Lysozyme Activity Research Articles

Quality characteristics, lysozyme activity, and albumen viscosity of fresh hatching duck eggs after a week's storage at various temperatures

The study aimed to analyze the qualitative features of Cherry Valley duck’ hatching eggs during storage at different temperatures. Eggs were divided into 3 equal groups with 30 eggs each: fresh egg and stored at 7 °C and 17 °C within one week. Qualitative analyses of duck eggs were carried out, considering the morphological composition, physicochemical characteristics, lysozyme activity, and albumen viscosity. The highest weight of yolk and its percentage was found in the 17 °C group. The weight and percentage of albumen were significantly the highest in the group of fresh eggs. Higher egg weight loss was observed in the group stored at higher temperatures. Higher thick albumen height and Haugh units were found in fresh eggs and eggs stored at 7 °C. Different temperatures of egg storage did not affect lysozyme activity in thick and thin albumen. Stored eggs were characterized by lower albumen viscosity only at a shear rate of 10 rpm. The higher viscosity of thick albumen compared to thin ones was demonstrated at 10 and 20 rpm shear rates. The presented research results indicate a large diversity of selected qualitative indicators of hatching duck eggs, which may affect their storage and suitability for incubation.

Enhancement of Skin Mucus Immunity, Carotenoid Content, Sexual Parameters, and Growth Response in Guppy Fish (Poecilia reticulata) Fed with Green Algae (Chaetomorpha aerea) Diets

The research aimed to analyze the influences of adding marine green algae Chaetomorpha aerea to the diet of guppy fish (Poecillia reticulate) on growth, immunological responses in skin mucus, total carotenoid content, and sexual characteristics. A total of 450 fish, with a mean body weight of 0.19 ± 0.1 g and 30 fish per tank (triplicate), were randomly fed into 15 experimental tanks, each containing 50 L. Five different diets with 0, 1, 2, 4, 8, and 10% of C. aerea g/kg diets were fed to P. reticulate for 30 days. After 30 days, growth, immunological responses in skin mucus, total carotenoid content, and sexual characteristics were investigated. The results observed that the feed conversion rate and fry output were significantly (p > 0.05) decreased in experimental groups compared to the control group. The results revealed that the dietary inclusion of C. aerea algal significantly increased (p < 0.05) in mucosal immunological parameters containing lysozyme activity, myeloperoxidase activity, total immunoglobulins, and alternative complement activity, which were the highest in the group with 4% of C. aerea g/kg. Additionally, lateral skin and the caudal fin of fish had higher total carotenoid levels from the dietary C. aerea algae diet than the control group, which were the highest in the groups with 4%. Among them, 4 and 8% of C. aerea g/kg diet resulted in better growth performance and feed conversion ratio. Thus, the study suggested that 4% of C. aerea g/kg diet has enrichment of immunity, total carotenoid concentrations, and skin mucus immunity of P. reticulate.

Dietary intakes of vitamin D promote growth performance and disease resistance in juvenile grass carp (Ctenopharyngodon idella).

Vitamin D3 (VD3) is an essential nutrient for fish and participates in a variety of physiological activities. Notably, both insufficient and excessive supplementation of VD3 severely impede fish growth, and the requirements of VD3 for fish vary considerably in different species and growth periods. The present study aimed to evaluate the appropriate requirements of VD3 for juvenile grass carp (Ctenopharyngodon idella) according to growth performance and disease prevention capacity. In this study, diets containing six supplemental levels of VD3 (0, 300, 600, 1200, 2400, and 4800IU/kg diet) were formulated to investigate the effect(s) of VD3 on the growth performance, antioxidant enzyme activities, and antimicrobial ability in juvenile grass carp. Compared with the VD3 deficiency group (0IU/kg), the supplementation of 300-2400IU/kg VD3 significantly enhanced growth performance and increased antioxidant enzyme activities in the fish liver. Moreover, dietary supplementation of VD3 significantly improved the intestinal health by manipulating the composition of intestinal microbiota in juvenile grass carp. In agreement with this notion, the mortality of juvenile grass carp fed with dietary VD3 was much lower than that in VD3 deficient group upon infection with Aeromonas hydrophila. Meanwhile, dietary supplementation of 300-2400IU/kg VD3 reduced bacterial load in the spleen and head kidney of the infected fish, and 1200IU/kg VD3 supplementation could decrease enteritis morbidity and increase lysozyme activities in the intestine. These findings strengthened the essential role of dietary VD3 in managing fish growth and antimicrobial capacity. Additionally, based on weight gain ratio and lysozyme activities, the appropriate VD3 requirements for juvenile grass carp were estimated to be 1994.80 and 2321.80IU/kg diet, respectively.

Influence of dysbiotic syndrome on the formation of experimental nepropathy

Kidneys are sensitive to damage by pathogenic factors of dysbiotic syndrome, among which lipopolysaccharide is the main one. However, the influence of the dysbiotic syndrome on the formation of nephropathy has not been studied enough. The aim of the study is to determine the effect on the kidneys of rats of an experimental dysbiotic syndrome, which was modeled using the antibiotic lincomycin in combination with the administration of epinephrine. Material and methods. Experimental dysbiotic syndrome (systemic dysbiosis) was reproduced in 21 Wistar rats (female, 11 months old, average weight 310±25 g) by administration of lincomycin with adrenaline hydrotartrate. The activity of urease and lysozyme was determined in blood serum and tissue homogenates and the degree of dysbiosis was calculated. The condition of the kidneys was assessed by the activity of elastase, urease, lysozyme, catalase, the content of malondialdehyde (MDA), creatinine, indicators of antioxidant protection. Results. The level of urease increases in the liver of rats with experimental dysbiosis by 2.3 times, in the mucous membrane of the stomach - by two times, and in blood serum - by 2.3 times. The activity of lysozyme decreases in rats with dysbiosis: in the liver - by 42%, in the stomach - by 36%, and in blood serum - by 32%. Urease activity in the kidneys increases by 76%, lysozyme activity decreases by 33%, which indicates a 2.6-fold increase in the degree of dysbiosis. As a result of dysbiosis in the kidneys, the level of biochemical markers of inflammation significantly increases: elastase by 79.5% and MDA by 18%, catalase activity significantly decreases (by 6%). Conclusions. The combined administration of lincomycin and adrenaline causes the development of a dysbiotic syndrome in experimental animals. Under the condition of modeling the dysbiotic syndrome, an inflammatory-dystrophic process develops in the kidneys, and the level of antioxidant protection significantly decreases. Under conditions of experimental dysbiosis, pathological processes in the kidneys are manifested to a lesser extent than in other organs, possibly due to the high initial level of lysozyme in the tissue.

Evaluation of palmatine as an immunostimulant in diet for genetically improved farmed tilapia (GIFT, Oreochromis niloticus)

Palmatine as a medicinal herbs extract has strong bacteriostatic ability and is a promising source of antibacterial additive. The objective of this study was to assess the feasibility of palmatine as an antibiotic alternative in diet for genetically improved farmed tilapia (GIFT, Oreochromis niloticus). Four experimental diets were formulated with 0, 200, 400, and 800 mg/kg palmatine. Tilapia with consistent size (initial body weight 3.43 ± 0.02 g) were fed the test diets twice daily for ten weeks. Dietary palmatine inclusion regardless of levels unaffected the growth performance and feed efficiency (P > 0.05). However, dietary 200–800 mg/kg palmatine inclusion reduced the levels of total cholesterol and low-density lipoprotein cholesterol as well as the ratios of low-density/high-density lipoprotein cholesterol and aspartate/alanine aminotransferase in serum (P < 0.05), but generally improved the antioxidant-related enzymes activities in intestine, plasma and liver, the immunoglobulin M, complement 3, and complement 4 contents and lysozyme activity in plasma, and the phagocytic activity of macrophages in head kidney. Bacterial challenge test demonstrated that dietary supplementation with 200–800 mg/kg palmatine enhanced the resistance of tilapia to Aeromonas hydrophila, and the relative percentage survival was the highest in fish fed diet with 400 mg/kg palmatine. In conclusion, dietary palmatine supplementation enhanced the antioxidant and innate immune responses as well as the resistance against A. hydrophila without compromising growth performance of tilapia, and the suitable inclusion level of palmatine was 400 mg/kg of diet for tilapia.

Toxic effect of chromium on nonspecific immune, bioaccumulation, and tissue structure of Urechis unicinctus.

The contamination of toxic heavy metals in aquatic environments has garnered significant global attention due to its detrimental effects on marine organisms and human health. Hexavalent chromium is a typical environmental and occupational heavy metal pollutant, identified as carcinogenic heavy metal. This study aimed to assess the impact of different Cr (VI) concentrations (0.05-2.5 mg/L) on Urechis unicinctus (U. unicinctus) by investigating bioaccumulation, antioxidant defense system, expression of resistance-related genes, and histological issues. A clear concentration-effect relationship was observed in the bioaccumulation of Cr (VI) in muscle tissues of U. unicinctus. Moreover, exposure to Cr (VI) can alter the activities of lysozyme (LSZ), catalase (CAT), and superoxide dismutase (SOD) to enhance cellular defense mechanisms in U. unicinctus. Likewise, maintained the normal protein structure and functional stability by regulating protein folding. The heat shock cognitive protein (HSC70) gene showed an upward and then downward trend after Cr (VI) exposure. At 12 h, the HSC70 gene expression reached the maximum values of 4.75 and 4.61-fold in the 0.1 and 1.5 mg/L groups, respectively. The organism produced a large number of free radicals, and elevated level of metallothionein (MT) was used to scavenge free radicals and alleviate oxidative stress. Additionally, histopathological examination revealed disorganization in the midgut, atrophic changes in intestinal connective tissue, uneven distribution in respiratory tissues, and irregular shape with a significant reduction in epithelial cells within the gastric cavity. These findings can serve as a valuable reference for elucidating the toxicity mechanisms of heavy metals towards marine benthic organisms and enhancing water environment monitoring strategies.

Effects of dietary bacterial culture of fengycin-producing Bacillus subtilis on growth performance, intestinal and hepatopancreas health, intestinal microbiota and disease resistance of common carp (Cyprinus carpio)

Probiotics are important for the normal physiological functions of fish and are used as feed additives in many aquatic animals. The purpose of this study is to explore the effects of dietary bacterial culture of fengycin-producing Bacillus subtilis (B. subtilis) on growth performance, intestinal and hepatopancreas health, intestinal microbiota and disease resistance of common carp (Cyprinus carpio). Three diets were formulated with supplementation of bacterial culture of fengycin-producing B. subtilis at 0, 0.3 and 0.5 g/kg levels, and these diets were defined as control, BS0.3 and BS0.5, respectively. After 6-week feeding, the infiltration of inflammatory cells was improved in the BS0.3 and BS0.5 groups. And the anti-inflammatory cytokines (interleukin (IL)-10 and tumor growth factor β (TGF-β) were prominently increased in the hepatopancreas of fish fed with dietary B.subtilis diets (P < 0.05). The activity of lysozyme, the levels of complement component (C) 3 and C4 were remarkably increased in the hepatopancreas and intestine of fish fed with B.subtilis supplemented diets (P < 0.05). Histological (intestinal villi height was increased in BS0.3 and BS0.5 groups) (P < 0.05) and the expression of the gut physical barrier-related genes (Zona occludens-1 (ZO-1) and occludin) were increased in the B.subtilis supplemented groups (P < 0.05). The expression of pro-inflammatory cytokines (IL-1β, IL-6 and tumor necrosis factor α (TNF-α) and anti-inflammatory cytokines (IL-10 and TGF-β) were remarkably up-regulated in the intestine of fish fed with BS0.3 diet (P < 0.05), while the expression of IL-6 was remarkably diminished in the intestine of fish fed with BS0.5 diet (P < 0.05). Besides, the intestinal microbiota analysis illustrated that in the intestine of the fish groups fed with BS0.3 and BS0.5 diets, the abundance of Fusobacteriota and Cetobacterium were prominently increased (P < 0.05). After challenged with Aeromonas veronii Hm091, the survival rate of fish fed with BS0.3 diet was markedly improved (P < 0.05). Based on these results, supplementation of bacterial culture of fengycin-producing B. subtilis at a level of 0.3 g/kg has beneficial effects on immunity, intestinal microbiota, and disease resistance of common carp.

Diet with optimal glutathione supplement improves growth, nonspecific immunity, intestinal microbiota, and antioxidant ability in Micropterus salmoides.

In this study, Micropterus salmoides were fed with dietary glutathione (GSH, 0, 100, 300, and 500 mg/kg) for 56 days to investigate its effects on growth performance, serum nonspecific immunity, liver antioxidant capacity, tissue morphology, and intestinal microbiota. The results showed that the survival rate, weight gain rate, and specific growth rate and condition factor increased, whereas the feed conversion ratio, hepato-somatic index, and viscerosomatic index decreased in the GSH groups. Compared with the control group, the serum total protein content significantly increased, whereas the triglyceride and total cholesterol significantly decreased in the 300-mg/kg dietary GSH group. The activities of lysozyme, alkaline phosphatase, and acid phosphatase were significantly higher in GSH-supplemented groups, peaking at 300-mg/kg GSH. GSH supplementation significantly increased total antioxidant capacity and decreased malondialdehyde content, with the most pronounced effects at 300-mg/kg GSH. Further antioxidant indicators showed that a dietary supplement of 300-mg/kg GSH significantly increased the activities of superoxide dismutase, glutathione transferase, endogenous glutathione, glutathione reductase, and catalase. At 300-mg/kg GSH, the liver exhibited improved characteristics with alleviated vacuolation and hepatocyte nuclear shift, and intestine showed enhanced structure with increased villus height and intestinal wall thickness. Additionally, a 300-mg/kg GSH supplementation improved the diversity of intestinal microbiota, increased the abundance of probiotics such as Bacillus, and inhibited the development of pathogenic bacteria such as Plesiomonas. Overall, the results suggest that the effect of GSH addition on improving growth performance, nonspecific immunity, antioxidant capacity, and intestinal microbiota of M. salmoides is best in the 300-mg/kg addition group. Based on second-degree polynomial regression analysis of weight gain, the optimum requirement of dietary GSH in M. salmoides is a 336.84-mg/kg diet.

Assessing safety, efficacy and residue depletion in golden mahseer, Tor putitora (Hamilton, 1822): biochemical and physiological responses to graded concentrations of oxytetracycline dietary supplementation.

The safety and effectiveness of oxytetracycline can potentially manage bacterial infections in fish. This, in turn, might reduce the concerns related to its use in aquaculture and human consumption, such as toxicity, antimicrobial resistance, and other associated risks. The primary objective of this study was to assess how adding oxytetracycline dihydrate to the diet affects its effectiveness, safety, and the presence of residues in T. putitora. T. putitora fingerlings, subjected to experimental infection with Aeromonas hydrophila at a concentration of 108 CFU mL- 1, received an oral administration of oxytetracycline dihydrate. The oxytetracycline dihydrate was added to the feed (corresponding to 2% of the fish body weight) at concentrations of 44.1, 88.2, 132.3 and 176.4mg Kg- 1 fish body weight per day. This treatment was carried out for 10 consecutive days. The biochemical and physiological responses of T. putitora and efficacy of oxytetracycline dihydrate were determined through estimation of microbial load (CFU mL- 1), haematogram, serum biomarkers, behavioral characteristics, non-specific immunity and residue depletion. Experimentally infected fish showed disease progression and induced histopathological conditions with highest microbial load (CFU mL- 1) in the muscle of both control and treated fish. The fish haematogram showed increased leucocyte and haemoglobin content, influenced by dietary oxytetracycline dihydrate. The fish demonstrated adaptive physiological response to oxytetracycline dihydrate at 44.1 to 88.2mg and resulted in increased albumin and globulin content. The serum-enzyme assay showed significant increase in aspartate aminotransferase (AST), alanine aminotransferase (ALT) and plasma alkaline phosphatase (ALP) activities in the test fish (< 0.05). Oxytetracycline dihydrate at 88.2 to 132.3mg Kg- 1 fish body weight per day recorded higher feed intake (75%), significant survivability (66-68%) and histopathological recovery. The suppressed immune response was manifested with decreased respiratory burst and lysozyme activity. The palatability, treatment of bacterial infection, histopathological changes and survivability by fingerlings of golden mahseer determined the safety and optimized the therapeutic potential of the oxytetracycline dihydrate at 88.2mg Kg- 1 fish body weight per day for 10 days to contain the infection by A. hydrophila. A withdrawal period of 8-d was recommended as oxytetracycline dihydrate concentration depleted below the legal maximum residue limit (MRL 2.0 mg g- 1) in the edible muscle of the golden mahseer reared at an average water temperature of 20°C. This is considered safe for human consumption.

Influence of in situ Biofloc Production on Bio Growth Performance, Physiological Immune Response, Digestive Enzyme Activity, Nutrient Composition and Disease Resistance of Etroplus suratensis

Backgroud: The present experiment was conducted to investigate the effects of insitu jaggery based biofloc on biogrowth performance, physiological immuno response, digestive enzyme activity, nutrient composition and disease resistance of Etroplus suratensis. Method: The fingerlings of pearlspot, E. suratensis with an average body weight of 5.67±0.08 g was stocked at the rate of 150/m3 in treatment and control system. The insitu biofloc was produced in 2.4 ton capacity outdoor tanks using jaggery as carbon source at two different C: N ratio of viz, 10: 1 (B1) and 20:1 (B2), where clear brackishwater system and without carbon source as control. Result: The levels of ammonia nitrogen, nitrite–nitrogen and nitrate–nitrogen were reduced in the insitu biofloc compared to control while TSS and TS were increased significantly in all insitu biofloc compared to control. The physiological immuno responses such as Respiratory burst test, Myeloperoxidase, serum lysozyme and antioxidant enzymes activity Superoxide dismutase and catalase were found to be significantly higher in B2 at end of the trial. Stress parameters glucose and cortisol were significantly lower in jaggery based biofloc system especially in (B2) C: N 20:1 compared to control. The fishes from the jaggery based biofloc groups possessed significantly (P less than 0.05) higher immune status as compared to control. The digestive enzymes activity such as amylase protease and lipase, of fish was higher in biofloc treatment and lower in control. Compare to control the nutrient composition of E. suratensis was significantly higher in the treatment groups. Moreover, jaggery based insitu fishes showed the lower cumulative mortality rate and Enhanced relative levels of protectio after experimental challenge with A. hydrophila compared to control.

The Photoperiod Significantly Influences the Growth Rate, Digestive Efficiency, Immune Response, and Antioxidant Activities in the Juvenile Scalloped Spiny Lobster (Panulirus homarus)

This study aimed to elucidate the effects of different photoperiods (0 L:24 D, 6 L:18 D, 12 L:12 D, 18 L:6 D, 24 L:0 D, “Light (L) and Dark (D)”) on the growth performance and physiological responses of the juvenile scalloped spiny lobster (Panulirus homarus). Over a period of 56 days, parameters such as growth rate, digestive enzyme, immune enzyme, and antioxidant enzyme were meticulously evaluated in 90 lobsters subjected to these varying light conditions. The present study found no significant differences in survival rate (SR), molting frequency (MF), and meat yield production (MYP) among the various photoperiod treatments (p &gt; 0.05). Notably, the highest weight gain rate (WGR) and specific growth rate (SGR) were observed under a 12 L:12 D photoperiod. In the continuous dark phase (0 L:24 D), pepsin (PEP) activity remained high in gastric tissues, while trypsin (TRYP) and chymotrypsin (CHT) activities reached the highest in hepatopancreas tissues. The α-amylase (AMS) activity in the hepatopancreas was most elevated under 18 L:6 D, and the optimal lipase (LPS) activity was recorded under 12 L:12 D. The activity of acid phosphatase (ACP) in the hepatopancreas was highest in the absence of light (0 L:24 D), whereas the activities of alkaline phosphatase (AKP) and lysozyme (LZM) were most effective under the 12 L:12 D photoperiod. The total antioxidant capacity (T-AOC), along with catalase (CAT) and superoxide dismutase (SOD) activities of the hepatopancreas reached the highest at 12 L:12 D. The highest activity of glutathione peroxidase (GSH-Px) was seen under 18 L:6 D. The concentration of malondialdehyde (MDA), a marker of oxidative stress, was found to be highest under 12 L:12 D. Consequently, this specific photoperiod is essential for achieving optimal growth and maintaining appropriate physiological balance in the scalloped spiny lobster during aquaculture. These findings provide a foundational guideline for establishing the lighting environment in the farming of the juvenile scalloped spiny lobster.

Δfur mutant as a potential live attenuated vaccine (LAV) candidate protects American eels (Anguilla rostrata) from Vibrio harveyi infection

The eel farming industry is highly susceptible to Vibriosis. Although various types of vaccines against Vibriosis have been investigated, there is limited research on decreasing the virulence of Vibrions through gene knockout and utilizing it as live attenuated vaccines (LAV). In this study, we aim to develop a LAV candidate against Vibrio harveyi infection in American eels (Anguilla rostrata) using a ferric uptake regulator (fur) gene mutant strain of V. harveyi (Δfur mutant). After the eels were administrated with the Δfur mutant at the dose of 4 × 102 cfu/g body weight, the phagocytic activity of the leucocytes, plasma IgM antibody titers, activity of lysozyme and Superoxide Dismutase (SOD) enzyme, and gene expression levels of 18 immune related proteins were detected to evaluate the protection effect of the LAV. Preliminary findings suggest that the LAV achieved over 60% relative percent survival (RPS) after the American eels were challenged by a wild-type strain of V. harveyi infection on 28 and 42 days post the immunization (dpi). The protection was mainly attributed to increased plasma IgM antibody titers, higher levels of lysozyme, enhanced activity of SOD and some regulated genes encoded immune related proteins. Together, the Δfur mutant strain of V. harveyi, as a novel LAV vaccine, demonstrates promising protective effects against V. harveyi infection in American eels, thus presenting a potential candidate vaccine for fish farming.

Evaluation of CpG-Oligodeoxynucleotides as Immunostimulants and Protecting Agents Against Aeromonas hydrophila in Freshwater Lobster (Cherax quadricarinatus)

MAS disease (Motyle Aeromonas hydrophila) is a prevalent bacterial infection that affects freshwater lobsters. This research aimed to evaluate the potential of CpG-ODN as an immunostimulant and protecting agent in Cherax quadricarinatus lobsters. The study was conducted at the Fish Health Laboratory, Pangkep State Polytechnic, Indonesia. The lobsters were divided into different groups and injected with three types of CpG-ODN (2133, 2006, 1668) or a control group using PBS. Parameters such as Total Hemocyte Count, Phagocytic Index, and Lysozyme Activity were measured at multiple time points, including before and after injection. CpG-ODN 2006 showed significant immunostimulant effects, as evidenced by a notable increase in total hemocyte count, phagocytic index, and lysozyme activity compared to the other CpG-ODN types. On the other hand, CpG-ODN 2133 exhibited potential as a protecting agent against Aeromonas hydrophila, as lobsters injected with this CpG-ODN demonstrated higher survival rates in the challenge test compared to the control group. These findings contribute to our understanding of immunostimulant strategies and protective mechanisms in freshwater lobsters. CpG-ODN, particularly CpG-ODN 2006, shows promise as an effective immunostimulant, while CpG-ODN 2133 exhibits potential as a protecting agent against A. hydrophila. Further exploration of CpG-ODN applications could lead to advancements in disease management in aquaculture.

Effects of dietary Spirulina platensis on growth performance, body composition, haematology, immune response, and gut microflora of stinging catfish Heteropneustes fossilis

The study was conducted to determine the inclusion effect of Spirulina platensis as a feed supplement on growth performance, feed efficiency, body composition, haematology, immune response, and gut microflora of the stinging catfish (Heteropneustes fossilis). Five iso-nitrogenous and iso-lipidic diets were prepared by replacing fish meal (FM) with S. platensis at a rate of 0, 2.5, 5, 7.5, and 10% and referred to as control, SP2.5, SP5, SP7.5, and SP10 diets, respectively. Three replicate groups of fish (average weight of 0.75 g) were reared in glass aquaria (180 L water capacity) and fed with each of the experimental diets up to satiation level twice a day, 7 days a week for 70 days. At termination, growth performance, carcass composition, immune parameters, and gut microflora were assessed. Moreover, ten fish from each aquarium were challenged with Aeromonas hydrophila and cumulative mortality was recorded for 15 days. Significantly (P < 0.05) higher weight gain, specific growth rate (SGR, % day−1), lower feed conversion ratio (FCR), and higher protein efficiency ratio (PER) were recorded in fish fed with SP7.5 and SP10 diets compared to control. Dietary S. platensis inclusion at 7.5–10% levels significantly (P < 0.05) boosted the whole-body protein and ash contents of stinging catfish, while decreasing the lipid and moisture contents compared to the control. Haematological parameters significantly (P < 0.05) improved in the S. platensis supplemented diet compared to the control. Serum lysozyme activity of fish fed with SP7.5 and SP10 diet was significantly (P < 0.05) higher compared to all other diets. Higher inclusion of S. platensis lowered post-challenge mortality and the lowest cumulative mortality (30%) was recorded in fish fed with the SP7.5 diet compared to the control (70%). Pseudomonas, Lactobacillus, and Lentibacillus were the prevalent genera in the gut microbiota found in the metagenomics study and the abundances of the above microbiota gradually increased with the level of dietary S. platensis inclusion. Based on the findings of the study, 7.5–10% inclusion of S. platensis was the most recommendable in stinging catfish diet to improve growth performance, feed utilization, body composition, haematology, immunological response, and gut microbiology. However, based on the regression analysis the optimum inclusion level of S. platensis as a feed supplement was calculated to be 9.3–9.9% in stinging catfish.

Effects of Zinc Compounds on the Enzymatic Activities of Lysozyme and Peroxidase and Their Antifungal Activities

This study aimed to investigate the effects of zinc compounds on the enzymatic activities of lysozyme, peroxidase, and the glucose oxidase–mediated peroxidase (GO-PO) system and their antifungal activities. Four different zinc compounds (zinc chloride, gluconate, lactate, and sulfate) were incubated with hen egg-white lysozyme (HEWL), bovine lactoperoxidase (bLPO), the GO-PO system, and human unstimulated whole saliva in solution and on a hydroxyapatite surface. Enzymatic activities of lysozyme, peroxidase, and the GO-PO system were measured through the hydrolysis of Micrococcus lysodeikticus, oxidation of fluorogenic 2′,7′-dichlorofluorescin, and glucose assay, respectively. Interactions between zinc and enzymes were analyzed by surface plasmon resonance (SPR). The minimum inhibitory concentration (MIC) and candidacidal activities of zinc compounds were examined against three Candida albicans strains. Zinc gluconate and sulfate significantly increased the enzymatic activities of salivary lysozyme in the solution assay and of HEWL and salivary lysozyme on the hydroxyapatite surface. However, all examined zinc compounds significantly decreased the enzymatic activities of bLPO and salivary peroxidase in solution and on the surface. SPR analyses revealed binding of zinc to lysozyme and peroxidase, with affinity differing according to the zinc compounds. The MIC of zinc compounds against C. albicans was 1.0–2.4 mM. Candidacidal activities were 17.7–38.8% and 23.7–47.0% at 1.0 and 10 mM concentrations, respectively. In conclusion, zinc compounds enhanced lysozyme activity but inhibited peroxidase activity. Zinc compounds exhibited concentration-dependent candidacidal activity against C. albicans. Zinc compounds are potential therapeutic agents for oral health, especially for geriatric patients.

Comparison of Thermal and High-Pressure Pasteurization on Immunoglobulins, Lysozyme and Microbial Quality of Donkey Colostrum

The effect of thermal pasteurization (TP, 62.5 °C/30 min—conditions similar to those used in milk banks/hospitals, known as Holder pasteurization) and high-pressure pasteurization (HPP: 400–625 MPa/2.5–30 min) was studied on immunoglobulin (IgG, IgM and IgA) content, lysozyme activity and microbial load of donkey colostrum (in this case, after 40 days at 4 °C). IgG level remained unchanged with HPP at 400 MPa, increased up to 4-fold at 625 MPa/10 min and decreased 90% with TP, while IgM decreased progressively with pressure treatment intensity increment to below the detection limit at 625 MPa and decreased 20% with TP. IgA decreased to below the detection limit after TP and all HPP treatments. Lysozyme activity presented overall a higher decrease after TP (37%) compared to HPP (decreasing from 20% at 400 MPa to about 40% at 600 MPa/10 and 30 min). Furthermore, both total aerobic mesophiles and Enterobacteriaceae remained below detection limits after 40 days of refrigerated storage for both TP and HPP. So, HPP can be considered a potential alternative to the conventional TP to preserve donkey colostrum, with overall equal to better retention, particularly for IgG and lysozyme activity. As far as the authors are aware, this is the first study evaluating the effects of HPP on donkey colostrum, and research in this field should be pursued.

A Comparative Study on Effects of Three Butyric Acid-Producing Additives on the Growth Performance, Non-specific Immunity, and Intestinal Microbiota of the Sea Cucumber Apostichopus japonicus.

The providers of butyric acid, Clostridium butyricum (CB), sodium butyrate (SB), and tributyrin (TB), have been extensively studied as aquafeed additives in recent years. However, no comparative study has been reported on the probiotic effects of CB, SB, and TB as feed additives on sea cucumber (Apostichopus japonicus). A 63-day feeding trial was performed to assess the effects of dietary live cells of C. butyricum (CB group, the basal diet supplemented with 1% CB), sodium butyrate (SB group, the basal diet supplemented with 1% SB), and tributyrin (TB group, the basal diet supplemented with 1% TB) on the growth, non-specific immunity, and intestinal microbiota of A. japonicus with a basal diet group as the control. Results indicated that all three additives considerably increased A. japonicus growth, with dietary CB having the optimal growth-promoting effect. Of the seven non-specific enzyme parameters measured in coelomocytes of A. japonicus (i.e., the activities of phagocytosis, respiratory burst, superoxide dismutase, alkaline phosphatase, acid phosphatase, catalase, and lysozyme), dietary CB, SB, and TB considerably increased the activities of six, five, and six of them, respectively. The immune genes (Aj-p105, Aj-p50, Aj-rel, and Aj-lys) expression in the mid-intestine tissues of A. japonicus was significantly increased by all three additives. The CB group had the highest expression of all four genes. Additionally, the relative expression of Aj-p105, Aj-p50, and Aj-lys genes was significantly up-regulated in the three additive groups after stimulation with inactivated Vibrio splendidus. Dietary CB enhanced the intestinal microbial diversity and richness in A. japonicus while dietary TB decreased them. Meanwhile, dietary CB, SB, and TB significantly enhanced the abundance of Firmicutes, unclassified_f_Rhodobacteraceae, and Proteobacteria, respectively, while dietary CB and SB reduced the abundance of Vibrio. Dietary CB and SB improved the stability of microbial ecosystem in the intestine of A. japonicus. In contrast, dietary TB appeared to have a negative effect on the stability of intestinal microbial ecosystem. All three additives improved the intestinal microbial functions associated with energy production and immunity regulation pathways, which may contribute directly to growth promotion and non-specific immunity enhancement in A. japonicus. Collectively, in terms of enhancing growth and non-specific immunity, as well as improving intestinal microbiota, dietary live cells of C. butyricum exhibited the most effective effects in A. japonicus.

Star anise extract supplementation improved growth performance, hepatic-antioxidant enzyme status, hematology, serum biochemistry, and survival against crowding stress in Catla catla

This study was conducted to evaluate the effectiveness of star anise extract (SAE) (prepared using ethanol as the solvent) on growth, proximate composition, hepatic-antioxidant enzyme status, blood-serum indices, and survival against crowding stress in Catla calta. Healthy fingerlings (N = 900) with an average initial weight of 14.16 ± 0.11 g were acclimatized under laboratory conditions and then placed in 5 hapas (60 fish in each) in triplicate. SAE was supplemented in basal feed at 0.0, 0.5, 1.0, 2.0 and 4.0 g/kg and fed to the experimental groups up to satiation twice daily for 90 days. After completion of the feeding trial, 40 fish from each treatment were subjected to crowding stress (60 kg/m3) in triplicate. In the feeding trial, orthogonal polynomial contrast analysis revealed significant enhancement in final weight, weight gain and specific growth in linear and quadratic trends with weight gain percentage, feed conversion ratio and protein efficiency in a quadratic trend observed in response to SAE supplementation. The optimum supplementation level of SAE based on weight gain % was estimated to be 0.93 g/kg using broken line regression analysis (R2 = 0.97) for C. catla. No significant differences in survival rate or proximate composition were observed during the feeding trial. Dietary supplementation with SAE improved the activities of antioxidant enzymes (catalase, superoxide dismutase and glutathione peroxidase) in linear and quadratic trends. The activities of hepatic enzymes, such as alanine transferase, alkaline phosphatase and aspartate transaminase, were also enhanced linearly and quadratically in response to SAE supplementation. Furthermore, hematological indices (red blood cells, white blood cells, hematocrit, hemoglobin and mean corpuscular hemoglobin concentration) were also significantly improved with SAE supplementation. The serum total protein, albumin and globulin concentrations increased linearly and quadratically. Furthermore, stress markers (glucose, cortisol) and triglycerides were significantly reduced linearly and quadratically in response to SAE supplementation. Lysozyme activities also increased linearly and quadratically, indicating a better immune response. After 7 days of crowding stress, the lowest cumulative mortality was observed for the fish fed a diet supplemented with 1.0 g/kg SAE. In conclusion, dietary SAE supplementation significantly improved growth, hepatic-antioxidant enzyme status, blood-serum indices and survival against crowding stress in C. calta.

Biofloc System with Different Carbon Sources Improved Growth, Haematology, Nonspecific Immunity, and Resistivity against the Aeromonas hydrophila in Common Carp, Cyprinus carpio

Biofloc technology (BFT) is one of the most sustainable aquaculture system, which is based on the principle of nutrient recycling and addition of carbon to enable heterotrophic microorganisms to the system. To evaluate the performance of the biofloc culture system for Cyprinus carpio fingerlings, a 60-day growth trial was conducted. The fingerlings (n = 600) of average body weight (4.92 g ± 0.14) were stocked in 12 circular fiberglass tanks (300 L, volume 10.59 cft) to form three biofloc treatments (T1, T2, and T3) along with one control group. The carbon sources for treatments were sugarcane molasses, tapioca, and wheat. The C/N ratio of 15 was maintained for all treatments. After 60 days of rearing, the fish were challenged with Aeromonas hydrophila, and the relative percentage survival (RPS) was observed over 14 days. A haematological, nonspecific immune, and stress parameters were analyzed using blood and serum samples collected at intervals of 20, 40, and 60 days. According to the results, the carbon sources affected the water quality parameters but were still adequate for fish welfare. An increased biofloc volume was observed with tapioca. Growth performance and better feed conversion ratio were recorded in biofloc with the tapioca group. The hematological parameters, including haemoglobin (Hb), hematocrit (HCT), white blood cells and lymphocytes were significantly (P &lt;0.05) higher in biofloc-based tapioca group than in other treatments and control. Further, the serum protein, globulin, albumin, total immunoglobulin, and respiratory burst activity were also found significantly (P &lt;0.05) higher in biofloc with tapioca as carbon source. However, the lysozyme activity was higher in biofloc with the wheat group. The RPS in tapioca was significantly higher, followed by biofloc with wheat. In conclusion, the tapioca-based biofloc can improve C. carpio growth, haematology, and nonspecific immune response under zero water exchange.

Efficacy of nutritional selenium nanoparticles on growth performance, immune response, antioxidant capacity, expression of growth and immune-related genes, and post-stress recovery in juvenile Sobaity seabream (Sparidentex hasta)

This study evaluated the impacts of nano-Se on the growth, immunity, antioxidant capacity, physiological parameters, gene expression, and stress resistance of fingerling Sobaity seabream (Sparidentex hasta). The fish with an average weight of 21.5 ± 0.1 g were divided into four treatment groups in triplicates that received one of the test diets supplemented with varying levels of nano-Se: 0 (control), 0.5 (Se-0.5), 1 (Se-1), and 2 (Se-2) mg/Kg for 60 days. The results showed that final weight, weight gain rate, specific growth rate, feed intake, and feed conversion ratio improved with significant linear and quadratic trends (P < 0.05) in response to nano-Se-supplemented diets, and the best values were measured in the Se-2 group. Superoxide dismutase activity level remained unaffected among the four groups (P > 0.05). Catalase activity increased in nano-Se-supplemented groups, with the highest level measured in fish fed the Se-0.5 diet. Glutathione peroxidase activity levels were not significantly different between the control and nano-Se groups, but the lowest malondialdehyde concentration was detected in the Se-2 group. Nano-Se had no marked effect on total plasma Ig levels; however, the highest lysozyme activity and alternative complement activity (ACH50) were observed in the Se-0.5 and Se-2 groups, respectively. No significant differences (P > 0.05) were observed in plasma total protein, albumin, globulin, triglyceride, and thyroid hormone (T3 and T4) contents among the groups. However, the lowest cholesterol and low-density lipoprotein values and the highest high-density lipoprotein concentration were measured in the Se-2 group. The Se-0.5 and Se-1 groups exhibited significantly lower levels of aspartate aminotransferase activity, and the lowest alkaline phosphatase activity level was detected in the Se-1 group. The expression level of insulin-like growth factor I gene in all nano-Se-fed groups was significantly higher than the control. Also, the expression of interleukin-1β and lysozyme genes was significantly upregulated in nano-Se-supplemented groups, with the highest values in the Se-2 group. Following acute crowding stress, plasma cortisol and lactate levels at all post-stress time intervals were not significantly different among the experimental groups. Fish fed the Se-0.5 and Se-2 diets tended to have lower plasma glucose concentrations than other groups. In conclusion, dietary nano-Se at 2 mg/kg is recommended to promote growth performance and enhance antioxidant and immune parameters in Sobaity juveniles.