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Articles by A.B. Abol-Munafi
Total Records ( 18 ) for A.B. Abol-Munafi
  E.A.T. Mubarak , M.A. Amiza , H.K. Bakhsh and A.B. Abol-Munafi
  A 3 weeks feeding trial was conducted to evaluate the Apparent Digestibility Coefficients (ADCs) of dry matter, protein, gross energy, nitrogen, crude lipid and fiber of Water Hyacinth (WH), Echhornia crassipes meal incorporated in pelleted test feeds (0, 10, 15, 20 and 25%, dry matter basis) for red tilapia fingerlings. Chromium dioxide (1%) was added as an inert bio-marker to iso-nitrogenous (35.00±0.20% crude protein) and iso-caloric (4.00±0.52 kcal kg-1) feeds. Results showed that the maximum value (p = 0.05) of ADCs for dry matter (68.09%) was recorded for the control feed with 0% WH while the minimum value of 50.36% was recorded for the test feed 4 including 20% WH. Similarly, the maximum ADCs values (p = 0.05) for crude protein, gross energy, ether extract, crude lipid and crude fiber were also found in control feed while the minimum values were observed in test feed 4 including 20% WH. The study was clearly indicated that red tilapia fingerlings efficiently digest the nutrients when only the maximum inclusion of WH 20% in their feed does not exceed 20% of WH meal (on dry basis).
  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.B. Abol-Munafi , N.H. Norazmi-Lokman , N.A. Asma , S. Sarmiza and M.Y. Abduh
  Standard histological protocol were performed on fifteen families of A. ocellaris collected at Pulau Bidong, Terengganu to describe and differentiate the gonad morphology between male, female and the non-breeders of protandrous Anemonefish (Amphiprion ocellaris). Based on the observations, non-breeder fish (3.5-5.8 cm TL) possess intermixed ovotestes with no clear boundaries between testicular and ovarian tissues. The ovotestes contained previtellogenic oocytes interspersed with fewer spermatocytes. Male fishes (5.3-6.9 cm TL) exhibited a different type of ovotestes. Male fish ovotestes were dominated by spermatogenic cells with fewer numbers of surrounding previtellogenic oocytes. In contrast, female fish gonadal tissue (6.0-9.2 cm TL) only contained ovarian tissues with no evidence of any testicular tissues. The ovarian was also larger and more mature than male and non breeders ovarian tissue with well organised ovigerous lamellae filled with oocytes visible at different developmental stages. The absence of the testicular tissues in the female gonad shows that the change of sex from male to female is an irreversible process.
  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.
  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.
  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.
  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.
  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 , 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.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.
 
 
 
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