Haemato‐immunological response of Labeo rohita (Hamilton) fingerlings fed leaf extracts and challenged by Aeromonas hydrophila

A 35 days feeding trial was conducted to assess the haemato‐immunological response of Labeo rohita fingerlings fed ethanolic leaf extracts of Psidium guajava and Mangifera indica, and infected with Aeromonas hydrophila. Six iso‐nitrogenous (354.6–361.6 g kg^?1) purified diets were prepared with graded level of leaf extracts viz., control (basal feed without any extract); TG‐5 (5 g kg^?1 guava extract); TG‐10 (10 g kg^?1 guava extract); TM‐5 (5 g kg^?1 mango extract); TM‐10 (10 g kg^?1 mango extract); and TGM (5 g kg^?1 guava extract +5 g kg^?1 mango extract). Haematological, immunological, biochemical, along with antioxidant enzyme activities were examined after a 35 day‐feeding trial and following a 7 day challenge with A. hydrophila. The haemoglobin, total leucocyte and erythrocyte counts, respiratory burst activity, lysozyme, total protein, albumin and globulin contents increased significantly (P < 0.05) in leaf extracts fed groups compared with the control in pre‐ and post‐challenge conditions. A significant (P < 0.05) decrease was observed in SOD (superoxide dismutase) and catalase activities of the treatment groups compared with the higher value in control. The trends in mortality indicated that groups of fish showing significantly elevated haemato‐immunological responses had the lowest mortality following challenge with A. hydrophila. The results showed that extracts of P. guajava and M. indica appear to be potential immunostimulant at an inclusion level of 5 g kg^?1 in the diet of rohu. But, mixing of both the extract at similar level did not show any synergistic effect, which needs to be tested at its lower level of inclusion.     

Effect of guava Psidium guajava (L.) aqueous extract diet on growth performance,intestinal morphology,immune response and survival of Oreochromis niloticus challenged with Aeromonas hydrophila

An 84‐day feeding trial was carried out to evaluate the effects of aqueous Psidium guajava leaf extract (PGE) on growth, intestinal morphology, physiology, immune response and susceptibility of Oreochromis niloticus to Aeromonas hydrophila. Diets containing 0% (P0), 0.25% (P1), 0.50% (P2), 0.75% (P3) and 1.00% (P4) PGE were fed to triplicate groups of fish (mean weight; 1.32 ± 0.04 g) for 84 days. After the 84‐day feeding trial, test fish were injected with pathogenic A. hydrophila and then fed for 14 days. More feed were consumed in groups of fish fed PGE‐treated diets and resulted in significantly higher weight gain and feed intake. Incidentally, there was an increase in the calculated area of absorption of fish fed PGE diets, as accounted for by marginally higher villi length and width. Antioxidant and immune response were improved with PGE inclusion in diets as total protein, superoxide dismutase, glutathione peroxidase and glutathione S‐transferase significantly increased (p < 0.05) in fish fed PGE diets. Results of the challenge test with A. hydrophila revealed that the highest mortality (100%) was recorded in P0. This study revealed that inclusion of P. guajava extract in the diet of O. niloticus improved growth, nutrient utilization, immune system and survival of O. niloticus fingerlings.     

Growth and immune response of giant freshwater prawn Macrobrachium rosenbergii (De Man) postlarvae fed diets containing Chlorella vulgaris (Beijerinck)

A 50‐day growth trial was conducted to evaluate the efficacy of Chlorella vulgaris (Beijerinck) as an ingredient in the diets of giant freshwater prawn Macrobrachium rosenbergii (De Man) postlarvae (PL30). Immune response (total haemocyte count and prophenoloxidase activity) was also assessed by subjecting postlarvae to a challenge test against Aeromonas hydrophila (Chester) for 14 days. Iso‐nitrogenous and iso‐lipidic test diets were prepared using a fishmeal‐based‐positive control diet (D0) and four basal diets with inclusion levels of 2% (D2), 4% (D4), 6% (D6) and 8% (D8) C. vulgaris. Postlarvae of M. rosenbergii were randomly stocked (mean initial body weight of 0.19 ± 0.02 g) in 30‐L tanks in three replicates per dietary treatment for evaluation of growth performance. Another set of postlarvae (mean initial body weight of 1.25 ± 0.02 g) was randomly distributed in 95‐L tanks in three replicates per dietary treatment for the assessment of immune response. Results showed that specific growth rate was significantly higher (P < 0.05) in postlarvae fed D4 and D6. Variations in values for carcass protein, lipid, moisture and ash were also evident. Postlarvae fed diets with Chlorella showed increased prophenol oxidase activity and total haemocyte counts. Moreover, survival rate after challenge with A. hydrophila was significantly increased (P < 0.05). Inclusion of C. vulgaris in diets enhanced immune response and resistance of M. rosenbergii postlarvae against A. hydrophila infection.     

Effect of Ocimum sanctum Linn. (Tulsi) extract on the immunity and survival of Labeo rohita (Hamilton) infected with Aeromonas hydrophila

A 60 days study was conducted to evaluate the efficacy of water extract of Ocimum sanctum Linn. leaf on the immune response and disease resistance of Labeo rohita fingerlings against the Aeromonas hydrophila infection. Ocimum sanctum extract was incorporated in the diets (at 0.0%, 0.05%, 0.1%, 0.2%, 0.5% and 1%) of Labeo rohita, rohu fingerlings (6.6 ± 0.013 g). After 42 days blood, plasma and serum were sampled to determine super oxide anion production, lysozyme activity, total immunoglobulin in plasma, blood glucose, serum total protein, albumin, globulin, albumin:globulin ratio, WBC, RBC, haemoglobin content. Fish were challenged with A. hydrophila after 42 days and mortalities were recorded over 18 days post infection. The results demonstrate enhanced super oxide anion production, lysozyme activity, total immunoglobulin in plasma, serum total protein, globulin, total RBC counts, total WBC counts and haemoglobin content (P < 0.05) in treatments group compared with control group. Dietary O. sanctum extracts of 0.2% showed significantly (P < 0.05) higher protection relative percentage survival (RPS 40.00 ± 5.773%) against A. hydrophila infection than control. These results indicate that O. sanctum leaf extract stimulates the immunity and makes L. rohita more resistant to bacterial infection (A. hydrophila).     

Growth performance,haematological and serum biochemical profiles in rainbow trout (Oncorhynchus mykiss) fed diets with varying levels of lupin (Lupinus albus) meal

A feeding experiment was conducted to study the response of rainbow trout juveniles fed different levels of lupin meal in diets for rainbow trout juveniles. Very limited information is available on the relationship between dietary lupin meal in rainbow trout health status. Therefore, this study aimed to determine the effects of lupin meal inclusion levels (0%, 15%, 30%, 45%, and 60%) on growth performance and health status of rainbow trout juveniles. The experimental diets (LM₀, LM₁₅, LM₃₀, LM₄₅, and LM₆₀) were formulated iso‐nitrogenous (41% crude protein) and iso‐calorific (18% crude lipid). The fish were fed twice a day. As a result, the best growth performance was observed in fish fed with LM₁₅ and LM₃₀ diets. No significant differences were detected among experimental groups in terms of body compositions. The haematological values showed significantly (p < 0.05) lower heamatocrit and mean cellular volume (MCV) in the group of LM₆₀ compared with the other groups. For the other haematological parameters such as haemoglobin, red blood cell and mean cellular haemoglobin studied in the present study no significant differences were observed (p < 0.05). The lupin meal included groups showed significant reduction in total protein (TPROT), triglyceride (TROG), cholesterol (CHOL), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) compared with the control group (p < 0.05). The inclusion of lupin meal did not cause any changes in glucose (GLU), glutamic oxaloacetic transaminase (GOT), and glutamic pyruvic transaminase (GPT) between the treatment groups (p < 0.05). In conclusion, lupin meal might be used in rainbow trout diets up to 30% without any malnutrition effect on growth performance, haemotological and serum biochemical parameters.     

Interactive effects of dietary protein and live bakery yeast, Saccharomyces cerevisiae on growth performance of Nile tilapia, Oreochromis niloticus (L.) fry and their challenge against Aeromonas hydrophila infection

Yeasts used as a probiotic in fish diets could stimulate fish resistance against bacterial infection and could enhance the activities of digestive enzymes in fish guts. In addition to yeast importance, dietary protein is another important part in fish diets that should be carefully optimized to meet fish requirement. It is proposed that the yeast supplementation may enhance the dietary protein turnover and reduce the protein requirement for fish. Therefore, the interactive effects of dietary protein and yeast levels on the growth performance of Nile tilapia, Oreochromis niloticus (L.) fry and their challenge against Aeromonas hydrophila infection was evaluated. In the present study, ten experimental diets were formulated to contain either 35% or 45% crude protein (CP). For each protein level treatment, bakery yeast (Saccharomyces cerevisiae) was supplemented at 0.0, 0.50, 1.0, 2.0, or 5.0 g/kg diet. Fish (0.25–0.48 g) were distributed at a rate of 25 fish per 140-L aquarium. For each diet, triplicate aquaria were fed twice a day, 5 days a week for 12 weeks. Fish growth and feed utilization were significantly affected by either dietary protein or yeast levels alone, while no significant effect of their interaction was observed. The highest fish growth was obtained at 1.0–5.0 g yeast/kg diet at both protein levels; however, the fish performance at 45% CP was better than that fed on 35% CP diets. The optimum feed conversion ratio (FCR) was obtained when fish fed on 1.0–5.0 and 2.0–5.0 g yeast/kg diet at 35 and 45% CP, respectively. The cumulative fish mortality, after interperitoneal injection with A. hydrophila for 10 days, and bactericidal activity was significantly higher in fish fed 35% CP diets than those fed 45% CP diets. Both variables decreased significantly with the increase in yeast levels. The lowest bacterial count and bactericidal activity were obtained in fish fed 5.0 g yeast/kg diet irrespective to dietary protein levels. It could be concluded that the inclusion of live bakery yeast in practical diets could improve the growth performances, feed utilization, and physiological status of Nile tilapia fry and their challenge against A. hydrophila infection. Moreover, fish performance when fed 45% CP diet was better than those fed 35% CP diet. Based on these results, the most suitable yeast level for maximum Nile tilapia growth was determined to be 2.0 g yeast/kg diet with 45% CP diet; however, this level was recommended to stimulate their productive performance and enhances their resistance against A. hydrophila infection.     

Effects of dietary levels of essential oil extracts from Moringa oleifera and Artemisia afra on kidney histology,haemato‐immunological parameters and disease resistance in Clarias gariepinus

This experiment investigated the effect of dietary supplementation with essential oils from Artemisia afra and Moringa oleifera on haemato‐immunological parameters and resistance in Clarias gariepinus (African Sharptooth catfish) (Burchell, 1822) challenged with Aeromonas hydrophila. The experiment lasted for 45 days. Prior to and postchallenge, white blood cells significantly increased (p < .05) from the 3% to 12% M. oleifera and A. afra supplementation. Pre‐challenge, no significant differences (p > .05) in red blood cells, haemoglobin and haematocrit were observed between the control and M. oleifera supplementation, while in the A. afra supplementation, RBC and HCT significantly increased from the 6% to 12%. Postchallenge, RBC, HGB and HCT were significantly lower (p < .05) in the control and the 3% and 6% M. oleifera and A. afra supplementation. Both pre‐ and postchallenge, nitro‐blue tetrazolium levels significantly increased (p < .05) from the 3% to 12% M. oleifera supplementation while no significant differences (p > .05) were observed in lysozyme activity between the control and treatment groups. In A. afra, NBT and lysozyme activities increased significantly (p < .05) from the 6% to 12% supplementation pre‐ and postchallenge. Survival rate increased from the 3% to 12% in M. oleifera and A. afra supplementation groups. Significant kidney histological alterations were noted in the control and the 3% and 6% supplementation of both plants postchallenge, while no major histological changes were observed between treatment groups and the control pre‐challenge. The results demonstrate that EO from M. oleifera and A. afra can improve immunity and enhance survival in C. gariepinus.     

Short-term baths with essential oils of Lippia sidoides,Ocimum gratissimum and Zingiber officinale influence blood parameters and survival of tambaqui (Colossoma macropomum) after infection with Aeromonas hydrophila

This study evaluated the effect of short-term baths with the essential oils (EOs) of Lippia sidoides, Ocimum gratissimum and Zingiber officinale on the survival, haematological, biochemical and histopathological parameters of native fish tambaqui (Colossoma macropomum), challenged with Aeromonas hydrophila. After challenge, the fish infected with A. hydrophila received therapeutic baths of 60 min, for five consecutive days. Treatments were as follows: challenged and not treated; challenged and treated with 10 mg L^?1 of gentamicin; challenged and treated with EO of L. sidoides (2.5 and 5.0 mg L^?1); challenged and treated with EO of O. gratissimum (5.0 and 10.0 mg L^?1); challenged and treated with Z. officinale EO (5.0 and 10.0 mg L^?1). The highest survival rates were 89.5% (fish treated with 5.0 mg L^?1 of O. gratissimum EO) and 75.0% (fish treated with 5.0 mg L^?1 of L. sidoides EO). Reductions in the haematocrit percentage, haemoglobin and erythrocyte number were observed in fish treated with L. sidoides and Z. officinale EOs compared with control fish. Mild to moderate damage to the liver tissue of fish after challenge and EO treatments were observed. It is suggested that baths containing O. gratissimum and L. sidoides EOs consist in a viable treatment to increase survival rates of C. macropomum infected with A. hydrophila.     

Dietary butyric acid improved growth,digestive enzyme activities and humoral immune parameters in Barramundi (Lates calcarifer)

A six‐week feeding trial was conducted to determine the effect of butyric acid (BA) inclusion in diet on performance of Barramundi (Lates calcarifer). Fish (12.0 ± 0.2 g) were fed with four experimental diets contained 0.0 (control), 2.5, 5.0 and 10.0 g BA/kg in triplicate. Fish fed on 5 and 10 g BA/kg diets had higher growth and total alkaline protease and lipase activities than the other treatments. Fillet protein content in BA‐supplemented groups was higher than the control. Liver catalase activity was highest in 2.5 g BA/kg group compared with the other groups. The values of liver superoxide dismutase activity and serum total protein concentration in 5 and 10 g BA/kg treatments were higher than those in the control (p < .05). Moreover, the highest and the lowest levels of serum lysozyme activity were noticed in the 10 g BA/kg and the control groups, respectively (p < .05). Also, the highest respiratory burst activity was observed in fish fed 5 g BA/kg feed. Supplementing diet with 2.5 or 5 g BA/kg resulted in higher serum haemolytic activity in fish compared with the other groups. The number of red and white blood cells increased in fish fed BA‐supplemented diets compared with the control (p < .05). In conclusion, the results of this study showed inclusion of BA at 5 g/kg diet in L. calcarifer juveniles improved growth and upgraded the general health condition by enhancing fish antioxidant enzyme activities and haemato‐immunological responses.     

An evaluation of replacing fish meal with cottonseed meal protein hydrolysate in diet for juvenile blunt snout bream (Megalobrama amblycephala): Growth,antioxidant, innate immunity and disease resistance

This study evaluated effects of fishmeal replacement by cottonseed meal protein hydrolysate (CPH) on growth, antioxidant and immunity of Megalobrama amblycephala and its resistance to Aeromonas hydrophila. Fish (average weight: 38.66 ± 0.08 g) were divided into five groups and fed with five isonitrogenous (320 g/kg crude protein), isolipidic (70 g/kg crude fat) and isocaloric (17.8 MJ/kg gross energy) diets replacing fishmeal with 0%, 1%, 3%, 5% and 7% CPH, respectively. The control diet contained 60 g/kg fishmeal. Final weight and weight gain of fish fed 5% and 7% CPH were significantly lower than that of fish fed control diet (p < 0.05). 3% CPH significantly increased total protein, globulin, acid phosphatase, C3 and C4 contents of plasma and total‐superoxide dismutase and catalase activities of liver (p < 0.05), as well as upregulated the antimicrobial peptides 1 and 2 (Leap‐1 and Leap‐2) expression levels in liver and spleen (p < 0.05). After challenge, the lowest mortality was observed in fish fed 3% CPH, and it was significantly lower than that in fish fed the 7% CPH (p < 0.05). In conclusion, dietary fishmeal replacement by 3% CPH could increase antioxidative capacity, as well as enhance immunity of fish.     

Effects of dietary soy isoflavones on growth,antioxidant status,immune response and resistance of juvenile grass carp (Ctenopharyngodon idella) to Aeromonas hydrophila challenge

The current study was conducted to evaluate the effect of dietary soy isoflavones (SI) on growth performance, antioxidant status, immune response and resistance to Aeromonas hydrophila in juvenile grass carp (Ctenopharyngodon idella). Six diets were formulated to contain 0 (control), 10, 50, 100, 500 or 1,000 mg SI per kg feed. Each diet was randomly allotted to triplicate net cages, and each net cage was stocked with 30 fish. The fish were fed one of the experimental diets to satiation twice per day for 60 days. The results showed that the WGR and DGC of the 500 mg/kg SI‐supplemented group were significantly higher than those of the non‐SI‐supplemented group (p < .05). Serum LZM and IgM activities in the SI‐supplemented groups were improved compared to the control group. SOD and GSH‐Px levels of fish fed the diet containing 500 mg/kg SI were significantly enhanced compared to those of fish fed the control diet (p < .05). Additionally, serum CAT, GSH‐Px and AKP activities in 50, 100 and 500 mg/kg SI‐supplemented groups were significantly higher than that in the control group (p < .05). The expression of most immune‐related genes (including IFN‐γ2, TNF‐α, M‐CSF2, IL‐6, IL‐12p40 and IL‐4) was significantly affected by dietary supplementation of SI. The group fed with 500 mg/kg SI had the highest 7‐day cumulative survival rate after challenge test (p < .05). The current results revealed that dietary inclusion of SI could improve the immune response and resistance against A. hydrophila and the supplementation level is suggested to be 500 mg/kg diet.     

Growth performance,immune response and disease resistance of rainbow trout (Oncorhynchus mykiss) fed Thymus vulgaris essential oils

Efficacy of Thymus vulgaris essential oils was assessed on growth, immune response and disease resistance of rainbow trout (Oncorhynchus mykiss). Fish weighing 10 g were fed with dietary supplemented of the oils at 0.5, 1.0 and 2.0 ml/kg feed for 2 months. Fish fed with the oils at 0.5 ml/kg feed demonstrated a better weight gain and specific growth rate, compared to other treatments (p < .05). Fish fed with 1.0 ml the oils showed the highest up‐regulation of complement component 3 (C3) and (cluster of differentiation 4) (CD4) genes expression (p < .05), while lysozyme gene expression level significantly increased in fish fed with 2.0 ml of the oils. In addition, at the end of the experiment, the expression of C3 and CD4 genes were significantly up‐regulated in fish fed with 1.0 and 2.0 ml of the oils, while IL‐1ß and lysozyme genes expression levels were significantly decreased in fish fed 2.0 ml oils, towards the end of the trial (p < .05). There was a fluctuation in the levels of Alanine aminotransferase, Aspartate aminotransferase and Alkaline phosphatase in all treatments during the experiment. When treated fish were challenged with Aeromonas hydrophila, the highest survival rate was observed in 0.5 ml/kg treatment, followed by 2 and 1 mg/kg treatments. Overall, these findings demonstrated that dietary administration of T. vulgaris oils especially at 0.5 mg/kg feed can be considered as a potential component for enhancing of the growth, immune responses and disease resistance of trout against motile Aeromonas septicemia caused by A. hydrophila.     

Effects of salinity level on the survival,growth, feed utilization,carcass composition,haematological and serum biochemical changes of juvenile Meagre (Argyrosomus regius) (Asso, 1801) grown in ground saltwater

A study was performed to examine the effects of salinity on water quality, fish performance, carcass composition and haemato‐biochemical parameters in juvenile meagre, Argyrosomus regius. Fish (5.0 g) were stocked in fibreglass tanks at four salinity levels: 8‰, 16‰, 24‰ and 32‰, and fed a pelleted diet (47/17 protein/lipid) for 56 days. Results indicated that the growth, feed utilization, carcass composition and haemato‐biochemical parameters of meagre gradually improved with the increase in salinity up to 24‰ and then significantly (p ≤ .05) decreased at 32‰. The survival per cent showed a significant decrease when A. regius exposed to 8‰ salinity. An improvement with 32%, 47% and 34.1% of FCR, protein productive value and energy utilization was detected at 24‰ compared with 8‰ salinity respectively. The highest content of protein and the lowest of lipids were recorded in fish carcass at 24‰ compared with the opposite trend at 8‰ salinity. The 24‰ salinity treatment exhibited the highest value of haemoglobin (4.9 g/dl) and the lowest ratio (0.73) of albumin/globulin. The serum total protein, albumin and globulin were significantly higher at 24‰ and 32‰ salinity than those at 8‰ and 16‰ salinity groups. These findings indicate that 24‰ salinity level might be the best for meagre.     

Microcystis aeruginosa (Kütz) incorporated diets increase immunity and survival of Indian major carp Labeo rohita (Ham.) against Aeromonas hydrophila infection

The aim of this study was to evaluate dietary dosages of Microcystis on the immune response and disease resistance against infections due to the opportunistic pathogen Aeromonas hydrophila in Labeo rohita fingerlings. Labeo rohita fingerlings were fed diet containing 0 (Control), 0.5, 1.0 and 5 g Microcystis powder kg^?1 dry diet for 90 days. Three replicate groups of fish averaging 20 ± 2 g were fed for 3 months daily. At 30 days interval, samples were assayed for different biochemical [serum total protein, albumin, globulin, albumin:globulin (A:G) ratio] and immunological (superoxide anion production, lysozyme and serum bactericidal activity) parameters. The results demonstrate that fish fed Microcystis showed increased levels of lysozyme, serum bactericidal activity, serum protein and albumin (P ≤ 0.05) as compared with the control group. After 90 days, fish were challenged with A. hydrophila and mortality (%) was recorded up to day 10 postchallenge. The group fed 1.0 g Microcystis kg^?1 dry diet showed the highest percentage survival (72%). These results indicate that Microcystis aeruginosa stimulates the immunity and makes L. rohita more resistant to infection by A. hydrophila when fed in dried form in feed.     

Immunomodulation and interferon gamma gene expression in sutchi cat fish,Pangasianodon hypophthalmus: effect of dietary fucoidan rich seaweed extract (FRSE) on pre and post challenge period

A 45‐days feeding trial was conducted to study the immunomodulatory effect and interferon gamma gene expression of dietary fucoidan rich seaweed extract (FRSE) from Sargassum wightii on Pangasianodon hypophthalmus fingerlings. One hundred and eighty fingerlings were distributed into six experimental groups in triplicates. Each group was stocked with 10 fish and fed to satiation with iso‐nitrogenous (34.96 ± 0.09–35.18 ± 0.03 CP%) and iso‐caloric (368.65 ± 0.86–375.09 ± 0.26 Kcal/100 g) purified diets containing either 0% FRSE (control), 1% FRSE (TF₁), 2% FRSE (TF₂), 3% FRSE (TF₃), 3% seaweed powder (TS₃) or 6% seaweed powder (TS₆) in the feed. After feeding trial the experimental fish were challenged with Aeromonas hydrophila. Immunological parameters like respiratory burst activity, lysozyme activity, phagocytic activity and total leukocyte count (TLC) were increased with the increasing level of dietary FRSE, whereas serum Albumin/Globulin (A/G) ratio and blood glucose level exhibited decreasing trend (P < 0.05). Increased TLC, blood glucose level, respiratory burst activity, serum A/G ratio, lysozyme and phagocytic activities were recorded during the post‐challenge period. Maximum expression of interferon gamma (IFN‐γ) gene was recorded in FRSE fed groups than the control group both in pre and post challenge condition. After challenged with A. hydrophila the highest survival was observed in TF₂ and TF₃ groups whereas lowest survival was observed in the control group. Hence, dietary supplementation of FRSE at an optimum level of 2% reduced the stress and improved the immune status of P. hypophthalmus fingerlings.     

Effect of using Spirulina and Chlorella as feed additives for elevating immunity status of Nile tilapia experimentally infected with Aeromonas hydrophila

The present trial was performed to evaluate the effect of dietary incorporation of dried Spirulina, Chlorella and their mixture on the immune status of Nile tilapia (Oreochromis niloticus) before and after artificial infection with pathogenic bacterium Aeromonas hydrophila (A. hydrophila). Two hundred and forty fish were divided into four groups: (a) a control group fed on a basal diet only, (b) a second group fed on a diet containing Spirulina 15%, (c) a third group fed on a diet containing Chlorella 15%, and (d) a fourth group fed on a diet containing a mixture of both Spirulina 15% and Chlorella 15%. At the end of the experiment (9 weeks), the four groups were experimentally infected with A. hydrophila for 7 days. Antioxidant enzymes, lysozyme and bactericidal activities and histopathological changes were determined just before the challenge test and 7 days post‐challenge. Significant (p ≤ 0.05) increases in fish body protein% before the challenge test and increases in serum antioxidant enzymes, lysozyme and bactericidal activity in the Chlorella and algal mixture groups before and after the challenge test were observed. Spirulina, Chlorella and their mixture groups significantly decreased serum malondialdehyde compared to the control group before and after the challenge test. Using Spirulina, Chlorella and their mixture mitigated the necrotic and degenerative changes induced by A. hydrophila and revealed well‐developed and multiple melanomacrophage centres. Thus, dietary Spirulina, Chlorella and their mixture inclusion in Nile tilapia fish proved to have a protective effect against A. hydrophila infection.     

Effects of dietary humic acid on growth performance,haemato‐immunological and physiological responses and resistance of Rainbow trout,Oncorhynchus mykiss to Yersinia ruckeri

This study investigated the effects of dietary humic acid sodium salt on growth performance, haemato‐immunological and physiological responses, and resistance of rainbow trout, Oncorhynchus mykiss to Yersinia ruckeri. The experimental fish were divided into four groups; three of them were fed with humic acid incorporated diets (0.3% H3, 0.6% H6, 1.2% H12) and an additive free basal diet served as the control. Growth performance and haematological parameters of rainbow trout were not affected by humic acid supplemented diets (p > 0.05). However, dietary humic acid especially with 0.6% incorporation significantly increased stomach pepsin, intestinal trypsin and lipase activities p < 0.05. Following 60 days of feeding trial, fish were challenged with Yersinia ruckeri for 20 days. At the end of the challenge period, significantly higher (p < 0.05) survival rates were found in the 6% humic acid group compared to all other experimental treatment. Thus humic acid might replace antibiotics in diets for rainbow trout to control yersiniosis.     

Dietary supplement of Enterococcus faecalis on digestive enzyme activities,short‐chain fatty acid production,immune system response and disease resistance of Javanese carp (Puntius gonionotus,Bleeker 1850)

This study investigated the effects of Enterococcus faecalis on digestive enzyme activities and short‐chain fatty acid production in fish intestine, resistance against Aeromonas hydrophila and humoral immunity response by 3 experiments on Javanes carp (Puntius gonionotus). The experiment 1 revealed that diet supplemented with E. faecalis significantly (P < 0.05) increased protease and lipase activities compared to control fed fish. Moreover, E. faecalis supplementation significantly enhanced the production of propionic and butyric acid in the intestine, while no significant difference (P > 0.05) in acetic acid production was observed. In the challenge study (experiment 2), fish were injected (intraperitoneal) with 10⁷ A. hydrophila per ml and survival was significantly improved when fish were fed diet supplemented with E. faecalis compared to control fish. In experiment 3, dietary E. faecalis affected immune system response as fish fed the probiont and exposed to 10⁶ A. hydrophila per ml displayed significantly elevated antibody levels compared to control fed fish. Fish fed diet supplemented with E. faecalis but not exposed to the pathogen revealed significantly higher antibody level than control fish (P < 0.05). Therefore, E. faecalis can be used as a probiotic in Javanese carp farming.     

Nanoparticles of Arthrospira platensis improves growth,antioxidative and immunological responses of Nile tilapia (Oreochromis niloticus) and its resistance to Aeromonas hydrophila

Several studies investigated the usage of spirulina (Arthrospira platensis) in aquaculture as a functional dietary additive or plant proteins replacers; however, more efforts still needed to test its nano form inclusion in aquafeed. In this study, Nile tilapia (Oreochromis niloticus) fed dietary spirulina (A. platensis) nanoparticles (SNP) at 0 (control), 0.1, 1 and 10 g/kg diet while the fifth diet was mixed with 10 g/kg of the normal form of A. platensis as a positive control. The final weight (FW), weight gain (WG) and feed conversion ratio of fish fed SNP at 10 g/kg displayed improved values comparing with the other groups (p < 0.05). Further, fish fed 1 g SNP had higher FW and WG than the control and 0.1 g SNP and 10 g normal A. platensis. The incorporation of SNP at 1 and 10 g/kg in Nile tilapia diets significantly (p < 0.05) enhanced total protein, albumin, globulin, lowered the values of serum glucose and activities of ALT and AST compared with control and spirulina groups. The SOD, CAT, GPx, lysozyme and respiratory burst activities were meaningfully promoted (p < 0.05) in fish received 1 and 10 g/kg diet concerning the control and spirulina groups. Nile tilapia challenged with Aeromonas hydrophila showed congestion of the kidney, liver and spleen tissues while SNP alleviated the inflammation induced by A. hydrophila. Different levels of SNP-incorporated group showed lower cumulative mortality than the other groups. In conclusion, the obtained results illustrated that spirulina in nano form is recommended to enhance Nile tilapia well-being.