Breeding and larval performance of novel hybrids from reciprocal crosses of Asian catfish Pangasianodon hypophthalmus (Sauvage, 1878) and African catfish Clarias gariepinus (Burchell, 1822) were investigated in this study. Spawning was by hormonal injection of brood fish, artificial fertilization, and incubation in triplicate aquarium tanks (0.5 × 0.5 × 0.5 m3) with continuous aeration. Reciprocal crosses (♀C. gariepinus × ♂P. hypophthalmus and ♀P. hypophthalmus × ♂C. gariepinus) had lower hatchability (≤50%) than their pure siblings (≥75%). Fish from all crosses survived until the juvenile stage but survival at 35 days post hatching (dph) was higher for pure C. gariepinus sib. ♀C. gariepinus × ♂P. hypophthalmus was observed to be less resistant to degradation of water quality than the other crosses, however it had higher body weight compared with the other crosses that showed similar performance. Morphological comparison of surviving juvenile at 35 dph, showed that all ♀P. hypophthalmus × ♂C. gariepinus and 13% of the ♀C. gariepinus × ♂P. hypophthalmus exhibited the very same morphology as that of their maternal parent species, while the other portion of the ♀C. gariepinus × ♂P. hypophthalmus cross exhibited morphological traits that were intermediate between those of both parent species. This study been the first successful attempt to hybridize both species and therefore, laid the groundwork for further studies on the aquaculture potentials of the novel hybrids.
Malaysian Mahseer (Tor tambroides) is considered as a good prospect for aquaculture in Malaysia. However, knowledge about Malaysian Mahseer-associated sperm microbiota is still limited, although some studies reported that sperm-related bacteria are a factor in the decline of sperm quality, as sperm may become the carrier of pathogenic bacteria to the egg. The goal of this study was to evaluate the sperm microbiota associated with Malaysian Mahseer from 3 different locations (Universiti Malaysia Terengganu [UMT], Ajil, and Pahang) using polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) fingerprinting and to compare location differences by cluster analysis. Our results showed that the UMT sample had different sperm microbiota composition and a different trend in its relationship with sperm quality. Correlation analysis showed a relationship between bacterial diversity and sperm quality. Phylogenetic analysis indicated that sperm microbiota was composed of diverse phyla, including Proteobacteria, Firmicutes, and Actinobacteria. Interestingly, bacteria such as Salinisphaera sp., Pelomonas sp., and Staphylococcus spp. were detected in all the locations, suggesting that these bacteria are indigenous bacterial members of the Malaysian Mahseer sperm microbiota, although their function is still unclear.
To obtain well spread chromosomes, the cytogenetic protocol for Pangasianodon hypophthalmus and Clarias gariepinus were optimized. This includes, the colchicine concentration (0.01%, 0.025%, 0.05%)/exposure duration (1, 3, and 5 h), hypotonic solution (distilled water or 0.075M KCl solution)/exposure duration (30 min, 1, and 2 h), the time of cell suspension preparation (at hypotonic treatment or before slide preparation) and chromosome aging period (0, 3, and 7 days in Carnoy's fixative). In addition, the type (i.e., fin, gill or kidney) and the amount of tissue (10, 50, 100 or 150 mg) were also investigated. Regardless of the species, the result obtained showed that well-spread chromosomes could be obtained using the following optimized protocol: Juveniles are injected with 0.05% colchicine (at one ml kg-1) and allowed to swim for 3 h. Then, 50 mg of gill tissue is made into cell suspension in 0.075M KCl for 1 h. The cell suspension is treated in Carnoy's fixative (changed three times at 20 min interval) and then aged for 3 days. Finally, chromosome slides are made and stained with 10% Giemsa for 1 h.
Twenty-five traditional and thirty-four geometric morphometric comparisons were carried out on pure and reciprocal crosses of Pangasianodon hypophthalmus (Sauvage, 1878) and Clarias gariepinus (Burchell, 1822). Thirty fish samples each of the C. gariepinus (CH), P. hypophthalmus (PH), Pangapinus (♀PH × ♂CG) and the two distinct morphotypes of the Clariothalmus (♀CG × ♂PH) (Clarias-like and Panga-like) between the ages of four and six months were used for this study. Phenotypically, the Clarias-like Clariothalmus and the Pangapinus progenies were indistinguishable from their maternal parents while the Panga-like Clariothalmus was a phenotypic intermediary of the putative parents but looks more closely to the paternal parent. Hence, both univariate proportion and multivariate analysis of the collected data successfully separated the different fishes into three multivariate spaces. The analysis of the dendrogram with complete linkage and Euclidean distance further showed the close relationship of the isolated Panga-like Clariothalmus progenies to the paternal parent, however, Clarias-like Clariothalmus and the Pangapinus were completely intermingled with their maternal parents. The most important index of discrimination of these fishes into different multivariate spaces was the fin characteristic which showed 100% exclusive ranges for the individual groups in many cases.
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 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
The development of sperm cryopreservation for Pangasius nasutus is necessary in order to serve the growing demand of this species through artificial fertilization and the preservation of valuable strains of male broodstocks. In the present study, the basic protocol of sperm cryopreservation for P. nasutus was established by identifying the optimal conditions for optimum cryoprotectant, toxicity of cryoprotectants, extenders, freezing condition and dilution ratio. Methanol (MeOH) at 10% concentration had the best post-thaw motility (26.3 ± 0.9%) and curvilinear velocity (VCL) compared to dimethyl acetamide and dimethyl sulfoxide. MeOH was the least toxic cryoprotectant; sperm suspended in 5 and 10% MeOH maintained motility up to 50 min. No significant differences were detected between the three types of extenders tested (0.9% sodium chloride, Calcium-free Hanks' Balance salt solution and ringer solution. P. nasutus sperm had a narrow range of optimal cooling rate. Significantly higher post-thaw motility was identified when cooling at 9.23 °C min-1, obtained by freezing at height of 14 cm above liquid nitrogen vapor for 7 min, showing lower cooling rate is suitable for this species. However, when cooling below and above the optimal cooling rate, post-thaw motility dropped drastically. There were no significant differences among the dilution ratios investigated, indicating the volume of cryodiluent at all tested ratios (1:9, 1:19 and 1:49) was sufficient for the protection of cells during the cryopreservation process. The development of the protocol for cryopreserved P. nasutus sperm will assist artificial seed production and provide an important tool for genetic and breeding research.
Discrimination of different fishes can be done through different means which includes morphological appearance. When two fishes are successfully hybridized, they produce progenies that have shared morphology between their pure parent, hence, making morphometric characterization an important aspect of hybrid discrimination. However, erythrocyte characterization is also a simpler method for characterization. The dataset presented in this article represents the traditional morphological data, truss network data and erythrocyte data of pure and novel hybrids from reciprocal crosses of African catfish Clarias gariepinus and Asian catfish Pangasianodon hypophthalmus. Breeding of the broodstocks was done to produce pure and hybrid progenies which were maintained for a period of four to six months. Based on the cross combinations and morphotypes, traditional measurement of twenty-five morphological characters and five meristic counts were recorded. Thereafter pictures of the different fish groups were used to determine values of thirty-six distances between ten landmark points. The morphological abnormality of the hybrids at market size is also presented in this data article for the very first time. Blood was then collected from the caudal peduncle of ten fish per group and smeared on a slide for observation under a compound microscope (at 100 × magnification). Data gotten included erythrocytes parameters such as cell major axis, cell minor axis, nucleus major axis, nucleus minor axis cell area, nucleus area, cell volume, and nucleus volume. Data recording was through the Microsoft excel spreadsheet; which was also used to process the data to get the exclusive ranges of values for paired progenies. The data as presented is associated with the research article "Morphological characterization of the progenies of pure and reciprocal crosses of Pangasianodon hypophthalmus (Sauvage, 1878) and Clarias gariepinus (Burchell, 1822)" [1]. The dataset presented in this article can be used for easy identification of the novel hybrid progenies of the African Catfish and Asian Catfish.
Dietary lipid manipulation in the feed of commercially cultured finfish is used not only to improve production and culture but also to enhance their reproductive performances. The inclusion of lipid in broodstock diet positively affects growth, immunological responses, gonadogenesis, and larval survival. In this review, existing literature on the importance of freshwater finfish species to aquaculture and the inclusion of dietary lipid compounds in freshwater fish feed to accelerate the reproduction rate is being summarized and discussed. Although lipid compounds have been confirmed to improve reproductive performance, only a few members of the most economically important species have reaped benefits from quantitative and qualitative lipid studies. There is a knowledge gap on the effective inclusion and utilization of dietary lipids on gonad maturation, fecundity, fertilization, egg morphology, hatching rate, and consequently, larval quality contributing to the survival and good performance of freshwater fish culture. This review provides a baseline for potential future research for optimizing dietary lipid inclusion in freshwater broodstock diets.