Anisakiasis dapat terjadi pada manusia setelah memakan ikan mentah atau setengah matang yang terinfeksi oleh Anisakis sp. Di Indonesia, khususnya di Sulawesi Selatan, informasi tentang Infeksi Anisakis ikan sering kali diabaikan dan tidak terdokumentasi dengan baik. Oleh karena itu, penelitian ini dimulai dengan mengumpulkan larva nematoda dari dua spesies ikan (Auxis thazard dan Rastrelliger kanagurta). Organ tubuh yang diperiksa meliputi rongga perut, organ internal dan jaringan otot. Larva nematoda yang diidentifikasi secara morfologis menunjukkan Anisakis spp kemudian dipisahkan untuk identifikasi secara molekuler lebih lanjut. Parasit difiksasi dalam etanol 70% dan dilanjutkan dengan PCR menggunakan primer ITS-1, 5.8S dan ITS-2 regions. Secara morfologi, Anisakis spp. dapat dibedakan dengan Anisakidae lain oleh adanya ventriculus yang jelas terlihat padaAnisakis spp. di bawah mikroskop stereo. Anisakis tipe I dan tipe II dibedakan oleh adanya mukron yang hanya terdapat pada Anisakistipe I dan tidak adanya di Anisakis tipe II. Spesies yang diidentifikasi menggunakan PCR-RFLP menunjukkan bahwa nematoda yang ditemukan pada ikan tongkol dan ikan kembung adalah Anisakis Typica

INFEKSI DAN PATOLOGI PARASIT Actinocleidus sp. (MONOGENEA) PADA INSANG IKAN LELE DUMBO, Clarias gariepinus

The aims of this research were to study level of infection of the monogenean parasite Actinocleidus sp. and its pathological effects on the gills of Catfish ( Clarias gariepinus). This research was conducted in April to June 2008 at several Catfish culture localities in Makassar and Maros, South Sulawesi. The fish were chategorized in three different ages group, i.e. small (5 < x ≤ 10 cm), middle (10 < x ≤15 cm) and large (x ≥15 cm). The totals of 270 fish were examined with 9 times frequency of sampling conducted every week. Prevalence of Actinocleidus sp. infection at the three culture facilities was very high (100%). Mean intensity at small, middle and large fish cultured in Adhyaksa were 85.9, 93.2 and 55.2, respectively. In Arief Rate, the mean intensity of infection at small, middle and large fish were 17.5, 15.1 and 7.2, respectively, whereas in Maros the mean intensity of infection were 56.7, 64.7 and 40.9, respectively. There was a high signifi cant difference of mean intensity between small, middle and large fish (P < 0.01). Pathological effects of the parasite Actinocleidus sp. on gills showed some damages, i.e. distal and basal hyperfl asia, fusion and overstimulated of mucus production. The high prevalence and mean intensity of the parasite infection indicating the worse culture management system applying by the Catfish farmers as well as by the high stocking density of the fish. The lower mean intensity of infection seen in the larger fish might be indicating that the larger fish had already developed immune system than the small ones

INFEKSI DAN PATOLOGI PARASIT Actinocleidus sp. (MONOGENEA) PADA INSANG IKAN LELE DUMBO, Clarias gariepinus = INFECTION AND PATHOLOGY OF PARASITE Actinocleidus sp. (MONOGENEA) ON THE GILLS OF CATFISH, Clarias gariepinus

The aims of this research were to study level of infection of the monogenean parasite Actinocleidus sp. and its pathological effects on the gills of Catfish (Clarias gariepinus). This research was conducted in April to June 2008 at several Catfish culture localities in Makassar and Maros, South Sulawesi. The fish were chategorized in three different ages group, i.e. small (5 < x S 10 cm). middle (10 < x 515 cm) and large (x 15 cm). The totals of 270 fish were examined with 9 times frequency of sampling conducted every week. Prevalence of Actinocleidus sp. infection at the three culture facilities was very high (100%). Mean intensity at small, middle and large fish cultured in Adhyaksa were 85.9, 93.2 and 552, respectively. In Arief Rate, the mean intensity of infection at small, middle and large fish were 17.5, 15.1 and 7.2, respectively, whereas in Maros the mean intensity of infection were 56.7, 64.7 and 40.9, respectively. There was a high significant difference of mean intensity between small, middle and large fish (P < 0.01). Pathological effects of the parasite Actinocleidus sp. on gills showed some damages, i.e. distal and basal hyperfiasia. fusion and overstimulated of mucus production. The high prevalence and mean intensity of the parasite infection indicating the worse culture management system applying by the catfish farmers as well as by the high stocking density of the fish. The lower mean intensity of infection seen in the larger fish might be indicating that the larger fish had already developed immune system than the small ones

TINGKAT INFEKSI PARASIT Thaparocleidus sp Jain, 1952 (MONOGENEA: Ancylodiscoididae) PADA INSANG IKAN PATIN (Pangasius sp)

Parasit monogenea adalah salah satu golongan parasit yang paling sering\ud menimbulkan masalah dalam budidaya ikan, karena siklus hidupnya yang langsung, reproduksi\ud tinggi dan patogenisitasnya tinggi, sehingga sangat sering menimbulkan kematian pada ikan.\ud Penelitian ini bertujuan untuk mengetahui tingkat infeksi parasit Thaparocleidus mahakamensis\ud yang menginfeksi benih ikan patin. Sampel yang digunakan dalam penelitian ini adalah benih\ud ikan patin yang berukuran 5 ??? 10 cm yang berasal dari tiga pengumpul ikan hias yang ada di\ud kota Makassar. Total ikan yang diperiksa ini adalah 90 ekor dengan frekuensi 9 kali\ud pengambilan sampel setiap minggunya. Sampel yang diambil dibawa ke laboratorium dengan\ud kantong plastik yang diberi oksigen, kemudian dimasukkan ke dalam aquarium yang diberi\ud aerasi sebelum dilakukan pemeriksaan parasit. Organ tubuh yang diperiksa adalah insang.\ud Berdasarkan hasil penelitian diketahui bahwa prevalensi parasit T. mahakemensis pada tiga\ud Lokasi sampling sama yaitu 100%. Nilai intensitas rata-rata parasit pada Lokasi I yaitu 98\ud ind/ekor, Lokasi II yaitu 58 ind/ekor dan pada Lokasi III adalah 70 ind/ekor. Tingginya tingkat\ud prevalensi dan intensitas parasit ini diduga sebagai salah satu masalah dalam pengembangan\ud budidaya ikan patin baik untuk kebutuhan sebagai ikan hias maupun untuk konsumsi

Keberadaan Anisakis Typica (Anisakidae) Dari Ikan Tongkol Dan Ikan Layang Dari Perairan Sulawesi Barat

Anisakis (L3-stage) is a parasitic nematode commonly found in marine fish or squid serve as intermediate or paratenic host. The purpose of this research was to identify Anisakis larvae using PCR-RFLP and sequencing methods. A total of 30 individuals of each species: bullet tuna (Auxis rochei) and Indian scad(Decapterus russeli) were examined for presence of Anisakis spp. Fish samples caught by fishermen around Mamuju waters were purchased at a fish market. The fish was necropsied and the organs: liver, intestine wall, heart and muscle were removed and put on separated petri dishes and examined for parasites under a stereo microscope. Anisakis found was cleaned and fixed in 70% alcohol. Initial identification was based on ventriculus shape and presence of mucron to distinguish Anisakis type I and Anisakis type II. All Anisakis found belong to Anisakis type I. Ten Anisakis type I were isolated, cleaned and stored in microfuge tubes containing 70% alcohol. The parasite gDNA was extracted using Wizard genomic DNA extraction and purification kit (Promega).Species of Anisakis was determined by PCR-RFLP at ITS1-5.8S-ITS2 region using two restriction enzymes: Taq I and Hinf I and PCR-sequencing in mtDNA cox2 region. Analysis by PCR-RFLP showed that all Anisakis type Iexamined was in the same pattern as Anisakis typica. Sequencing in mtDNAcox-2 region and the phylogenetic analysis showed that all samples were in the same cluster as A. typica. Based on PCR-RFLP and sequencinganalysis, all Anisakis found in this study belong to A. typica

The Biology and Pathology of Pseudocaligus uniartus Ho, 2004 (Copepoda: Siphonostomatoida: Caligidae) Infecting Siganids (Siganus sp) in Indonesia

Siganids are marine fish which are potentially cultured in ponds or floating net cages. In attempt to produce spawners from wild fish stock, the Caligid Pseudocaligus uniartus appeared in culture facilities causing mild to heavily damage on infected fish. The purposes of this research were to study the mode of infection, biology and pathology of P. uniartus. Egg strings were collected from ovigerous females infecting Siganus guttatus. The parasite posses two nauplius stages, one copepodid, four chalimus stages and young adult. Hatching rates reached 96% at 20 ppt, and 94% and 90% at 25 and 30 ppt, respectively. The eggs started to hatch theree hours after incubation and most eggs had hatched within seven hours. Nauplius stages were estimated for about three hours. Copepodid remained alive after two days without host and started to die within three days and all died within six days. Newly attached copepodid as well as chalimus was mainly found on posterior part of dorsal and anal fins and pectoral fin. Young adult and/or adult parasite were found on the surface of the body, caudal and pectoral fins of host. The estimated time needed from copepodid to adult was seven days. All larvae have become adults on day 10. The complete life cycle was estimated within 7.6 - 10.6 days. Heavily infected fish showed lesion on skin and eroded fins. Large number of inflammatory cells was infiltrated on the infected areas

The Use of Red ginger (Zingiber officinale var. rubrum) Extract to Control Ectoparasite Monogenean in Catfish (Clarias gariepenus Bruchell, 1822)

Monogenea ectoparasites are one of the most pathogenic parasites in aquaculture that need to control. The use of mediciinal plants to overcome diseases related to aquaculture can be applied and. relatively have no side effect.  One of the plants that can be potentially used as a treatment for curing purpose is red ginger. The effects of ethanol extracts of red ginger on the intensity of monogenetic ectoparasites in catfish were investigated in this study. About 390 experimental fish was used in the study. The experimental animals were catfish seedlings (total length of 3-5 cm). This research was conducted with 4 treatments and 3 replications consisting of 0 ppm, 5 ppm, 10 ppm and 15 ppm bath treatments within 24 hours. Determination of the dose applied was based on the results of the LC50 test that has been carried out previously. Based on the research results, the monogenetic ectoparasites found were Quadriacanthus clariadis and Gyrodactylus sp. with mean intensity of 16.4 and 4.47, respectively. LC50 value of the red ginger extract on carp seeds was 27.37 ppm. Red ginger extract can be used to cure infections at a concentrations of 15 ppm for Q. clariadis and Gyrodactylus sp. Concentration of 15 ppm for Q. clariadis and 5 ppm for Gyrodactylus sp. can be used as a preventive measure and improve resistance of a catfish seed

DETEKSI WHITE SPOT SYNDROME VIRUS (WSSV) DAN MONODON BACULO VIRUS (MBV) SECARA SIMULTAN PADA INDUK UDANG WINDU (Penaeus monodon) DARI PERAIRAN MAKASSAR DAN SEKITARNYA DENGAN TEKNIK DUPLEX PCR

Penyakit bintik putih pada udang windu akibat infeksi virus WSSV telah menyebabkan kematian yang sangat signifikan dalam budidaya udang di seluruh dunia dimana kegiatan budidaya udang dilakukan, sedangkan virus MBV merupakan salah satu virus penyebab kekerdilan udang di tambak. Kedua virus ini masih sangat sering ditemukan di tambak sampai saat ini. Virus secara umum dapat menular secara horizontal maupun vertical dari induk ke benih pada saat pemijahan. Salah satu factor keberhasilan budidaya udang adalah penggunaan induk yang bebas virus. Penelitian ini dilakukan untuk menentukan kondisi infeksi virus pada induk udang di perairan Makassar dan sekitarnya sebagai salah satu sumber induk yang digunakan dalam pembenihan, dengan teknik duplex PCR. Tahap awal PCR adalah optimasi terhadap konsentrasi MgCl2, suhu annealing, dan konsentrasi templat DNA, dan selanjutnya digunakan dalam mendeteksi keberadaan virus tersebut dari induk. Sebanyak 171 induk diperoleh dari pengumpul, yang diperoleh dari sekitar perairan Makassar, Takalar dan Pangkep. Udang diperiksa secara individu dengan dupleks PCR. Hasil penelitian menunjukkan bahwa prevalensi WSSV di Makassar, Takalar dan Pangkep adalah masing-masing 15%, 12% dan 19%. Prevalensi MBV di Makassar, Takalar dan Pangkep adalah masing-masing 29%, 22% dan 19%. Persentase induk yang terinfeksi oleh virus WSSV dan MBV secara bersamaan adalah masing-masing 15%, 12% dan 19% di Makassar, Takalar dan Pangkep. Prevalensi infeksi kedua jenis virus lebih tinggi pada induk betina dari pada induk jantan. Kesimpulan penelitian ini adalah virus WSSV dan MBV telah menginfeksi induk udang pada ketiga perairan tersebut dengan prevalensi yang tinggi, sehingga skrining induk sebelum digunakan dalam pembenihan mutlak diperluka