A B S T R A C TFailure of adaptation in feed utilization can result in larval mortality. Post-larval stage is a critical period of fish development in which transition from internal feeding to external feeding occurs. This study aimed to determine the growth and survival of hard-lipped barb (Osteochilus hasselti C.V.) post-larvae fed with different feeding patterns. The experiment was conducted on an experimental basis with a completely randomized design (CRD). The treatments consisted of (P1) 100% Spirulina sp., (P2) 75% Spirulina sp. + 25% pellet powder, (P3) 50% Spirulina sp. + 50% powder pellet, (P4) 25% +75% sp Spirulina pellet powder, and (P5) 100% powder pellets. Day 1 posthatching larvae were reared in aquaria with density of 17 fish / L for 8 weeks. The larvae were fed as much as 5% of the tota l weight of biomass. Post-larval development was evaluated by observing the development of the larvae morphology, the body length (every week), the biomass weight (every 2 weeks), and calculating their survival rate at week 8. Morphological data were analyzed descripti vely. Data on body length, biomass weight and survival were analyzed using two-way ANOVA followed by LSD test. The results showed that the feeding pattern significantly affected the development of post-larvae (p<0.05). The most advance post-larval development was obtained from the post-larvae fed with 100% pellet powder. In this group, morphological characters of post-larvae in the first week were the mouth start opening, the skin was pigmented, and gall bladder in bi-lobes shaped, caudal fin and dorsal fins were differentiated. Completion of fins development was achieved by the third week when the post-larvae have adult morphological character. The highest body length and biomass weight of the post-larvae was obtained in the group fed with100% of pellets and the highest survival rate was found in the group fed with combination of 50% Spirulina sp. + 50% pellet powder (P3).KEY WORDS: Osteochilus hasselti C.V., post-larvae development, survival rate, Spirulina platensis Penulis korespondensi: DIDI HUMAEDI YUSUF |
Identification and expression of two types of chicken GnRH-II genes in mature hard-lipped barb, Osteochilus hasselti. Biodiversitas 17: xxx. Gonadotropin-releasing hormone (GnRH) is synthesized in the brain and acts in the anterior pituitary to stimulate the release of gonadotropins in fishes as well as in other vertebrates. Genomic DNAs and cDNAs of two chicken-type GnRH-II genes of hard-lipped barb, namely cGnRH-II type 1 and type 2, were cloned. The length of cloned genomic DNA of cGnRH-II type 1 was 580 bp and cDNA was 206 bp. The length of cloned genomic DNA of cGnRH-II type 2 was 570 bp and cDNA was 196 bp. The cGnRH-II type 1 and type 2 cDNAs encode precursors of 68 and 63 amino acids, respectively. Those precursors consist of a signal peptide, cGnRH-II decapeptide and a GnRH-associated peptide (GAP) linked by a Gly-Lys-Arg proteolytic site. Using quantitative Real Time-PCR, expression levels of these two cGnRH-II genes were detected in the brain, liver and gonad of hard-lipped barb. Expression of the GnRH-II type 1 gene was found only in the brain and liver, on the other hand, expression of the cGnRH-II type 2 gene was found in the gonad, in addition to the brain and liver. The expression of the cGnRH-II genes outside the brain suggested that cGnRH-II might act as an autocrine or paracrine regulator.
A B S T R A C TThis paper describes a simple protocol of paraffin-embedded histological section for fish eggs, embryo and larvae of the hard-lipped barb and the giant gourami. The specimens were fixed in Bouin solution, washed in 70% ethanol, then were dehydrated in a series of ethanol solution of increasing concentration until absolute ethanol was reached. The specimens were cleared in graded xylene and were infiltrated with liquid paraffin then were embedded in pure paraffin. Upon sectioning, at 4-5 µm thick the specimens were attached to the gelatin-coated glass slide and let to dry at room temperature or 37°C overnight. The specimens were deparaffinized in xylene, rehydrated then were stained with hematoxylin and eosin. After being dehydrated in graded ethanol, the specimens were cleared in xylene and were mounted with an organic mounting agent. Any step in preparing histological section including samples collection, fixation, dehydration, infiltration and embedding might contribute to the quality of histological features. A proper knowledge of the tissues beeing processed, fixative solution and the histological techniques is essential to gain good results. Bouin fixative is preferable to fix fish larvae and produce a good histological feature. Decalcification is necessary to produce a good histological section on the specimens containing bone. KEY WORDS: paraffin embedded, fish embryo, fish larvae, Osteochilus vitatus, Osphronemus goramyCorresponding Author: Gratiana E Wijayanti | email: 1.bugrat@gmail.com I N T R O D U C T I O NThe most routinely employed techniques of histological preparation is the paraffin embedded sections. Since it first publication by Professor Edwin Klebs in early 1860s (Sanderson et al., 2013), paraffin embedded sections had served as an important tool in the field of biomedical (Grave, 2007;Treuting et al., 2012) as well as developmental biology (e.g. Balogh and Sótonyi, 2003;Habashy et al., 2012;Mathew and Appel, 2016). Using histological examination scientists can detect a stable condition or early sign of diseases not easily recognized by a morphological examination. The paraffin-embedded section in combination with immunoreaction has developed into a powerful technique to study tissue lineage and fate map, the immunohistochemistry.The paraffin-embedded technique has been widely used in fish research to study general structure of various organs as well as their pathological examination (Takashima and Hibiya, 1985;Palikova et al., 2004). Protocols for processing various fish organs are readily available for example eyes (Nuckel and Gross, 2007), gills, liver, kidney (Carmago andMartinez, 2007) and gonad (Wijayanti et al., 2009). However, paraffin embedded protocol for egg, embryo, and larvae are less. One of the most comprehensive protocols for fish larvae was developed by Sabaliaukas et al. (2006) enabling to process a large number of zebrafish larvae at the same time. This protocol, however, needs more sophisticated equipment such as acrylic mold and automated-closed reagent...
The cell lines play an important role in the development of basic research and applied research. Most of the available cell lines derived from mammalian tissue and only 5% of cell lines derived from fish tissue. This experimental study was conducted to evaluate the potency of caudal fin, gill lamella, hepatopancreas and spleen explant of the Osteochilus vittatus as resources for cell line development. The explants were culture in DMEM/-Ham’s 12 with 15mM HEPES supplemented with 5% L-glutamine, 5% penicillin-streptomycin, and 10% Fetal Bovine Serum for 9 days at 25°C. The results showed that the caudal fin, hepatopancreas, spleen and gill filament cell outgrowth formed confluent monolayer. In addition to monolayer cells, floating life cells were found in the culture media of spleen (1.74×106 cells) and gill lamella (1.54×105 cells). The time needed to reach confluent was significantly different among explant type (p<0.05) with the fastest growth was observed in spleen explant culture. These results indicated that the four tested explants have good potency as resources for cell line development in Osteochilus vittatus.
A B S T R A C TThe availability of fish seed is very important in fish propagation. Good quality of fish seeds were produced by a good brooder which characterized a large number of egg production at spawning. The aims of this research were to evaluate the oocytes development in the hard-lipped Barb (Osteochilus hasselti C.V.) given 17β-estradiol, different percentage of protein in the diet, and their combination. This research was conducted experimentally applying Factorial Completely Randomized Design. The first factor was percentage of protein in the diet consisted of 4 levels namely 25% (P1), 30% (P2), 35% (P3), and 40% (P4), the second factor was dose of 17β-estradiol benzoate (EB) consisted of 3 levels namely 0 µg/kg of body weight (D0), 126 µg/kg of body weight (D1), and 210 µg/kg of body weight (D2), thus there were 12 combinations of treatment with 3 replicates. The results showed that neither of protein proportion or 17β-estradiol affected the proportion of oocytes of any developmental stage (p>0,05) within the first two weeks post spawning. The treatments, however, significantly increased the proportion of oocytes at V3-V5, V6-V7, and post-vitellogenic stages (p<0,05) started at week 4 th post spawning. The GSI increased in correlation to the proportions of oocytes at stage ≥ V6 (r=0,701; p<0,01) and the dose of 17β -estradiol (r = 0.357, p <0.05). There was no significant different on GSI amongst the experimental groups. The HSI tend to decrease as the dose of 17β-estradiol increased (r = -0.210, p> 0.05). In conclusion, the percentage of protein in the diet and different doses of 17β-estradiol improved oocyte development of Hard-Lipped Barb (Osteochilus hasselti C.V.).KEY WORDS: Osteochilus hasselti, 17β-estradiol, protein content of diet, oocyte diameter, oocyte proportion Penulis korespondensi: GRATIANA E WIJAYANTI | email: bugrat_1@yahoo.co.uk P E N D A H U L U A NKeberadaan benih penting dalam usaha budidaya perikanan. Benih yang baik harus dihasilkan dari induk yang baik pula. Induk ikan yang baik memiliki gonad yang mampu menghasilkan telur dalam jumlah optimum. Maka dari itu, salah satu usaha yang dilakukan untuk mendapatkan induk dengan gonad yang baik adalah dengan pemberian pakan berprotein tinggi dan pemberian hormon untuk membantu memacu perkembangan gonad ikan.Kualitas telur sangat ditentukan oleh kualitas dan kuantitas kandungan nutrien dalam pakan. Pakan yang diberikan pada ikan ada dua macam, yaitu pakan dengan protein dominan dari sumber hewani dan pakan dengan protein dominan dari sumber nabati. Batubara (2009), menjelaskan bahwa kuantitas dan kualitas pakan, yakni protein, lemak dan vitamin yang diberikan kepada induk merupakan faktor penting yang memiliki hubungan erat dengan kematangan gonad, jumlah telur yang diproduksi, dan kualitas telur serta larva. Halver dan Hardy (2002), menjelaskan bahwa intake protein atau asam amino sangat dibutuhkan oleh ikan karena digunakan secara langsung untuk pertumbuhan dan reproduksi serta mengganti jaringan yang rusak.Kebutuhan protein pada ika...
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