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Articles by M. Ikhwanuddin
Total Records ( 27 ) for M. Ikhwanuddin
  M.Z. Ihwan , W. Wahidah , M.A. Ambak , M. Ikhwanuddin and H. Marina
  Seventy three sample of wild mud crab genus Scylla were collected from setiu wetland, Terengganu coastal waters, Malaysia with the measured of Carapace Width (CW) and Body Weight (BW) 8.9±0.1 and 103.9±3.8, respectively. The percentage prevalence of parasites from these studies is 81.9% and the mean intensity was 13. The total number of 887 parasites that were collected includes barnacle (Octolasmis spp.), ciliates, sessile protozoan (Epistylis sp.), nematode and copepod. The higher number of parasites that were found is barnacle with prevalence of 71.1% and mean intensity 14.4 followed by ciliates, nematode, copepod and sessile protozoan with prevalence of 18.1, 8.4, 2.4 and 1.2% and mean intensity 1.1, 2.1, 2.0, 1.0, respectively. Results show that pedunculate barnacle genus Octolasmis show the most abundance species of parasites, compared to others.
  A.D. Talpur , A.J. Memon , M.I. Khan , M. Ikhwanuddin , M.M. Danish Daniel and A.B. Abol-Munafi
  Vibrio harveyi, Vibrio parahaemolyticus, Pseudoalteromonas piscicida, Staphylococcus epidermidis and Micrococcus luteus were isolated from the gut of blue swimming crab, Portunus pelagicus captured from Strait of Tebrau Johor Malaysia and studied for pathogenicity against the Zoea-1 (Z1 stage) of P. pelagicus. Pathogenic isolates V. harveyi and P. piscicida resulted in 100% mortality at 106 cfu mL-1 and 105 cfu mL-1 after 24 h and 72 h post dose. Conversely, V. parahaemolyticus produced 100% deaths at inoculation 106 cfu mL-1 after 72 h post dose. Cumulative mortality was observed rising with the increase in dose potency of pathogens. S. epidermidis and M. luteus detected with feeble pathogenic characteristics. The LD50 of V. harveyi was 1.2x103 cfu ML-1 (24 h), V. parahaemolyticus was 9.6x105 cfu mL-1 (72 h), P. piscicida was 9.8x103 cfu mL-1 (24 h) and S. epidermidis was 9.8x105 cfu mL-1 (72 h). The mean differences among various pathogenic doses were statistically significant (p<0.05). Susceptibility tests of total 662 isolates were under taken including V. harveyi (n = 180), V. parahaemolyticus (n = 180) and P. piscicida (n = 119), isolates showed mixed trend as multiple resistance and sensitive to antimicrobial agents tested while S. epidermidis (n = 88) and M. Luteus (n = 95) were sensitive to all antibiotics tested. V. harveyi, V. parahaemolyticus and P. piscicida did not show 100% resistance to any of the antibiotics tested. From the results of 14 antibiotics tested, we observed that the highest frequency of single drug resistance in V. harveyi was Streptomycin (89.44%) and sensitive to chloramphenicol (70.55%). Similarly, the highest frequency of single-drug resistance in V. parahaemolyticus was to kanamycin (92.78%) and sensitive to chloramphenicol (93.33%) and P. piscicida was to penicillin (80.67+19.33% intermediate but no sensitive) and sensitive to gentamicin (98.32%). Infections caused by antibiotic resistant pathogens have serious consequences and therapeutic use of tested antibiotic is questionable in larviculture of P. pelagicus.
  A.J. Memon , A.D. Talpur , M.I. Khan , M.O. Fariddudin , J. Safiah , A.B. Abol-Munafi and M. Ikhwanuddin
  The objectives of this study were to determine the effects of different cryoprotectants on sperm viability and optimization of spermatophore cryopreservation protocol for durable storage of Banana shrimp (Penaeus merguiensis). Spermatophore suspended for 15 min in Calcium-Free saline (Ca-F saline), used CPA MgCl2 and with concentration (15%), thawing temperature was 27°C. Use 15 min equilibration in room temperature (25°C) overall. Exposure and cooling rate selected as 25, 20, 16, 4, 2, -4, -20, -80, -150°C/10 min. Examination of sperm viability used a modified eosin-nigrosin staining technique. The smallest reductions in apparent sperm viability occurred with MgCl2, however freezing protocol was developed using Ca-F saline containing 15% MgCl2. Spermatophores were cryopreserved using above exposure/cooling rate and -196°C in liquid nitrogen up to 180 days. Mean sperm viability for fresh (93.8±1.3%) and cryopreserved spermatophore held for 24 h and 60 days was 83.5±0.6 and 61±1.2 did not differ (p>0.05), however that for spermatophore stored in liquid nitrogen between 90 and 180 days were lower (p<0.05) and varied from 55.4±0.3-16.4±1.2. Spermatophores earlier held in liquid nitrogen for 60 and 90 days. However, storage beyond 90 days caused a significant decline (p<0.05) in sperm viability. Spermatophores kept for 120 and 150 days had viabilities of 48.9±0.9 and 32.4±0.9%, respectively. Cryopreserved spermatophore stored in liquid nitrogen from 150-180 days had low viabilities (<35%). Mean fertilization rate of P. merguiensis females artificially inseminated with cryopreserved spermatophore that had been stored in liquid nitrogen for 7-30 days and for 60-90 days were 73.9±1.5-66.7±3.1 and 67.3±3-64.1±2.1%, respectively whereas that of fresh spermatophore was 88.2±1.5%. Hatching rates of eggs fertilized with cryopreserved spermatophore kept for 7-30 days and for 60-90 days were 77.6±2.5-72.7±3.5 and 81.5±12.1-62.5±1.5 which were not different (p>0.05) from those of the control group 76.2±13.5%, respectively. In conclusion, Cryopreserved spermatophore held in liquid nitrogen l<90 days revealed high sperm viability although, for longer periods, sperm viability declined at 180 days.
  A.D. Talpur , A.J. Memon , M.I. Khan , M. Ikhwanuddin , M.M. Danish Daniel and A.B. Abol-Munafi
  Blue swimming crab, Portunus pelagicus has not anchored the roots in aquaculture due to non availability of commercial seed production. Letdown of seed production is owing to microbial infections. To combat with microbes, study was aimed to isolate and screen probiotics from the gut of female crab for larviculture. Based on characteristics of inhibitory activity against pathogenic V. harveyi, V. parahaemolyticus and P. piscicida, bile, acid, salt tolerances and survival in sea water, isolates were identified as L. plantarum, L. salivarius, L. rhamnosus W. confusa and W. cibaria and evaluated for probiotics. A new model small scale in vivo validation was developed for conformity of the isolates as probiotics for P. pelagicus larviculture. The LAB isolates were administrated as water additive at concentrations 102, 104 and 106 cfu mL-1 for one day and five days in vivo validation experiments and positive control was inoculated with same concentrations of V. harveyi while negative control employed with larvae and no inoculation. Highest larval survival achieved at concentration 106 cfu mL-1 and L. plantarum, L. salivarius and L. rhamnosus did show significant larval survival. W. confusa and W. cibaria did not demonstrate as probiotics. L. plantarum showed highest survival 49.45±4.80% and 54.44±6.74% in both inoculations, respectively and no survival was observed in five days+ve control. Water quality degradation was not evident but improvement in pH was noticed. Based on results of small-scale in vivo test three LAB probiotics, L. plantarum, L. salivarius and L. rhamnosus were selected for larviculture of P. pelagicus.
  S. Noor Baiduri , S. Nurul Akmal and M. Ikhwanuddin
  Scylla olivacea and Scylla tranquebarica are two mud crabs species that live associated in the wild. This study was done to investigate the mating success between the two species by determination of the duration and recording the process of mating activity. Wild mud crab samples were collected from Terengganu coastal water, Malaysia. Limb autotomy was subjected to female crabs to induce molting before the introduction of male crab for mating trials. Ten pairs of each mating trial were observed. Mating success of control trials of S. olivacea (T1) and S. tranquebarica (T2) is 60 and is 50%, consecutively. Mating success of hybrid trials for male S. olivacea with female S. tranquebarica (T3) is 40% and male S. tranquebarica with female S. olivacea (T4) is 30%. The highest mean duration of pre-copulatory guarding is T3 (12,240 min±1,859.0) while the lowest is T2 (8,064 min±1,287.9). As for copulation, the highest mean duration is T4 (59.3 min±18.0) while the lowest is T1 (59.3 min±18.0). Meanwhile, the highest mean duration of post-copulatory guarding is T2 (312.0 min±50.2) and the lowest is T3 (82.5 min±28.7). Longer duration of pre-copulatory guarding and copulation were observed on hybrid trials (T3, T4) compare to the controls (T1, T2). It has been observed that all successful mating trials in the present study show general mating activity, involving pre-copulatory guarding, copulation and post-copulatory guarding. Result of the present study shows that hybridization can occur in captivity and there are also possibilities of hybridization between the two mud crab species to occur in the wild.
  M. Ikhwanuddin , M.N. Azra , Y.Y. Sung , A.B. Abol- Munafi and M.L. Shabdin
  The study were aim to demonstrate the affects of live food type’s i.e., mixed diatom, Artemia nauplii and rotifer on survival rate and molt time of larvae stage till 1st day juvenile crabs (C1) of P. pelagicus. Three types of feeding regimes given to the crab larvae through out the study trials are with and without mixed diatom, with and without Artemia nauplii and with and without rotifer. The study shows that zoea fed with rotifer alone was not enough to sustain survival in the next zoea stages and to promote metamorphosis up to megalopa stage. Survival of zoea fed with Artemia nauplii alone shows that this type of food is not suitable for the very early zoea stages. The study also shows that the adding of mixed diatom to larvae rearing system where rotifer and Artemia nauplii is main food items did not produced high survival rate as compared to larvae rearing fed on rotifer and Artemia nauplii alone. The results of the study demonstrated that the food types not only effect survival rate but also the growth of crab larvae. The study generally ended that the combination diet of rotifer and Artemia nauplii alone is enough to produced C1.
  M. Ikhwanuddin , M.N. Azra , A. Redzuari , Z.A. Aizam and A.B. Abol-Munafi
  The experiment was conducted to determine the ingestion rate of Artemia sp. nauplii and Brachionus sp. by individual blue swimming crab, Portunus pelagicus larvae from zoea 1 until megalopa stages after 24 h. The study also to determined, if the presence of Brachionus sp. influences the ingestion of Artemia sp. nauplii by the individual P. pelagicus larvae. This involved three different feeding treatments, with Artemia sp. nauplii only (Treatment 1), Brachionus sp. only (Treatment 2) and with both Artemia sp. nauplii and Brachionus sp. (Treatment 3) in culture tank. Results indicates that ingestion rates of Artemia sp. nauplii and Brachionus sp. after 24 h by P. pelagicus larvae are 0 Artemia sp. nauplii and 35-36 Brachionus sp. for zoea 1, 1-2 Artemia sp. nauplii and 37-38 Brachionus sp. for zoea 2; 8-15 Artemia sp. nauplii and 38-40 Brachionus sp. for zoea 3; 12-18 Artemia sp. nauplii and 27-37 Brachionus sp. for zoea 4 stage; and finally 32-35 Artemia sp. nauplii and 16-30 Brachionus sp. for Megalopa stages. The individual P. pelagicus larvae ingested more Artemia sp. nauplii during the late larval stages (zoea 3, zoea 4 and megalopa stage) as compared to the initial larval stages (zoea 1 and zoea 2) but ingested more Brachionus sp. during the initial larval stages compared to the late larval stages. However, the presence of Brachionus sp. did not influence the consumption of Artemia sp. nauplii by the individual P. pelagicus larvae at each larval stage.
  M. Ikhwanuddin , S. Noor Baiduri , W.I. Wan Norfaizza and A.B. Abol-Munafi
  This study was conducted to determine the effects of salinity on the mating success of orange mud crab, Scylla olivacea. Male crabs and immature female crabs were reared at the salinity of 15, 20, 25 and 30 ppt in 30 days study period. Brood stocks were fed with squids and maintained until the pre-copulation observed. The pre-copulated crab pairs were isolated from the other brood stock and observation was done for duration of pre-copulation, copulation and post-copulation. The mating duration was recorded. Sperm deposition in the female’s spermathecae was also examined. Mating was successful in all tested salinities and mean pre-copulation and copulation period for S. olivacea was 22.4 and 2.9 h, correspondingly. The mating success was 50, 30, 20 and 10% for salinity of 25, 15, 20 and 30 ppt, respectively. However, there were no significant relationship between the salinity level and mating success of S. olivacea where 25 ppt salinity was the optimum salinity preferable by mud crab pairs to achieve successful mating with sperm deposition.
  A.B. Noor- Hidayati , M.S. Shahreza , A.B. Abol- Munafi and M. Ikhwanuddin
  Gynogenesis was considered as powerful tools for producing all female larvae. One of the most critical point in gynogenesis was studied in banana shrimp, Fenneropenaeus merguiensis. The aim of this study was to determine the effect of different UV induction on F. merguiensis sperms quality for initial development of gynogenesis protocol. Fresh sperms taken from male’s broodstock were diluted 1:10 with Ringer’s solution and UV irradiated with doses at 254 and 365 nm from 20-80 sec. Sperms viability was determined using modified eosin-nigrosin staining method. Irradiation for 20 and 40 sec of 254 and 365 nm exposures were optimum doses to achieve haploid gynogenesis. Microscopy image showed destruction of the sperm in the UV irradiated sperms. The sperms abnormalities could be identified by damage on malformed bodies or by a bent, short or missing spike. The results indicated that viability of sperms generally decreased with increased UV dose and time of exposure. Although, the sperm abnormalities present in all treatments with morphologically missing spike, bent spike and malformed body but still the number of abnormal sperm present was less critical. The application of comet assay for detecting the DNA damage was applied on ultraviolet irradiated sperm of F. merguiensis. Sperms DNA damage was highly scored at 60 and 80 sec of ultraviolet irradiation exposures.
  S. Noorbaiduri , A.B. Abol-Munafi and M. Ikhwanuddin
  Orange mud crab, Scylla olivacea is the most distributed mud crab species along the Malaysian coastal water. Most crab farmers targeting the seed from the wild for stocking in the ponds since the seed production of mud crab in captivity is unavailable for commercialization and still at the laboratory stage. The sperm activation also known as Acrosome Reaction (AR), is one of the important aspects for fertilization to occur. Thus, this study aimed to describe the sperm activation stages of acrosome in male and female mud crab, S. olivacea mating in wild and in captivity and to describe the relationship between post-mating periods and the acrosome stages. Life specimens were dissected and sperm were collected from vas deference of males and spermathecae of females. The AR in S. olivacea has four main stages. Stage 1 is an inactive sperm, Stage 2 is the activation of AR where the extrusion of acrosomal materials began. Stage 3 is the intermediate stage between Stage 2 and 4. Lastly, Stage 4 is the ejection of acrosomal filament. Sperm from males were mainly in inactive Stage 1 (59.30%). While, sperm in wild-mated and captive-mated females were mostly in Stage 4 (71.80%) and Stage 2 (53.50%), respectively. The AR activation and development in females can be influenced by the ovarian maturation. The AR description in the present study can be used to manipulate artificial seed production in captivity.
  M.Z. Ihwan , M. Ikhwanuddin and H. Marina
  Samples were collected from Setiu Wetland (5°40'47.93"N, 102°42'45.04"E) of Terengganu coastal water, Malaysia. Sixty wild mud crab genus Scylla spp. were measured for carapace length (7.9±1.06 cm) and weight (90.8±34.0 g) before further examined. The 473 of barnacle species Octolasmis angulata were collected from three species of wild mud crab i.e., Scylla olivacea, S. tranquebarica and S. paramamosain. The basic morphological features and terminology are applied for easily identified the species such as body shape and size, the presence or absence of calcareous plates or scutum, the variations in plate size, shape and disposition. The external and internal parts of pedunculate barnacle, Octolasmis angulata were draw and describe the morphological characteristic i.e., capitulum, peduncle, scutum and carina. The entire characteristics were measured for comparison by referring to the previous study. The structures of mandible, maxilla, maxillule, cirri, setae, penis, ovary, teeth, etc. were recognized.
  S.N. Fatihah , J. Safiah , A.B. Abol-Munafi and M. Ikhwanuddin
  The study was aimed to determine the biochemical changes of total protein, glucose, lactate dehydrogenase (LDH) and total lipid in the cryopreserved sperm of mud spiny lobster, Panulirus polyphagus. The mean sperm viability, total protein, glucose, LDH and lipid for control and cryopreserved sperm that stored in liquid nitrogen from control, 6, 12, 24 h, 7, 30 and 60 days were 94.91±2.17% (control), 80.58±1.33, 77.08±2.53, 76.49±3.10, 63.32±7.08, 62.93±3.72 and 60.91±10.78% (sperm viability), 21.44±3.52, 18.82±0.23, 17.79±0.80, 16.98±0.48, 16.65±0.07, 15.03±2.53 and 12.16±2.02 mg mL-1 (total protein), 0.28±0.012, 0.26±0.004, 0.20±0.035, 0.15±0.062, 0.11±0.036, 0.07±0.036 and 0.04±0.013 mg mL-1 (glucose), 72.65±7.88, 64.38±6.40, 64.24±4.80, 62.84±7.20, 62.11±3.84, 49.94±2.78, 38.31±3.29 IU L-1 (LDH) and 8.77±0.05, 4.87±0.25, 4.69±0.29, 4.37±0.02, 4.16±0.17, 2.87±0.29 and 2.83±0.26 mg mL-1 (total lipid). There were significant differences in sperm viability, total protein, glucose, LDH and total lipid between cryopreserved periods (p<0.05). As a conclusion, the sperm viability and biochemical changes of total protein, glucose, LDH and total lipid were determined in the cryopreserved sperm for control, 6, 12, 24 h, 7, 30 and 60 days. The storage duration in P. polyphagus cryopreserved sperm was also determined until 60 days and suitable for further breeding program.
  S. Noorbaiduri and M. Ikhwanuddin
  Crablets production in mud crab, genus Scylla is highly depends on the depleted wild crablets supply. Artificial crablets production through in-vitro fertilization technique can be an alternative for the crablets production of this mud crab. Therefore, this study was conducted to explore the artificial crablets production through in-vitro fertilization technique on orange mud crab, Scylla olivacea through the determination of (1) Fertilization rate and (2) Embryonic developments from the manipulation of sperm mass with Stage 4 ovary. The sperm mass were collected from the spermathecae of mature females, as well as from the vas deferens of males and allowed to fertilize the Stage 4 ovary in the Ca-F saline medium. Fertilized ovary were incubated in artificial seawater with controlled salinity (28-30 ppt) and temperature (28-30°C in water bath). Observation on fertilization rate and embryonic development were observed daily. Fertilization Membrane (FM) was developed when the ovary was fertilized. However, the embryos could only develop until Day 5 at most. Subsequently, FM could not be maintained and started to deteriorate resulting the embryos to lose their form. Further development could not be observed after Day 5. Manipulation of sperm from vas deferens (male) shows lower fertilization rate than spermathecae (female). Present study has proven that artificial crablets production S. olivacea through in-vitro fertilization technique from the manipulation of sperm mass with Stage 4 ovary was a success.
  M. Ikhwanuddin , S.N. Fatihah , A.H. Nurfaseha , R. Ruhinisa , Z.Z. Tira , A. Shamsudin , A. Siti Aishah and A.B. Abol- Munafi
  A study was carried out to determine the effect of salinity on ovarian maturation stages (external morphology and histology characteristics of the ovary) and embryonic development (time of embryonic development and size of egg) of mud spiny lobster, Panulirus polyphagus. For the ovarian maturation stages, there were three treatments; treatment 1 for 20 ppt (E1T1), treatment 2 for 40 ppt (E1T2) and control treatment for 30 ppt (E1C2) and conducted within 100 days. There was no significant difference (p>0.05) between control in 30 ppt at day 1 (E1C1), E1C2, E1T1and E1T2 noted in the Oocyte Size Index (OSI). Mean oocyte diameter and Gonadosomatic Index (GSI) had significantly differences (p<0.05) between E1C1, E1C2, E1T1 and E1T2. For the embryonic development, the berried female was placed in tank with 20 ppt for treatment 1 (E2T1), 30 ppt for control treatment (E2C1) and 40 ppt for treatment 2 (E2T2) and 20-30 eggs were taken for each salinity to observe the embryonic development. Stage 1, 2 and 3 for 20 ppt and stage 1 for 40 ppt cannot be observed the development of embryo. At low salinity of 20 ppt (E2T1), the shortest duration from stage 4 until hatched was about 4 days while at high salinity of 40 ppt (E2T2), the longest duration from stage 4 until hatched was about 8 days. Thus, the low salinity was the shortest time to hatch. The effect on the sizes changes was stronger in the salinities of 30 and 20 ppt than 40 ppt. Therefore, the high salinity was the lowest egg size compared to low and control treatments. In conclusion, the different salinity affected the ovarian maturation stages and embryonic development of P. polyphagus.
  M. Ikhwanuddin , A.B. Noor-Hidayati , N.M.A. Aina-Lyana , H. Zulaikha , H. Muhd-Farouk and A.B. Abol-Munafi
  The aim of the study was to develop an appropriate basis for the optimization of in vitro fertilization of Fenneropenaeus merguiensis using three different culture medium including Natural Sea Water (NSW) as control medium, Artificial Sea Water (ASW) and Calcium Free saline (Ca-F saline). The unfertilized mature eggs were collected from the broodstock ovaries during spawning. The non-motile sperm of F. merguiensis activated as natural spawning. In NSW medium, ASW medium and Ca-F saline medium, cortical rods were released and hatching envelope formation took place in which the eggs activation events were reported. The Ca-F saline and ASW solution induced a slow egg activation contradict with the sequence of event for natural spawning of F. merguiensis. Fertilization was successfully obtained in all treatments with 8.67±4.04% in ASW, 19.67±7.38 and 4.33±4.04 in Ca-F saline. Although, the hatching rate were not successfully obtained by ASW and NSW culture medium treatment, hatching yield in Ca-F saline medium was obtained with 3.00±2.65. Overall, these findings will contribute to the development of F. merguiensis breeding technology and further understanding on sperm biology, cryobiology and reproductive biology in shrimp.
  M. Ikhwanuddin , I.O. Khairil , M.N. Azra and K. Waiho
  Biological features of size at maturity, fecundity of berried female, size distribution, relationship of Carapace Width (CW) and Body Weight (BW) and sex ratio of sentinel crab, Podophthalmus vigil from Terengganu coastal water, Malaysia of South China Sea were studied. Sexual maturity (CW50) was achieved at 11.51 cm for male and 9.53 cm for female. The mean fecundity was 312613.50±89835.08 eggs. Size distribution of male and female was slightly different with males have size range from 8.0 to 18.9 cm CW, while females ranged between 7.0 to 11.4 cm CW. The mean CW and BW of P. vigil males were significantly greater than that of females. The CW-BW relationship for males P. vigil was attained as BW = 46.892e0.0719CW (R2 = 0.1155) and for females was estimated as BW = 6.3117e0.2665CW (R2 = 0.5523). Finally, the sexual ratio of P. vigil for male to female was 1: 0.65. The data obtained from this study can be used as the baseline data for future study on P. vigil fishery and aquaculture management in Malaysia coastal water especially and within the Indo-pacific region generally.
  M. Ikhwanuddin , M.N. Azra , H. Siti-Aimuni and A.B. Abol-Munafi
  Blue swimming crab, Portunus pelagicus is widely study and research throughout the Indo-West Pacific, but little is known of its reproductive biology in Malaysia. The present study describes the fecundity, embryonic development and ovarian development stages of the P. pelagicus from Johor coastal water, Malaysia. Carapace width range of berried crabs sampled was from 9.64 to 13.32 cm, while the body weight range was from 75 to 235 g. The mean number of egg produced by females in different sizes ranged from 105443.333±35448.075 per eggs batch. Mean egg size during embryonic development at stage 1 was 0.307±0.037, while 0.386±0.039 and 0.396±0.033 for stage 2 and stage 3, respectively. Study showed that there was significant (p<0.05) relationship between the number of eggs and carapace width/body weight. Mean diameter oocyte during ovarian development at stage 1 was 97.732±12.391 while for stage 2 was 149.516±23.287. Stage 3 showed increasingly of size with mean diameter was 158.506±27.616 and 181.013±24.339 for stage 4.
  S.N. Fatihah , J. Safiah , A.B. Abol-Munafi and M. Ikhwanuddin
  The present study aimed to determine the effect of testosterone undecanoate hormone on sperm quality (sperm viability) and sperm quantity (sperm counts) and its levels in the hemolymph of male mud spiny lobster, Panulirus polyphagus. Male P. polyphagus was injected laterally in fifth abdominal segment of pure hormone, Testosterone Undecanoate (TU) and ethanol at days 1, 8, 15, 22 and 29. Hemolymph of P. polyphagus was taken every two weeks and checked with Enzyme-linked immunosorbent assay (ELISA) to measure hormone levels. The mean sperm quality and quantity were increased due to increase the TU dose and TU levels also increase. The sperm quality, quantity and hormone levels were relevance each others. These findings indicate that TU injection should be evaluated as a practical way of improving sperm quality and quantity in commercial operations.
  M. Ikhwanuddin , H. Muhd-Farouk , A.J. Memon , W. Wendy and A.B. Abol-Munafi
  The aim of this study was to evaluate how long the fresh sperm maintained at 2°C would be utilized for fishery management. The study was conducted every 2 h to assess the sperm viability of orange mud crab Scylla olivacea. Evaluations were conducted as 3 treatments; T1, T2 and T3. In T1, the live specimens were sacrificed; for T2, only spermatophores were extracted and for T3 spermatophore extraction followed by homogenization to create a sperm suspension. All samples were stored with ice in an insulated box was keep fresh longer at 2°C. The time ‘0’ referred the immediate collection of sperm after the specimen was sacrificed. Spermatophore viability was determined using the sperm suspension by eosin-nigrosin staining method. Sperm viability for the fresh sample at time zero was 97.36±0.53%. Viability of the sperm significantly decreased in the 2nd h in all treatments, T1 was 44.66±0.54 to 4.2 ±0.22% at 16 and 18th h, T2 was 36.56±0.5 to 2.69±0.06% at the 12 and 14th h and T3 was 33.69±1.26 to 6.4±0.29% at 8 and 10th h. In comparison, T1 showed significantly higher than other treatments (p<0.05). Extremely low viability percentages were recorded in T3. This study also proved that the time elapse had significant impact on the percentage of viable sperm count.
  S.N. Fatihah , J. Safiah , A.B. Abol-Munafi and M. Ikhwanuddin
  The present study aimed to determine the effect of 17α-hydroxyprogesterone (17α-OHP) and 17α-hydroxypregnenolone (17α-OHPL) on sperm quality and sperm quantity in male mud spiny lobster (Panulirus polyphagus). The mean of sperm quality was increased in 17α-OHP and 17α-OHPL treated hormones. In 17α-OHP injected animals, the mean of sperm quantity of dose 0.01 μg g-1 b.wt. was increased than 17α-OHPL. Meanwhile, 17α-OHP and 17α-OHPL concentrations were lower when injected with the hormones but 17α-OHP was higher at only day 15 (dose 0.01 and 0.1 μg g-1 b.wt.). For 17α-OHPL, the hormone was a prohormone in the body of P. polyphagus and only required smaller to increase the sperm quantity. Besides, when the higher dose of 17α-OHPL (0.1 μg g-1 b.wt.) was used in P. polyphagus, the development of P. polyphagus was inhibited and decreased the sperm quantity and 17α-OHPL concentration in hemolymph was lower. Injection of 17α-OHP in P. polyphagus has increased the sperm quality and quantity for both 17α-OHP dosage of 0.01 and 0.1 μg g-1 b.wt. However, injection of 17α-OHPL in P. polyphagus has decreased the sperm quantity only, also for both dosage of 0.01 and 0.1 μg g-1 b.wt. and lower on hormone concentration.
  A. Amin-Safwan , H. Muhd-Farouk , M. Nadirah and M. Ikhwanuddin
  Background and Objective: Mud crab from the genus Scylla are considered as one of the most demanded seafood items nowadays as their flesh has high quality, tasty and higher growth rate thus support and boosted expansion in aquaculture sector especially in Malaysia. Present study was designed to focus on the effect of water salinity on the ovarian maturation of orange mud crab, Scylla olivacea based on morphological characteristics. Methodology: Samples were collected from Setiu wetlands, Terengganu, Malaysia from July-September, 2015. Ovarian maturation of S. olivacea was classified into four stages based on previous study which were: Immature (Stage 1), early mature (Stage 2), late mature (Stage 3) and fully mature (Stage 4). Results: Morphologically as the ovary develop the colouration start to change from translucent or whitish in colour and sometimes creamy to pale yellow, follow by light orange and lastly reddish orange. Stage 1 ovary was translucent and whitish in colour, stage 2 ovary was pale yellow in colour, stage 3 was light orange and stage 4 ovary was reddish orange in colour. Gonad Somatic Index (GSI) of S. olivacea remained low at stage 1 and 2 and began to increase started at stage 3. This present study involved three different salinities treatments, which treatment 1 (10 ppt), treatment 2 (20 ppt) and treatment 3 (30 ppt). Treatment 2 produce the highest number of stage 4 ovarian maturation based on colouration and the highest GSI recorded, follow by treatment 1 and lastly treatment 3. Conclusion: This present study proved that salinity does affected the ovarian maturation of S. olivacea in captivity and provides important information regarding the effect of water salinity on ovarian maturation for further studies on reproductive biology of this species.
  N. Hasyima Ismail , A. Amin Safwan , N. Fairuz Fozi , F.H. Megat , H. Muhd Farouk , S.A. Kamaruddin , M. Ikhwanuddin and M.A. Ambak
  Background: Orange mud crab Scylla olivacea is one of the most important fisheries resources. A new development in ageing technique of crustaceans has been introduced. The detection of growth band deposited in hard structure of gastric mill in the cardiac stomach are found retained after moulting process can be used as age indicator and growth estimation. Objective: This study was carried out to determine the comparison between carapace width and growth band count of S. olivacea in Malaysia. Materials and Methods: Samples were collected from Setiu Wetlands, Terengganu, Malaysia from February until August, 2016. Samples were categorized based on their morphological measurements. The mesocardiac and zygocardiac ossicles in the gastric mill of S. olivacea was dissected out and preserved in solutions and underwent a cross sectioning process. A total of 76 of wild S. olivacea ranging from 6.56 to 12.84 cm in carapace width were analysed. The growth band counts were examined for each individual and ranging from 1 to 3 band counts. Results: A positive linear relation was observed between CW and GBC with r2 = 0.5178, p<0.01. Overall, there was a strong, positive correlation between CW and GBC. Increase in CW were correlated with increases in GBC respectively for this species. Conclusion: Therefore, the carapace width, growth band counts and body weight can be used to improve data on growth, recruitment, maturation and mortality. Thus, this study would able to improve new ageing technique and contribute greatly to improve the conservation and management of S. olivacea in Setiu Wetlands, Terengganu, Malaysia.
  A.D. Talpur , A.J. Memon , M.I. Khan , M. Ikhwanuddin , M.M. Danish Daniel and A.B. Abol-Munafi
  The cause of mass mortality of Portunus pelagicus larvae reared in a hatchery system was investigated. The gut content of 180 female crabs and egg specimen of 24 female were studied for pathogenic microbes. The gut of female crabs were harboring fish pathogenic bacteria includes Staphylococcus epidermidis, Vibrio harveyi, Vibrio parahaemolyticus, Micrococcus luteus and Pseudoalteromonas piscicida and eggs were found associated with fish pathogens include Vibrio harveyi, Micrococcus luteus and Pseudoalteromonas piscicida. A causative transmitting pathogen V. harveyi through the feces of adult female crab and responsible for heavy mortality during larval rearing was determined by examining samples associated with the gut, hatching tanks, eggs, larvae rearing tanks, live and dead larvae of P. pelagicus. All isolates were identified by 16S rRNA gene sequencing. Vibrio harveyi was the major pathogen associated with all sources brought under study. Larvae were found to harbor a higher number of bacteria than larvae rearing tank. Experimental challenges with various doses indicated that the V. harveyi isolates were highly pathogenic. Doses 105 cfu mL-1 produced upto 96.67% mortality and 106 cfu mL-1 resulted in 100% mortality within 24 h post challenge. The differences among various doses of pathogen were statistically significant (p<0.05). The presence of these pathogens in P. pelagicus beyond the consequence for larval rearing is of epidemiological and health significance to humans.
  A.J. Memon , M. Ikhwanuddin , A.D. Talpur , M.I. Khan , M.O. Fariddudin , J. Safiah and A.B. Abol-Munafi
  The study was conducted to every hour assess the sperm viability of banana shrimp (Penaeus merguiensis) collected from Kedah water, West Malaysia (5°39'N; 100°19'E). Evaluations were done on three ways: group A: whole fresh specimens maintained at 2°C prior to extraction, group B: whole spermatophore maintained at 2°C and C: sperm suspension maintained at 2°C. Spermatophores counts were determined by sperm suspension using modified eosin-nigrosin staining method. Percentage of mean sperm viability for the fresh sample in group A at time zero was 93.96±3.74%, B was 98.8±0.7% and C was 99.02±0.7%. In group A, viability of the sperm considerably decreased after the first 60 min was 76.4±5.96% but in group B and C was gradually decreased at 91.4±0.9 and 97.6±1.2%. These were 67.9±5.13 and 62.5±5.1% in group A, 82.3±1.4% and 75.9±2.2% in group B and in group C was 93.3±1.9 and 88.2±3.1%, respectively after 1st-2nd h times elapsed. At 7th h in all groups the viability decreased significantly to <60% (p<0.05). Following, mean sperm viability were considerably decreased to group A was 50.7±4.79 and 37±6.52%, group B was 51.2±1.6 and 31.6±1.4% and group C was 56.2±1.9 and 41.5±2.3%, respectively after 7th and 9th h. However, for spermatophores in group A after 7th h fertilization and hatching rate was 44.3 and 64.4% in group B was 61.9 and 67.7% and group C was 42.9 and 61.3%, respectively at rates comparable to control 88.2 and 76.2%, respectively. There was no significant relationship was observed between biomass of the spermatophore to the body weight of the shrimp (p>0.05). The present study also revealed that specimens, spermatophore or sperm suspension maintained at 2°C could be utilized for fishery management through artificial insemination process till 7th h.
  A.J. Memon , N. Hidayati , A.D. Talpur , M.I. Khan , M.O. Fariddudin , J. Safiah , M. Ikhwanuddin and A.B. Abol-Munafi
  In aquaculture industry, spermatophore transfer technique such as artificial insemination in particular with reference to banana shrimp P. merguiensis is challenging. The aim of this study is to examine a novel technique for artificial insemination in P. merguiensis using SHDAI (Shrimp Holder Device for Artificial Insemination). In order to transfer the spermatophore properly into thelycum, an appropriate shrimp holder with continuous aeration system has been developed. During the process of manual spermatophore transfer (artificial insemination) also, a protective device to keep the female P. merguiensis shrimps alive and under minimum stress is necessary. The artificial insemination process carried out using SHDAI showed no signs of stress and/or mortality of the broodstock. During the experiments, female shrimps were 100% alive and active. Altogether, 78 female shrimps were tested of which 63 successfully accepted the spermatophore by using SHDAI. The accepted spermatophore percentage was significantly higher and achieved as 80.76%. Accepted spermatophore mass in the frozen, control and mean of both were recorded as 48.36±10.45, 45.0±15.28 and 46.68±2.38% which indicated no significant differences (p<0.05) However, frozen sperm at -196°C LN up to 90 days was 64.1±4.3% which indicated no significant differences between the SHDAI and control (p<0.05). Whereas frozen sperm at 196°C up to 90 days in LN was 62.5±2.9% there was no significant difference between the SHDAI and control (p<0.05). In the present study, overall quality of sperm in terms of the fertilization rate and hatching rate were almost similar between inseminated (using SHDAI) and control (natural mating) broodstock.
  A.J. Memon , A.D. Talpur , M.I. Khan , M.O. Fariddudin , J. Safiah , A.B. Abol-Munafi and M. Ikhwanuddin
  The spermatophore morphology of the P. merguiensis from Kedah, Malaysia is described. About 10 cryopreserved groups as 6, 12 and 24 h, 7, 30, 60, 90, 120, 150 and 180 days and fresh as control was examined from each of three replication were evaluated for sperm gross morphology evaluations. A fully mature male’s broodstock of P. merguiensis was taken with fresh spermatophores and evaluated for sperm morphologically. Cryopreserved spermatophore after the thawing 27°C/2 min (fresh and frozen) individually transferred into glass homogenizer (High speed variable speed reversible, Glas-col, Terre Havte In USA) with 200 μL of Ca-F saline. Fixation, dehydration by series of alcohol, Critical Point Dry (CPD) and mount specimens on to stubs using or carbon dots as well as using Auto Fine Coater and Sputter Coater moreover scanning by Model JEOL 6360LA scanning electron microscope. The cryopreserved spermatophore shows similarities with those of fresh, there were no significant differences (p>0.05) between the freshly spermatophore and spermatophorestored up to 90 days at -196°C liquid nitrogen.
  M.Z. Ihwan , M. Ikhwanuddin and H. Marina
  More than one species of pedunculate barnacle genus Octolasmis are found attached on the gill of wild mud crab in this study. For the gill preference study, Octolasmis cor prefers to attach on the gill G3 to G6. The areas of site infection are still in the middle part of the mud crab gill. These results are different with previous studies which prefer G7 as attachment. For the identification study, twenty good specimens, that have been mounted on glass slide, were taken for identification of morphological part. The general part of shape, carina and scutum were captured and drawn using lucida camera (Leica). Shape of carina (1.44±0.29), capitular plates (3.04±0.53), capitulum (1.74±0.35) and peduncle (2.32±0.39) of Octolasmis cor were measured, respectively. Data analysis using one-way ANOVA show that part of carina and scutum show significant difference (p>0.05) and can be used as identification tools. Studies on genus Octolasmis were not well-published. Most of previous literatures just explain the genus of Octolasmis but not into species because of no standard method for identification of this genus.
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