Inorganic nitrogen control,growth, and immunophysiological response of <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis> (Boone, 1931) in a biofloc system and in clear water with or without commercial probiotic

A 5-week feeding trial was conducted to evaluate the effects of biofloc in situ and commercial probiotic supplementation on white shrimp (1.87?±?0.03 g) inorganic nitrogen control, growth, and immunophysiological response. For this purpose, four treatments were conducted: clear water with no probiotic application (CW), clear water with probiotic application (CW+P), biofloc with no probiotic application (FLOC), and biofloc with probiotic application (BFT+P); each group had three replicates. Growth parameters (final body weight, daily weight gain, specific growth rate) were significantly higher in the two biofloc systems (P?<?0.05), and the FLOC and FLOC+P group did not have a significant difference (P?>?0.05). The immune responses (total hemocyte count, complement component protein, and lysozyme) and antioxidant status (glutathione, catalase) in the CW+P, FLOC, and FLOC+P groups were increased significantly at the end of the experiment compared with the CW group (P?<?0.05), and the FLOC and FLOC+P groups did not have a significant difference (P?>?0.05). Results of a 10-day Vibrio harveyi challenge test show that the survival rates in CW+P, FLOC, and FLOC+P groups were significantly higher (P?<?0.05), and the FLOC and FLOC+P groups did not have a significant difference (P?>?0.05). These results suggest that probiotic addition in the biofloc system had little advantage, but probiotics can improve the immune status of the shrimp in the clear water system. Further, cost-effectiveness analysis showed that the biofloc system was an efficient and economical option for the production of white shrimp.     

The effect of different alkalinity levels on <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis> reared with biofloc technology (BFT)

The initial stages of rearing marine shrimp using biofloc technology (BFT) involve the biofloc formation process. At the same time, there is an increase in the levels of total suspended solids and a decrease in alkalinity and pH. This reduction of alkalinity and pH occurs due to the consumption of inorganic carbon by the autotrophic bacteria present in the bioflocs and biofilms. The aim of this study was to evaluate the effects of different alkalinities on water quality and the zootechnical performance of the marine shrimp Litopenaeus vannamei in a BFT system. The experiment consisted of four treatments, with three replicates each: 75, 150, 225 and 300 mg CaCO₃/L. To maintain the alkalinity at the established level, sodium bicarbonate was applied. For the experiments, twelve experimental units (area = 0.20 m²) with an effective volume of 50 L were stocked with 30 juvenile L. vannamei (0.20 ± 0.07 g), to achieve a stocking density of 150 shrimps/m² and were maintained for an experimental period of 49 days. The 75 treatment presented the highest levels of ammonia and nitrite throughout the study, compared to the 150 and 300 treatments. The results showed that higher alkalinity favors biofloc formation and the establishment of nitrifying bacteria.     

Effects of biofloc on growth performance,digestive enzyme activities and liver histology of common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.) fingerlings in zero-water exchange system

Biofloc technology is considered as a method that degrades organic waste by microorganisms and produces microbial flocs. A 30-day experiment was performed to investigate the effects of partial replacement of daily feeding intake with biofloc on the growth performances, digestive enzymes activity and liver histology of the common carp Cyprinus carpio L. fingerlings. Two hundred and eight healthy fingerlings (58.6 ± 0.2 g) were randomly distributed in 12 tanks (30 L) at a density of 25.4 kg m^?3 and fed experimental treatments (100 % daily feeding rate as a control, biofloc + 75 % daily feeding rate, biofloc + 50 % daily feeding rate, biofloc + 25 % daily feeding rate). At the end of experiment, the results indicated that the highest weight gain was observed in the fish fed BFT 75 % and control which differed significantly from those fed BFT 25 % (P < 0.05). Diet BFT 75 % improved total protease and pepsin activity compared with BFT 25 and 50 % (P > 0.05). No significant difference was observed in case of lipase, amylase and alkaline phosphatase activity between the treatments. In the liver, histological alterations were found in the treatments, and feeding the fish with BFT 75 % significantly improved hepatocellular quantification and qualification than the other groups. The results obtained in this experiment suggest that the biofloc improves growth performances, digestive enzyme activity and liver condition of the common carp fingerlings when 25 % of daily feeding rate (BFT 75 %) was replaced with one carbohydrate such as molasses in zero-water exchange system.     

A growth comparison among three commercial tilapia species in a biofloc system

Despite a worldwide boom in tilapia aquaculture, South Africa has failed to follow suit, resulting in a small, very much undeveloped local industry. Much of the justification for this reality lies on the inadequacy of the species used and the stringent legislation preventing the use of the Nile tilapia, Oreochromis niloticus. With momentum gained in recent years, the local industry lobbying for the legalization of the O. niloticus has made great progress and its use in certain systems has recently been facilitated. This study was conducted to compare the growth performance between the indigenous Mozambique tilapia, Oreochromis mossambicus, another African alternative gaining momentum in Zambia, the Three Spot tilapia, Oreochromis andersonii, and the globally reputable O. niloticus. The study was conducted in a biofloc technology (BFT) system, showing great potential as an environmentally friendly technology. The trial was run for 10 weeks with 60 fish/tank stocked into four repeats tanks per species. Fish were sampled on six occasions with a regression fitted to the mass data. Oreochromis niloticus showed a significantly higher growth rate with an average daily gain of 0.693 ± 0.018 g/day and the lowest feed conversion ratio (FCR) of 1.01 ± 0.05, followed by O. mossambicus with average daily gain of 0.405 ± 0.025 g/day and FCR of 2.24 ± 0.16, and then O. andersonii with an average daily gain of 0.185 ± 0.025 g/day and an FCR of 2.53 ± 0.28, respectively. From this study, O. niloticus therefore appears to be the most adequate species for use in BFT systems in South Africa.     

Performance of <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis> postlarvae reared in indoor nursery tanks under biofloc conditions at different salinities and zero-water exchange

Shrimp farming at low salinity is a trend that will continue to grow globally. Performance of Litopenaeus vannamei postlarvae in the nursery at different salinities with a biofloc technology (BFT) system needs to be explored further, as the nursery is currently used as a transitional stage between the hatchery and grow-out ponds. Hence, this study evaluated the effect of seven salinity levels (2, 4, 8, 12, 16, 25, and 35 ‰) on the performance of L. vannamei postlarvae reared with a BFT system and zero-water exchange at 2000 org/m³. Additionally, this study evaluated the water quality of all salinity treatments. After 28 days of culture, the findings showed that, under biofloc conditions, salinity affected the performance of some variables of water quality in some cases, but only the combination of a high nitrite-N concentration (>4 mg/l) and low salinity (2 and 4 ‰) caused up to 100 % shrimp mortality in the first 2 weeks. In the rest of the treatments (8, 12, 16, 25, and 35  ‰), shrimp survival was >72 %. Shrimp mortality was affected by salinity, especially when it decreased from 35–25–16 to 12 and 8 ‰. The organisms reared at low salinities presented lower final weights and specific growth rate than those reared at higher salinities. An inverse relationship was shown between the ion concentration and the final weight of shrimp.     

Genetic,nutritional and pathological investigations on the effect of feeding low protein diet and biofloc on growth performance,survival and disease prevention of Indian white shrimp Fenneropenaeus indicus

This experiment was carried out to investigate the effect of feeding low protein diets (^~20BFd_20.13; ^~22BFd_22.20; ^~24BFd_24.32, and ^~26BFd^26.44) in the presence of the biofloc on the growth performance and feed utilization of (Fenneropenaeus indicus) in comparison with commercial feed of 35% CP as control. The present study was based on application of genetic, nutritional and pathological tools. The biofloc was developed in the low protein diet using molasses as a carbon source. Fifty juveniles F. indicus with an average body weight of 0.52?±?0.03 g were stocked in 50 L plastic tanks. Low protein diets and the control were tested in three replicates over a 112 days feeding trial. There were significant differences in protein, amino acids profile, lipids and fatty acids profile between the control and low diets group in the presence of the biofloc. However, less variation was noticed in the whole body composition (protein, amino acid profile, lipids and fatty acid profiles) of the shrimp between the control and low diets groups in the presence of the biofloc. There were no significant differences in final weight between control and low diets groups. Same trend was noted in the SGR, which did not vary significantly between low protein diets and control. However, the utilization of the biofloc by shrimp as a feed source was evident by the significant differences (P<?0.05) in FCR between low protein diets and control. There was a significant difference in the shrimp survival (%) (P<?0.05) between the low protein diets and the control which ranged between 86.66 % and 66.66 %, respectively. The development of biofloc significantly reduced the TAN, nitrate NO₃-N and nitrite NO₂-N levels in the low protein diets tanks. Diagnostic of mortality cases revealed that biofloc can serve as a biosecurity system for shrimp farmimg. RAPD-PCR were used to study the epidemiology of Vibrio parahaemolyticus responsible for early mortality syndrome (EMS) isolated from the water of culture system. The OPC5 (GATGACCGCC) primer produced bands ranged from 1 to 8 with sizes from 0.2–5.0 kb.     

Effects of <Emphasis Type="Italic">Euphorbia hirta</Emphasis> plant leaf extract on growth performance,hematological and organosomatic indices of hybrid catfish, <Emphasis Type="Italic">Clarias macrocephalus </Emphasis>× <Emphasis Type="Italic">C. gariepinus</Emphasis>

This study was conducted to evaluate the effects of Euphorbia hirta leaf extract on the growth performance, hematological and organosomatic indices of hybrid catfish, Clarias macrocephalus?×?C. gariepinus. The fish were treated with 0 (control), 300, 500 and 800 mg/kg (ppm) for 90 days. The weight gain, average daily growth rate, and specific growth rate were at significantly higher levels in fish from all the treatment groups on days 75 and 90, while the feed efficiency and protein efficiency ratio were consistently higher in fish from all the treatment groups from day 60 up until day 90. The feed conversion rate significantly decreased from day 60 until day 90 in all treatment groups when compared with the control group, and the survival rate was significantly different from day 30 until day 90; a consistently higher rate was observed in fish fed 800 mg/kg. The highest viscerosomatic index and intraperitoneal fat were observed in the group fed 500 mg/kg (p?<?0.05). The hepatosomatic index was significantly increased alongside increasing levels of E. hirta extract. The total white blood cell count in the control group was significantly higher on day 30, but on day 90 all the treatment groups showed higher levels. Hematocrit percentage was significantly different on days 30 and 90. Lymphocyte, eosinophil and thrombocyte levels were shown to be significantly different (p?<?0.05) when different groups of fish were compared. In conclusion, 300 mg/kg of E. hirta leaf extract could improve growth performance, hematological and some organosomatic indices in hybrid catfish.     

Complete replacement of fish meal by other animal protein sources on growth performance of <Emphasis Type="Italic">Clarias gariepinus</Emphasis> fingerlings

To completely replace the fish meal by a mixture of earthworm and maggot meals, experimental diets were tested during 42 days on Clarias gariepinus fingerlings. Five isoproteic and isoenergetic diets (40 % crude protein and 17.9 ± 0.3 kJ g^?1) including the control diet (D1) based on fish meal, were formulated. All these diets satisfied the essential amino acids requirements of C. gariepinus fingerlings. These diets were tested on triplicate groups of 50 fishes (initial body weight: 3 ± 0.1 g) bred in tank (0.5 m³). The approximate ratios 2:5; 1:4; 1:12 and 0:1 between the earthworm meal and the maggot meal were used, respectively, to formulate four diets D2, D3, D4 and D5 without fish meal. After the feeding period, significant differences (P < 0.05) were observed on growth, feed utilization between control diet (D1) and test diets (D2–D5). Fish fed earthworm- and maggot-based diets were grown better than those fed the control diet. Survival and feed utilization were not significantly affected by the ratio between earthworm meal and maggot meal in the test diets. Lipid content was higher in carcass and fillet of fishes fed earthworm- and maggot meals-based diets than that of those fed fish meal-based diet. This study indicates that when the ratio 2:5 between the earthworm meal and the maggot meal is used to entirely replace fish meal and the ratio lysine/arginine of the diet is inferior to 1, the growth performances and feed utilization of Clarias gariepinus fingerlings are improved.     

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.     

<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.     

Dietary ascorbic acid reduced micronucleus and nuclear abnormalities in <Emphasis Type="Italic">Clarias gariepinus</Emphasis> (Burchell 1822) exposed to hospital effluent

Hospital effluents contain myriad of mutagens and genotoxins capable of increasing DNA damage in aquatic biota. African mudfish, Clarias gariepinus, are exposed to genotoxins when cultured in swamps and derelict water bodies often contaminated by effluents. Moreover, its DNA is susceptible to xenobiotic-induced lesions since it lacks l-gulonolactone oxidase and hence cannot synthesize l-ascorbic acid. This study investigated 96-h acute toxicity and protective effects of dietary ascorbic acid (AA) against micronucleus (MN) and abnormal nuclear (NAs) formation in C. gariepinus exposed to sub-lethal concentrations of hospital effluent. Six concentrations (0.5–3.0%) of the effluent were selected to determine the 96-h acute toxicity of the effluent in C. gariepinus, after range finding test. Fish were exposed to sub-lethal concentrations (0.08–1.30%) of the 96 h LC₅₀. Two other groups were exposed to the 96 h LC₅₀ (1.30%) of the effluent +50 and +100 mg/kg of dietary ascorbic for 7 days, and MN and NAs assessed in peripheral erythrocytes. The 96 h LC₅₀ (1.30%) was 1.18 times more toxic than the 24 h LC₅₀ (1.54%), indicating that the toxicity of the effluent increased with exposure duration. MN, nuclear bud, enucleated, fragmented nucleus (apoptosis), and necrotic erythrocytes significantly increase in effluent treated fish. Dietary AA reduced MN from 6.35-fold (1.30% treated group) to 3.72-fold (1.30% + 50 mg AA) and 3.54-fold (1.30% + 100 mg AA). Also, AA reduced total NAs from 2.26-fold (1.30%) to 1.40-fold (1.30% + 50 mg AA) and 1.06-fold (1.30% + 100 mg AA) compared to the control. Heavy metals and physicochemical parameters analyzed in the tested effluent possibly induced the mortality and cytogenotoxicity in C. gariepinus, and this was ameliorated by dietary AA.     

Alternative microalgal diets for cultivation of the fairy shrimp <Emphasis Type="Italic">Branchinella thailandensis</Emphasis> (Branchiopoda: Anostraca)

Fairy shrimp is known as a nutritional food for fish and crustaceans in aquaculture. In most hatcheries, the microalga Chlorella sp. appears to be the most common, suitable, and nutritious food to feed fairy shrimp. In this study, we attempted to determine other alternative algal diets for cultivation of fairy shrimp Branchinella thailandensis. Seven experimental diets including three treatments of dried Spirulina sp. at 0.75 (S1), 1.5 (S2), and 3.0 mg dry weight individual^?1 (S3); three treatments of Chlorococcum humicola at 5 × 10⁵ (Ch1), 1 × 10⁶ (Ch2), and 2 × 10⁶ cells mL^?1 (Ch3); and a control diet (Chlorella vulgaris at 1 × 10⁶ cells mL^?1) were fed to 5-day-old shrimp for 15 days. Evaluation of growth performance, egg production, survival percentage, and nutritional and carotenoid content of the experimental fairy shrimp revealed that Ch3 is the most suitable algal diet. Our results suggest that C. humicola is the best alternative food source for the cultivation of B. thailandensis. In addition, dried Spirulina powder is also a good choice when live algae are not available and can be used as an alternative feed in fairy shrimp cultures.     

Effects of low-voltage constant direct current on plasma biochemical profiles and gene expression levels in crucian carp <Emphasis Type="Italic">Carassius carassius</Emphasis>

This study investigated the plasma biochemical profiles and gene expression levels of crucian carp Carassius carassius subjected to anesthetization with low-voltage constant direct current (DC) and tricaine methanesulfonate (MS222). Fish in the MS222 treatments and the DC treatments were exposed to either two concentrations of MS222 (100 and 200 mg/l) or constant DC with a voltage of 26 V, at a voltage gradient of 0.68 V/cm, respectively. The results showed that low-voltage constant DC immobilized fish more quickly, resulting in significant elevations of plasma cortisol levels compared to exposure of high or low doses of MS222. Electronarcosis led to an increased expression of heat shock proteins 70 (HSP70) as well as HSP90 compared to the treatment group with MS222. Also, electronarcosis enhanced mitochondrial respiratory chain gene (atp6, cox1 and nd5) and antioxidant enzyme gene (CAT, SOD and GSH-px1) expression levels. In conclusion, low-voltage constant DC can disturb fish plasma biochemical profiles and gene expression levels.     

<Emphasis Type="Italic">Curcuma longa</Emphasis> as additive in the diet for <Emphasis Type="Italic">Astyanax aff. bimaculatus</Emphasis>

The aim of the present study was to evaluate the effect of turmeric (Curcuma longa) as additive in the diet for Astyanax aff. bimaculatus. Fish (0.83 ± 0.04 g) were fed, for 60 days, with six diets containing 0.0, 20.0, 40.0, 60.0, 80.0, and 100.0 g turmeric kg^?1 feed. There was an increasing linear effect of turmeric on the thickness of the muscular layer, and height and width of the folds of the intestine. In the liver, a quadratic effect was observed of turmeric on the percentage of hepatocyte cytoplasm and a decreasing linear effect on the percentage of sinusoid capillaries. A quadratic effect was also observed of turmeric on the liver glycogen. There was no effect of turmeric on the antioxidant activity in the liver, carcass composition or productive performance of the fish. Thus, we concluded that Curcuma longa has trophic effects on the epithelium and the muscular layer of the intestine of A. aff. bimaculatus. Additionally, low levels of Curcuma longa cause increased deposition of liver glycogen and high levels cause reduction.     

Pre-sedation and transport of <Emphasis Type="Italic">Rhamdia quelen</Emphasis> in water containing essential oil of <Emphasis Type="Italic">Lippia alba</Emphasis>: metabolic and physiological responses

The effects of transporting silver catfish (Rhamdia quelen) for 6 h in plastic bags containing 0 (control), 30 or 40 µL/L of essential oil (EO) from Lippia alba leaves were investigated. Prior to transport, the fish in the two experimental groups were sedated with 200 µL/L of EO for 3 min. After transport, dissolved oxygen, carbon dioxide, alkalinity, water hardness, pH, temperature and un-ionized ammonia levels in the transport water did not differ significantly among the groups. However, total ammonia nitrogen levels and net Na⁺, Cl^? and K⁺ effluxes were significantly lower in the groups transported with EO of L. alba than those in the control group. PvO₂, PvCO₂ and HCO₃ ^? were higher after transporting fish in 40 µL/L of EO of L. alba, but there were no significant differences between groups regarding blood pH or hematocrit. Cortisol levels were significantly higher in fish transported in 30 µL/L of EO of L. alba compared to those of the control group. The metabolic parameters (glycogen, lactate, total amino acid, total ammonia and total protein) showed different responses after adding EO to the transport water. In conclusion, while the EO of L. alba is recommended for fish transport in the conditions tested in the present study because it was effective in reducing waterborne total ammonia levels and net ion loss, the higher hepatic oxidative stress in this species with the same EO concentrations reported by a previous study led us to conclude that the 10–20 µL/L concentration range of EO and lack of pre-sedation before transport are more effective.     

Effect of dietary chitosan on growth performance,hematological parameters,intestinal histology and stress resistance of Caspian kutum (<Emphasis Type="Italic">Rutilus frisii kutum</Emphasis> Kamenskii, 1901) fingerlings

This study was conducted to investigate the effects of dietary chitosan on growth performance, hematological parameters, intestinal histology, stress resistance and body composition in the Caspian kutum (Rutilus frisii kutum, Kamenskii, 1901) fingerlings. Fish (1.7 ± 0.15 g) were fed diets containing chitosan at different levels (0, 0.25, 0.5, 1 and 2 g kg^?1 diet) for a period of 60 days. Results showed that the feed conversion ratio significantly decreased in fish fed diet containing 1 g kg^?1 of chitosan compared to the other groups (P < 0.05), but there were no significant differences between treatments in terms of specific growth rate and condition factor (P > 0.05). Leukocyte increased in fish fed diet containing 2 g kg^?1 of chitosan compared to the other groups (P < 0.05). Lymphocytes, eosinophils and neutrophils did not significantly change among dietary treatments (P > 0.05). Also, the chitosan supplementation did not affect the whole-fish body composition (P > 0.05). Light microscopy demonstrated that the intestinal villus length increased in fish fed diet containing 1 g kg^?1 of chitosan compared to control group (P < 0.05). While 11 and 13 ‰ salinity and 30 °C thermal stress had no effect, 1 g kg^?1 of chitosan (P < 0.05) showed highest survival rate (70 %) in 34 °C thermal stress. The results showed that chitosan in the diet of the Caspian kutum fingerlings could improve feed conversion ratio, the nonspecific defense mechanisms and resistance to some of the environmental stresses.     

Monitoring of <Emphasis Type="Italic">Francisella noatunensis</Emphasis> subsp. <Emphasis Type="Italic">orientalis</Emphasis> in farmed Nile tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>) in Brazil

Francisella noatunensis orientalis is a bacterium that causes emerging bacteriosis in Nile tilapia (Oreochromis niloticus) in many parts of the world, including Brazil. It is a non-motile, Gram-negative, strictly aerobic, facultative intracellular coccobacillus. This species of bacteria is responsible for low to high mortality in fish farms, causing economic losses for fish farmers. This study aimed to detect the presence of F. noatunensis orientalis using qPCR (real-time polymerase chain reaction) and to describe lesions caused by the bacterium in O. niloticus in Brazilian aquaculture. For this purpose, 360 fish from six fish farms (30 per farm) were sampled at two time points (n = 180 per sampling). Necropsies and histopathology were performed for lesion observation, in addition to qPCR and sequencing for detection and identification of Francisella species. Environmental data were collected using a multiparameter sonde YSI EXO2. All measured limnological variables were within the optimum range for cultivation of Nile tilapia. The major lesions present were melanization of the skin, splenomegaly, granulomas, and inflammatory cell responses. The prevalence of francisellosis varied from 0 to 86.66% between time periods and fish farms analyzed, and an outbreak was observed during the second sampling period. This study describes the prevalence of francisellosis in O. niloticus and reports that the lesions found are not exclusively associated with this bacterial disease.     

Growth and fatty acid composition of <Emphasis Type="Italic">Octopus vulgaris</Emphasis> paralarvae fed with enriched <Emphasis Type="Italic">Artemia</Emphasis> or co-fed with an inert diet

The rearing of Octopus vulgaris paralarvae during its planktonic life stage is a major challenge, as mortality is currently very high and unpredictable. In this study, we examined the survival and growth rates, as well as the fatty acid composition, of O. vulgaris paralarvae fed on three different dietary treatments: group ArDHA was offered juvenile Artemia enriched with a lipid emulsion (Easy DHA-Selco^®); group ArMA was fed with juvenile Artemia enriched with a mixture of microalgae (Rhodomonas lens and Isochrysis galbana); and group ArMA+ID received the same Artemia as group ArMA complemented with an inert diet. Dietary treatments were tested in triplicate with homogenous groups of paralarvae (25 individuals l^?1) established in 50-l tanks, and the experiment was conducted for 15 days. The survival rate of 15-day post-hatch (-dph) paralarvae from groups ArMA (20 ± 8%) and ArMA+ID (17 ± 4%) tended to be higher than in group ArDHA (13 ± 5%), though these differences were not statistically different. The dry weight (DW) of 15-dph paralarvae increased by almost 60% in groups ArMA and ArMA+ID, and nearly 40% in group ArDHA, with respect to hatchlings. The fatty acid (FA) composition of paralarvae revealed a remarkable drop of docosahexaenoic acid (22:6n-3, DHA) from hatchlings to 15-dph paralarvae of all groups (P < 0.05). However, paralarvae from group ArDHA contained higher levels of DHA than those from ArMA and ArMA+ID (P < 0.05). Despite Artemia enriched with DHA-Selco^® contained three-times more DHA than Artemia enriched with microalgae, no beneficial effects of this dietary treatment were observed on the performance of paralarvae.     

Influence of the dietary inclusion of <Emphasis Type="Italic">Gracilaria cornea</Emphasis> and <Emphasis Type="Italic">Ulva rigida</Emphasis> on the biodiversity of the intestinal microbiota of <Emphasis Type="Italic">Sparus aurata</Emphasis> juveniles

Partial substitutions of fish meal by 5, 15, or 25 % of Gracilaria cornea or Ulva rigida in experimental diets were evaluated to study their effects on biodiversity of intestinal microbiota composition in gilthead seabream (Sparus aurata) juveniles. The diets were offered to duplicate groups of 15 juvenile fish (14.0 ± 0.5 g) for 70 days, and at the end of the experiment the intestinal microbiota from four specimens of each treatment was analysed by denaturing gel gradient electrophoresis. Results showed that the substitution of fish meal by algae meal induced important modifications in the intestinal microbiota community, as a big reduction of the biodiversity when the highest percentage (25 %) of U. rigida was included. On the contrary, an increase on the number of species was detected when a 15 % of algae was included. Various Lactobacillus delbrueckii subspecies were selectively stimulated when G. cornea was included in the feed, and other bacterial species, such as those included in the Vibrio genus, were reduced.     

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.