Larval Quality Research Articles

Environmental microplastics compromise reproduction of the marine invertebrate Mytilus galloprovincialis: A holistic approach

The extensive presence of microplastics (MPs) in marine ecosystems constitutes a major threat to aquatic environments. The gametes of the marine invertebrate Mytilus galloprovincialis, which is essential for coastal ecosystems, are released directly into the water, potentially exposing them to environmental microplastics (EMPs). This study examined the effects of exposing M. galloprovincialis gametes to 50 or 100 µg/L EMP for 1 h on fertilization rates, larval quality, and the molecular mechanisms underlying the induction of apoptosis and shell growth. Our findings show that increased EMP concentrations correlate with reduced fertilization success and higher rates of larval malformations, indicating negative impacts on embryonic development. Additionally, DNA degradation in larvae is related to the EMP concentration. The apoptosis-associated proteins Bax, P53, and Cas-3 are upregulated, whereas Bcl-2 and DNA-ligase are downregulated with increasing EMP concentrations. Prothymosin-ɑ (PTMA), which is crucial for cell proliferation, also decreases with increasing EMP concentrations, contributing to impaired cell proliferation and growth imbalances. Reduced HRG gene expression is correlated with decreased shell growth and larval malformations. This study underscores the detrimental impact of EMPs on bivalve gametes, which impacts fertilization success and larval quality and highlights the potential risks to species survival and marine ecosystem stability.

Ocean conditions influence the quality of recruiting benthic marine invertebrate larvae—Insights from fatty acids

AbstractMany marine benthic invertebrates have a pelagic life stage, during which larvae need to accumulate enough reserves to complete metamorphosis to a settled benthic juvenile. Currently, very little is known about how ocean conditions affect quality of the larvae. We studied this for three settlement seasons (2017–2019) by collecting returning Dungeness crab (Metacarcinus magister) megalopae at the Oregon coast and analyzing them for fatty acid biomarkers. We found that the larvae are omnivorous and have versatile diets. The daily larval abundance was positively correlated with larval quality. Despite the relatively high day‐to‐day variation, we found pronounced seasonal and inter‐annual differences in the body condition (size and total fatty acid content) and biomarker composition of megalopae. Especially, the early season recruits of 2017 had lower content of lipids and polyunsaturated fatty acids known to be beneficial to crustaceans. This is likely related to lingering effects of the eastern Pacific marine heat wave (2014–2016) on pelagic communities. The larvae were rich in docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) with lower levels in benthic juveniles, indicating an ontogenetic diet shift and likely lower availability of DHA and EPA in the benthic environment. The pulsed megalopa recruitment may provide substantial carbon and lipid inputs to the nearshore ecosystem as a form of pelagic subsidy. Our results reveal that ocean conditions may have an effect on the quality of returning larvae, which likely influence their recruitment performance and early juvenile success, and thus potentially also the population size and commercial catch 4 yr later.

The AutoSpawner system - Automated ex situ spawning and fertilisation of corals for reef restoration

The restoration of reefs damaged by global and local pressures remains constrained by the scale of intervention currently feasible. Traditional methods for ex situ sexual propagation of corals produce limited materials, typically of limited genetic diversity and only sufficient for small field trials. The development and validation of new technologies to upscale and automate coral propagation is required to achieve logistically and financially feasible reef restoration at ecologically relevant scales. To address the need for upscaled production of genetically diverse material for use in reef restoration we designed an automated system (the AutoSpawner) for harvesting, fertilising and washing gametes from tropical broadcast-spawning corals. The system includes a novel high density dynamic fertilisation process, which enables the production of large numbers of fertilised coral eggs (>7 million per night for highly fecund species) without any downstream negative effects on larval quality. The functionality of the system and the quality of the produced larvae was assessed using multiple species from two coral families (Acroporidae and Merulinidae) across a range of spawning and gamete characteristics. We present the schematics and protocols required for automated sexual propagation of high-quality coral larvae using this novel system; and demonstrate that the time demands, and labour costs, associated with traditional manual-based sexual propagation of corals can be reduced by up to 113-fold using the AutoSpawner.

Cultivating black soldier fly (Hermetia illucens) larvae on coconut endosperm and soybean curd residue: Impact on growth performance, waste reduction efficiency and larval nutritional composition

AbstractBlack soldier fly larva (BSFL), Hermetia illucens (L.) (Diptera: Stratiomyidae), presents an attractive waste management solution that not only addresses the issue of waste but also generates alternative proteins and fats. Substrate quality and pre‐treatment by fermentation may play a crucial role in waste reduction efficiency and larval quality. The aim of this study was to investigate the growth performance, waste reduction efficiency and nutritional composition of BSFL rearing using different ratios of coconut endosperm (CE) and soybean curd residue (SC) with pre‐treatment by self‐fermentation (F), fermentation with baker's yeast (Y) and without pre‐treatment (C). Seven‐day‐old larvae were randomly separated into 15 experimental groups with three replicates. The experiment ended when the prepupae appeared. Based on the results, the larvae fed a diet with a high percentage of fresh SC exhibited the highest weight and growth rate, whereas the shortest development time was observed in larvae reared on Y. Pre‐treatment of the substrate with fermentation proved effective in waste reduction. The nutritional composition of the larvae showed that the highest crude protein was produced in larva reared on fresh 100% SC, and the highest ether extract was found when fed CE at 100% with pre‐treatment with F or Y. In conclusion, it is recommended to employ a fresh diet with a high SC content to optimise protein production, whereas a high level of CE with F or Y may be performed to obtain a high ether extract content. However, incorporating yeast fermentation with an SC content exceeding 25% provided high efficiency in waste reduction. Therefore, the selection of the ratio between CE and SC and of pre‐treatment techniques depends on the producer's objective.

Optimizing initial stocking densities of wild coral spawn slicks for mass production of larvae and settled corals for restoration

Mass culturing coral larvae collected from spawn slicks during spawning events and rearing them directly on reefs is an important method for coral reef restoration as it utilizes high genetic diversity and is readily upscaled. Understanding optimal initial stocking densities in mass cultures is, therefore, central to optimizing larval production efficiency. Yet, no studies have focused on optimizing initial stocking densities while coral embryos are still floating, despite the influence of density‐dependent effects during larval culturing. This study aimed to (1) explore initial stocking densities based on slick coverage and (2) determine the effects of initial densities on embryo development to coral settlement and early post‐settlement survival. Wild coral spawn slicks were collected at Lizard Island and reared at different combinations of slick coverage and density based on tank volume. Immediate density‐dependent effects were observed in treatments with 45% slick coverage, which had lower abundances of embryos 12 hours post‐spawning. The 12‐hour embryo densities ranged from 1.7 to 10.7 mL−1 and continued to display a negative relationship between density and survival of developing larvae. Overall, a density of approximately 5.1 embryos mL‐1 was the optimal stocking density for larval survival to competency (16.5%). The proportion of settlement and early post‐settlement survival was negatively correlated with higher initial densities, while a positive relationship was observed between the abundance of competent larvae and those settled, including initial settlers and 2‐month‐old recruits. These results highlight the importance of both larval quality and supply, and opportunities for site and system‐specific optimization of larval production in future work.

Early postovulatory aging reveals the first proteomic markers of egg quality in pikeperch

This study explored the molecular mechanisms underlying egg quality deterioration due to in vivo postovulatory aging in pikeperch (Sander lucioperca L.), a key species in aquaculture. We employed tandem mass tag (TMT) peptide labeling coupled with LC–MS/MS quantitative proteomics to analyze eggs collected at various postovulation intervals. Our research revealed four distinct proteomic markers (Gins4, Atrx, DnaJB14, and Mrpl10) that are differentially expressed in response to early aging, shedding light on their potential roles in DNA replication, chromatin organization, protein folding, and mitochondrial function. The study confirmed that eggs maintain morphological integrity up to 5 h postovulation but exhibit compromised fertilization capacity, underscoring the importance of timely egg utilization in aquaculture practices. These findings enhance the understanding of egg aging at the molecular level, offering insights for improving reproductive success and larval quality in pikeperch aquaculture. The data are available via ProteomeXchange with the identifier PXD048349.

Comparative transcriptome analysis reveals molecular damage associated with cryopreservation in Crassostrea angulata D-larvae rather than to cryoprotectant exposure

BackgroundThe Portuguese oyster Crassostrea angulata, a bivalve of significant economic and ecological importance, has faced a decline in both production and natural populations due to pathologies, climate change, and anthropogenic factors. To safeguard its genetic diversity and improve reproductive management, cryopreservation emerges as a valuable strategy. However, the cryopreservation methodologies lead to some damage in structures and functions of the cells and tissues that can affect post-thaw quality. Transcriptomics may help to understand the molecular consequences related to cryopreservation steps and therefore to identify different freezability biomarkers. This study investigates the molecular damage induced by cryopreservation in C. angulata D-larvae, focusing on two critical steps: exposure to cryoprotectant solution and the freezing/thawing process.ResultsExpression analysis revealed 3 differentially expressed genes between larvae exposed to cryoprotectant solution and fresh larvae and 611 differentially expressed genes in cryopreserved larvae against fresh larvae. The most significantly enriched gene ontology terms were “carbohydrate metabolic process”, “integral component of membrane” and “chitin binding” for biological processes, cellular components and molecular functions, respectively. Kyoto Encyclopedia of Genes and Genomes enrichment analysis identified the “neuroactive ligand receptor interaction”, “endocytosis” and “spliceosome” as the most enriched pathways. RNA sequencing results were validate by quantitative RT-PCR, once both techniques presented the same gene expression tendency and a group of 11 genes were considered important molecular biomarkers to be used in further studies for the evaluation of cryodamage.ConclusionsThe current work provided valuable insights into the molecular repercussions of cryopreservation on D-larvae of Crassostrea angulata, revealing that the freezing process had a more pronounced impact on larval quality compared to any potential cryoprotectant-induced toxicity. Additionally, was identify 11 genes serving as biomarkers of freezability for D-larvae quality assessment. This research contributes to the development of more effective cryopreservation protocols and detection methods for cryodamage in this species.

Experimental Infection Models and Their Usefulness for White Spot Syndrome Virus (WSSV) Research in Shrimp

White spot syndrome virus (WSSV) is marked as one of the most economically devastating pathogens in shrimp aquaculture worldwide. Infection of cultured shrimp can lead to mass mortality (up to 100%). Although progress has been made, our understanding of WSSV’s infection process and the virus–host–environment interaction is far from complete. This in turn hinders the development of effective mitigation strategies against WSSV. Infection models occupy a crucial first step in the research flow that tries to elucidate the infectious disease process to develop new antiviral treatments. Moreover, since the establishment of continuous shrimp cell lines is a work in progress, the development and use of standardized in vivo infection models that reflect the host–pathogen interaction in shrimp is a necessity. This review critically examines key aspects of in vivo WSSV infection model development that are often overlooked, such as standardization, (post)larval quality, inoculum type and choice of inoculation procedure, housing conditions, and shrimp welfare considerations. Furthermore, the usefulness of experimental infection models for different lines of WSSV research will be discussed with the aim to aid researchers when choosing a suitable model for their research needs.

Recent insights into egg quality and larval vitality of the European eel Anguilla anguilla

To date, the eel industry still depends on wild-caught juveniles that are grown to marketable size. There is an urgent need to close the eel life cycle in captivity to make aquaculture independent of the natural population. With this artificial reproduction protocol, yolk-sac larvae can be produced but egg quality may be impaired. Low survival rates and high deformity rates are frequently observed during the first week after hatching. Over the past four years, we have conducted studies with the aim to optimize the artificial reproduction protocol, thereby focussing on increasing egg and larval quality. Weekly carp or salmon pituitary extract (PE) treatment was successfully replaced with recombinant gonadotropins (rGTHs) to mature female eels and produce larvae. 17α,20β-dihydroxy-4-pregnen-3-one (DHP) was replaced with upstream precursor progesterone (P) to induce the endogenous production of DHP by the female eel. DHP and P were found equally potent in inducing oocyte maturation and ovulation. The effects of antibiotics on larval survival and the occurrence of deformities were investigated. Antibiotic treatment increased survival and decreased the occurrence of deformities indicating bacterial infection as an important cause. A deformity determination key for young eel larvae has been developed that provides a framework of reference for larval deformities which will be instrumental with gaining insights on the reasons behind each larval deformity. These improvements of the artificial reproduction protocol and hatchery practices will contribute to the production of robust eel larvae that survive, grow and metamorphose into juveniles that will later be able to reproduce in captivity.

Natural Spawning of Pigfish Orthopristis Chrysoptera in Captivity and Predictors of Egg and Larval Quality

Pigfish Orthopristis chrysoptera are a popular aquaculture species for use as bait in the southeastern United States.

Broodstock nutrition in Nile tilapia and its implications on reproductive efficiency

Broodstock nutrition is the most essential constituent of sustainable aquaculture production. Its impact on the reproductive performance and profitability of commercial aquaculture has been the subject of considerable research. The present paper reviews the literature available on the subject, examining the implication of Nile tilapia broodstock nutrition on parameters including maturation, fecundity, fertilization, embryo development, larval quality, and survival rate. The provision of a nutritive diet composed of essential macro- and micronutrients including proteins, lipids, carbohydrates, vitamins, minerals, and functional additives, such as prebiotics, enzymes, hormones, and probiotics, in different proportions is discussed. Special emphasis is given to literature dealing with the effects of dietary protein and lipid on vitellogenesis and ovarian maturation, fecundity, egg hatching rate, larval quality, and fry survival number. The impact of the feeding regime on reproductive performance is described.

Intensification of Amazon River Prawn Hatchery

The effects of the intensification on the performance of the Amazon River prawn hatchery carried out in a simple recirculation system were investigated. Newly hatched larvae were stocked in 120 L tanks at 80, 100, 120 and 140 larvae L−1 in a closed recirculating system. The experiment used a randomized block design with five replicates. An exponential equation was adjusted to express the relationship between the stocking density and productivity (postlarvae L−1). The development, larval quality, survival and postlarval (PL) dry weight did not significantly differ among the treatments (p > 0.05). When 80 larvae were stocked, the productivity (54 ± 11 PL L−1) was lower than those at higher densities (p < 0.05). Stocking 120 and 140 larvae L−1 resulted in higher productivities (75 ± 18 and 80 ± 17 PL L−1, respectively) with a lower use of Artemia nauplii to produce each postlarvae (~1200 Artemia nauplii PL−1). The maximum mean M. amazonicum postlarval production estimated by the exponential model was 93 PL L−1. This means that despite the increase in stocking density, productivity tends to stabilize. The results showed that M. amazonicum tolerates high intensification in recirculating hatchery systems based on a crushed shell bed biofilter, and the intensification optimizes Artemia use.

Observation of Egg and Larval Quality of Milkfish (Chanos-chanos Forskall) G1 and G0 Broodstock in Controlled Tanks Supports The Development and Cultivation of Milkfish in The Community

Milkfish, a key aquaculture commodity, is vital for industrialization and food security. Obtaining milkfish seeds from nature is challenging, but hatcheries, especially in Buleleng north coast, can produce them in large quantities. However, the varying quality of milkfish eggs has led to a decline in seed quality. Research on G1 and G0 milkfish broodstock development is necessary. The aim is to assess the performance and spawning of G1 and G0 broodstock reared in controlled tanks to aid community cultivation. The study used a descriptive method to evaluate the spawn, eggs, and fry of G0 and G1 broodstock. Results showed that G1 and G0 broodstock have similar spawning and egg quality. G1 broodstock can produce 42,940,000 eggs, with 60-82% fertilization and SAI 2.50-4.12, while G0 can spawn 55,940,000 eggs, with 50-80% fertilization and SAI 2.00-4.40. The G1 derivative’s performance and egg quality are consistent, and the eggs are like the G0 natural broodstock. Milkfish fry production and pond culture cultivation continue to develop in the archipelago.

Reproductive Performance and Larval quality of Freshwater Prawn Broodstock of different water resources

The present study was conducted to compare the reproductive performance and offspring quality of adults of the giant freshwater prawn, Macrobrachium rosenbergii broodstock from four different sites (1) Ganga riverine wild breeders (GW) (2) Ganga riverine pond-reared (GP) breeders (3) Gandak riverine wild breeders (gp); and (4) Gandak riverine pond-reared breeders (gw) were grown out in culture ponds and collected as broodstock at the end of grow-out culture period. M. rosenbergii females were individually stocked for 120 days in three 1500L freshwater recirculation system tanks and fed a commercial diet. Ovarian development, molting and spawning events were checked daily. In addition a number of egg and larval quality parameters were determined. The breeding frequency, fecundity, egg laying success rate, egg dimensions and egg hatchability were not significantly different between animals from the four different sources. However, there were significant differences in terms of offspring quality between the different broodstock sources. Individual dry weight, larval development rate, time to reach the post-larval stage, post-larval survival and tolerance to ammonia toxicity were all better in offspring originating from all broods did not significantly differ in terms of breeding frequency, fecundity and egg dimensions. However, larval quality of Ganga riverine wild and Ganga riverine pond-reared breeders was markedly better than that of Gandak riverine wild reared stock and Gandak riverine pond-reared stock in terms of larval development, survival and post larval production. These results indicate that broodstock sourcing deserves proper attention in hatchery operations of M. rosenbergii. It furthermore proves that domesticated (pond-reared) animals are not necessarily inferior as breeders as compared to wild-sourced animals. The results may also point out the potential to selectively breed stocks with improved characteristics adapted to the local culture environment.

Diet–egg relationships and equilibration of fatty acids in eggs of southern flounder, Paralichthys lethostigma

There is increasing interest in the transfer of nutrients from broodstock diets to eggs due to the consequences for egg and larval quality. Analyses were conducted with southern flounder Paralichthys lethostigma to identify which fatty acids (FAs) in eggs are influenced by broodstock diet and how long it takes for the FA composition of eggs to equilibrate to the broodstock diet. Data were obtained from a laboratory experiment and from hatchery production operations. For each source, two populations of adult flounder were given one or the other of two diet treatments. In the laboratory experiment, the diets differed markedly in FA composition. In the hatchery, a base diet was supplemented on two different schedules. Each of the four diet treatments produced eggs with significantly different mean FA compositions. Of the 27 FAs measured, 19 differed significantly between diets and had corresponding significant differences in eggs (direct diet–egg relationships). Combining results with data from a prior study showed that equilibration of eggs to a change in adult diet takes 8–16 weeks for southern flounder. These findings will be useful for improving broodstock diets and feeding protocols for captive flounder being used by stock-enhancement programs that are designed to mitigate the long-term and widespread population declines of this species.

Performance of Orbicella faveolata larval cohorts does not align with previously observed thermal tolerance of adult source populations.

Orbicella faveolata, commonly known as the mountainous star coral, is a dominant reef-building species in the Caribbean, but populations have suffered sharp declines since the 1980s due to repeated bleaching and disease-driven mortality. Prior research has shown that inshore adult O. faveolata populations in the Florida Keys are able to maintain high coral cover and recover from bleaching faster than their offshore counterparts. However, whether this origin-specific variation in thermal resistance is heritable remains unclear. To address this knowledge gap, we produced purebred and hybrid larval crosses from O. faveolata gametes collected at two distinct reefs in the Upper Florida Keys, a nearshore site (Cheeca Rocks, CR) and an offshore site (Horseshoe Reef, HR), in two different years (2019, 2021). We then subjected these aposymbiotic larvae to severe (36°C) and moderate (32°C) heat challenges to quantify their thermal tolerance. Contrary to our expectation based on patterns of adult thermal tolerance, HR purebred larvae survived better and exhibited gene expression profiles that were less driven by stress response under elevated temperature compared to purebred CR and hybrid larvae. One potential explanation could be the compromised reproductive output of CR adult colonies due to repeated summer bleaching events in 2018 and 2019, as gametes originating from CR in 2019 contained less storage lipids than those from HR. These findings provide an important counter-example to the current selective breeding paradigm, that more tolerant parents will yield more tolerant offspring, and highlight the importance of adopting a holistic approach when evaluating larval quality for conservation and restoration purposes.

Assessment of Larval Health and Metamorphosis of the Giant Freshwater Prawn Macrobrachium rosenbergii Larvae during Acute Exposure to Imidacloprid

An acute toxicity study of imidacloprid on the larval health and metamorphosis of the giant freshwater prawn Macrobrachium rosenbergii larvae were carried out during the months of January–August 2021 at the aquatic animal raring and experimenting facility in the Cochin University of Science and Technology, Kerala, India. Imidacloprid is a broad-spectrum neonicotinoid insecticide used against sucking pests infecting agricultural crops as both contact poison and stomach poison. The present study investigated the adverse effects of imidacloprid on the survival rate, feeding behaviour, larval quality and incidence of metamorphosis in Macrobrachium rosenbergii larvae. The 48 h median lethal concentrations (LC50 values) and safe concentration of imidacloprid determined for different larval stages (Zoea I–XI) of M. rosenbergii ranged between 0.000635–0.001388 mg l-1 and 0.000055–0.000191 mg l-1, respectively. Larvae exposed to imidacloprid for 48 h showed decrease in survival rate with increase in concentration of imidacloprid. The results of larval condition index (LCI) that indicates the larval quality showed that imidacloprid reduced the larval health significantly (p<0.05) at high concentrations and is potent to delay the incidence of metamorphosis. The feeding rate of larvae exposed to different concentrations of imidacloprid were found to be significantly (p<0.05) reduced due to paralytic effect of imidacloprid. The present work provides relevant data beneficial for the risk assessment of imidacloprid in M. rosenbergii.

Examination of female energy dynamics and larval quality in the southern king crab, Lithodes santolla: Annual and interannual variability

The southern king crab, Lithodes santolla, constitutes one of the most important fishery resources in the Beagle Channel. In this study, we considered that Lithodes santolla females showing better physiological conditions may produce larvae with higher survival rates. To determine whether this quality may be useful for the enhancement of natural crab stocks, in the present study, we aimed to evaluate several biochemical parameters considering the size and ovigerous condition of the female parent. Primiparous (<80 mm CL) and multiparous (>80 mm CL) females, as well as ovigerous and non-ovigerous females, were evaluated. The patterns of accumulation or use of several macromolecules (glycogen, lipids and proteins) in different organs and development stages (midgut gland, ovary, eggs, larval stages and first crabs) were also evaluated along two reproductive seasons. The possibility that some females could be producing larvae with high survival was evaluated through the relationship between the female midgut gland and ovarian quality and the larvae. The results obtained indicate that the energy content of L. santolla females is highly variable according to the year, season, and month, and between primiparous and multiparous females. We hypothesized that this variability in the energetic quality is explained by differences in the female's diet: food availability or food quality during ovary maturation. Most of the energy reserves of the ovary are transferred to the offspring. The only female parent-brood connection is through the energy reserves transferred to the eggs 10 months earlier, and we hypothesize that this depends on the previous environmental and female feeding conditions during ovarian development. Moreover, zoea I larvae showed high variability in survival, not influenced by their own energy reserves or by the reserves of the eggs or the ovary. The physiological variables here analyzed did not allow identifying better females that can produce larvae with higher survival.

Quality Characteristics of Black Soldier Flies Produced by Different Substrates

Black soldier fly (BSF; Hermetia illucens) has a high capacity for amino acids and fatty acid accumulation. This study was conducted to assess the effectiveness of tofu by-products, food waste, and vegetables for BSF growth and conversion efficiency. BSFs under tofu by-product treatments showed the greatest weight at d 12 and the harvest period. Moreover, BSF larval weight was greater in the food waste treatment compared with the vegetable treatment at d 12 and harvest. The larva yield result was greater in the vegetable treatment compared with the tofu by-product. The bioconversion rate was higher in the tofu by-product treatment compared with the food waste and vegetable treatments. The protein conversion rate and lipid conversion rate were the highest in the vegetable treatment. The protein yield and lipid yield were greatest in the tofu by-product treatment. The lauric acid content was increased in BSFs fed tofu by-products compared with the food waste treatment. The concentration of C16:1 was the highest in the tofu by-product treatment. The content of oleic acid and α-linolenic acid was higher in BSFs fed tofu by-products compared with the vegetable treatment. In conclusion, the tofu by-products show benefits for larval growth and nutrient accumulation, which can improve larval quality for livestock feed ingredients.

Inclusion of live mud polychaete (Marphysa iloiloensis) in the feeding regime improved the hatchery performance of domesticated Indian white shrimp (Penaeus indicus)

A 60-day feeding trial was conducted to evaluate the effect of inclusion frequency of hatchery-bred mud polychaete, Marphysa iloiloensis, as part of a combination diet on the reproductive performance and larval quality of domesticated P. indicus. The two treatments were fed with a combination of formulated diet, mussel, squid, and mud polychaete with once (T1) and twice (T2) a week inclusion in the feeding regime. In contrast, the control group was fed only a diet combination of mussels, squid, and formulated diet. There was no significant difference in the mean survival of male and female broodstock in all treatments upon termination (p>0.05). Shrimp in T1 and T2 performed significantly better than the control in terms of spawning frequency (p=0.037) and fertilization rate (p=0.001), while T1 had significantly high re-maturation (p=0.038). The survival, spawning, hatching rate, egg, and nauplii production did not differ significantly among treatments (p>0.05). Also, no difference in the gonado-somatic index (GSI), hepato-somatic index (HSI), spermatophore weight, and sperm count was found among diet treatments (p>0.05). Post larvae from T1 and T2 performed significantly better than the control after 25 (p=0.008) and 30 (p=0.009) minutes of abrupt exposure to freshwater, respectively. Therefore, even once weekly, mud polychaete inclusion is sufficient to improve the reproductive performance of P. indicus broodstock and produce offspring with boosted resistance in the hatchery.