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Fulltext Characterization of the complete mitochondrial genome of Xanthid crab, Atergatis integerrimus from China (Decapoda: Brachyura) and its phylogenetic analysis

Xie Z, Tan H, Lin F, Guan M, Waiho K, Fang S, et al.

Mitochondrial DNA B Resour, 2018 Mar 27;3(1):397-398.
PMID: 33474181 DOI: 10.1080/23802359.2018.1456374

The complete mitochondrial genome sequence of Atergatis integerrimus from China has been amplified and sequenced in this study. The mitogenome assembly was found to be 15,924 bp in length with base composition of A (32.88%), G (10.58%), C (20.87%), T (35.66%), A + T (68.54%), and G + C (31.46%). It contained 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes and a control region. The phylogenetic position was constructed and the A. integerrimus was closely clustered with Pseudocarcinus gigas and Leptodius sanguineus. The complete mitochondrial genome sequence would be useful for further understanding the evolution of A. integerrimus.
Fulltext The whole mitochondrial genome of the mangrove crab, Metopograpsus frontalis (Miers, 1880) (Decapoda, Grapsidae) and its phylogenetic relationship

Guan M, Liu X, Lin F, Xie Z, Fazhan H, Ikhwanuddin M, et al.

Mitochondrial DNA B Resour, 2018 Mar 14;3(1):368-369.
PMID: 33490509 DOI: 10.1080/23802359.2018.1450685

In this study, we sequenced and analyzed the whole mitochondrial genome of Metopograpsus frontalis Miers, 1880 (Decapoda, Grapsidae). The circular genome is 15,587 bp in length, consisting of 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, as well as a control region. Both atp8/atp6 and nad4L/nad4 share 7 nucleotides in their adjacent overlapping region, which is identical to those observed in other Grapsidae crabs. The genome composition and gene order follow a classic crab-type arrangement regulation. The phylogenetic analysis suggested that Grapsidae crabs formed a solid monophyletic group. The newly described mitochondrial genome may provide genetic marker for studies on phylogeny of the grapsid crabs.
Gonadal microRNA Expression Profiles and Their Potential Role in Sex Differentiation and Gonadal Maturation of Mud Crab Scylla paramamosain

Waiho K, Fazhan H, Zhang Y, Zhang Y, Li S, Zheng H, et al.

Mar Biotechnol (NY), 2019 Jun;21(3):320-334.
PMID: 30835008 DOI: 10.1007/s10126-019-09882-1

Although the sexual dimorphism in terms of gonadal development and gametogenesis of mud crab has been described, the internal regulating mechanism and sex differentiation process remain unclear. A comparative gonadal miRNA transcriptomic study was conducted to identify miRNAs that are differentially expressed between testes and ovaries, and potentially uncover miRNAs that might be involved in sex differentiation and gonadal maturation mechanisms of mud crabs (Scylla paramamosain). A total of 10 known miRNAs and 130 novel miRNAs were identified, among which 54 were differentially expressed. Target gene prediction revealed a significant enrichment in 30 KEGG pathways, including some reproduction-related pathways, e.g. phosphatidylinositol signalling system and inositol phosphate metabolism pathways. Further analysis on six differentially expressed known miRNAs, six differentially expressed novel miRNAs and their reproduction-related putative target genes shows that both miRNAs and putative target genes showed stage-specific expression during gonadal maturation, suggesting their potential regulatory roles in sex differentiation and reproductive development. This study reveals the sex-biased miRNA profile and establishes a solid foundation for understanding the sex differentiation and gonadal maturation mechanisms of S. paramamosain.
Fulltext Amino acid compounds released by the giant freshwater prawn Macrobrachium rosenbergii during ecdysis: a factor attracting cannibalistic behaviour?

Juneta-Nor AS, Noordin NM, Azra MN, Ma HY, Husin NM, Ikhwanuddin M

J Zhejiang Univ Sci B, 2020 10 13;21(10):823-834.
PMID: 33043647 DOI: 10.1631/jzus.B2000126

Ecdysis is a common phenomenon that happens throughout the life phase of the giant freshwater prawn Macrobrachium rosenbergii. It is vital to better understand the correlation between cannibalism and biochemical compound that exists during the moulting process. The objective of the present study was to determine the amino acid profile released by M. rosenbergii during the ecdysis process that promotes cannibalism. To accomplish this, changes in amino acid levels (total amino acid (TAA) and free amino acid (FAA)) of tissue muscle, exoskeleton, and sample water of culture medium from the moulting (E-stage) and non-moulting (C-stage) prawns were analysed using high-performance liquid chromatography (HPLC). Comparison study revealed that among the TAA compounds, proline and sarcosine of tissues from moulting prawn were found at the highest levels. The level of FAA from water that contains moulting prawns (E-stage) was dominated by tryptophan and proline. Significant values obtained in the present study suggested that these amino acid compounds act as a chemical cue to promote cannibalism in M. rosenbergii during ecdysis. The knowledge of compositions and compounds that were released during the moulting process should be helpful for better understanding of the mechanism and chemical cues that play roles on triggering cannibalism, and also for future dietary manipulation to improve feeding efficiencies and feeding management, which indirectly impacts productivity and profitability.
Fulltext mRNA profile provides novel insights into stress adaptation in mud crab megalopa, Scylla paramamosain after salinity stress

Zhang Y, Wu Q, Fang S, Li S, Zheng H, Zhang Y, et al.

BMC Genomics, 2020 Aug 14;21(1):559.
PMID: 32795331 DOI: 10.1186/s12864-020-06965-5

BACKGROUND: Mud crab, Scylla paramamosain, a euryhaline crustacean species, mainly inhabits the Indo-Western Pacific region. Wild mud crab spawn in high-salt condition and the salinity reduced with the growth of the hatching larvae. When the larvae grow up to megalopa, they migrate back to estuaries and coasts in virtue of the flood tide, settle and recruit adult habitats and metamorphose into the crablet stage. Adult crab can even survive in a wide salinity of 0-35 ppt. To investigate the mRNA profile after salinity stress, S. paramamosain megalopa were exposed to different salinity seawater (low, 14 ppt; control, 25 ppt; high, 39 ppt).
RESULTS: Firstly, from the expression profiles of Na+/K+/2Cl- cotransporter, chloride channel protein 2, and ABC transporter, it turned out that the 24 h might be the most influenced duration in the short-term stress. We collected megalopa under different salinity for 24 h and then submitted to mRNA profiling. Totally, 57.87 Gb Clean Data were obtained. The comparative genomic analysis detected 342 differentially expressed genes (DEGs). The most significantly DEGs include gamma-butyrobetaine dioxygenase-like, facilitated trehalose transporter Tret1, sodium/potassium-transporting ATPase subunit alpha, rhodanese 1-like protein, etc. And the significantly enriched pathways were lysine degradation, choline metabolism in cancer, phospholipase D signaling pathway, Fc gamma R-mediated phagocytosis, and sphingolipid signaling pathway. The results indicate that in the short-term salinity stress, the megalopa might regulate some mechanism such as metabolism, immunity responses, osmoregulation to adapt to the alteration of the environment.
CONCLUSIONS: This study represents the first genome-wide transcriptome analysis of S. paramamosain megalopa for studying its stress adaption mechanisms under different salinity. The results reveal numbers of genes modified by salinity stress and some important pathways, which will provide valuable resources for discovering the molecular basis of salinity stress adaptation of S. paramamosain larvae and further boost the understanding of the potential molecular mechanisms of salinity stress adaptation for crustacean species.
Fulltext Fatty Acid Composition of Hepatopancreas and Gonads in Both Sexes of Orange Mud Crab, Scylla olivacea Cultured at Various Water Flow Velocities

Taufik M, Shahrul I, Mohd Nordin AR, Ikhwanuddin M, Abol-Munafi AB

Trop Life Sci Res, 2020 Jul;31(2):79-105.
PMID: 32922670 DOI: 10.21315/tlsr2020.31.2.5

Nutritional quality of the hepatopancreas and gonads of orange portunid mud crab, Scylla olivacea was evaluated for each gender under four treatment of different water velocities (0, 20, 40 and 60 cm s-1), in terms of nutrient reserve and nutrient for reproduction. About 56 crabs were used in this study in which fatty acids composition was analysed using gas chromatography mass-spectrometry (GC-MS). For hepatopancreas analysis, monounsaturated fatty acids (MUFAs) were present in the highest fatty acids concentration, followed by polyunsaturated fatty acids (PUFAs) and, saturated fatty acids (SFAs). However, long-chain polyunsaturated fatty acids (LC-PUFAs) were displayed in low concentration in the hepatopancreas. Total fatty acid (TFAs) composition was significantly higher at moderate velocity of 20 cm s-1 compared to other water velocity treatments. For gonad analysis, 20 cm s-1 showed the highest TFA concentration of 93.34 mg g-1 while, the lowest concentration of 3.90 mg g-1 occurred at 0 cm s-1. There were significant differences in male and female crab's fatty acids contents of gonads at all flow velocities challenged (p < 0.05). PUFAs and MUFAs were dominant while, SFAs were observed at low concentration. This study revealed that, concentration of PUFAs increased as gonad maturation increased. The decreasing concentration of hepatopancreas fatty acids over the culture period indicated that nutrient was shifted from the hepatopancreas, to be used as energy reserved to gonads for further growth of eggs and offspring. The linkages between water flow strength, hepatopancreas, and gonad fatty acids concentrations, is fundamental knowledge useful in establishing efficient habitat velocities selection which will improve aquaculture production of mud crabs with high quality broodstock.
The significant sex-biased expression pattern of Sp-Wnt4 provides novel insights into the ovarian development of mud crab (Scylla Paramamosain)

Farhadi A, Fang S, Zhang Y, Cui W, Fang H, Ikhwanuddin M, et al.

Int J Biol Macromol, 2021 Jul 31;183:490-501.
PMID: 33957197 DOI: 10.1016/j.ijbiomac.2021.04.186

The wingless-type MMTV integration site family member-4 (Wnt4), a member of the wingless-related integration site (Wnt) family, is widely accepted as a key regulator of ovarian development in mammals. In this study, a full-length cDNA of Wnt4 (designated as Sp-Wnt4) was cloned, characterized, and functionally studied in mud crab (Scylla paramamosain). The full-length cDNA of Sp-Wnt4 consists of 2659 bp with an open reading frame (ORF) encoding 359 amino acids, a 907 bp 5'-UTR and a 672 bp 3'-UTR. Sp-Wnt4 contains 25 cysteine (Cys) residues and three potential N-glycosylation sites. Sp-Wnt4 protein shared the highest identity (98.9%) to the Wnt4 protein of Portunus trituberculatus. The phylogenetic tree showed that Sp-Wnt4 and Wnt4 protein of Malacostracan crustaceans clustered together, indicating that they had a close genetic distance. Sp-Wnt4 was expressed at a higher level in the ovary compared to other tissues, with the highest expression level at the third stage (O-III) of the ovarian development (P < 0.05). A downward trend was observed in the expression level of Sp-Wnt4 from the embryo stage to crablet stages (P < 0.05). After unilateral eyestalk ablation, the expression level of Sp-Wnt4 significantly increased in testis (14-fold) and downregulated (3.1-fold) in the gill (P < 0.05) of females. In situ hybridization (ISH) assay revealed that Sp-Wnt4 transcripts were mainly localized in the cytoplasm of oocyte cells. These findings showed that Sp-Wnt4 play crucial roles in the ovarian development of S. paramamosain. In conclusion, our study provides novel insights into the evolution and roles of the Wnt4 gene.
Fulltext Use of Pelleted Diets in Commercially Farmed Decapods during Juvenile Stages: A Review

Aaqillah-Amr MA, Hidir A, Azra MN, Ahmad-Ideris AR, Abualreesh MH, Noordiyana MN, et al.

Animals (Basel), 2021 Jun 12;11(6).
PMID: 34204676 DOI: 10.3390/ani11061761

The increasing market demand for decapods has led to a considerable interest in cultivating decapod species at a larger scale. Following the development of hatchery technologies, most research has focused on the development of formulated feeds for commercially farmed decapods once they enter the juvenile stages. The use of formulated feed for decapods at a commercial scale is still in the early stages. This is probably because of the unique feeding behavior that decapods possess: being robust, slow feeders and bottom dwellers, their feeding preferences change during the transition from pelagic larvae to benthic juveniles as their digestive systems develop and become more complex. The current practice of decapod aquaculture involves the provision of juveniles with food such as natural diet, live feed, and formulated feed. Knowledge of nutrient requirements enables diets to be better formulated. By manipulating the levels of proteins and lipids, a formulated feed can be expected to lead to optimal growth in decapods. At the same time, the pellet's physical characteristics are important factors to be considered upon formulating commercially farmed decapod feeds, considering the unique feeding behavior of the decapod. However, most published studies on decapod nutrition lack data on the physical characteristics of the feed types. Thus, it is difficult to establish a standard feed formulation that focuses on the physical pellet properties. Moreover, careful consideration must be given to the feeding behavior of species, as decapods are known as bottom feeders and are robust in terms of handling feed. Information on the pellet forms, diet composition, and unique feeding behaviors in commercially farmed decapods is gathered to suggest potential better formulated diets that can optimize growth and reproduction. Thus, the purpose of this review is to summarize the information that has been published to date and to come up with suggestions on ways to improve the feed formulation in decapods that comply with their feeding behavior and nutrient requirements. Further research is needed to explore the potential of the pelleted feed at the adult stage so the decapod can take full advantage of the nutrients present in the pellets.
Fulltext The final spawning ground of Tachypleus gigas (Müller, 1785) on the east Peninsular Malaysia is at risk: a call for action

Nelson BR, Satyanarayana B, Moh JHZ, Ikhwanuddin M, Chatterji A, Shaharom F

PeerJ, 2016;4:e2232.
PMID: 27547542 DOI: 10.7717/peerj.2232

Tanjung Selongor and Pantai Balok (State Pahang) are the only two places known for spawning activity of the Malaysian horseshoe crab - Tachypleus gigas (Müller, 1785) on the east coast of Peninsular Malaysia. While the former beach has been disturbed by several anthropogenic activities that ultimately brought an end to the spawning activity of T. gigas, the status of the latter remains uncertain. In the present study, the spawning behavior of T. gigas at Pantai Balok (Sites I-III) was observed over a period of thirty six months, in three phases, between 2009 and 2013. Every year, the crab's nesting activity was found to be high during Southwest monsoon (May-September) followed by Northeast (November-March) and Inter monsoon (April and October) periods. In the meantime, the number of female T. gigas in 2009-2010 (Phase-1) was higher (38 crabs) than in 2010-2011 (Phase-2: 7 crabs) and 2012-2013 (Phase-3: 9 crabs) for which both increased overexploitation (for edible and fishmeal preparations) as well as anthropogenic disturbances in the vicinity (sand mining since 2009, land reclamation for wave breaker/parking lot constructions in 2011 and fishing jetty construction in 2013) are responsible. In this context, the physical infrastructure developments have altered the sediment close to nesting sites to be dominated by fine sand (2.5Xφ ) with moderately-well sorted (0.6-0.7σφ), very-coarse skewed (-2.4SKφ), and extremely leptokurtic (12.6Kφ) properties. Also, increased concentrations of Cadmium (from 4.2 to 13.6 mg kg(-1)) and Selenium (from 11.5 to 23.3 mg kg(-1)) in the sediment, and Sulphide (from 21 to 28 µg l(-1)) in the water were observed. In relation to the monsoonal changes affecting sheltered beach topography and sediment flux, the spawning crabs have shown a seasonal nest shifting behaviour in-between Sites I-III during 2009-2011. However, in 2012-2013, the crabs were mostly restricted to the areas (i.e., Sites I and II) with high oxygen (5.5-8.0 mg l(-1)) and moisture depth (6.2-10.2 cm). In view of the sustained anthropogenic pressure on the coastal habitats on one hand and decreasing horseshoe crabs population on the other, it is crucial to implement both conservation and management measures for T. gigas at Pantai Balok. Failing that may lead to the loss of this final spawning ground on the east coast of P. Malaysia.
Fulltext Transcriptome Analysis and Differential Gene Expression on the Testis of Orange Mud Crab, Scylla olivacea, during Sexual Maturation

Waiho K, Fazhan H, Shahreza MS, Moh JH, Noorbaiduri S, Wong LL, et al.

PLoS One, 2017;12(1):e0171095.
PMID: 28135340 DOI: 10.1371/journal.pone.0171095

Adequate genetic information is essential for sustainable crustacean fisheries and aquaculture management. The commercially important orange mud crab, Scylla olivacea, is prevalent in Southeast Asia region and is highly sought after. Although it is a suitable aquaculture candidate, full domestication of this species is hampered by the lack of knowledge about the sexual maturation process and the molecular mechanisms behind it, especially in males. To date, data on its whole genome is yet to be reported for S. olivacea. The available transcriptome data published previously on this species focus primarily on females and the role of central nervous system in reproductive development. De novo transcriptome sequencing for the testes of S. olivacea from immature, maturing and mature stages were performed. A total of approximately 144 million high-quality reads were generated and de novo assembled into 160,569 transcripts with a total length of 142.2 Mb. Approximately 15-23% of the total assembled transcripts were annotated when compared to public protein sequence databases (i.e. UniProt database, Interpro database, Pfam database and Drosophila melanogaster protein database), and GO-categorised with GO Ontology terms. A total of 156,181 high-quality Single-Nucleotide Polymorphisms (SNPs) were mined from the transcriptome data of present study. Transcriptome comparison among the testes of different maturation stages revealed one gene (beta crystallin like gene) with the most significant differential expression-up-regulated in immature stage and down-regulated in maturing and mature stages. This was further validated by qRT-PCR. In conclusion, a comprehensive transcriptome of the testis of orange mud crabs from different maturation stages were obtained. This report provides an invaluable resource for enhancing our understanding of this species' genome structure and biology, as expressed and controlled by their gonads.
De novo assembly of genome and development of polymorphic microsatellite loci in the blue swimming crab (Portunus pelagicus) using RAD approach

Wu Q, Miao G, Li X, Liu W, Ikhwanuddin M, Ma H

Mol Biol Rep, 2018 Dec;45(6):1913-1918.
PMID: 30203240 DOI: 10.1007/s11033-018-4339-9

The blue swimming crab (Portunus pelagicus) is a valuable marine fishery resource in Indo-West Pacific Ocean. So far, rare genetic resource of this species is available. In this report, the restriction-site associated DNA (RAD) approach was employed to mine the genomic information and identify molecular markers in P. pelagicus. A total of 0.82 Gbp clean data were generated from the genome of individual "X2A". De novo assembly produced 85,796 contigs with an average length of 339 bp. A total of 45,464 putative SNPs and 17,983 microsatellite loci were identified from the genomes of ten individuals. Furthermore, 31 pairs of primers were successfully designed, with 16 of them exhibiting polymorphism in a wild population. For these polymorphic loci, the expected and observed alleles per locus ranged from 1.064 to 7.314 and from 2 to 11, respectively. The expected and observed heterozygosity per locus ranged from 0.0615 to 0.819 and from 0.0626 to 1.000, respectively. Nine loci showed high informative with polymorphism information content (PIC) > 0.5. Five loci significantly deviated from Hardy-Weinberg equilibrium in the samples analyzed. No linkage disequilibrium was found among the 16 polymorphic microsatellite loci. This study provided massive genetic resource and polymorphic molecular markers that should be helpful for studies on conservation genetics, population dynamics and genetic diversity of P. pelagicus and related crab species.
Experimental data on glucose and energy levels of orange mud crab, Scylla olivacea at different water velocities

Taufik M, Amin-Safwan A, Mohd Nordin AR, Shahrul I, Abol-Munafi AB, Ikhwanuddin M

Data Brief, 2020 Apr;29:105232.
PMID: 32099875 DOI: 10.1016/j.dib.2020.105232

The present datasets were conducted to investigate glucose concentration in hemolymph, energy levels at selected body parts (hepatopancreas, muscle, gonad), and feces among different sexes of crabs cultured at four different water velocities (0, 20, 40, and 60 cm/s) during a 60-day culture period. A total of 102 immature crabs (51 males, and 51 females) were sampled from Kuala Muda, Kedah coastal water, Peninsular Malaysia (5°39'N 100°19'E) from April to November of 2018. Results indicated that glucose concentration was the highest at water velocity of 60 cm/s for both male and female crabs (♂: 3.76 ± 0.08 mmol/L; ♀: 3.63 ± 0.06 mmol/L), whereas at 0 cm/s, the lowest levels of glucose concentration (♂: 0.13 ± 0.08 mmol/L; ♀: 0.19 ± 0.06 mmol/L) were recorded. As for energy analysis in hepatopancreas, results showed that both male and female crabs recorded the highest levels at 0 cm/s (no flow) with 37.919 ± 0.07 KJ/g and 34.636 ± 0.50 KJ/g, respectively. Energy for locomotion (muscle) of male crabs recorded the highest at 0 cm/s (♂: 26.823 ± 0.06 KJ/g), meanwhile for females, the highest was recorded at 20 cm/s (26.607 ± 0.34 KJ/g). Energy for reproduction of males could not be compared due to an insufficient available amount of testes/vas deferens, whereas female crabs recorded the highest energy usage at 20 cm/s water velocity (♀: 37.895 ± 0.08 KJ/g). For feces, both male and female crabs recorded the lowest energy at 60 cm/s (♂: 5.841 ± 0.03 KJ/g; ♀: 5.393 ± 0.01 KJ/g). Glucose assessment showed a direct relationship between increased velocity and glucose secretion in hemolymph at high velocity of 60 cm/s (stress condition) compared to other treatments. Regarding energy analysis, this research improved the mechanism of hepatopancreas, gonad, muscle and feces functions in development and reproduction, while it shed light on the influence of velocity on energy metabolism of S. olivacea.
Comparative profiling of ovarian and testicular piRNAs in the mud crab Scylla paramamosain

Waiho K, Fazhan H, Zhang Y, Li S, Zhang Y, Zheng H, et al.

Genomics, 2020 01;112(1):323-331.
PMID: 30807818 DOI: 10.1016/j.ygeno.2019.02.012

PIWI-interacting RNAs (piRNAs) are abundantly found in germ cells and involved in gametogenesis and gonadal development. Information on the regulatory roles of piRNAs in crustacean reproduction, however, is scarce. Thus, we identified gonadal piRNAs of mud crab Scylla paramamosain. Of the 115,491 novel piRNAs, 596 were differentially expressed. Subsequently, 389,887 potential piRNA-target genes were predicted. The expression of 4 piRNAs and 9 genes with high piRNA interactions were validated with the inclusion of additional immature specimens, including LRP2 that is involved in growth and reproduction, MDN1 in ribosome biogenesis pathway and gametogenesis, and PRKDC, a DNA repair gene involved in gonadal differentiation and maturation. KEGG analysis further revealed the involvement of predicted piRNA target genes in gametogenesis- and reproduction-related pathways. Our findings provide baseline information of mud crab piRNAs and their differential expression between testes and ovaries suggests that piRNAs play an essential role in regulating gametogenesis and gonadal development.
Successful sperm cryopreservation of the brown-marbled grouper, Epinephelus fuscoguttatus using propylene glycol as cryoprotectant

Yusoff M, Hassan BN, Ikhwanuddin M, Sheriff SM, Hashim F, Mustafa S, et al.

Cryobiology, 2018 04;81:168-173.
PMID: 29355519 DOI: 10.1016/j.cryobiol.2018.01.005

This study developed the cryopreservation of brown-marbled grouper spermatozoa for practical application. We examined 32 cryodiluents, developed from four types of cryoprotectants [propylene glycol (PG), dimethyl-sulphoxide (Me2SO), dimethyl-acetamide (DMA) and ethylene glycol (EG)] at four concentrations of 5, 10, 15 and 20% in combination with two extenders [Fetal bovine serum (FBS) and artificial seminal plasma (ASP). Cooling rates were examined by adjusting the height of straws (2.5-12.5 cm) from the liquid nitrogen (LN) vapor and cooled for 5 min before immersion into LN. DNA laddering was used to detect DNA damage in cryopreserved sperm. In fertilization trials, 0.5 g of eggs was mixed with cryopreserved sperm stored for 30 days in LN. The best motility of post-thaw sperm was achieved using 15% PG + 85% FBS (76.7 ± 8.8%); 10% PG + 90% FBS was also effective as cryodiluent. Generally, FBS gave better post-thaw motility compared to ASP. The optimum cooling rate was at 17.6 °C min-1 obtained by freezing at the height of 7.5 cm surface of LN. The results obtained showed that cryopreserved sperm of brown-marbled grouper suffered slight DNA fragmentation, which resulted in significantly lower motility. However, the fertilization (90.9 ± 0.5%), hatching (64.5 ± 4.1%) and deformity rates (3.8 ± 0.2%) obtained from cryopreserved sperm showed no significant difference with fresh sperm. These findings show that the developed protocol for cryopreservation of brown-marbled grouper sperm was viable and will be useful for successful breeding and seed production of brown-marbled grouper.
Ovarian lavage method as an alternative route for hormonal administration and short-term sperm storage in Clarias gariepinus

Okomoda VT, Amighty RO, Bem TM, Amaantimin J, Nurizzati I, Koh ICC, et al.

Theriogenology, 2023 Mar 01;198:203-209.
PMID: 36592519 DOI: 10.1016/j.theriogenology.2022.12.037

Ovarian lavage is a term used to describe the injection of fish with a catheter through the oviduct into the ovary. In this study, the efficacy of this technique was evaluated as a route for hormone administration and sperm preservation in the African catfish Clarias gariepinus. Firstly, the effects of hormone injection routes (namely, intramuscular, intraperitoneal, and ovarian lavage) were evaluated on breeding and haematological parameters. In the second study, the fish's spermatozoa were stored in the ovaries for 1, 2, 3, and 4 days before stripping, sperm activation with freshwater, and fertilization. The breeding performance was then compared with eggs fertilized using spermatozoa refrigerated for the same duration. The study showed that the administration of synthetic hormone (ovaprim®) through the ovaries was comparable to the intramuscular route, while those injected intraperitoneally had the least values (P 0.05) the fertilization (92-93%) and hatching (81-83%) of the eggs when compared to the control (91% and 82%). Beyond this 24hr threshold, breeding performances were significantly reduced in the ovarian lavage treatments compared to those fertilized with refrigerated sperm (P
Fulltext Effects of Annona muricata extraction on inhibition of polyphenoloxidase and microbiology quality of Macrobrachium rosenbergii

Ibrahim A, Ibrahim MSC, Bakar K, Bakar J, Ikhwanuddin M, Karim NU

J Food Sci Technol, 2022 Mar;59(3):859-868.
PMID: 35153318 DOI: 10.1007/s13197-021-05081-w

Giant freshwater prawn (Macrobrachium rosenbergii) is one of the important aquaculture species and quickly expanding in many countries. High demand and mass commercialization on M. rosenbergii regulating 18% of the international seafood business. Seafood products contend with various level across the supply chains and time to reach the consumers depending upon the marketing and delivery channels after harvesting. Therefore, these may cause biodeterioration such as melanosis (dark pigmentation) and microbial changes that limit the shelf life. This studies reveal the antioxidant properties from Annona muricata leaves extract and their effectiveness in inhibiting the polyphenoloxidase (PPO) activity and delaying the bacterial accumulation during 20 days of chilled storage. Five metabolites including coumarins, flavonoid, glycoside, terpenoids and steroid compound were found in A. muricata leaves extract. Total phenolic content and total flavonoid content of A. muricata were recorded at 191.24 ± 0.03 mgGAEg-1 and 1777.48 ± 1.08 mgQEg-1, respectively. Sixteen percent (16%) of A. muricata leaf extract effectively inhibit 82.41% PPO. Furthermore, 15% of A. muricata leaves extracts showed a significant reduced (p M. rosenbergii. These conclude that the present of listed secondary metabolites and at approximately ~ 15-16% of A. muricata leaves extracts were effectively inhibiting the melanosis and prolong the shelf life for up to 8 days of M. rosenbergii stored at chilled condition. Therefore, A. muricata leaves extract is potential used as natural preservative agent in obtaining high quality seafood products.
Fulltext Aquaponics production system: A review of historical perspective, opportunities, and challenges of its adoption

Okomoda VT, Oladimeji SA, Solomon SG, Olufeagba SO, Ogah SI, Ikhwanuddin M

Food Sci Nutr, 2023 Mar;11(3):1157-1165.
PMID: 36911833 DOI: 10.1002/fsn3.3154

The aquaponics production system integrates hydroponics and recirculatory aquaculture system for the simultaneous production of plants and fish. At a time, such as the postpandemic era, the aquaponics system represents an efficient green farming and eco-friendly alternative to sustainable agricultural production. In this review, the history and development of the production systems were traced vis-a-vis its pros and cons. Although there has been much dispute about the origin of the system, the numerous records of developmental attempts in history have all led to the current complexity of the systems and their efficiency. Water conservation, improved performance, food security, less pollution, and low energy consumption are some of the advantages identified in the use of aquaponics systems for food production. Challenges to the domestication of the system, however, include moderately high start-up capital, the need for stable electricity to operate the system, nutrient availability, as well as treatment of diseases in the system. Although the aquaponics production system could be a panacea for food security in Africa, modalities for the domestication of this technology are largely not in place, hence the need for some government interventions in this regard.
Fulltext Assessment on Water Quality Parameter and Nutrients Level of Nyatuh River in Relations with Macrobrachium Rosenbergii Prawn Populations

Kasan NA, Ikhwanuddin M, Manan H, Zakaria NS, Kamaruzzan AS, Rahim AIA, et al.

Trop Life Sci Res, 2023 Mar;34(1):51-66.
PMID: 37065803 DOI: 10.21315/tlsr2023.34.1.4

In order to determine the safety level of water parameters and nutrients in the natural environment of fish and freshwater prawn in Nyatuh River, Terengganu, Malaysia, it is necessary to conduct an assessment of water quality parameters. Due to its important, a study was conducted to assess the water quality parameter and nutrients contents from Nyatuh River of Setiu Terengganu in relations to the population of freshwater prawn, Macrobrachium rosenbergii caught along the Nyatuh River basin. Total of four expeditions and five stations at different tidal condition for the water quality parameter were assessed during the study. From the results achieved, the overall temperature varied between (26.56°C-29.30°C), dissolved oxygen, DO (3.59 mg/L-6.50 mg/L), pH (4.99-7.01), salinity (0.01ppt-4.22ppt), depth (2.71 m-5.54 m) while for ammonia (0.01 mg/L-0.24 mg/L), nitrite (0.01 mg/L-0.05 mg/L) and phosphate (0.01 mg/L-0.12 mg/L). While the number of prawns caught are 176, 160, 102 and 68 for Expeditions 1, 2, 4 and 3, respectively. Possibly, the heterogeneous number of prawns caught is a result of significant differences in water level depth during high tides and low tides, as well as a fluctuation in the ammonia concentration levels in each of the stations and expeditions. For statistical analysis, the temperature showed no significant difference between the expedition, stations and tidal. That is p = 0.280, p > 0.05 and F = 1.206, respectively. While dissolved oxygen, DO, showed no significant difference as well, that is p = 0.714, p > 0.05 and F = 0.737. However, the level of water depth was significantly different between expedition, station and tidal, that is p = 0.000, p < 0.05 and F = 3.120. Ammonia, on the other hand shows no significant difference between expedition, station and tidal, that is p = 0.476, p > 0.05 and F = 0.973. The same goes for nitrite and phosphate concentration. There was no significant difference between expedition, stations and tidal, that is p = 0.569, p > 0.05 and F = 0.879 and p = 0.247, p > 0.05, F = 1.255, respectively. In Expedition 1, the good water quality parameter and very low ammonia concentration resulted in a larger prawn population as compared to other expeditions. The distribution or mixture of prawns caught is heterogeneous at different stations due to the significant differences in water depth and also to the fluctuation in water quality due to varying ammonia levels. In conclusion, the water quality in Nyatuh River fluctuated across expeditions, stations, and tides, as well as significant differences in water level depths between high and low tides. Due to the rapid growth and importance of industrial and aquaculture operations along the river, extra attention should be devoted to avoid the impact of excessive pollutant in order to protect the ecosystem.
Transcriptome-seq provides insights into sex-preference pattern of gene expression between testis and ovary of the crucifix crab ( Charybdis feriatus)

Zhang Y, Miao G, Fazhan H, Waiho K, Zheng H, Li S, et al.

Physiol Genomics, 2018 05 01;50(5):393-405.
PMID: 29570432 DOI: 10.1152/physiolgenomics.00016.2018

The crucifix crab, Charybdis feriatus, which mainly inhabits Indo-Pacific region, is regarded as one of the most high-potential species for domestication and incorporation into the aquaculture sector. However, the regulatory mechanisms of sex determination and differentiation of this species remain unclear. To identify candidate genes involved in sex determination and differentiation, high throughput sequencing of transcriptome from the testis and ovary of C. feriatus was performed by the Illumina platform. After removing adaptor primers, low-quality sequences and very short (<50 nt) reads, we obtained 80.9 million and 66.2 million clean reads from testis and ovary, respectively. A total of 86,433 unigenes were assembled, and ~43% (37,500 unigenes) were successfully annotated to the NR, NT, Swiss-Prot, KEGG, COG, GO databases. By comparing the testis and ovary libraries, we obtained 27,636 differentially expressed genes. Some candidate genes involved in the sex determination and differentiation of C. feriatus were identified, such as vasa, pgds, vgr, hsp90, dsx-f, fem-1, and gpr. In addition, 88,608 simple sequence repeats were obtained, and 61,929 and 77,473 single nucleotide polymorphisms from testis and ovary were detected, respectively. The transcriptome profiling was validated by quantitative real-time PCR in 30 selected genes, which showed a good consistency. The present study is the first high-throughput transcriptome sequencing of C. feriatus. These findings will be useful for future functional analysis of sex-associated genes and molecular marker-assisted selections in C. feriatus.
Histological Characteristics on the Testes of Mud Spiny Lobster, Panulirus polyphagus (Herbst, 1793)

Nor Fatihah S, Muhd-Farouk H, Amin-Safwan A, Hafiz Mahsol H, Ikhwanuddin M

Pak J Biol Sci, 2017;20(7):365-371.
PMID: 29023069 DOI: 10.3923/pjbs.2017.365.371

BACKGROUND AND OBJECTIVE: Mud spiny lobsters, Panulirus polyphagus (P. polyphagus) are one of the most important fisheries resources now-a-days due to quality that it's possessed. However, there is still lack of in-depth study about this species mainly on males testicular characteristics and sexual maturity size. Therefore, the present study was carried out to investigate the histological characteristics on the testes and sexual maturity sizes of mud spiny lobster, Panulirus polyphagus.
MATERIALS AND METHODS: The testes were dissected out and fixed in 10% buffered formalin solution for 11 h, dehydrated in 70% alcohol and lastly placed in tissue processor for 18±1 h at 60°C. The tissues blocks were cut at the thickness of 4 μm on a rotary microtome. Stained tissues were taken under Advance Microscope (Nikon Eclipse 80i Nomarski DIC). Collected data were analyzed using Microsoft Excel 2013. Data were presented as mean±standard deviation. Statistical analyses were done using one-way ANOVA using SPSS (Version 22).
RESULTS: These lobules of mature P. polyphagus were formed via different germinative lineage cells such as spermatogonia, spermatocytes, spermatids and spermatozoa. The histological characteristics of testes showed that the process of spermatogenesis went through the stages of four testes maturation which were spermatogonia I and II, spermatocytes I and II, spermatids and spermatozoa stages within different body weight of P. polyphagus. It was found that there were significant difference between body weight and carapace length to the testicular maturation stages (one-way ANOVA and p = 0.000).
CONCLUSION: The results of this experiment indicated that males P. polyphagus have four stages of testes maturation and can be considered to have fully mature testes that ready for fertilization at 452 g body weight (BW) and 107 mm carapace length (CL) or more.

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