Rh proteins and H+ transporters involved in ammonia excretion in Amur Ide (Leuciscus waleckii) under high alkali exposure.

High alkaline environment can lead to respiratory alkalosis and ammonia toxification to freshwater fish. However, the Amur ide (Leuciscus waleckii), which inhabits an extremely alkaline lake in China with titratable alkalinity up to 53.57 mM (pH 9.6) has developed special physiological and molecular mechanisms to adapt to such an environment. Nevertheless, how the Amur ide can maintain acid-base balance and perform ammonia detoxification effectively remains unclear. Therefore, this study was designed to study the ammonia excretion rate (Tamm), total nitrogen accumulation in blood and tissues, including identification, expression, and localization of ammonia-related transporters in gills of both the alkali and freshwater forms of the Amur ide. The results showed that the freshwater form Amur ide does not have a perfect ammonia excretion mechanism exposed to high-alkaline condition. Nevertheless, the alkali form of Amur ide was able to excrete ammonia better than freshwater from Amur ide, which was facilitated by the ionocytes transporters (Rhbg, Rhcg1, Na+/H+ exchanger 2 (NHE2), and V-type H+ ATPase (VHA)) in the gills. Converting ammonia into urea served as an ammonia detoxication strategy to reduced endogenous ammonia accumulation under high-alkaline environment.

Sweet orange peel waste as a feed additive in growth promoting and protective effect against Aeromonas hydrophila of juvenile bagrid catfish Mystus nemurus.

Sweet orange Citrus sinensis peel is a phytobiotic agricultural waste with bioactive compounds that have potential functional properties as a growth promoter and immune stimulator. This study aims to evaluate the dietary effects of sweet orange peel (SOP) as a feed additive on growth enhancement of juvenile bagrid catfish Mystus nemurus and their disease resistance ability against Aeromonas hydrophila infection. Four experimental diets were formulated to contain 0 (SOP0, control), 4 (SOP4), 8 (SOP8) and 12 g/kg (SOP12) SOP. After 90 d of the feeding experiment, improvement in weight gain, specific growth rate, feed conversion ratio, and protein efficiency ratio were observed in the fish fed with SOP4. While fish survival was not significantly affected, hepatosomatic and viscerosomatic indices were significantly higher in fish fed with SOP12. Muscle protein was higher in fish fed with SOP4, SOP8, and SOP12 than in control but muscle lipids showed an opposite trend. A 14-d post-challenge test against A. hydrophila revealed no significant effect on the fish survival. Nevertheless, fish fed SOP4 encountered delayed bacterial infection compared to other treatments and fish fed with SOP0 and SOP4 performed numerically better survival. Infected fish showed skin depigmentation, haemorrhagic signs at the abdomen and anus, internal bleeding, and stomach and intestine enlargement. In conclusion, SOP4 could be recommended as a growth promoter while slightly delaying A. hydrophila infection in M. nemurus.

Aquaculture wastewater-raised Azolla as partial alternative dietary protein for Pangasius catfish.

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) improvement in weight gain (WG), specific growth rate (SGR), protein efficiency ratio (PER) and feed conversion ratio (FCR) in Pangasius catfish fed 10 g kg-1 Azolla protein. Beyond that, WG, SGR, PER and FCR decreased to the lowest value when fed with 30 g kg-1 Azolla protein. No 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.

High environmental temperature and low pH stress alter the gill phenotypic plasticity of Hoven's carp Leptobarbus hoevenii.

Climate warming and low pH environment are known to negatively impact all levels of aquatic organism from cellular to organism and population levels. For ammonotelic freshwater species, any abiotic factor fluctuation will cause disturbance to the fish, specifically at the gills which act as a multifunctional organ to support all biological processes. Therefore, this study was designed to investigate the effect of temperature (28 vs. 32°C) and pH (7.0 vs. 5.0) stress on the gill plasticity of Hoven's carp after 20 days of continuous exposure. The results demonstrated that high temperature and low pH caused severe changes on the primary and secondary lamellae as well as the cells within lamellae. An increasing trend of the proportion available for gas exchange was noticed at high temperature in both pH exposures, which resulted from a reduction of the primary lamellae width with elongated and thinner secondary lamellae compared to fishes at ambient temperature. Following exposure to high temperature and acidic pH, Hoven's carp experienced gill modifications including aneurysm, oedema, hypertrophy, curling of secondary lamellae, epithelial lifting, hyperplasia and lamellae fusion. These modifications are indicators of the coping mechanism of Hoven's carp to the changing environment in order to survive.

Diel osmorespiration rhythms of juvenile marble goby (Oxyeleotris marmorata).

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.

Common carp, Cyprinus carpio, prefer branchial ionoregulation at high feeding rates and kidney ionoregulation when food supply is limited: additional effects of cortisol and exercise.

This study aims to examine ionoregulatory parameters during exercise and cortisol elevation in common carp fed different food rations. Fish subjected to two different feeding regimes (0.5 or 3.0% body mass (BM) daily) received no implant or an intraperitoneal cortisol implant (250 mg/kg BM) or sham, and were monitored over a 168-h post-implant (PI) period under resting, low aerobic swimming or exhaustive swimming conditions. Plasma osmolality was maintained at relatively stable levels without much influence of feeding, swimming or cortisol, especially in low feeding groups. Nevertheless, a transient hyponatremia was observed in all low feeding fish implanted with cortisol. The hyponatremia was more pronounced in fish swum to exhaustion but even in this group, Na+ levels returned to control levels as cortisol levels recovered (168 h-PI). Cortisol-implanted fish also had lower plasma Cl- levels, and this loss of plasma Cl- was more prominent in fish fed a high ration during exhaustive swimming (recovered at 168 h-PI). Cortisol stimulated branchial NKA and H+ ATPase activities, especially in high ration fish. In contrast, low ration fish upregulated kidney NKA and H+ ATPase activities when experiencing elevated levels of cortisol. In conclusion, low feeding fish experience an ionoregulatory disturbance in response to cortisol implantation especially when swum to exhaustion in contrast to high feeding fish.

Nitrogen handling in the elasmobranch gut: a role for microbial urease.

Ureotelic elasmobranchs require nitrogen for both protein growth and urea-based osmoregulation, and therefore are probably nitrogen-limited in nature. Mechanisms exist for retaining and/or scavenging nitrogen in the gills, kidney, rectal gland and gut, but as yet, the latter are not well characterized. Intestinal sac preparations of the Pacific spiny dogfish shark (Squalus acanthias suckleyi) incubated in vitro strongly reabsorbed urea from the lumen after feeding, but mucosal fluid ammonia concentrations increased with incubation time. Phloretin (0.25 mmol l-1, which blocked urea reabsorption) greatly increased the rate of ammonia accumulation in the lumen. A sensitive [14C]urea-based assay was developed to examine the potential role of microbial urease in this ammonia production. Urease activity was detected in chyme/intestinal fluid and intestinal epithelial tissue of both fed and fasted sharks. Urease was not present in gall-bladder bile. Urease activities were highly variable among animals, but generally greater in chyme than in epithelia, and greater in fed than in fasted sharks. Comparable urease activities were found in chyme and epithelia of the Pacific spotted ratfish (Hydrolagus colliei), a ureotelic holocephalan, but were much lower in ammonotelic teleosts. Urease activity in dogfish chyme was inhibited by acetohydroxamic acid (1 mmol l-1) and by boiling. Treatment of dogfish gut sac preparations with acetohydroxamic acid blocked ammonia production, changing net ammonia accumulation into net ammonia absorption. We propose that microbial urease plays an important role in nitrogen handling in the elasmobranch intestine, allowing some urea-N to be converted to ammonia, which is then reabsorbed for amino acid synthesis or reconversion to urea.

A field and laboratory study of the responses of cytoprotection and osmoregulation to salinity stress in mosquitofish (Gambusia affinis).

The mosquitofish (Gambusia affinis) naturally inhabits freshwater (FW; 1-3‰) and seawater (SW; 28-33‰) ponds in constructed wetland. To explore the physiological status and molecular mechanisms for salinity adaptation of the mosquitofish, cytoprotective responses and osmoregulation were examined. In the field study, activation of protein quality control (PQC) mechanism through upregulation of the abundance of heat shock protein (HSP) 90 and 70 and ubiquitin-conjugated proteins was found in the mosquitofish gills from SW pond compared to the individuals of FW pond. The levels of aggregated proteins in mosquitofish gills had no significant difference between FW and SW ponds. Furthermore, the osmoregulatory responses revealed that the body fluid osmolality and muscle water contents of the mosquitofish from two ponds were maintained within a physiological range while branchial Na+/K+-ATPase (NKA) expression was higher in the individuals from SW than FW ponds. Subsequently, to further clarify whether the cellular stress responses and osmoregulation were mainly induced by hypertonicity, a laboratory salinity acclimation experiment was conducted. The results from the laboratory experiment were similar to the field study. Branchial PQC as well as NKA responses were induced by SW acclimation compared to FW-acclimated individuals. Taken together, induction of gill PQC and NKA responses implied that SW represents an osmotic stress for mosquitofish. Activation of PQC was suggested to provide an osmoprotection to prevent the accumulation of aggregated proteins. Moreover, an increase in branchial NKA responses for osmoregulatory adjustment was required for the physiological homeostasis of body fluid osmolality and muscle water content.

Kinematics of mouthbrooding in Oreochromis niloticus (Cichlidae).

Many species from several different families of fishes perform mouthbrooding, where one of the sexes protects and ventilates the eggs inside the mouth cavity. This ventilation behaviour differs from gill ventilation outside the brooding period, as the normal, small-amplitude suction-pump respiration cycles are alternated with actions including near-simultaneous closed-mouth protrusions and high-amplitude depressions of the hyoid. The latter is called churning, referring to its hypothetical function in moving around and repositioning the eggs by a presumed hydrodynamic effect of the marked shifts in volume along the mouth cavity. We tested the hypothesis that churning causes the eggs located posteriorly in the mouth cavity to move anteriorly away from the gill entrance. This would prevent or clear accumulations of brood at the branchial basket, which would otherwise hinder breathing by the parent. Dual-view videos of female Nile tilapias (Oreochromis niloticus) during mouthbrooding showed that churning involves a posterior-to-anterior wave of expansion and compression of the head volume. Flow visualisation with polyethylene microspheres revealed a significant inflow of water entering the gill slits at the zone above the pectoral fin base, followed by a predominantly ventral outflow passing the ventrolaterally flapping branchiostegal membranes. X-ray videos indicated that particularly the brood located close to the gills is moved anteriorly during churning. These data suggest that, in addition to mixing of the brood to aid its oxygenation, an important function of the anterior flow through the gills and buccal cavity during churning is to prevent clogging of the eggs near the gills.

Cortisol affects metabolic and ionoregulatory responses to a different extent depending on feeding ration in common carp, Cyprinus carpio.

Interacting effects of feeding and stress on corticoid responses in fish were investigated in common carp fed 3.0% or 0.5% body mass (BM) which received no implant, a sham or a cortisol implant (250 mg/kg BM) throughout a 168 hour post-implant period (168 h-PI). At 12h-PI, cortisol implants elevated plasma cortisol, glucose and lactate. Plasma osmolality and ions remained stable, but cortisol increased gill and kidney Na(+)/K(+) ATPase (NKA) and H(+) ATPase activities. Gill NKA activities were higher at 3%-BM, whereas kidney H(+) ATPase activity was greater at 0.5%-BM. Cortisol induced liver protein mobilization and repartitioned liver and muscle glycogen. At 3%-BM, this did not increase plasma ammonia, reflecting improved excretion efficiency concomitant with upregulation of Rhesus glycoprotein Rhcg-1 in gill. Responses in glucocorticoid receptors (GR1/GR2) and mineralocorticoid receptor (MR) to cortisol elevation were most prominent in kidney with increased expression of all receptors at 24 h-PI at 0.5%-BM, but only GR2 and MR at 0.5%-BM. In the liver, upregulation of all receptors occurred at 24 h-PI at 3%-BM, whilst only GR2 and MR were upregulated at 0.5%-BM. In the gill, there was a limited upregulation: GR2 and MR at 72 h-PI and GR1 at 168 h-PI at 3%-BM but only GR2 at 72 h-PI at 0.5%-BM. Thus cortisol elevation led to similar expression patterns of cortisol receptors in both feeding regimes, while feeding affected the type of receptor that was induced. Induction of corticoid receptors occurred simultaneously with increases in Rhcg-1 mRNA expression (gill) but well after NKA and H(+) ATPase activities increased (gill/kidney).

The combined effect of hypoxia and nutritional status on metabolic and ionoregulatory responses of common carp (Cyprinus carpio).

In the present study, the combined effects of hypoxia and nutritional status were examined in common carp (Cyprinus carpio), a relatively hypoxia tolerant cyprinid. Fish were either fed or fasted and were exposed to hypoxia (1.5-1.8mg O2L(-1)) at or slightly above their critical oxygen concentration during 1, 3 or 7days followed by a 7day recovery period. Ventilation initially increased during hypoxia, but fasted fish had lower ventilation frequencies than fed fish. In fed fish, ventilation returned to control levels during hypoxia, while in fasted fish recovery only occurred after reoxygenation. Due to this, C. carpio managed, at least in part, to maintain aerobic metabolism during hypoxia: muscle and plasma lactate levels remained relatively stable although they tended to be higher in fed fish (despite higher ventilation rates). However, during recovery, compensatory responses differed greatly between both feeding regimes: plasma lactate in fed fish increased with a simultaneous breakdown of liver glycogen indicating increased energy use, while fasted fish seemed to economize energy and recycle decreasing plasma lactate levels into increasing liver glycogen levels. Protein was used under both feeding regimes during hypoxia and subsequent recovery: protein levels reduced mainly in liver for fed fish and in muscle for fasted fish. Overall, nutritional status had a greater impact on energy reserves than the lack of oxygen with a lower hepatosomatic index and lower glycogen stores in fasted fish. Fasted fish transiently increased Na(+)/K(+)-ATPase activity under hypoxia, but in general ionoregulatory balance proved to be only slightly disturbed, showing that sufficient energy was left for ion regulation.

Female tilapia, Oreochromis sp. mobilised energy differently for growth and reproduction according to living environment.

This study was conducted to investigate the energy mobilisation preference and ionoregulation pattern of female tilapia, Oreochromis sp. living in different environments. Three different treatments of tilapia as physiology compromising model were compared; tilapia cultured in recirculating aquaculture system (RAS as Treatment I-RAS), tilapia cultured in open water cage (Treatment II-Cage) and tilapia transferred from cage and cultured in RAS (Treatment III-Compensation). Results revealed that tilapia from Treatment I and III mobilised lipid to support gonadogenesis, whilst Treatment II tilapia mobilised glycogen as primary energy for daily exercise activity and reserved protein for growth. The gills and kidney Na+/K+ ATPase (NKA) activities remained relatively stable to maintain homeostasis with a stable Na+ and K+ levels. As a remark, this study revealed that tilapia strategized their energy mobilisation preference in accessing glycogen as an easy energy to support exercise metabolism and protein somatogenesis in cage culture condition, while tilapia cultured in RAS mobilised lipid for gonadagenesis purposes.

Hoven's carp Leptobarbus hoevenii strategized metabolism needs to cope with changing environment.

Current water warming and freshwater acidification undoubtedly affect the life of aquatic animals especially ammonotelic teleost by altering their physiological responses. The effect of temperature (28 °C vs 32 °C) and pH (7 vs. 5) on the metabolic compromising strategies of Hoven's carp (Leptobarbus hoevenii) was investigated in this study. Fishes were conditioned to (i) 28 °C + pH 7 (N28°C); (ii) 32 °C + pH 7 (N32°C); (iii) 28 °C + pH 5 (L28°C) and (iv) 32 °C + pH 5 (L32°C) for 20 days followed by osmorespiration assay. Results showed that feeding performance of Hoven's carp was significantly depressed when exposed to low pH conditions (L28°C and L32°C). However, by exposed Hoven's carp to L32°C induced high metabolic oxygen intake and ammonia excretion to about 2x-folds higher compared to the control group. As for energy mobilization, Hoven's carp mobilized liver and muscle protein under L28°C condition. Whereas under high temperature in both pH, Hoven's carp had the tendency to reserve energy in both of liver and muscle. The findings of this study revealed that Hoven's carp is sensitive to lower water pH and high temperature, thereby they remodeled their physiological needs to cope with the environmental changes condition.

An in vitro study of urea, water, ion and CO2/HCO3- transport in the gastrointestinal tract of the dogfish shark (Squalus acanthias): the influence of feeding.

In vitro gut sac preparations made from the cardiac stomach (stomach 1), pyloric stomach (stomach 2), intestine (spiral valve) and colon were used to examine the impact of feeding on transport processes in the gastrointestinal tract of the dogfish shark. Preparations were made from animals that were euthanized after 1-2 weeks of fasting, or at 24-48 h after voluntary feeding on a 3% ration of teleost fish (hake). Sacs were incubated under initially symmetrical conditions with dogfish saline on both surfaces. In comparison to an earlier in vivo study, the results confirmed that feeding caused increases in H(+) secretion in both stomach sections, but an increase in Cl(-) secretion only in stomach 2. Na(+) absorption, rather than Na(+) secretion, occurred in both stomach sections after feeding. All sections of the tract absorbed water and the intestine strongly absorbed Na(+) and Cl(-), regardless of feeding condition. The results also confirmed that feeding increased water absorption in the intestine (but not in the colon), and had little influence on the handling of Ca(2+) and Mg(2+), which exhibited negligible absorption across the tract. However, K(+) was secreted in the intestine in both fasted and fed preparations. Increased intestinal water absorption occurred despite net osmolyte secretion into the mucosal saline. The largest changes occurred in urea and CO2/HCO3(-) fluxes. In fasted preparations, urea was absorbed at a low rate in all sections except the intestine, where it was secreted. Instead of an increase in intestinal urea secretion predicted from in vivo data, feeding caused a marked switch to net urea absorption. This intestinal urea transport occurred at a rate comparable to urea reabsorption rates reported at gills and kidney, and was apparently active, establishing a large serosal-to-mucosal concentration gradient. Feeding also greatly increased intestinal CO2/HCO3(-) secretion; if interpreted as HCO3(-) transport, the rates were in the upper range of those reported in marine teleosts. Phloretin (0.25 mmol l(-1), applied mucosally) completely blocked the increases in intestinal urea absorption and CO2/HCO3(-) secretion caused by feeding, but had no effect on Na(+), Cl(-) or water absorption.

Modulation of Rh glycoproteins, ammonia excretion and Na+ fluxes in three freshwater teleosts when exposed chronically to high environmental ammonia.

We investigated relationships among branchial unidirectional Na(+) fluxes, ammonia excretion, urea excretion, plasma ammonia, plasma cortisol, and gill transporter expression and function in three freshwater fish differing in their sensitivity to high environmental ammonia (HEA). The highly ammonia-sensitive salmonid Oncorhynchus mykiss (rainbow trout), the less ammonia-sensitive cyprinid Cyprinus carpio (common carp) and the highly ammonia-resistant cyprinid Carassius auratus (goldfish) were exposed chronically (12-168 h) to 1 mmol l(-1) ammonia (as NH4HCO3; pH 7.9). During HEA exposure, carp and goldfish elevated ammonia excretion (JAmm) and Na(+) influx rates ( ) while trout experienced higher plasma ammonia (TAmm) and were only able to restore control rates of JAmm and . All three species exhibited increases in Na(+) efflux rate ( ). At the molecular level, there was evidence for activation of a 'Na(+)/NH4(+) exchange metabolon' probably in response to elevated plasma cortisol and TAmm, though surprisingly, some compensatory responses preceded molecular responses in all three species. Expression of Rhbg, Rhcg (Rhcg-a and Rhcg-b), H(+)-ATPase (V-type, B-subunit) and Na(+)/K(+)-ATPase (NKA) mRNA was upregulated in goldfish, Rhcg-a and NKA in carp, and Rhcg2, NHE-2 (Na(+)/H(+) exchanger) and H(+)-ATPase in trout. Branchial H(+)-ATPase activity was elevated in goldfish and trout, and NKA activity in goldfish and carp, but NKA did not appear to function preferentially as a Na(+)/NH4(+)-ATPase in any species. Goldfish alone increased urea excretion rate during HEA, in concert with elevated urea transporter mRNA expression in gills. Overall, goldfish showed more effective compensatory responses towards HEA than carp, while trout were least effective.

Cortisol emphasizes the metabolic strategies employed by common carp, Cyprinus carpio at different feeding and swimming regimes.

The objective of this study was to investigate the interaction between feeding, exercise and cortisol on metabolic strategies of common carp over a 168h post-implant period. Feeding provided readily available energy and clearly increased muscle and liver protein and glycogen stores. Swimming, feeding and cortisol all induced aerobic metabolism by increasing oxygen consumption, and stimulated protein metabolism as demonstrated by the increased ammonia and urea excretion and ammonia quotient. Hypercortisol stimulated ammonia self-detoxifying mechanisms by enhancing ammonia and urea excretion, especially during severe exercise. At high swimming level, higher branchial clearance rates in cortisol treated fish succeeded in eliminating the elevation of endogenous ammonia, resulting in reduced plasma Tamm levels compared to control and sham implanted fish. Carp easily induced anaerobic metabolism, both during routine and active swimming, with elevated lactate levels as a consequence. Both feeding and cortisol treatment increased this dependence on anaerobic metabolism. Hypercortisol induced both glycogenesis and gluconeogenesis resulting in hyperglycemia and muscle and liver glycogen deposition, most likely as a protective mechanism for prolonged stress situations and primarily fuelled by protein mobilization.

Feeding and swimming modulate iono-and-hormonal regulation differently in goldfish, Carassius auratus and common carp, Cyprinus carpio.

Feeding and swimming can influence ion balance in fish. Therefore we investigated their impact on ionoregulation and its hormonal control in goldfish and common carp. As expected due to the osmorespiratory compromise, exhaustive swimming induced increases in gill Na(+)/K(+) ATPase (NKA) activity in both species, resulting in stable levels of plasma ions. In contrast to our expectations, this only occurred in fed fish and feeding itself increased NKA activity, especially in carp. Fasting fish were able to maintain ion balance without increasing NKA activity, we propose that the increase in NKA activity is related to ammonia excretion rather than ion uptake per se. In goldfish, this increase in NKA activity coincided with a cortisol elevation whilst no significant change was found in carp. In goldfish, high conversion of plasma T4 to T3 was found in both fed and fasted fish resulting in low T4/T3 ratios, which increased slightly due to exhaustive swimming. In starved carp the conversion seemed much less efficient, and high T4/T3 ratios were observed. We propose that thyroid hormone regulation in carp was more related to its role in energy metabolism rather than ionoregulation. The present research showed that both species, whether fed or fasted, are able to sufficiently adapt their osmorepiratory strategy to minimise ions losses whilst maintaining gas exchange under exhaustive swimming.

Sustainable aquafeed development: Incorporating select fruit wastes into Zebrafish diets using mathematical model-based approach.

Expensive aquafeed is a major problem in aquaculture, creating the need for a low-cost feed that provides ideal nutritional requirement to maximize growth performances. This study aims to formulate and evaluate two new optimized Zebrafish feeds (F1 and F2) using linear programming mathematical model, one of which incorporates two pigment rich fruit wastes (Pitaya peel and Roselle calyx) in the formulation. The model represents nutritional content and cost of each ingredient into linear equations, with the goal of finding ideal combination that satisfies the specific nutrient requirements. By systematically evaluating ingredient proportions, linear programming ensures that Zebrafish receives adequate nutrients at the lowest possible cost, making the feed development process more efficient and cost-effective. The novel feed formulations derived from the mathematical model were tested on the growth and pigmentation of Zebrafish in comparison to a commercial feed (control). Feed intake of F1 and F2 were generally found to be similar to the control feed, indicating the acceptability of the formulated feeds by the Zebrafish. Body weight and length of Zebrafish fed with F1 and F2 were comparable to Zebrafish fed with control feed (p > 0.05). Similarly, Zebrafish fed with F1 and F2 showed no significant differences in pigment intensity compared to Zebrafish fed with control feed (p > 0.05). The survival rate of fishes in all feeding groups were greater than 70 % with no significant differences (p > 0.05). Results obtained in this study illustrated the potential of mathematic linear programming and effectiveness of utilising pigment-rich fruit wastes in formulating an optimized economic aquafeed.

Sensory-mediated feeding behaviour in the larvae of marble goby (Oxyeleotris marmorata).

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.

Evaluation of crystalline amino acids as potent stimulatory chemoattractants for the slipper lobster Thenus orientalis.

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.