The effect of weaning diet type on grey mullet (Mugil cephalus) juvenile performance during the trophic shift from carnivory to omnivory

In captive grey mullet (Mugil cephalus) juveniles, the weaning stage overlaps the period where there are changes in the ontogeny of digestive enzymes as the fry transit from carnivory to omnivory. The aim of this study was to evaluate growth, survival, weight distribution and the activity of pancreatic and brush border digestive enzymes when fry are fed a carnivorous, herbivorous or omnivorous weaning diet. Fifteen 17-L aquaria in a flow through system with 40‰, UV treated, temperature (24.5 ± 0.5 °C) controlled seawater were stocked with eighty-five 23 dph grey mullet larvae per aquarium. This allowed the testing of three weaning dietary treatments, differing in their protein and carbohydrate content, in 5 replicate aquaria per treatment from 24 to 53 dph. Diet 1 was the dried macroalgal species Ulva lactuca and was designated as a low protein: high carbohydrate herbivorous diet. Diet 2 was a commercial microencapsulated starter feed designated as a high protein: low carbohydrate carnivorous diet. Diet 3 was a 1:1 ww mixture of diets 1 and diet 2 representing an omnivorous feeding regime. The average final weight of the omnivorous feeding fish was significantly (P .05). The activity levels of brush border alkaline phosphatase and intracellular leucine alanine peptidase were similar in grey mullet fry fed the carnivorous and omnivorous diets, but were higher than those in fish fed the herbivorous diet (P < .05). The intestinal maturation index exhibited the highest and lowest values in mullet fry fed the carnivorous and herbivorous diets, respectively, whereas those from the omnivorous group showed intermediate values (P = .03). This study broadly suggests that aquaculture feeds for juvenile grey mullet should be designed for omnivorous feeding habits.info:eu-repo/semantics/acceptedVersio

The key neuroendocrine regulators of the onset of puberty in the Atlantic bluefin tuna (Thunnus thynnus)

Recently, significant progress on spawning induction in captive bluefin tuna (BFT, Thunnus thynnus), has been achieved providing the basis for the species' domestication. To further promote the development of a self- sustained BFT aquaculture, we investigated first sexual maturity in BFT reared from an immature stage in captivity. Accordingly, our major objectives were to evaluate: (i) maturational status of the brain-pituitary-gonadal (BPG) axis, and (ii) responsiveness of the BPG to exogenous hormones. Special emphasis was given to characterize the gonadotropins follicle stimulating hormone (FSH) and luteinizing hormone (LH) that act as central regulators of gonadal development and gamete maturation. The growth parameters recorded for the captive BFT juveniles are consistent with the length-weight relationship established for wild Mediterranean BFT stocks. The histological analyses of the gonads indicate advanced sexual maturation in BFT males compared to females, yet it is not yet clear whether this phenomenon typifies wild stocks or is induced due to the culture conditions. The hormone measurements show expression and accumulation of both gonadotropins in the pituitaries of immature and mature BFT. The pituitary LH content increased concomitantly with the age of the fish, exhibiting sex dimorphic patterns (i.e. 3-fold higher levels in females) in adult but not in juvenile BFT. The pituitary FSH levels, however, were elevated in 2Y immature males and in fully mature adults. Comparable to mammals, the intra-pituitary FSH/LH ratio was found to be higher (>1) in sexually immature than in maturing or pubertal BFT. Nevertheless, in the 3Y BFT females, which were all immature, the onset of puberty appears to require some other prerequisites, such as a rise in the LH storage above a minimal threshold. Our in vitro trials further demonstrated the capacity of rFSH and to a lesser extent that of rLH to stimulate cell proliferation in the immature ovarian and testicular fragments. Both rFSH and rLH have failed to stimulate steroidogenesis, yet pre-treatment with KiSS containing EVAc implants appeared to potentiate FSH-stimulated steroidogenesis in the immature testes. On the other hand, the expression levels of both the GtH-R and IGF I genes in the testicular fragments, derived from BFT juveniles and further exposed to the rLH treatment, showed dose-dependent pattern. Future studies testing the effects of captivity and hormone-based treatments on precocious maturity at relatively small body size are expected to facilitate the handling in confined environments, and to greatly improve the cost-efficiency of BFT farming.Postprin

Genetic Stock Identification of the Flathead Grey Mullet (Mugil cephalus) in Lake Tiberias Based on ParentOffspring Relationship

The Lake Tiberias (Sea of Galilee) grey mullet (Mugil cephalus) population originates from translocated wild-caught, or hatchery-reared, fish. The aim of the study was to identify the taxonomic status and stock origin of a sample of 32 mullet individuals caught in the Sea of Galilee, based on the mitochondrial Cytochrome Oxidase I (COI) sequence analysis and nuclear microsatellite markers. A total of 13 microsatellite markers were selected from nine different linkage groups with a number of alleles ranging from 5-23, with an average of 9.85 alleles/locus. By using COI sequences and the Barcode of Life Data (BOLD) identification system, seven individuals were taxonomically classified as Thinlip grey mullet (Liza ramda) and the remaining 25 as M. cephalus. Of 663 nucleotides, 122 (18.4%) differed between the COI sequences of the two distinct mullet species. A preliminary parentage analysis of the hatchery-reared stocking batch, based on 13 markers, assigned them as progeny of three mating pairs. These three mating pairs only were tested as potential parents of the 25 M. cephalus individuals captured from the lake. Marker-based comparison showed that a parent-offspring relationship was rejected for 17 M. cephalus individuals by at least four genetic markers. Eight of 25 M. cephalus individuals (32%), were identified as progeny of the three parental pairs with the overall probability of 3.88 x 10-7 for correspondence by chance to any of these three parental pairs. The 13 markers used has high statistical power to reject a putative parent-offspring relationship obtained by chance thus resulting in familial identification. This approach represents an accurate method of genetic stock identification and should also be applicable to populations of other species. The results confirm that the mullet hatcheryreared fingerlings survived in the Sea of Galilee when released alongside wildcaught fingerlings

The effect of weaning diet type on grey mullet (Mugil cephalus) juvenile performance during the trophic shift from carnivory to omnivory

In captive grey mullet (Mugil cephalus) juveniles, the weaning stage overlaps the period where there are changes in the ontogeny of digestive enzymes as the fry transit from carnivory to omnivory. The aim of this study was to evaluate growth, survival, weight distribution and the activity of pancreatic and brush border digestive enzymes when fry are fed a carnivorous, herbivorous or omnivorous weaning diet

Assessment of the effects of captivity on first sexual maturity of wild-caught and hatchery produced mullet (Mugil cephalus)

INTRODUCTION In fish, puberty is susceptible to multiple, interacting environmental cues. As a result, farming conditions may have varying influences on the age of first sexual maturity, causing precocious puberty in some species or delayed and even completely blocked puberty in others. Therefore, this study aimed at characterizing pubertal development in grey mullet (Mugil cephalus), an important fish candidate for domestication and aquaculture production. MATERIALS AND METHODS Two stocks of grey mullet were compared: wild-caught (WC; Apulia, Italy) and hatchery produced (HP; National Center for Mariculture, Eilat, Israel) grey mullet, both subjected to captive conditions consist of ambient seawater salinity (40 ppt; Gulf of Eilat, Red Sea) and photo-thermal regime. The age of the fish was estimated based on their scales. Growth performance and gonadal development were monitored in 2- and 3- year old fish (2y and 3y). RESULTS AND CONCLUSIONS Our results revealed that all 2y fish had immature gonads. The majority of 2y females exhibited late perinucleolar oocytes as the most advanced oocyte stage. Yet, HP females had significantly larger oocytes than WC specimens. Testis from an immature age 2y HP specimen showed small seminiferous lobules. Only spermatogonia, along with somatic cells were visible. Gonad section from a 2y HP intersex showed the presence of all stages of spermatogenesis. Scattered perinucleolar stage oocytes were visible, indicating that both WC and HP, had immature testes. Following 3y age category revealed that WC females were larger than cognate males. Although not significant, the HP mullets appear to exhibit a similar trend. Gonadosomatic index (GSI) values in 3y HP females and males were significantly higher than those of the WC of the same age. Undifferentiated gonads were found in 20% of WC fish compared to 5% in the HP fish. While only 33% had reached vitellogenic oocytes level, 54% of the HP females reached this level of development. Males exhibited a significant difference in development: 100% of the WC males showed first stages of spermatogenesis, while 67% of the male population in the HP group produced mature sperm. HP females, sampled during natural spawning season, reached maturity and ovulation. Their GSI exceeded 15%. Interestingly, approximately 50% of the HP females’ GSI exhibited markedly developed gonads with GSI values ranging between 10-20% while all the others were lagging far behind, having GSI values between 0.2 to 0.3 %. GSI values in WC females were also divided into 2 sub groups and found to be significantly lower than those of the hatchery produced females (1-6% and &lt;1% respectively). Males also exhibited two GSI groups pattern, higher GSI values were exhibited by HP fish (1.5%-2.75% and &lt;1% compared to 0.2-0.5% and &lt;0.15%). Suggesting hierarchy plays a major role affecting gonadal development in mullets. In summary, the 3 yr old HP mullet females and males exhibited enhanced gonadal maturation as compared to that of the WC captive-reared fish, probably as a result of domestication. The project received funding from the European Union 7FP (GA 603121, DIVERSIFY)

Gonadotropin expression, pituitary and plasma levels in the reproductive cycle of wild and captive-reared greater amberjack (Seriola dumerili)

We compared the endocrine status of the pituitary-gonad axis of wild and captive-reared greater amberjack (Seriola dumerili) during the reproductive cycle (April – July), reporting on the expression and release of the two gonadotropins for the first time in the Mediterranean Sea. Ovaries from wild females were characterized histologically as DEVELOPING in early May and SPAWNING capable in late May-July, the latter having a 3 to 4-fold higher gonadosomatic index (GSI). SPAWNING capable wild females exhibited an increase in pituitary follicle stimulating hormone (Fsh) content, plasma testosterone (T) and 17,20β-dihydroxy-4-pregnen-3-one (17,20β-P), while almost a 10-fold increase was observed in pituitary luteinizing hormone (Lh) content. An increasing trend of plasma 17β-estradiol (E2) was also recorded between the two reproductive stages in wild females. Captive-reared females sampled during the reproductive cycle exhibited two additional reproductive categories, with REGRESSED females having extensive follicular atresia and fish in the REGENERATING stage having only primary oocytes in their ovaries. Pituitary content of Fsh and Lh, fshb and lhb expression and plasma levels of Fsh and Lh remained unchanged among the four reproductive stages in captive females, in contrast with plasma E2 and T that decreased in the REGENERATING stage, and 17,20β-P which increased after the DEVELOPING stage. In general, no significant hormonal differences were recorded between captive-reared and wild DEVELOPING females, in contrast to SPAWNING capable females, where pituitary Lh content, plasma Fsh and T were found to be lower in females in captivity. Overall, the captive females lagged behind in reproductive development compared to the wild ones and this was perhaps related to the multiple handling of the sea cages where all the sampled fish were maintained. Between wild males in the DEVELOPING and SPAWNING capable stages, pituitary Lh content, plasma T and 17,20β-P, and GSI exhibited 3 to 4-fold increases, while an increasing trend of pituitary Fsh content, lhb expression levels and plasma 11-ketotestosterone (11-KT) was also observed, and an opposite trend was observed in plasma Lh. Captive males were allocated to one more category, with REGRESSED individuals having no spermatogenic capacity. During the SPAWNING capable phase, almost all measured parameters were lower in captive males compared to wild ones. More importantly, captive males showed significant differences from their wild counterparts throughout the reproductive season, starting already from the DEVELOPING stage. Therefore, it appears that captivity already exerted negative effects in males prior to the onset of the study and the multiple handling of the cage where sampled fish were reared. Overall, the present study demonstrated that female greater amberjack do undergo full vitellogenesis in captivity, albeit with some dysfunctions that may be related to the husbandry of the experiment, while males, on the other hand, may be more seriously affected by captivity even before the onset of the study

First summary of studies carried out on the grey mullet (Mugil cephalus) in the EU project: diversify

The deliverable (D7.1) of WP7 “Establishment of a Computer Assisted Sperm Analysis (CASA) for the evaluation of grey mullet sperm” has been submitted. Preliminary analyses show that mullet sperm motility pattern is not similar to that of sea bass (Dicentrarchus labrax) or rainbow trout (Oncorhynchus mykiss) in terms of velocity as well as motility duration (ca 1 min; Fig. 1a). Our preliminary results, indicate that the major bio-technical settings, dilution of sperm, quality of activation and mastering of video recordings have been determined and can now be applied to experimental protocols. Since fish sperm quality is usually assessed through subjective observation, in terms of motility classes and duration, statistical analysis becomes less precise. The current results will help improve assessment of the effect of different treatments on reproductive performances of mullet males

Intra-pituitary relationship of follicle stimulating hormone and luteinizing hormone during pubertal development in Atlantic bluefin tuna (Thunnus thynnus)

As part of the endeavor aiming at the domestication of Atlantic bluefin tuna (BFT; Thunnus thynnus), first sexual maturity in captivity was studied by documenting its occurrence and by characterizing the key hormones of the reproductive axis: follicle stimulating hormone (FSH) and luteinizing hormone (LH). The full length sequence encoding for the related hormone β-subunits, bftFSHβ and bftLHβ, were determined, revealing two bftFSHβ mRNA variants, differing in their 5' untranslated region.A quantitative immuno-dot-blot assay to measure pituitary FSH content in BFT was developed and validated enabling, for the first time in this species, data sets for both LH and FSH to be compared. The expression and accumulation patterns of LH in the pituitary showed a steady increase of this hormone, concomitant with fish age, reaching higher levels in adult females compared to males of the same age class. Conversely, the pituitary FSH levels were elevated only in 2Y and adult fish. The pituitary FSH to LH ratio was consistently higher (&gt;1) in immature than in maturing or pubertal fish, resembling the situation in mammals. Nevertheless, the results suggest that a rise in the LH storage level above a minimum threshold may be an indicator of the onset of puberty in BFT females. The higher pituitary LH levels in adult females over males may further support this notion.In contrast three year-old (3Y) males were pubertal while cognate females were still immature. However, it is not yet clear whether the advanced puberty in the 3Y males was a general feature typifying wild BFT populations or was induced by the culture conditions. Future studies testing the effects of captivity and hormonal treatments on precocious maturity may allow for improved handling of this species in a controlled environment which would lead to more cost-efficient farming