Ovarian lavage method as an alternative route for hormonal administration and short-term sperm storage in Clarias gariepinus.

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) for breeding performance. The trend of the haematological parameters also suggests the intraperitoneal administration procedure elicited more stress compared to intramuscular or ovarian lavage routes (P < 0.05). In the second experiment, the use of sperm stored in the ovaries or refrigerated for 24hr didn't affect (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 < 0.05). The loss of fertilizing capacity observed herein was justified by the reduction in sperm quality over time (P < 0.05). It was concluded that ovarian lavage is a viable route for hormonal administration and 24hr sperm storage in C. gariepinus.

Environmental effects on the oxygen consumption rate in juvenile Epinephelus fuscoguttatus (Forsskal, 1775).

This study was designed to optimize the culture conditions of juvenile Epinephelus fuscoguttatus (Forsskål, 1775) under laboratory conditions. To this effect, the rate of oxygen consumption was monitored as an index of stress under different temperature, salinity, pH, photoperiod, and urea concentrations. The result obtained after 12 h of exposure suggests the preference of the juvenile E. fuscoguttatus to a temperature range of 15-25 °C and salinity of 30 ppt. Based on this study, temperature was found to be the most lethal as 100% mortality was observed after 6 h in fish exposure to temperatures above the optimal (≥ 30 °C). However, the oxygen consumption rate was similar under the different pH, photoperiod, and urea concentration tested. It was concluded that water temperature was most critical in terms of respiration physiology of the juvenile E. fuscoguttatus given the range and levels of environmental factors tested in this study.

Diet and internal physiological changes of female orange mud crabs, Scylla olivacea (Herbst, 1796) in different ovarian maturation stages.

Recently, there has been a growing interest in the ovarian maturation of mud crabs, genus Scylla. Studies regarding the factors that affect ovarian maturation in mud crabs, however, are still lacking. This study, therefore, evaluates the relationship between diet and internal physiological changes of female orange mud crabs, Scylla olivacea. Sixty female adult S. olivacea were sampled from Setiu Wetland, Malaysia. Foreguts were sampled to study fullness and content. The hepatopancreas was sampled to study digestive enzyme activity, biochemical composition, and histology. Ovaries were sampled to study ovarian biochemical composition and histology. Foregut fullness data suggest that there is an increase in feeding with advancing ovarian maturation in mud crabs. Data for foregut contents indicated that when the ovary was in Stage 3 of maturation the diet was animal-based. The activity of amylase, cellulase, lipase, and trypsin during Stages 1 and 4 of ovarian maturation were less than during Stage 3 of ovarian maturation. Biochemical composition (protein and lipid) was greater during Stage 3 of ovarian maturation. Histological analysis of the hepatopancreas indicated an increase in hepatopancreas tubules, B and R cells during Stage 3 as compared with Stage 1 of ovarian maturation. Histological analysis of the ovary indicated increases in oocyte diameter due to concentrated large yolk globules. Based on these results, it is concluded that vitellogenesis mainly occurred during Stage 3 of ovarian maturation, based on evidence of increased feeding with the diet being predominantly animal-based. Likewise, digestive enzyme activities, proteins, lipids, B cells, and R cells were all greater during Stage 3 of ovarian maturation.

Study on Carapace Width Growth Band Counts Relationship of Orange Mud Crab, Scylla olivacea (Herbst, 1796) from Terengganu Coastal Waters, Malaysia.

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.

Effect of Water Salinity on the External Morphology of Ovarian Maturation Stages of Orange Mud Crab, Scylla olivacea (Herbst, 1796) in Captivity.

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.

Effect of 17α-hydroxyprogesterone and 17α-hydroxypregnenolone on sperm quality and sperm quantity in male mud spiny lobster (Panulirus polyphagus).

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.

Sperm Viability Assessment over elapsing time maintained at 2 degrees C of orange mud crab, Scylla olivacea (Herbst, 1796).

The aim of this study was to evaluate how long the fresh sperm maintained at 2 degrees 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 degrees 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.

Effect of testosterone undecanoate hormone on sperm and its level in the hemolymph of male mud spiny lobster, Panulirus polyphagus.

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.

Fecundity, embryonic and ovarian development of blue swimming crab, Portunus pelagicus (Linnaeus, 1758) in coastal water of Johor, Malaysia.

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.