This study was conducted to determine the taste preference of hybrid grouper (Epinephelus fuscoguttatus × Epinephelus lanceolatus) for nucleoside (inosine - INO) and nucleotides (inosine-5’-monophosphate – IMP and guanosine-5’- monophosphate - GMP), which are commonly used as feeding stimulants in fish diets. Behavioural assay was conducted by feeding the fish with the agar gel pellets that contained the taste substances and observing their feeding response through
video recording. Preference Index (PrfInd; minimum = 0, maximum = 1) was calculated to evaluate the fish preference for the taste substances tested. Data so collected suggested that INO, IMP, and GMP were generally preferable by the hybrid grouper. However, INO was the most potential FS for both S and L sizes (19.7 ± 1.4 g and 162.3 ± 50.5 g in BW, mean ± S.D.) hybrid grouper based on its functional dosage. In the S size fish, INO attained high PrfInd at both 0.1% and 1% (0.84 and 1) concentration levels, while IMP and GMP attained high PrfInd at 1% (both 1) but low PrfInd (0.6 and 0.13,respectively) at 0.1%. In the L size fish, INO also attained very high PrfInd at 0.01% (0.87) but IMP and GMP were not (0.13 and 0). In addition, improvement in the taste acceptance to INO of the hybrid grouper was evident in this study. The S size fish rejected INO at 0.01% (PrfInd = 0) but the L size fish accepted it very well (0.87). These results suggested that the supplementation level of INO in the practical diets for the hybrid grouper can be decreased with the fish growth during the grow-out period.
Combining feeding appendage morphology and behavioural observation of the motion pattern of the feeding appendages clarified many aspects underlying the feeding processes of the giant freshwater prawn (Macrobrachium rosenbergii) and the marine whiteleg shrimp (Litopenaeus vannamei) in aquaria. The food intake behaviour was video recorded during eating pellet food, and pieces of fresh squid and fish. While M. rosenbergiitook pellet one by one, L. vannamei picked up many pellets at one time and held them at the mouth with the 3rd maxilliped endopods and the 1st walking legs. Both species used the right chelate walking legs rather than the left walking legs to pick up the food. The 3rd walking legs of L. vannameiwere longest and heaviest among the chelate walking legs but their major role was not for feeding but for feeding contests often in the form of aggression. While M. rosenbergii easily crunched pellets by the mandibles, L. vannamei did not crunch pellets due to the softer and not so strong mandibles and frequently spat out them, indicating that the present hard pellets are not suitable for L. vannamei. Both the species kept a piece of elastic fresh squid or fish flesh at the mouth and tore the food into small pieces with the help of repeated pulling down motion of the 3rd maxilliped endopods. However, the mandible teeth of the two spices were not sharp enough to gnaw off the fibrous muscle in one bite. The 2nd and 3rd maxilliped endopods were used for holding food at the mouth and did not contribute to mastication of food. The 2nd and 3rd maxilliped exopods exhibited the horizontal fanning motion, which caused a unidirectional water flow moving backwards in the gill chamber (visualized with milk). The maxilliped exopods were found to contribute not to feeding but ventilation. Based on the results obtained, development of softer pellets was recommended for L. vannamei.
Tieshangang Bay in the Beibu Gulf, Guangxi of China, is a strategic location for pearl farming. Although water pollution has been reported in this bay but the general health of the pearl oyster, Pinctada fucata martensii, farmed there has never been assessed. The present study examined the condition of P. fucata martensii farmed in the Tieshangang Bay by analyzing its length-weight relationship (LWR) and relative condition factor (RCF). A total of 111 specimens were sampled for measuring their shell height and total weight for determining the LWR and RCF. The coefficient of correlation of the LWR was high (R2 = 0.93), significant at 0.01 level. Negative allometric growth (b = 2.7048) was observed. However, P. fucata martensii achieved the expected growth in terms of weight, as determined through the RCF (mean 1.13). Negative allometric growth is commonly reported on the wild Pinctada spp. collected from different regions. Apparently, the water pollution in the Tieshangang Bay did not compromise the general health of the pearl oyster cultured there. Nevertheless, further study on the farm’s surrounding water quality and plankton availability is necessary to investigate the interaction between the growth of the oyster and its culture environment. In conclusion, the P. fucata martensii farmed in the Tieshangang Bay was considered healthy and the bay is still suitable for pearl oyster farming.
The present study was aimed at evaluating the growth performance and feed utilization of marble goby (Oxyeleotris marmorata) juveniles fed with the acidified diets (AD). In feeding trial I, five fish meal-based diets were prepared [control (pH 6.0), AD 5.3, 4.3, 3.2 and 2.5]. Each diet was fed to triplicate batches of wild-caught O. marmorata (19 fish/ tank; total length, TL = 4.72±0.46 cm) for 8 weeks. The control, AD 5.3, and AD 4.3 treatments were terminated at week 4, due to drastic decline in total feed intake (TFI=0–0.05 g) and weight loss (weight gain, WG = -15.3 to -16.9%) in the fish. The AD 3.2 and AD 2.5 treatments were continued until week 8. Fish fed with the AD 3.2 showed significantly higher (P < 0.05) TFI (0.98 g) compared to those fed with the AD 2.5 (0.73 g) at the end of the trial. Feeding trial II was done to assess the long-term effects of AD for another 7 weeks. Thirty fish specimens were randomly selected from each of AD 3.2 and AD 2.5 treatments and stocked individually in 7 L aquaria to eliminate the territorial behaviour that was observed in the first trial. At the end of the experiment, fish fed AD 3.2 attained significantly higher (P < 0.05) WG (34%) than those fed AD 2.5 (13%). However, the growth performance and feed utilization results of the present study were very poor compared to those fed the normal fishmeal-based diet as is evident from synthesis of data from the literature review. Feeding of acidified diets was, therefore, not recommended for an extended period (> 3 weeks). Nevertheless, based on the strong preference of the fish for AD, it is worth trying to develop a weaning protocol using these diets as the starter feed and then slowly replacing them with the normal diet.
A 25-week feeding trial was conducted to assess the growth performance, organoleptic quality, and to estimate the viability of nourishing hybrid grouper (Epinephelus fuscoguttatus x Epinephelus lanceolatus) with low-cost fish (LCF) and commercially compound feed (CCF). A group of 3600 juvenile fish (182g) were released in four sea cages and fed with either LCF or CCF in duplicate. At the end of the trial, the hybrid grouper provided LCF attained a significantly higher (P0.05). Although technicalities of fish fed with LCF suggest that LCF is more efficient than CCF, feeding LCF to high-value fish is an unsustainable practice as LCF is usually obtained through trawling – a destructive fishing method for the marine ecosystem. Therefore, feeding with CCF without the use of LCF as the source of protein for its fishmeal will contribute to sustainable aquaculture. In order to convince the local farmers in Sabah to adopt the practice of feeding CCF, future research should focus on completing the species-specific diet formulation to promote optimum growth, and find ways to reduce the CCF local selling price.
The previous behavioural studies on vision of decapod crustaceans were often based on the assumption that the visual field of the test animals was all around and there was little or no blind area above or to the rear of the animals. In the present study, we determined the visual field of the wild captured purple mud crab (Scylla tranquebarica) and the farmed whiteleg shrimp (Litopenaeus vannamei) by eliminating the directions in which vision is anatomically blocked in all directions around the eyes. The mud crab had the visual field covering the entire visual world except for the ventral-most blind area. The whiteleg shrimp has the visual filed with a 66˚ binocular field and can see all around but is morphologically blocked by the scaphocerite extended forward between the eyes. While the transparent scaphocerites transmit 80 % of light from 400 to 700 nm wavelengths, an object seen through the scaphocerites is faded due to the light refraction, indicating that the morphological blocking is not always negligible. The trait of these visual fields should be taken into consideration in the design of visual behaviour experiments.
Feeding attractiveness of 21 fresh herbs was examined for the giant freshwater prawn (Macrobrachium rosenbergii) and the whiteleg shrimp (Litopenaeus vannamei) at different growth stages in tanks. The feeding attractant and enhancement were assessed by behavioural observations of the test animals in response to pelleted feeds incorporated with the herbal extracts in different concentrations. For M. rosenbergii, peppermint (Mentha piperita) and dokudami (Houttuynia cordata) were the most attractive herbs at all stages, and garlic (Allium sativum) strongly attracting to the juveniles but only weekly to the adults, postlarvae and larvae. Peppermint significantly enhanced feeding of M. rosenbergii at all stages except for the larvae. Garlic was a significant feeding enhancer for M. rosenbergii juveniles and postlarvae. The inclusion of the herbal extract at high concentrations had a negative effect on the feeding. As far as L. vannamei is concerned, the red chilli (Capsicum annuum) was the best feeding attractant and ginger (Zingiber officinale) and peppermint were moderate attractants for adults and juveniles. While galangal (Alpinia galangal), ginger and yellow onion (Allium cepa sp.) enhanced feeding in L. vannamei adults, but red chilli, garlic and peppermint did not enhance feeding. Evidently, the strong feeding attractants were not necessarily effective feeding enhancers. The incorporation of several herb extracts at higher concentrations caused a negative effect on the feeding of the test animals. This could be due to the presence of feeding deterrents in the herb such as saponins which are known to lower food palatability in insect, decapod crustaceans and fishes. More long-term work is warranted to determine if inclusion of feeding enhancing herbs in artificial feed promotes growth performance of the shrimp.
Oxyeleotris marmorata is an ambush predator. It is known for slow growth rate and high market demand. Farming of O. marmorata still remains a challenge. In order to establish a proper feeding practice to stimulate growth, knowledge of its metabolic processes and cost should be examined. Therefore, this study was designed to investigate the diel osmorespiration rhythms of O. marmorata in response to feeding challenge by using an osmorespirometry assay. The results have shown that oxygen consumption rate of the fed fish was approximately 3 times higher than that of the unfed fish in early evening to support specific dynamic action. Digestion and ingestion processes were likely to be completed within 18-20 h in parallel with the ammonia excretion noticeable in early morning. Under resting metabolism, metabolic oxygen consumption was influenced by diel phase, but no effect was noted in ammonia excretion. As a nocturnal species, O. marmorata exhibited standard aerobic metabolic mode under dark phase followed by light phase, with high oxygen consumption rate found in either fed or unfed fish. It can be confirmed that both the diel phase and feeding have a significant interactive impact on oxygen consumption rate, whereas ammonia metabolism is impacted by feeding state. High metabolic rate of O. marmorata supports the nocturnal foraging activity in this fish. This finding suggested that feeding of O. marmorata should be performed during nighttime and water renewal should be conducted during daytime.
The effect of low pH on the tactile sense of Macrobrachium rosenbergii postlarvae was determined in the laboratory by means of two behavioural assays: shelter (netting) occupancy and jumping response to touch stimuli (taps) by a glass micropipette. The postlarvae were acclimated to pH 4, pH 5, pH 6 and pH 7.5 (control) in 45 L aquaria 5-7 d before the experiments. Shelter occupancy decreased with pH and was significantly lower at pH 4 and pH 5 than at pH 6 and in the control. The jumping response instantly followed a tap 93-98% of the time in the control, pH 6 and pH 5 treatments. However, the postlarvae showed significantly lower jumping response (65%) at pH 4, indicating an impaired tactile sense. Low pH 4-5 probably degrades the chitin of the sensory setae and inhibits the surface mechanoreceptors of the prawn postlarvae.
Carotenoids are biologically active pigments that are well-known to enhance the defense and immunity of the vertebrate system. However, in invertebrates, the role of carotenoids in immunity is not clear. Therefore, this study aims to review the scientific evidence for the role of carotenoids in invertebrate immunization. From the analysis of published literatures and recent studies from our laboratory, it is obvious that carotenoids are involved in invertebrate immunity in two ways. On the one hand, carotenoids can act as antioxidant enzymes to remove singlet oxygen, superoxide anion radicals, and hydroxyl radicals, thereby reducing SOD activity and reducing the cost of immunity. In some organisms, carotenoids have been shown to promote SOD activity by up-regulating the expression of the ZnCuSOD gene. Carotenoids, on the other hand, play a role in the expression and regulation of many genes involved in invertebrate immunity, including thioredoxins (TRX), peptidoglycan recognition receptor proteins (PGRPs), ferritins, prophenoloxidase (ProPO), vitellogenin (Vg), toll-like receptor (TLRs), heat shock proteins (HSPs), and CuZnSOD gene. The information in this review is very useful for updating our understanding of the progress of carotenoid research in invertebrate immunology and to help identify topics for future topics.
Azolla is a freshwater floating aquatic fern found in the tropical, subtropical and temperate regions with a high nitrogen-fixing rate from the result of symbiotic relationship with the blue-green cyanobacterium, Anabaena azollae. Azolla can effectively remediate aquaculture wastewater owing to its high production capacity and the ability to absorb nutrients and toxic compounds. The Azolla biomass generated as a by-product is currently underutilized and could potentially benefit the aquafeed industry in replacing the unfeasible and expensive fishmeal protein at a certain level. This study evaluates the incorporation of red tilapia wastewater-raised Azolla as a dietary protein for the growth performance, feed efficiency, survival, body indices, body composition and nutrient utilization of Pangasius catfish Pangasianodon hypophthalmus during a 90-days feeding experiment. Dried Azolla was incorporated into four isonitrogenous (30 g kg-1) and isolipidic (12 g kg-1) practical diets containing 0 g kg-1 (Control), 10 g kg-1 (A10), 20 g kg-1 (A20) and 30 g kg-1 (A30) fishmeal protein replacement. One hundred and twenty juveniles with an initial mean weight of 45 ± 15 g were distributed into 12 tanks representing four dietary treatments in triplicates. Results showed significant (p 0.05) effects were recorded for feed intake, survival, body indices and nutrient utilization amongst all dietary treatments. In conclusion, Azolla raised from red tilapia aquaculture wastewater can replace fishmeal protein up to 10 g kg-1 in the diet of Pangasius catfish juveniles having better growth, feed efficiency and nutrient utilization without affecting its survival, body indices and body composition.
Intensive research on the effectiveness of chemoattractants has been widely explored to improve the feed qualities in expanding crustacean farming. Taste preferences in slipper lobster remained unknown despite their significant contribution to the lobster fisheries. Chemoattractants allow better performance in aquaculture species by increasing food attractiveness and palatability. Amino acids (AA) have been leading in previous research on crustacean feeding behavior. Given that slipper lobster possesses chemoreceptors to detect and orient towards food, this study investigated an approach to identify the AA with the most potent chemoattractant in eliciting a response from slipper lobster. Behavioral assays were performed to evaluate the responses of slipper lobster Thenus orientalis (carapace length, 52.34 ± 1.52 mm) on 15 crystalline AA and three derivatives of AA (DAA) at three concentrations between 10-1 and 10-3 M as test substances (TS). Meretrix sp. extract was used as a positive control and clean filtered seawater as a negative control. The behavioral responses of 14 T. orientalis were evaluated based on their antennular flicking rate, third maxillipeds activity, and substrate probing by the pereiopods. T. orientalis responded to the solutions of single AA down to a concentration of 10-3 M, excluding histidine and serine. The behavioral activity displayed by T. orientalis increased with the TS concentrations. L-glutamic acid monosodium salt monohydrate, betaine, and glycine solutions elicited the most behavioral responses, whereas histidine exhibited the lowest behavioral responses. Conclusively, L-glutamic acid monosodium salt monohydrate, betaine, and glycine can be potential chemoattractants for T. orientalis.
Swimming crabs have a characteristic fifth pair of legs that are flattened into paddles for swimming purposes. The dactyl of these legs bears a thick seta along its edge. The chemoreceptive and feeding properties of the seta are supported with scientific evidence; however, there is no available data on the sensitivity of the setae in portunid crabs. The underlying mechanisms of the chemo- and mechano-sensitivity of appendages and their involvement in feeding activities of the mud crab (Scylla paramamosain) were investigated using electrocardiography and behavioural assay, which focused on the responses of the mud crab to chemical and touch stimulus. Electrocardiography revealed the sensory properties of the appendages. The dactyls of swimming legs and the antennules were chemosensitive, but not mechanosensitive and vice versa for the antennae. However, the mouthparts, claws, and walking legs were chemo- and mechanosensitive. Only the chemosensitive appendages, including the swimming legs, were directly involved in feeding. The flattened dactyls of the swimming legs were more efficient than the pointed dactyls of the walking legs in detecting the food organism crawling on the substrate. The structural features enhanced the capacity of the crab in coming into contact with scattered food items. This study revealed that the swimming legs are important appendages for feeding in the mud crab.
Inadequate gonadal maturation and poor spawning performance increasingly threaten the sustainability of shrimp aquaculture. Unraveling the mechanisms regulating ovarian development and maturation hence is critical to address industry challenges. Vitellogenin (Vtg), a precursor of yolk protein found in the hepatopancreas and ovary of shrimp, plays a key role in facilitating shrimp's oocyte maturation and embryonic development after oviposition. This study found that FpVtg was specifically expressed in F. penicillatus hepatopancreas and ovary. FpVtg was localized predominantly in the oocyte cytoplasm and distributed uniformly in the hepatopancreas tissue. Silencing FpVtg led to apoptosis in both hepatopancreas and ovary tissues. Furthermore, FpVtg depletion upregulated the expression of ovarian peritrophin 1, ovarian peritrophin 2, serine proteinase inhibitor 6, and juvenile hormone esterase-like carboxylesterase 1, while downregulated that of vitellogenin, delta-9 desaturase, and insulin-like receptor. KEGG pathway analysis implicated such as PI3K-AKT signaling, RNA transport, ECM-receptor interaction, hippo signaling, oocyte meiosis, and apoptosis were enriched and involved in ovarian development. These findings have provided insights into the FpVtg's reproductive role and the associated regulatory genes and pathways in F. penicillatus. This knowledge can contribute to establishing strategies to improve the breeding and aquaculture production of F. penicillatus by elucidating its vitellogenesis regulation in redtail prawn and other penaeid species. Further characterization of the implicated pathways and genes will clarify the intricacies underlying ovarian maturation.
This study was conducted to determine the senses that facilitate prey detection in the marble goby (Oxyeleotris marmorata) larvae. The ingestion ratios of live (generate chemical and mechanical stimuli) or frozen Artemia nauplii (generate chemical but no mechanical stimuli) by the intact or free neuromast (mechanoreceptor)-ablated O. marmorata larvae (11 mg/L streptomycin treatment before feeding) under the light or dark (fish vision was obstructed) condition were examined. Vision, mechano-, and chemoreceptions were all found to be essential in prey detection of the O. marmorata larvae. Prey movement has a significant influence as a visual stimuli on the O. marmorata larval feeding as the Artemia nauplii ingestion ratio was approximately 40% higher with significant (p = 0.001, d = 3.0), when the intact larvae were fed with the live (78.1 ± 1.5%), rather than the frozen (40.9 ± 2.8%) Artemia nauplii, under the light condition. This result was assured when no significant difference (p = 0.572, d = 0.2) was found between the ingestion ratios of frozen Artemia nauplii by the intact O. marmorata larvae under light and dark conditions. These findings demonstrate that prey detection in the O. marmorata larvae was facilitated by multi-modal senses, allowing O. marmorata larvae to survive in their natural habitats.
The vitellogenin present in the bloodstream undergoes internalization into developing oocytes through the vitellogenin receptor (VgR), a process mediated by receptor-mediated endocytosis. VgR plays a crucial role in facilitating the accumulation of vitellogenin and the maturation of oocytes. In this study, we characterized a Tachypleus tridentatus vitellogenin receptor (TtVgR) gene from the tri-spine horseshoe crab, revealing a length of 1956 bp and encoding 652 amino acid residues with 12 exons. TtVgR has a molecular weight of 64.26 kDa and an isoelectric point of 5.95. Predictions indicate 85 phosphorylation sites and 7 glycosylation sites within TtVgR. Transcriptional analysis demonstrated specific expression of TtVgR in the ovary and yellow connective tissue. TtVgR was identified and distributed in the plasma membrane of oocytes. The siRNA-mediated TtVgR knockdown significantly reduced the transcriptional activity of TtVgR. This depletion induced excessive ROS production, resulting in DNA damage in ovarian primary cells. TUNEL and flow cytometry analyses confirmed ovarian cell apoptosis following TtVgR knockdown, indicating DNA damage in ovarian primary cells. These findings underscore the importance of TtVgR in ovarian cell development, suggesting its potential involvement in vitellogenesis and oocyte maturation. This knowledge may inform innovative breeding strategies and contribute to the sustainable management and conservation of the tri-spine horseshoe crab.
Horseshoe crabs are living fossils. In recent decades, the population of horseshoe crabs, especially the tri-spine horseshoe crab Tachypleus tridentatus, has decreased significantly and was listed as an 'endangered species' under the IUCN Red List in 2019. In order to improve the reproduction of T. tridentatus to facilitate stock enhancement, it is important to understand their ovarian development. In this study, a novel TtVtg2-like gene from T. tridentatus was cloned and functionally characterized. The total legth of TtVtg2-like was 5469 bp, encoding a protein consisting of 1822 amino acid with a pI value of 6.51 and a molecular weight of 208.68 KDa. The TtVtg2-like was highly expressed in the ovary and yellow connective tissues, mainly localized in cytoplasm and endoplasmic reticulum vesicles of oocytes and yellow connective tissues, respectively. RNA interference of TtVtg2-like caused the accumulation of ROS, DNA damage, and apoptosis of ovarian primary cells. The results of this study provide useful baseline information for future studies on ovarian development in horseshoe crabs.
The Bokashi leachate (BL) is a by-product from the anaerobic digestion of food waste, following the Bokashi composting method. Bokashi leachate is acidic and it contains effective microorganisms hence it has potential to be a functional feed additive to the plant proteins based diets for fish farming. This study evaluated the growth performance and feed utilization of the red tilapia (Oreochromis sp.) fingerlings fed with the BL supplemented soybean meal (SBM) based diets. After an 8-week feeding trial, fish fed with the 5% BL supplemented SBM diet attained the highest weight gain. This result was significantly higher (p 0.05) to those fed with the control full fish meal (FM) diet. Generally, dietary inclusion of BL enhanced the fish feed intake on the SBM diet but it did not show clear sign of improvement in their feed utilization. In addition, no significant difference was found across the hepatosomatic index and viscerosomatic index from all dietary treatments. These outcomes concluded that dietary inclusion of BL can enhance the feed intake and growth performance of the red tilapia fingerlings fed with the SBM based diet without compromising their health, and the optimum BL inclusion level was 5%. Nevertheless, further study on the properties and substances content of the BL produced from different types and ratios of food waste is strongly recommended. In this study, BL was also discovered to be capable of reducing the crude fiber content in the SBM diets. Such observation deserves a further exploitation on the application of BL to manipulate the crude fiber content in the plant proteins based diets in fish farming.
Microplastic pollution in our environment, especially water bodies is an emerging threat to food security and human health. Inevitably, the outbreak of Covid-19 has necessitated the constant use of face masks made from polymers such as polypropylene, polyurethane, polyacrylonitrile, polystyrene, polycarbonate, polyethylene, or polyester which eventually will disintegrate into microplastic particles. They can be broken down into microplastics by the weathering action of UV radiation from the sun, heat, or ocean wave-current and precipitate in natural environments. The global adoption of face masks as a preventive measure to curb the spread of Covid-19 has made the safe management of wastes from it cumbersome. Microplastics gain access into aquaculture facilities through water sources and food including planktons. The negative impacts of microplastics on aquaculture cannot be overemphasized. The impacts includes low growth rates of animals, hindered reproductive functions, neurotoxicity, low feeding habit, oxidative stress, reduced metabolic rate, and increased mortality rate among aquatic organisms. With these, there is every tendency of microplastic pollution to negatively impact fish production through aquaculture if the menace is not curbed. It is therefore recommended that biodegradable materials rather than plastics to be considered in the production of face mask while recycle of already produced ones should be encouraged to reduce waste.
Vitellogenin (Vtg) serves as the precursor of yolk protein and exhibits widespread distribution in tissues, including in the ovary of both vertebrates and invertebrates. Vtg plays a critical role in facilitating oocyte maturation and embryonic development following oviposition. In this study, we have successfully elucidated the complete transcript sequence of TtVtg6-like from an ancient chelicerate Tachypleus tridentatus. The TtVtg6-like transcript encompassed a length of 4887 bp and encoded 1629 amino acids residues. Notably, TtVtg6-like was found to contain 25 exons. Furthermore, the molecular weight and isoelectric point of TtVtg6-like were determined to be 191.6 KDa and 6.73, respectively. Subsequent mRNA expression analysis demonstrated the specific expression of TtVtg6-like in ovary and yellow connective tissue. In addition, TtVtg6-like was located and distributed in both ovary and yellow connective tissue. Intriguingly, employing an siRNA approach to silence TtVtg6-like resulted in a decrease in TtVtg6-like transcription levels. Concomitantly, TtVtg6-like silencing led to increase production of ROS, ultimately resulting in DNA damage and cell apoptosis within the ovarian primary cell. The induction of apoptosis ovarian primary cells due to TtVtg6-like silencing was further corroborated through TUNEL assay and flow cytometry analysis. Overall, our findings underscore the significance of TtVtg6-like in ovarian cell development, revealing its potential association with ovarian cell apoptosis. Consequently, the insights gained from this study contribute to the future exploration of vitellogenesis and ovarian development in T. tridentatus.