Response of gastrointestinal melatonin,antioxidants, and digestive enzymes to altered feeding conditions in carp (<Emphasis Type="Italic">Catla catla</Emphasis>)

The purpose of present study was to ascertain whether the response of gastrointestinal (gut) melatonin to altered feeding conditions was related to the levels of different antioxidants and digestive enzymes in the same gut tissues of a sub-tropical carp (Catla catla). Accordingly, the fish were subjected to food deprivation for 4 or 8 days and separately to re-feeding for 4 or 8 or 12 days after deprivation of food for 8 days, and their gut tissue homogenates were used to measure the levels of melatonin, both enzymatic [superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione S-transferase (GST)] and non-enzymatic [reduced glutathione (GSH)] antioxidants, as well as different digestive enzymes (α-amylase, cellulase, protease, and lipase). Notably, the gut levels of melatonin, SOD, CAT, GPx, and GST underwent gradual increase with the progress of food deprivation, but a sudden fall after restoration of food supply for 4 days and a rise thereafter. Conversely, the activity of all the digestive enzymes significantly decreased after deprivation of food, but started increasing when food supply was reinforced. Gut melatonin concentrations by showing a positive correlation with the titers of different antioxidants (in both food-deprived and re-fed fish groups) and a negative (in food-deprived fish) or a positive (in re-fed fish) correlation with the activity of each digestive enzyme underlined possible physiological interplay between them. Collectively, our findings lend support to the hypothesis that gut melatonin response to altered feeding conditions in carp might be associated with the oxidative status as well as the digestive functions of the gastrointestinal tissues itself.     

Immunogene expression in head kidney and spleen of common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.) following thermal stress and challenge with Gram-negative bacterium, <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

To evaluate the effect of thermal and microbial stress on the immune response of common carp (Cyprinus carpio L.), relative mRNA expression level of pro-inflammatory cytokines [tumor necrosis factor alpha (TNF-α) and interleukin (IL)-1β] and other genes related to immune or stress response [inducible nitric oxide synthase (iNOS), heat shock protein 70 (Hsp70), superoxide dismutase one (SOD1), and glucocorticoid receptor (GR)] was measured by quantitative PCR (qPCR). In addition, total protein and total immunoglobulin level in blood plasma of experimental common carp was also assayed. All the above parameters were estimated 24 h post-challenge with Gram-negative bacterium, Aeromonas hydrophila. Common carp (54.89?±?6.90 g) were initially exposed to 20 °C (control group) and 30 °C (thermal stress group) water temperature for 30 days, followed by experimental challenge with 2.29?×?10⁸ colony forming unit/mL (CFU/mL; LD₅₀ dose) of A. hydrophila. Exposure of fish to thermal stress and subsequently challenge with A. hydrophila significantly (P?<?0.05) increases the IL-1β mRNA expression in head kidney and spleen of common carp by ~?39.94 and ~?4.11-fold, respectively. However, TNF-α mRNA expression in spleen decreased ~?5.63-fold in control fish challenged with A. hydrophila. Thermal stress and challenge with bacterium suppresses the iNOS and GR mRNA expression in spleen of common carp. Moreover, significant (P?<?0.05) increase in total protein content of blood plasma (~?43 mg/g) was evident in fish exposed to thermal stress and challenged with A. hydrophila. In conclusion, our study highlights the importance of elevated temperature stress and microbial infection in differential regulation of expression of several immunogenes in common carp.     

A study of the damage of the intestinal mucosa barrier structure and function of <Emphasis Type="Italic">Ctenopharyngodon idella</Emphasis> with <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

The aim of this study is to explore the effect of Aeromonas hydrophila on the intestinal mucosal barrier structure and intestinal permeability in grass carp (Ctenopharyngodon idella). Histopathological examinations showed that A. hydrophila induced severe intestinal lesions, including inflammatory cell infiltration and intestinal villus fusion and swelling. Messenger RNA (mRNA) expression of the inflammatory cytokines TNF-α, IL-1β, IL-8, IL-10 and MyD88 was significantly increased after infection with A. hydrophila. The permeability of intestinal mucosa was determined using Evans blue (EB) and d-lactic acid. The results indicated that the levels of EB and serum d-lactic acid were significantly increased after infection with A. hydrophila (p < 0.05). Our results also indicated that the intestinal mucosal barrier injury induced by A. hydrophila infection was closely associated with the expression of the tight junction (TJ) protein zonula occludens-1 (ZO-1), occludin, claudin b and claudin c as well as the activity of Na⁺, K⁺-ATPase and Ca²⁺, Mg²⁺-ATPase. Lower mRNA levels of occludin and lower Na⁺, K⁺-ATPase and Ca²⁺, Mg²⁺-ATPase activity in the intestines were observed after challenge. ZO-1 and claudin c were significantly increased 24 h after infection with A. hydrophila. The most interesting finding was that claudin b also significantly increased 24 h after challenge and then decreased to lower levels at 72, 120 and 168 h post-infection compared to the PBS-treated control group. The results demonstrated that grass carp infection with A. hydrophila induced intestinal inflammation and impaired the structure and function of the intestinal mucosal barrier.     

<Emphasis Type="Italic">Virgibacillus proomii</Emphasis> and <Emphasis Type="Italic">Bacillus mojavensis</Emphasis> as probiotics in sea bass (<Emphasis Type="Italic">Dicentrarchus labrax</Emphasis>) larvae: effects on growth performance and digestive enzyme activities

This study examined the effects of two probiotics (Virgibacillus proomii and Bacillus mojavensis) on the digestive enzyme activity, survival and growth of Dicentrarchus labrax at various ontogenetic stages in three separate experiments. These probiotics were incorporated as single or mixed into fish feed for a period of 60 days. The growth parameters, proximate composition of whole body, digestive enzymes and gut microbiology were monitored at regular. The increments in length and weight and the survival were significantly higher (P < 0.05), and the values of food conversions were significantly lower (P < 0.05) in fishes fed the probiotic. The administration of V. proomii and B. mojavensis in diet resulted in an increase (P > 0.05) in body ash and protein content and in the specific activity of phosphatase alkaline and amylase in the digestive tract of all the fishes. V. proomii and B. mojavensis persisted in the fish intestine and in the feed in high numbers during the feeding period (group 1: 5.8 × 10⁴ CFU/ml, group 2: 9.6 × 10⁴ CFU/ml, and group 3: 9.8 × 10⁴ CFU/ml day 60). The two probiotics V. proomii and B. mojavensis were adequate for improved growth performance and survival and for healthy gut microenvironment of the host.     

Environmental factors on virulence of <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

Aeromonas hydrophila are known for being opportunistic pathogens, harboring various virulence factors and triggering lesions and death in fish. The disease caused by bacteria can make fish inappropriate for human consumption, besides representing a risk to public health. The pathogenesis can be influenced by environmental variables, affecting fish productivity and mortality. The present study aimed to determine whether A. hydrophila harbor the virulence genes aerolysin, hydrolipase, elastase, lipase, cytotonic enterotoxin (ast), lateral flagellum (laf), and polar flagellum (fla) and to evaluate the influence of environmental variables on in vitro growth, in vivo virulence and expression of some of these genes. Polymerase chain reaction (PCR)-based screening for the presence of these virulence genes was performed on 35 isolates. Six isolates containing different profiles of virulence genes were tested for in vitro growth under different conditions of pH, temperature, and ammonia and for in vivo virulence under these same environmental conditions. RT-qPCR was used to quantify the expression of aerolysin, lipase, and fla genes. All the tested environmental factors influenced the growth of A. hydrophila, while pH and ammonia concentrations influenced the bacterial virulence. The expression of the fla gene increased when bacteria were grown in higher ammonia concentration. The mortality established by Aeromonas is influenced by several environmental factors pinpointing the importance of its control in fish farming to avoid higher economic loses associated to bacterial disease outbreaks.     

Stimulatory effect of dietary taurine on growth performance,digestive enzymes activity,antioxidant capacity,and tolerance of common carp, <Emphasis Type="Italic">Cyprinus carpio</Emphasis> L., fry to salinity stress

The present study was carried out to evaluate the effect of dietary taurine (Tau) on performance, digestive enzymes, antioxidant activity, and resistance of common carp, Cyprinus carpio L., fry to salinity stress. Fish (0.97?±?0.033 g) were fed on different taurine levels of 0.0 (control), 5, 10, 15, or 20 g/kg diet up to satiation twice daily for 8 weeks. At the end of the feeding trial, fish were stressed by exposure to 10 ppt salinity for 3 days during which fish mortality was observed. Fish performance was significantly (P?<?0.05) improved by dietary taurine up to 15 g Tau/kg diet after which fish growth and feed intake were almost the same. Also, taurine supplementation significantly (P?<?0.05) elevated activities of intestinal amylase, lipase, and protease resulting in an improving in feed intake giving better performance. Furthermore, Tau-stimulated antioxidant activity of common carp was observed in a dose-related manner, where activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were significantly (P?<?0.05) higher, but malondialdehyde (MDA) value was significantly (P?<?0.05) lower in Tau-fed fish groups than those fed the control diet. In salinity stress experiment, highest survival rate was observed at fish fed Tau-supplemented diets without significant (P?>?0.05) differences over fish fed the control diet. It appears that taurine could be used as a feed supplement to confer better growth and health of common carp fry with optimal level of 15 g/kg diet.     

Molecular cloning and expression analysis of <Emphasis Type="Italic">Megalobrama amblycephala</Emphasis> transferrin gene and effects of exposure to iron and infection with <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

Transferrin (Tf) plays an important function in iron homeostasis and metabolism of organisms. In this study, we identified and characterized the Tf gene in Megalobrama amblycephala and evaluated its expression in basal conditions as well as after iron overload and experimental infection with Aeromonas hydrophila. Furthermore, we studied the iron binding properties of recombinant Tf. The full-length M. amblycephala Tf complementary DNA (cDNA) (GenBank accession no.: KX698308) of 2245 bp was cloned and contained a 1953 bp open reading frame (ORF) encoding 650 amino acid residues and flanked by a 68 bp 5′ and a 204 bp 3′ untranslated regions (UTR). Predicted conservative structure illustrated that M. amblycephala Tf consisted of two conservative Tf domains. Amino acid sequence alignment revealed that M. amblycephala Tf had high similarity with that of cyprinids deposited in Genbank, and phylogenetic analysis showed that M. amblycephala Tf clustered with Ctenopharyngodon idella and Hypophthalmichthys molitrix. Tissue expression pattern analyses demonstrated that the liver was the main Tf mRNA expressing organ, being significantly higher than other tissues (p < 0.05). In the liver, Tf mRNA expression in fish artificially injected with the pathogenic bacteria A. hydrophila was significantly upregulated, reaching a peak at 12 h post injection (hpi) and then decreasing afterward. The expression in FeCl₃-injected fish showed a similar tendency, but reached a peak at 8 hpi. Meanwhile, fish serum iron significantly decreased following A. hydrophila injection, but increased to peak at 4 hpi and then decreased in FeCl₃-injected fish. The recombinant M. amblycephala Tf showed iron binding capacity using CAS analysis. These results are helpful to understand the structure and regulation of expression of Tf, as well as the specific function of Tf for both immune responses and iron homeostasis.     

Development of a simple and rapid monoclonal antibody-based flow through immunogold assay (FIA) for detection of <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

In this study, we developed a simple, low-cost, and rapid flow through immunogold assay (FIA) for detection of Aeromonas hydrophila in fish tissues and validated. The developed assay relied on colloidal gold conjugated highly specific monoclonal antibody (B8E3) against 20 kDa protein of A. hydrophila. The assay can be completed within 5 min including antigen (sample) preparation and exhibited no cross-reaction with other major aquatic pathogens such as Vibrio anguillarum, V. parahaemolyticus, V. harveyi, white spot syndrome virus (WSSV), and Aphanomyces invadans. A wider range of Aeromonas species were tested including A. caviae, A. dhakensis, A. sobria, A. veronii, A. culicicola, A. sharmana, and A. schubertii and no cross-reactivity could be observed. The developed technique FIA could detect 10³ CFU/ml of aeromonad cells. In addition, the FIA is as sensitive as the first-step polymerase chain reaction (PCR) and 1000 times sensitive than the immunodot blot method. The validation study revealed the accuracy of FIA by comparing with the standard first-step PCR. The proposed FIA is low-cost, easy-to-use, and compatible for field-level analysis of A. hydrophila in fish tissues. Our developed FIA could successfully detect A. hydrophila at early stage of the infection and could be easily availed by farmers, fish producers, non-technicians, and technicians in the aquaculture industry.     

<Emphasis Type="Italic">Streptococcus agalactiae</Emphasis> from tilapia (<Emphasis Type="Italic">Oreochromis</Emphasis> sp.) transmitted to a new host,bighead carp (<Emphasis Type="Italic">Aristichthys nobilis</Emphasis>), in China

Several outbreaks of Streptococcus agalactiae infection of bighead carp (Aristichthys nobilis) were observed in China. The molecular epidemiology and pathogenicity of S. agalactiae in bighead carp and tilapia (Oreochromis sp.) is poorly understood. In the present study, we identified S. agalactiae strains isolated from diseased bighead carp using the API 20 Strep kit and 16S rDNA sequencing and determined whether these strains came from tilapia. Of the 46 identified S. agalactiae strains, 24 strains were successfully isolated from diseased bighead carps, 20 S. agalactiae strains were isolated from tilapia, and two S. agalactiae strains were isolated from tiger frog (Hoplobatrachus chinensis). The results of molecular typing, including multilocus sequence typing, molecular serotyping, surface protein gene detection, and virulence-related gene detection showed that the 44 strains from bighead carp and tilapia were highly similar, whereas different from tiger frog GBS strains. Remarkably, the bighead carp strain Hn1404 showed high virulence in bighead carp and zebrafish. Moreover, this strain was pathogenic to Nile tilapia (Oreochromis niloticus). In addition, comparative genomic analysis showed that isolate Hn1404 had a close relationship with the bighead carp and tilapia S. agalactiae strains. All the analyses of the genetic characteristics of bighead carp and tilapia strains showed that tilapia S. agalactiae strains could be transmitted to other fish species such as bighead carp.     

Effects of potential probiotic <Emphasis Type="Italic">Lactococcus lactis</Emphasis> subsp. <Emphasis Type="Italic">lactis</Emphasis> on digestive enzymatic activities of live feed <Emphasis Type="Italic">Artemia franciscana</Emphasis>

In the present study, cellulase, protease, lipase and amylase activities were performed to investigate the effects of Lactococcus lactis subsp. lactis CF4MRS bioencapsulation of Artemia franciscana. Our results show that cellulase activities (total cellulase—FPase activity, exoglucanase and endoglucanase—CMCase activity) were significantly higher (P < 0.05) in A. franciscana tissue homogenates compared to those in the control group after 8 h of L. lactic bioencapsulation. Notably, an exception case was found in β-D-glucosidase activity, whereby the cellulase activity was not significantly different (P > 0.05) compared to the control group. Administrations of L. lactis at cell concentration of 10⁸ CFU mL^?1 showed considerable improvement of other important enzymatic activities such as amylase, protease and lipase in A. franciscana. The amylase/protease ratio in probiotic-treated A. franciscana was recorded at 0.343, approximately two times higher than those without probiotic administration (0.184). In contrary, amylase/lipase ratio showed half of a reduction (0.330) in L. lactis-administrated A. franciscana compared to the control (0.614). Our study suggests that important digestive enzymes, e.g., cellulase, amylase, protease and lipase, can be enhanced through bioencapsulation of A. franciscana with L. lactis subsp. lactis, which could in turn lead to further stimulation of endogenous enzymes in the fish and shrimp larvae.     

Cloning and characterization of <Emphasis Type="Italic">Bax1</Emphasis> and <Emphasis Type="Italic">Bax2</Emphasis> genes of <Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis> and evaluation of transcript expression in response to grass carp reovirus infection

Multidomain proapoptotic Bcl-2-associated X (Bax) protein is an essential effector responsible for mitochondrial outer membrane permeabilization, resulting in cell death via apoptosis. In this study, two Bax genes of grass carp (Ctenopharyngodon idellus), designated as CiBax1 and CiBax2, were isolated and analyzed. The obtained CiBax1 cDNA is 2058 bp long, with a 579 bp open reading frame (ORF) coding a protein of 192 amino acid residues. The full-length cDNA of CiBax2 is 1161 bp, with a 618 bp ORF coding 205 amino acids. Both CiBax1 and CiBax2 are typical members of Bcl-2 family containing conserved Bcl and C-terminal domains, and they share conserved synteny with zebrafish Bax genes despite the grass carp Bax mapping to different linkage groups. Phylogenetic analysis showed that CiBax1 was clustered with Bax from most teleost fish, and CiBax2 was close to Bax2 from teleost fish but far separated from that of Salmo salar. Quantitative real-time PCR analysis revealed broad expression of CiBax1 and CiBax2 in tissues from healthy grass carp, but the relative expression level differed. The mRNA expression of CiBax1 and CiBax2 was both upregulated significantly and peaked in all examined tissues at days 5 or 6 post-infection with grass carp reovirus. Subcellular localization indicated that CiBax1 protein was localized in both nucleus and cytosol, while CiBax2 protein only in cytosol. Moreover, CiBax2, but not CiBax1 was colocalized with mitochondrion under normal condition. Taken together, the findings would be helpful for further understanding of the function of Bax in teleost fish.     

Comparative nutritional value of <Emphasis Type="Italic">Jatropha curcas</Emphasis> protein isolate and soy protein isolate in common carp

Jatropha seed cake (JSC) is an excellent source of protein but does contain some antinutritional factors (ANF) that can act as toxins and thus negatively affect the growth and health status of fish. While this can limit the use of JSC, detoxified Jatropha protein isolate (DJPI) may be a better option. An 8-week study was performed to evaluate dietary DJPI to common carp Cyprinus carpio. Five iso-nitrogenous diets (crude protein of 38%) were formulated that consisted of a C _ontrol (fish meal (FM) based protein), J ₅₀ or J ₇₅ (50 and 75% of FM protein replaced by DJPI), and S ₅₀ or S ₇₅ (50 and 75% of FM protein replaced by soy protein isolate, SPI) and fed to triplicate groups of 75 carp fingerlings (75; av. wt. ± SD; 11.4 ± 0.25 g). The growth, feeding efficiencies, digestibility, plasma biochemistry, and intestinal enzymes were measured. Results showed that growth performance of fish fed the S ₇₅- or DJPI-based diets were not significantly different from those fed the C _ontrol diet, while carp fed the S ₅₀ had significantly better growth than the J ₇₅ diet. Fish fed the J ₇₅ diet had significantly lower protein and lipid digestibility as well as significantly lower intestinal amylase and protease activities than all other groups. However, all plant protein-based diets led to significantly higher crude protein, crude lipid, and gross energy in the body of common carp compared to the control treatment. Plasma cholesterol and creatinine significantly decreased in the plant protein fed groups, although plasma triglyceride as well as the red blood cells count, hematocrit, albumin, globulin, total plasma protein, and lysozyme activity were higher in plant protein fed groups compared to FM fed group. White blood cells, hemoglobulin concentration, alkaline phosphatase and alanine transaminase activities, and glucose level in blood did not differ significantly among treatments. The results suggest that the DJPI is non-toxic to carp and can be used to replace FM in the diets of common carp up to 75%, but further research to potentially reduce some inherent ANF within this protein source, such as non-starch polysaccharides, may improve nutrient utilization.     

The effect of live feeds bathed with the red seaweed <Emphasis Type="Italic">Asparagopsis armata</Emphasis> on the survival,growth and physiology status of <Emphasis Type="Italic">Sparus aurata</Emphasis> larvae

Larval rearing is affected by a wide range of microorganisms that thrive in larviculture systems. Some seaweed species have metabolites capable of reducing the bacterial load. However, no studies have yet tested whether including seaweed metabolites on larval rearing systems has any effects on the larvae development. This work assessed the development of Sparus aurata larvae fed preys treated with an Asparagopsis armata product. Live prey, Brachionus spp. and Artemia sp., were immersed in a solution containing 0.5% of a commercial extract of A. armata (Ysaline 100, YSA) for 30 min, before being fed to seabream larvae (n = 4 each). In the control, the live feed was immersed in clear water. Larval parameters such as growth, survival, digestive capacity (structural-histology and functional-enzymatic activity), stress level (cortisol content), non-specific immune response (lysozyme activity), anti-bacterial activity (disc-diffusion assay) and microbiota quantification (fish larvae gut and rearing water) were monitored. Fish larvae digestive capacity, stress level and non-specific immune response were not affected by the use of YSA. The number of Vibrionaceae was significantly reduced both in water and larval gut when using YSA. Growth was enhanced for YSA treatment, but higher mortality was also observed, especially until 10 days after hatching (DAH). The mortality peak observed at 8 DAH for both treatments, but higher for YSA, indicates larval higher susceptibility at this development stage, suggesting that lower concentrations of YSA should be used until 10 DAH. The application of YSA after 10 DAH onwards promotes a safer rearing environment.     

Molecular evolution of myoglobin in the Tibetan Plateau endemic schizothoracine fish (<Emphasis Type="Italic">Cyprinidae</Emphasis>, <Emphasis Type="Italic">Teleostei</Emphasis>) and tissue-specific expression changes under hypoxia

Myoglobin (Mb) is an oxygen-binding hemoprotein that was once thought to be exclusively expressed in oxidative myocytes of skeletal and cardiac muscle where it serves in oxygen storage and facilitates intracellular oxygen diffusion. In this study, we cloned the coding sequence of the Mb gene from four species, representing three groups, of the schizothoracine fish endemic to the Qinghai-Tibetan Plateau (QTP), then conducted molecular evolution analyses. We also investigated tissue expression patterns of Mb and the expression response to moderate and severe hypoxia at the mRNA and protein levels in a representative of the highly specialized schizothoracine fish species, Schizopygopsis pylzovi. Molecular evolution analyses showed that Mb from the highly specialized schizothoracine fish have undergone positive selection and one positively selected residue (81L) was identified, which is located in the F helix, close to or in contact with the heme. We present tentative evidence that the Mb duplication event occurred in the ancestor of the schizothoracine and Cyprininae fish (common carp and goldfish), and that the Mb2 paralog was subsequently lost in the schizothoracine fish. In S. pylzovi, Mb mRNA is expressed in various tissues with the exception of the intestine and gill, but all such tissues, including the liver, muscle, kidney, brain, eye, and skin, expressed very low levels of Mb mRNA (<?8.0%) relative to that of the heart. The trace levels of Mb expression in non-muscle tissues are perhaps the major reason why non-muscle Mb remained undiscovered for so long. The expression response of the Mb gene to hypoxia at the mRNA and protein levels was strikingly different in S. pylzovi compared to that found in the common carp, medaka, zebrafish, and goldfish, suggesting that the hypoxia response of Mb in fish may be species and tissue-specific. Notably, severe hypoxia induced significant expression of Mb at the mRNA and protein levels in the S. pylzovi heart, which suggests Mb has a major role in the supply of oxygen to the heart of Tibetan Plateau fish.     

Aquaculture produces wholesome food: cultured fish as a valuable source of n-3 fatty acids

Long-chain unsaturated n-3 fatty acids are of remarkable significance in human nutrition. They have antiatherosclerotic efficacy and other beneficial health effects too. Aquaculture fish, e.g. cyprinids, such as silver carp (Hypophthalmichthys molitrix), bighead carp (Aristichthys nobilis), common carp (Cyprinus carpio), tench (Tinca tinca), and salmonids like Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss) as well as European catfish (Silurus glanis), Baltic whitefish (Coregonus maraena), European seabass (Dicentrarchus labrax), gilthead seabream (Sparus aurata), and Nile tilapia (Oreochromis niloticus), are rich in these fatty acids. When fed on suitable diets, the fatty acid composition of cultured fish can be influenced advantageously. Several clinical tests proved the effectiveness of the consumption of farmed fish in promoting human health. Thus, aquaculture fish can be recommended as wholesome food.     

Minimal water volume for intensively producing male Siamese fighting fish (<Emphasis Type="Italic">Betta splendens</Emphasis> Regan, 1910)

Water volume is a key parameter affecting the individual rearing of male Siamese fighting fish (Betta splendens Regan, 1910). In this study, minimization of water volume was pursued by assessing growth, feed utilization, digestive enzyme activities, color coordinates, muscle quality, and carcass composition. One-month-old solid-red male fish (0.97?±?0.01 g initial body weight) were distributed individually into glass aquaria with five alternative water volumes (100, 150, 200, 250, and 300 mL), comprising 15 fish per treatment (n?=?15), over 8 weeks duration. No mortality of the reared fish was found during the study. Growth performance and feed utilization of the fish reared in 150 mL water were superior to the other treatments. The water volume significantly affected specific activities of the digestive enzymes (P ? 0.05), except for amylase, and no differences in enzyme activities were observed between fish reared in 150 and in 300 mL water. The preferred treatment maintained skin lightness (L*) and had the highest redness (a* and a*/b*) among the treatments. Protein synthesis (RNA concentration) and its turnover rate (RNA/protein ratio) and myosin and actin in muscle also benefited from this treatment. Carcass composition, in terms of moisture, crude protein, and crude ash, was maintained, but the amount of crude lipid fluctuated with water volume. Based on our experiments, the preferred minimal water volume for individual rearing of male Siamese fighting fish should be about 150 mL.     

Effect of <Emphasis Type="Italic">Lactococcus lactis</Emphasis> K-C2 on the growth performance,amino acid content and gut microflora of amberjack <Emphasis Type="Italic">Seriola dumerili</Emphasis>

This study aimed to evaluate the effect of Lactococcus lactis K-C2 on the growth performance, microbial diversity, and release of free amino acids in the intestinal tract and the edible parts of young amberjack, Seriola dumerili. Fish were fed a diet with or without strain K-C2 (2?×?10¹⁰ cfu/g feed) for 25 days. The results indicated that the growth performance of fish in the treated group was significantly higher than those in the control group (p?<?0.05). The amount of five amino acids (aspartate, sarcosine, taurine, alanine, and arginine) in the gut content and 13 of 21 amino acids in the edible parts of fish in the treated group were significantly higher (p?<?0.05) than those in the control group. Sphingomonas, Propionibacterium, and Mycobacterium were observed in gut microflora of fish in both the control and treated groups. Staphylococcus and Kocuria were detected in one sample from the control and treated groups; Acinetobacter and Acidobacteria were found in one sample from the control group. L. lactis was only found in one sample in the treated group. In conclusion, the dietary administration of probiotic L. lactis stimulated growth, reduced feed consumption, and improved the nutritional value of cultured amberjack.     

Correlation between C-reactive protein level,immunology, and hematology of a <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> infected with <Emphasis Type="Italic">Lactococcus garvieae</Emphasis>

The aim of this experiment was to evaluate the symptoms, variation, and correlation of hematological factors and some parameters of the innate immune response of the infected rainbow trout (Oncorhynchus mykiss) fish with Lactococcus garvieae bacteria. A total of 270 fish were divided into three groups including infected fish with a high and low concentration of L. garvieae and a control group without infection. The blood and tissue (brain, head, kidney, and spleen) samples were collected from each group (n?=?6 fish) at 0, 3, 14, and 21 days after treatment. The most observed symptoms during infection were lethargy, exophthalmia, and ascites. The mortality rates for the high and low dose-infected fish were defined as 60 and 25%, respectively. The infected groups had significantly (p?<?0.05) higher activity of serum lysozyme, myeloperoxidase, classical and alternative pathways of complement, serum bactericidal effects, and the specific antibody titer. Also, a significant (p?<?0.05) lower hematocrit and hemoglobin levels and higher white blood cell numbers were observed in the infected groups. A significant correlation was observed between the CRP levels and some of the hematological and immunological indices as bactericidal effects, classical complement pathways, lysozyme activity myeloperoxidase activity, white blood cell numbers, and hematocrit levels. The clinical symptoms, immune responses, and hematological indices variation among L. garvieae-infected fish are dependent on the duration and bacterial dose of the infection.