ISOLATION AND IDENTIFICATION OF PARTIAL GROWTH HORMONE (GH) mRNA OF TIGER SHOVELNOSE CATFISH Pseudoplatystoma fasciatum (LINNAEUS, 1766)

The growth of fish is regulated by growth hormones secreted in a limited amount by its pituitary glands. Tiger shovelnose catfish has a relatively faster growth indicating that some growth hormones suspectedly play roles in the process and could be used in improving other farmed fish species. This study aimed to isolate and identify the growth hormone (GH) mRNA gene in Tiger shovelnose catfish (Pseudoplatystoma fasciatum Linnaeus, 1766). The sample was isolated from the pituitary, the fish measuring 602 g and 43 cm body length; Total RNA was extracted using the Tri Reagent Kit, followed with cDNA synthesis. The success of the extraction was confirmed by quantification and PCR using the β-actin gene as an internal control. GH mRNA gene was isolated by RT-PCR method, with degenerated primers from seven catfish species sequence data in the NCBI gene bank. The single band from GH gene was cloned and sequenced. Total RNA quantification with a concentration of 227 ng/µL and purity of 1.821. The successful isolation of mRNA from the pituitary gland was confirmed by amplifying the β-actin gene generated at 300 bp. This isolation of the GH mRNA gene had a sequence length of 234 bp. Tiger shovelnose catfish GH gene consists of 17 amino acid residues. The GH gene of Tiger shovelnose catfish was close to that of striped catfish (P. hypophthalmus) and Indian catfish (Clarias batrachus) with almost similar homology value of 90.6 %. Partial GH gene from tiger shovelnose catfish can be used a molecular marker in revealing the role of growth hormone on fish development, fish biology, growth gene expression, selective breeding and other mechanisms related to the aquaculture

Typification and taxonomic notes of Smilacaceae species in Java

Smilacaceae is known as a taxa with wide phenotypic variation and their taxonomical complexities remain unsolved. The three species of Smilacaceae housed in Java, are given nomenclature history. Since all the potential sources of original material have been investigated, but nothing has been identified, three neotypifications were designated here. Herbarium specimen of Koorders 34990β in Herbarium Bogoriense (BO) was chosen as the neotype of Smilax klotzschii. The de Groot & Wehlburg RD52 herbarium specimen in BO was chosen as the neotype of S. nageliana and Blume 463 herbarium specimen in L was chosen as the neotype of S. odoratissima

The Intracellular Cryoprotectant Effects in Preserving Goramy Spermatozoa after Two Days Sub-Zero Freezing

The spermatozoa quality of goramy after 2 d sub-zero freezing was examined. The quality of spermatozoa examined included motility, viability, and abnormality. We aimed to determine the optimum concentration of glycerol protecting spermatozoa during preservation. We used 0%, 1%, 3%, 5%, 7%, and 9% of glycerol, respectively. Sperms were diluted by the combination of glycerol and fish ringer (1 part of sperm + 3 part of solvent). The dilute sperms were then equiliberated at 4°C for 45 min, and were freezed at -34°C for 2 d. Thawing was then carried out at 30°C for 2 min. Based on Dunnet test, 5% of glycerol was the optimum concentration maintaining spermatozoa motility (75.95±4.76)%

Effect of Ethylene Glycol on Structural Integrity at Each Stage of Preantral Follicle Development Post Vitrification of Rat Ovary-Histological Analysis

The structure of follicular tissue affects the ability to maintain the structural integrity of follicles against cryoinjury post-vitrification. Histological analysis was conducted on the structural integrity of each stage of preantral follicles post-vitrification using 7.5% and 15.0% doses of ethylene glycol (EG), and ovarian sections with HE staining were observed using an Olympus CX21 microscope connected to Optilab 3.0 lens and Image Raster software. Analysis was conducted on the ovarian cortex in the tracing line area using polygon measure tools to obtain follicle density (follicles/mm2) and follicle index (%) data. The result showed that the EG group 7.5% (KP1) increased follicle density compared to the vitrified group (KKV) in primordial (15.83±1.77) and primary (22.94±8.51) stages. Meanwhile, KP2 (EG 15%) was in primordial (41.92±6.45), primary (11.69±1.95), secondary (33.48±3.63), and tertiary (5.93±0.69) stages. KP1 increased grade 3 follicle index compared to KKV in primary (27.66±2.34), secondary (32.41±6.99), and tertiary (25.00±5.00) stages. Meanwhile, KP2 was in primary (26.87±6.68) and tertiary (25.00±5.00) stages. Both doses of 7.5% and 15.0% EG were able to maintain structural integrity at certain stages of preantral follicles. Secondary and tertiary follicles are the best stages in maintaining grade 3 follicular integrity with the addition of 7.5% EG

Peranan Dimetil Sulfoksida Sebagai Krioprotektan Dalam Mempertahankan Kualitas Spermatozoa Ikan Gurami, Osphronemus Goramy Lacepede, 1801, Dua Puluh Empat Jam Pascakriopreservasi

Penelitian dilakukan dengan tujuan mengetahui pengaruh berbegai konsentrasi DMSO (0%, 5%, 7%, 10%, 13%, 15%, dan 17%) sebagai krioprotektan terhadap kualitas spermatozoa ikan gurami, Osphronemus goramy Lacepede, 1801, dua puluh empat jam pascakriopreservasi. Penelitian dilakukan di Balai Besar Pengembangan Budidaya Air Tawar (BBPBAT), Sukabumi. Semen dikoleksi dengan cara pengurutan (stripping) dan dievaluasi secara makroskopis (warna, pH, dan volume) dan mikroskopis (persentase motilitas, viabilitas, abnormalitas, dan konsentrasi spermatozoa) baik sebelum maupun sesudah kriopreservasi. Semen diencerkan dengan pengencer yang mengandung ekstender 189M dan krioprotektan DMSO (0%, 5%, 7%, 10%, 13%, 15%, dan 17%), disimpan pada cryotube 2 ml dan dikriopreservasi selama 24 jam. Peningkatan konsentrasi DMSO memengaruhi kualitas spermatozoa ikan gurami pascakriopresevasi. Konsentrasi DMSO yang terbaik berdasarkan hasil pengamatan adalah DMSO konsentrasi 13% (P > 0,05) dengan persentase motilitas sebesar 68,58%; abnormalitas sebesar 29%; dan viabilitas sebesar 63,5%

DNA Barcode Characterization of Chocolate Hind Grouper (Cephalopholis boenak) in Several Indonesia Waters with the New Sequences Record from Madura Island

This study aims to describe the molecular characteristics of DNA barcodes in chocolate hind grouper (Cephalopholis boenak) in Indonesian waters with new sequence records from Madura Island waters. Partial sequences of the cytochrome c oxidase 1 (CO1) gene were successfully obtained from two samples of C. boenak in Madura Island waters. In contrast, the other sequences were obtained from the BOLD system database, covering the Islands of Bali, Lombok, and Ambon waters. The overall molecular analysis involved 10 C. boenak sequences with a length of 625 bp. The results indicated that the population of C. boenak in Indonesia waters has high genetic diversity, as evidenced by the value of haplotype diversity (Hd) = 0.956 and nucleotide sequence diversity (Pi) = 0.01746, in which the population is distributed into eight haplotypes. Results of phylogenetic tree reconstruction of neighbor-joining and maximum-likelihood indicate similar topology. The branching of NJ and ML phylogenetic trees of C. boenak in Indonesia waters is grouped into two geographical clades. Clade 1 with the subpopulations of the waters from Madura, Bali, and Lombok Islands. Clade 2 with the subpopulations of the waters from Ambon Island. The results of the median-joining network reconstruction depicted a similar topology with both phylogenetic trees

The impact of late follicular progesterone level on in vitro fertilization-intracytoplasmic sperm injection outcome: Case-control study

Background: Studies have been conducted to improve the pregnancy rate through the in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) program. In recent years, researchers have been focusing on finding impact of high progesterone level on endometrial receptivity. However, data on whether progesterone level also affects the quality of the embryo is still limited. Objective: The aim is to assess the effect of late follicular progesterone level on the outcome of in vitro fertilization-intracytoplasmic sperm injection (IVF-ICSI). Materials and Methods: This was a case-control of 245 women who underwent in vitro fertilization cycle at Halim Fertility Center, Indonesia. The outcomes assessed were number of oocytes retrieved (OR), maturation rate (MR), fertilization rate (FR), number of good embryos (GE), number of fair embryos (FE), and number of poor embryos (PE). The progesterone (P4) and estradiol (E2) levels were analyzed on the day of human chorionic gonadotropin injection. Serum progesterone level was divided into three groups: 1. low progesterone (≤ 0.50 ng/ml), 2. normal progesterone (0.51-1.50 ng/ml), and 3. high progesterone (> 1.50 ng/ml). All outcomes were compared amongst the groups. Results: Significant differences occurred between progesterone level on the day of human chorionic gonadotropin administration. The number of OR in group 1, 2, and 3 were 8.41 ± 5.88 vs. 12.99 ± 8.51 vs. 17.58 ± 9.52, respectively. Conclusion: Progesterone level on the day of human chorionic gonadotropin injection may have an impact on the outcome of IVF-ICSI. Key words: Intracytoplasmic sperm injection, Embryo, Progesterone, In vitro fertilization

Genetic Diversity Of Giant Mantis Shrimp Harpiosquilla raphidea Fabricius, 1798 From Banten Bay and Other Indonesian Waters

The Harpiosquilla raphidea Fabricius, 1798 is one of the essential mantis shrimps in Banten Bay that has unique waters characteristic. Nucleotide sequence analysis of mtDNA COI region of 4 mantis shrimp population dendrogram obtained in 3 main clusters; first cluster consists of Cirebon and Jambi population, Pontianak in the second cluster population and Banten Bay in the third cluster population. Farthest genetic distance value based on mtDNA COI sequences are amongst Banten Bay and Jambi population, while the closest population is Pontianak

The Effect of Cryopreservation on Cytochrome Oxidase1 (CO1) Gene and the Relationship with Spermatozoa Motility of Albino Pangasius catfish (Pangasionodon hypophthalmus)

Cryopreservation is a technique for storing cells and tissues at very low temperatures for the possible usage of the stored cells and tissues throughout the year. Sperm cell cryopreservation in some species causes a decrease in sperm quality and DNA damage. Inappropriate cryopreservation protocols can cause changes in sperm physiology. Mitochondria are organelles that play a role in producing energy for sperm motility. Mitochondria have DNA molecules with small sizes compared to structures from nuclear DNA. This study analyzed the effect of cryopreservation on sperm motility and the Cytochrome Oxidase1 (COI) gene. The CO1 gene in mitochondrial DNA plays a role in energy production for spermatozoa motility. The cryopreservation was performed using skim milk and 10% methanol cryoprotectant, and the temperature in the equilibration process was 4-5°C for 10 minutes. Cryopreservation took place for 14 days in the freezer at -80°C. In addition, the thawing process was performed for 1-2 minutes at 40°C. This study found that the number of lesions per 10 kb in the CO1 gene in post-equilibration spermatozoa was (9.24±3.74), and post-thawing spermatozoa (was 10.26±7.54). Spermatozoa motility was obtained in fresh spermatozoa, i.e., 87±1.5%, post-equilibration spermatozoa 79±4.5%, and post-thawing spermatozoa 30±3.2%. This study concluded that cryopreservation of spermatozoa causes CO1 gene lesions and that in cryopreserved spermatozoa, there is a decrease in spermatozoa motility compared to fresh spermatozoa

Length-weight relationship and condition factor in giant harpiosquillid Mantis shrimp, Harpiosquilla raphidea (Crustacea: Stomatopoda) in Banten Bay waters, Indonesia

Length-weight relationship of Harpiosquilla raphidea from Banten Bay waters was studied from 146 males and 186 females ranging from 38 to 354 and from 37 to 348 mm, respectively. The following equations were obtained Ln W = 5.164 + 2.478 Ln X (for males), Ln W = 5.333 + 2.596 Ln X (for females) and Ln W = 5.264 + 2.549 Ln X, Ln X (pooled). The ‘b’ values (2.5497) found not near to 3 indicating allometric growth being matched up with the cube law. The ANCOVA indicated not much variation between the ‘b’ values for males and females. In general, the growth rate of the mantis shrimp H. raphidea almost the same or not much different from other mantis shrimp species of the order Stomatopoda