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The nutritional status of broodstock profoundly affects their reproductive performance and offspring survival. Studies on lipids and essential fatty acids in broodstock diets highlight their importance in cell structure, fecundity, fertilization, egg and larval quality, and providing metabolic energy for reproduction. Long-chain polyunsaturated fatty acids (Lc-PUFA) like DHA (22:6 n-3) and EPA (20:5 n-3) are vital for egg and larval development, while arachidonic acid (ARA) produces eicosanoids essential for reproduction. The fatty acid requirements vary by habitat; freshwater fish typically lack ∆12 and ∆15 desaturase enzymes to convert oleic acid into vital polyunsaturated fatty acids like linoleic and linolenic acids but can synthesize linoleic (18:2 n-6) and linolenic (18:3 n-3) into Lc-PUFAs such as EPA, DHA, and ARA through desaturation and elongation, whereas marine teleost cannot. Hence, broodstock feed fatty acid composition must be tailored by incorporating ingredients with a specific fatty acid composition to enhance reproductive performance. This review provides updated information on fatty acid supplementation in broodstock diets to improve reproductive outcomes in commercially important finfish, offering valuable insights for researchers, academicians, hatchery owners, and fish farmers to produce better-quality seeds.

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This study evaluates the effect of different diets supplemented with vitamin solutions on Pangasius catfish broodstock reproduction, growth performances, and the survival rates of fish larvae and fingerling. The growth and reproductive performances of breeders fed with different test diets showed significant differences among the six tested diets (p < 0.05). The highest final body weight (FBW), weight gain (WG), daily weight gain (DWG), specific growth rate (SGR) of broodstock, and survival rate of Pangasius fingerlings were found in Treatment 5, which contained 0.6% H-OVN mixed with 12.6% algal oil, and Treatment 3, which contained 0.6% vitamin premix H-OVN. The average gonadosomatic index (GSI), relative fecundity index (RFI), fertilized eggs, hatching rates of eggs, and survival rate of fingerlings was 9.1 ± 2.8 (6.7-12.8%), 133,224 ± 39,090 (104,267-199,512 eggs/kg), 77.9 ± 22.2 (62.2-93.6%), and 45.3 ± 17.4 (22.0-66.3%), respectively. The findings of this study showed that the diet containing 35% CP contents supplemented with 0.6% vitamin premix H-OVN mixed with algal oils showed the highest results in terms of growth, reproductive performance indices, and survival rates of Pangasius catfish fingerlings.

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Pangasius catfish, a significant player in the global whitefish market, encounters challenges in aquaculture production sustainability. Quality broodstock maintenance and seed production are impeded by growth, maturation, and fecundity issues. This review investigates the efficacy of strategic nutrient composition and molecular strategies in enhancing broodstock conditions and reproductive performance across various fish species. A notable knowledge gap for Pangasius catfish hampers aquaculture progress. The review assesses nutrient manipulation's impact on reproductive physiology, emphasizing pangasius broodstock. A systematic review analysis following PRISMA guidelines was conducted to identify research trends and hotspots quantitatively, revealing a focus on P. bocourti and fertilization techniques. Addressing this gap, the review offers insights into dietary nutrients manipulation and genetic tool utilization for improved seed production, contributing to pangasius catfish aquaculture sustainability.

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The use of sterile recipients is crucial for efficiently producing donor-derived offspring through surrogate broodstock technology for practical aquaculture applications. Although knockout (KO) of the dead end (dnd) gene has been used in previous studies as a sterilization method, it has not been reported in marine fish. In this study, nibe croaker was utilized as a model for marine teleosts that produce small pelagic eggs, and the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9) system was utilized to produce dnd KO fish. The F1 generation, which carried a nonsense mutation in the dnd gene, was produced by mating founder individuals with wild-type counterparts. Subsequently, the F2 generation was produced by mating the resulting males and females. Among the F2 generations, 24.0% consisted of homozygous KO individuals. Histological analysis revealed that primordial germ cells (PGCs) were present in homozygous KO individuals at 10 days post-hatching (dph), similar to wild-type individuals. However, by 20 dph, PGCs were absent in KO individuals. Furthermore, no germ cells were observed in the gonads of both sexes of homozygous KO individuals at 6 months old, which is the typical maturity age for wild-type individuals of both sexes. In addition, when cryopreserved donor nibe croaker testicular cells were transplanted, only donor-derived offspring were successfully obtained through the spontaneous mating of homozygous KO recipient parents. Results indicate that dnd KO nibe croaker lacks germ cells and can serve as promising recipients, producing only donor-derived gametes as surrogate broodstock.

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Atlantic salmon (Salmo salar) broodstock recruits are normally fed a specialized diet with a higher content of essential nutrients for a limited time period prior to fasting and transfer to freshwater. Typically, this period lasts for about six months, but may vary among producers. Reduced use of marine ingredients in commercial salmon diets during the last decades has affected the content of essential nutrients, such as n-3 long chained polyunsaturated fatty acids (LC-PUFA), minerals and vitamins. Furthermore, to minimize the risk of losses and implement new breeding achievements faster, breeding companies have shortened the production cycle of broodstock from 4 to 3 years, which may affect the number of fish that are large enough to mature. In the present study, we have extended the broodstock feeding period from 6 to 15 months prior to the freshwater transfer giving a higher content of n-3 LC-PUFA (higher inclusion of marine oils) from February to December (Phase 1), and thereafter a diet with a higher energy content to ensure growth towards the spring and maturation (Phase 2). Four sea cages with approximately 80.000 salmon postsmolt, two sea cages with males and two with females, were given a control diet and an experimental diet. Samples were taken in Phase 1 at start (1.7 kg), mid (3.4 kg) and end Phase 1/start of Phase 2 (8.3 kg), and end of Phase 2 (13.4 kg). The fish were thereafter fasted, and selected fish transferred to landbased freshwater tanks where light and temperature were used to manipulate the spawning time of the fish in two groups (early or late). Due to disease in the facility, measures of egg quality and hatching were only obtained from the early group. During the trial and spawning period, biometrical measurements were recorded, and samples of liver, gonad, fillet and red blood cells (RBC) were collected for fatty acid composition and blood plasma for analysis of lipid and health-related parameters. Samples were also collected for gonadal transcriptomic analysis by microarray and qPCR (end Phase 2) and plasma steroids (end Phase 2, mid maturation and spawning). Males fed the test diet had a larger body size compared to the control group at the end of Phase 2, while no differences were observed between dietary groups for the females. Total mortality in the trial was lower in the test group compared to the control, losses were caused mainly by sea lice treatments, loser fish or cardiomyopathy syndrome (CMS). The dietary LC-PUFA levels in the test diet were reflected in the tissues particularly during Phase 1, but only different in the fillet samples and eggs at the end of Phase 2 and at spawning. Plasma sex steroids content increased at mid maturation and showed lower levels of androgens and estrogens in females fed the test diet compared to the control. At the end of Phase 2, transcriptional analysis showed upregulation of steroidogenic enzymes, although not reflected in changes in plasma steroids in Phase 2, indicating changes to come during maturation. The differences in LC-PUFA content in tissues and plasma steroids did not appear to affect fecundity, sperm quality, egg survival or hatching rate, but the test group had larger eggs compared to the control in the early spawner-group. Prolonged feeding of n-3 LC-PUFA to pre-puberty Atlantic salmon broodstock appears to be important for higher survival in challenging sea cage environments and has an effect on sex steroid production that, together with high energy diet during early maturation, cause the test group to produce larger eggs.

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Background: Mystacoleucus padangensis living in Lake Singkarak, Indonesia, has high potential market demand but is threatened by overfishing and has not been successfully cultured. This study describes the first broodstock development, induced breeding, and larval rearing of M. padangensis. Methods: A total of 1,000 female and 1,000 male broodfish were collected from the wild and reared in two concrete ponds (128 m 2) at the Centre for Biodiversity Conservation, P.T. Semen Padang, Indonesia. The broodfish were fed commercial feed to satiation at 09:00 and 17:00 h. The females (average weight 7.56 ± 0.85 g) and males (4.86 ± 1.20 g) were selected at a ratio of 1:4 (female:male), and gonad maturation was induced with a single dose of GnRH analogue (Ovaprim) of 0.1 ml/fish. At 16 h after hormone injection, eggs were collected individually into a plastic vessel. Spermatozoa were collected with sterile syringes. Eggs were fertilized using the "dry" method, and 0.5 ml samples (equal to 100 eggs) were taken. The eggs were incubated in a plastic strainer with a water volume of 1.57 litres and placed in a tarpaulin pond with a volume of 150.72 litres. Results: The overall hatching rate was 78.93 ± 4.13%. The newly hatched larvae were 3900.81 µm long, with a yolk sac of 82881.480 µm 2. The mouth opened at 72 days post hatching (DPH) with a gape measuring approximately 61.880 µm. The protocol of larval feeding started with artificial feed, followed by Artemia nauplii up to 30 DPH. Weaning of larvae started at 4 DPH. Larvae started metamorphosis by 15 DPH and ended by 22 DPH when the larvae reached 7430.27 µm. Larval rearing resulted in an average survival rate of 28.4 ± 3.04%. Conclusions: Its successful spawning induction and high larval hatching and survival rates make M. padangensis an excellent aquaculture candidate.

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The unprecedented loss of global biodiversity is linked to multiple anthropogenic stressors. New conservation technologies are urgently needed to mitigate this loss. The rights, knowledge and perspectives of Indigenous peoples in biodiversity conservation-including the development and application of new technologies-are increasingly recognised. Advances in germplasm cryopreservation and germ cell transplantation (termed 'broodstock surrogacy') techniques offer exciting tools to preserve biodiversity, but their application has been underappreciated. Here, we use teleost fishes as an exemplar group to outline (1) the power of these techniques to preserve genome-wide genetic diversity, (2) the need to apply a conservation genomic lens when selecting individuals for germplasm cryobanking and broodstock surrogacy and (3) the value of considering the cultural significance of these genomic resources. We conclude by discussing the opportunities and challenges of these techniques for conserving biodiversity in threatened teleost fish and beyond.

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Long-whiskered catfish Sperata aor is a freshwater catfish known for its supreme flesh quality and fast growth, whose captive-reared broodstock denotes a difficult challenge for aquaculture. The reproductive dysfunctions in long-whiskered catfish raised in tank conditions were observed by comparing tissue biochemical composition and ovarian histology of wild female broodstock. Sixty (60) female broodstocks were used in the current study, consisting of 30 reared at sandy-muddy soil tank bottoms in captive conditions and 30 wild individuals collected from the haor basin during the breeding season. The fish reproductive state was investigated using the biometric and reproductive parameters, biochemical composition and levels of amino acids in the different tissues, and histological analysis of ovarian development. Results revealed that the biometrical parameters of wild and captive female broodstocks exhibited no remarkable difference (p > 0.05). Nevertheless, the wild fish had remarkably higher (p < 0.05) GSI (8.73%), oocyte weight (0.45 mg/egg), and ripeness (27.08%) in comparison with captive-reared broodstock. The total length and body weight, body weight and ovary weight, ovipositor diameter and ovary weight, and GSI and HSI displayed a positive relationship with R2 = 1, R2 = 1, R2 = 0.993, and R2 = 0.973, respectively, for wild broodstock, while R2 = 0.994, R2 = 0.806, R2 = 0.804, and R2 = 0.896, respectively, for captive broodstock. Additionally, the proximate composition in oocytes and liver tissues in both broodstocks did not differ significantly (p > 0.05). However, two essential amino acids (EAA), i.e., lysine and phenylalanine, and two non-essential amino acids, i.e., glutamic acid and glycine, were highly significant differences (p < 0.05) in the oocytes and liver of wild broodstock compared to the captive-reared broodstock. On the other hand, the EAA, e.g., isoleucine, threonine, leucine, and arginine, were highly dominated in both wild and captive female brood oocytes and liver. The ovarian histological slides from each fish group showed three oocytes developmental stages that indicated the asynchronous-reproductive ovarian oocytes of this fish. This study may be useful to fully understand the factors affecting the spawning and reproduction of S. aor broodstock, crucial for management in captive conditions as well as conservation and protection for sustainable aquaculture management of S. aor.

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The current status of the barbels inhabiting the Kazakhstan section of the Syrdarya River needs special study, as has been noted since the second half of the 20th century. Stocks of commercial fish valuable in the past as barbels in the Aral-Syrdarya basin are now catastrophically reduced as a result of anthropogenic impact on the ichthyofauna and natural environment of the Aral Sea region. The study of the condition; abundance and distribution range is necessary to determine measures for their restoration in natural habitats and their breeding in fish farms. Research on the biotechnology of barbel breeding with subsequent acclimatization and reacclimatization of these fish species will not only improve the composition of the ichthyofauna of the Aral-Syrdarya basin, but also preserve the genetic potential of natural populations. At present, the only way to restore the population of the Aral barbel is stocking hatchery reared juveniles in natural environments. Under the current situation, a way forward is seen in the creation of domesticated replacement-broodstocks of barbels. As a result of the influence of anthropogenic factors, the populations of this species have come to almost complete degradation and depletion and require the development and adoption of urgent measures for their conservation and restoration through reintroduction, which is a priority and urgent task for the fisheries of the republic.

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The present experiment aims to study the effects of dietary multi-strain probiotics on semen quality, seminal plasma compositions, and fertilization ability of male rainbow trout. For this purpose, a total of 48 broodstocks with an average initial weight of 1366.1 ± 33.8 g were divided into 4 groups and 3 replicates. Fish were fed with diets containing 0 (control), 1 × 109 (P1), 2 × 109 (P2), and 4 × 109 (P3) CFU probiotic kg-1 diet for 12 weeks. Results showed that dietary supplementation of probiotics significantly increased plasma testosterone level, motility time of spermatozoa, sperm density, and spermatocrit value in P2 and P3 treatments and Na+ level in P2 treatment compared to the control group (P < 0.05). The activities of aspartate aminotransferase and lactate dehydrogenase had significantly decreased in the P2 treatment compared to the control group (P < 0.05). No considerable variations were observed between control fish and treatment groups (P > 0.05) in semen biochemical parameters, percentage of motile spermatozoa, osmolality, and pH of seminal plasma. Based on the results, the highest fertilization rate (97.2 ± 0.9%) and eyed egg survival (95.7 ± 1.6%) were observed in the P2 treatment, and those values showed remarkable differences with the control group (P < 0.05). The results indicated that multi-strain probiotics have potential efficacy on semen quality and fertilization ability of rainbow trout broodstock spermatozoa.

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The longest Baltic River, the Vistula, historically hosted numerous sea trout populations. However, dam construction in the twentieth century drastically reduced the spawning migration rate. Reduced natural reproduction has resulted in a population collapse and, consequentially, a substantial reduction in catches. In response, like other Baltic countries, Poland has initiated an intensive stocking program, mainly involving smolt. Initially, stocking was conducted primarily with offspring of sea trout caught during spawning migration. Currently, due to difficulties in obtaining fish, most stocking involves fish from breeding stocks. Therefore, determining the proportion of fish derived from stocking has become an important issue. Experiments based on traditional tagging did not provide sufficient material for analysis; hence, we decided to use genetic methods based on analysis of relatedness. In this study, we performed parentage-based tagging and an analysis of the origins of parent animals used for artificial spawning in 2013, and offspring returning to the Vistula in subsequent years. We based the analysis on three different algorithms and compared the results, showing that the presented methods were effective for estimating mass stocking success. The study also indicated that a certain level of natural reproduction in the Vistula continues to occur. The proportion of sea trout from spawning in 2013 in Vistula sea trout catches from 2017 to 2018 was approximately 30%.

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Acquiring immunocompetence is essential in the development of fish embryos, as they are exposed to environmental pathogens even before they are fertilized. Despite the importance of the antimicrobial function as the first line of defense against foreign microorganisms, little knowledge is available about its role in larval development. In vertebrates, transgenerational immune priming influences the acquisition of immunocompetence of specimens, regulating the selective allocation of nongenetic resources to their progeny and modulating their development. In this work, we primed teleost European sea bass broodstock females with a viral protein expression vector in order to evaluate the innate immunity development of their offspring. Several antimicrobial functions, the pattern of expression of gene coding for different antimicrobial peptides (AMPs), and their protein levels, were evaluated in eggs and larvae during development. Our data determined the presence of antimicrobial proteins of maternal origin in eggs, and that female vaccination increases antimicrobial activities and the transcription and synthesis of AMPs during larval development.

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European grayling (Thymallus thymallus) is an endangered salmonid fish species and its populations have decreased considerably in European waters in recent decades. One of the conservation measures applied to grayling is stocking with hatchery-reared fish. To advance the knowledge of grayling reproduction in the hatchery, husbandry data were systematically collected in the period from 2006 to 2015 during grayling spawning seasons and analyzed to quantify grayling female reproductive parameters such as fecundity, egg size, and embryo survival to the eyed egg stage. Data was collected for first time spawners (3 years old) and repeat spawners reared in the hatchery. Statistically significant differences were found for the number of eggs between first time spawners, producing on average 3600 eggs, and repeat spawners, producing on average 8000 eggs. Absolute fecundity was positively correlated to grayling female total length and weight for both first time and repeat spawners. However, relative fecundity did not correlate with these parameters. A positive correlation was observed between female total length and mean egg weight for repeat spawners, while the correlation was negative for first time spawners. Heavier eggs tended to have a higher rate of survival until the eyed egg stage, though the correlation was stronger for first time spawners than for repeat spawners. Due to the higher reproductive success of repeat spawners, it could be concluded that larger and older grayling females should be included in hatchery reproduction.

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The meagre Argyrosomus regius (Asso, 1801) is a promising aquaculture species that shows reproductive dysfunctions when reared in tanks. The aim of this study was to assess the capacity of meagre, reared in cages under routine farming conditions, to mature gonads and reproduce spontaneously. Meagre adults, reared in a fish farm located in the Gulf of Taranto (Italy), were sampled from March to July 2021. The gonadosomatic index and sex steroid plasma concentrations increased from March-April to June, and then decreased in July. In March-April, most of the females showed perinucleolar or cortical alveoli oocytes as the most advanced stages in the ovaries, and most of the males had testes at early spermatogenesis stage. In June, most of the sampled females had oocytes at late vitellogenesis or early post-vitellogenesis stages, and males had seminiferous tubules filled with spermatozoa. In July, most of the females had signs of previous spawning, and males showed scarce amounts of luminal spermatozoa. The present study demonstrated the capacity of meagre, reared in sea cages under commercial conditions, to carry out gametogenesis and spontaneously release gametes. Meagre reproduction, under routine farming conditions, may represent an opportunity for the expansion of meagre aquaculture production.

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In this study, a practical protocol for artificial reproduction was developed for the rudd, Scardinius erythrophthalmus, to help to produce this fish for aquaculture industries and also for restocking programs. For this purpose, the efficiency of different hormonal agents and their combinations for spawning induction of female breeders of rudd was evaluated. Ovulations were stimulated using different treatments as: two injections of Ovaprim (Ova), two injections of Ovopel (Ovo), a priming dose of Ovopel with a resolving dose of Ovaprim (Comb1), a priming dose of Ovaprim with a resolving dose of Ovopel (Comb2), and two injections of sterile 0.9% NaCl solution as a control group. During the study, ovulation success, egg developmental competence as well as the performance of the freshly hatched larvae were recorded. There was no ovulation in females from the control group. In hormone-treated groups, 84-100% of fish ovulated, but no differences were observed in ovulation success (P > 0.05). The shortest latency period in treated fish was recorded in the Ovo group (431.6 degree-hour, P < 0.05). The best results in terms of working fecundity (number of obtained eggs per fish), relative fecundity, fertilization success, hatching rate, and embryo survival up to the eyed stage were achieved in the Ova and Comb1 groups (P < 0.05). Moreover, better survival rates up to the absorbing yolk sac stage and the lowest abnormality of larvae were observed in Ova and Comb1 groups (P < 0.05). There was no significant difference in the incubation time among different groups (P > 0.05). The obtained results indicate that two injections of Ovaprim and/or a priming dose of Ovopel with a resolving dose of Ovaprim are suitable for the artificial reproduction of rudd female breeders. Due to the proper effects and ease of use, two injections of Ovaprim could be recommended for the production of this species for restocking or aquaculture programs.

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A study was carried out to determine the maturation period, quality and quantity of sperms production in Banana shrimp male broodstock, Penaeus merguiensis fed different natural diets. The three different natural diets namely; squid, fish and shrimp flesh used in this study were obtained from known sources and fed to the tested shrimp in triplicate groups. Based on the results obtained, squid seem to be the most effective natural diet as it enhances the sperms maturation within 20 days. Feeding fish and shrimp flesh as diets for the Banana shrimp broodstock resulted in the observation of maturation in 22 and 24 days, respectively. Similarly, squid diet also recorded the highest (p = 0.002; p < 0.05) sperms count (58.6 to 74.5) as compared to fish diet (44.0 to 61.3) or shrimp diet (28.0 to 42.8). Also, feeding squid diet resulted in a higher percentage of live sperms with ranged between 97.75% to 98.80%. On the other hand, broodstocks fed fish and squid flesh was observed with ranges of between 96% to 97.86% and 92.54% to 96.06%, respectively. It was therefore concluded that squid diet was most effective to improve sperm quality, quantity and maturation period in male broodstock of P. merguiensis. Further study should be carried on the reproductive performance of broodstock fed with squid as diets and it effect on the performances of post larvae obtained.

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Mud crabs (genus Scylla) are gaining attention as potential aquaculture species due to their lucrative market price and high demand. One of the essential components of mud crab culture is broodstock quality. The optimisation of mud crab broodstock culture currently focused on its nutritional aspects and common rearing parameters, including stocking density and temperature. The use of sandy substrate to induce egg extrusion in female Scylla broodstock is common; however, its optimisation has never been conducted. This study investigated (1) the substrate association of two Scylla species (S. olivacea, S. paramamosain) during broodstock conditioning until egg extrusion when the choices of fine (66.99 ± 14.48 µm) and coarse (656.17 ± 38.13 µm) sands were given; (2) the female reproductive output when S. olivacea females were individually exposed to either fine sand, coarse sand, or no sand treatments. Females, spawners and non-spawners, of S. olivacea and S. paramamosain were associated with fine sand and none was observed to bury in the coarse sand tray. The occurrence of egg extrusion was not significantly different between species but moderately associated with the duration of visits (stayed in sand for 1 d, 2 d, ≥3 d). The final incubation period in the sand tray was more than 2 days in all cases, except for one female S. paramamosain (buried in the sand for 1 day prior to egg extrusion). When no choice was available, the highest percentage (58.3%) of females extruded eggs in fine sand treatment, followed by coarse sand treatment (33.3%), and no sand treatment (8.3%). Sand type influenced the weight of egg clutch, total egg number, fecundity, and clutch size. These results suggest that fine sand (<70 µm) substrate should be incorporated into Scylla broodstock rearing to maximise female reproductive output.

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Generally, successful testis transplantation has been considered to require immune suppression in the recipient to avoid rejection of the transplanted tissue. In the present study, we demonstrate in medaka that allogeneic adult testicular tissue will engraft in adult recipients immediately after partial castration without the use of immunosuppressive drugs. The allografted testes are retained in the recipient's body for at least 3 months and are able to produce viable sperm that yield offspring after natural mating. Some recipients showed a high frequency (over 60%) of offspring derived from spermatozoa produced by the transplanted testicular tissue. Histological analyses showed that allografted testicular tissues included both germ cells and somatic cells that had become established within an immunocompetent recipient testis. The relative simplicity of this testis transplantation approach will benefit investigations of the basic processes of reproductive immunology and will improve the technique of gonadal tissue transplantation.

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The wallago catfish (Wallago attu) is a new potential fish for aquaculture in Vietnam. Data related to the reproductive cycle of W. attu in captivity are, however, not available. To provide reliable indicators for oocyte maturation (OM) and the spawning season of the captive W. attu, this study investigated the temporal variation in hepatosomatic and gonadosomatic indices, oocyte diameter and color (greenish vs yellowish), germinal vesicle migration, and plasma concentrations of estradiol-17ß (E2) and vitellogenin (Vtg) in female broodstock in association with changes in light density, temperature and amount of rainfall during the reproductive cycle. The results of this study displayed a clear seasonality in all the investigated parameters. The highest concentration of E2 (2.6 ± 3.5 ng/mL) was found in April, followed by a peak of Vtg (543 ± 43 ng/mL) in June. Meanwhile, the largest mean oocyte diameter (1.70 ± 0.02 mm) was observed in June. The shortest distance between the germinal vesicle and the edge of the oocyte (0.20 ± 0.01 mm) was recorded in July. Correspondingly, the amount of rainfall increased remarkably in July from 43.9 mm to over 200 mm in August. Taken together, we conclude that OM and the onset of the spawning season of captive W. attu occur in July and August, respectively. The percentage of greenish oocytes increased significantly over sampling time points. The changes in the color of oocytes combined with oocyte diameter could, therefore, be considered as promising indicators to predict the OM and spawning season of captive W. attu.

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