Respiratory Burst Activity Research Articles

Retinoic Acid-Induced HL-60 Cell Differentiation Is Augmented by Copper Supplementation1–4

The objective of this study was to determine how copper influences the ability of HL-60 cells to differentiate into cells of the granulocytic lineage. We hypothesized that granulopoiesis requires copper because copper-deficient humans become neutropenic. Differentiation of HL-60 cells along the granulocytic lineage with retinoic acid was enhanced by copper. The results showed a greater number of cells were more differentiated when copper was added to the medium for 96 h. The respiratory burst activity of retinoic acid-induced cells was increased by copper supplementation, but intracellular superoxide anion generation was not affected. Supplementation with copper resulted in more cell-associated copper in both noninduced and induced cells; however, the induced cells accumulated three times more copper than the noninduced cells. Even though the amount of copper associated with retinoic acid-treated cells was greater than in untreated cells, the activity of a copper-requiring enzyme, copper/zinc superoxide dismutase, was significantly lower. Copper supplementation increased the activity of this enzyme in both retinoic acid-treated and untreated cells. Cytochrome c oxidase activity was not affected by retinoic acid treatment or by copper supplementation. Copper seems to play a specific role during the early stages of granulocyte differentiation.

Evaluation of largemouth bass (Micropterus salmoide) fed selenium yeast diets: Growth, histopathology, antioxidant ability, and apoptosis

The objective of this study was to investigate the effects of selenium yeast on the growth, histomorphology, antioxidant capacity and apoptosis of largemouth bass (Micropterus salmoide). The five iso-nitrogen and iso-lipid feeds were supplemented with selenium yeast levels of 0, 0.5, 1.0, 1.5 and 2.0 mg/kg, respectively. The 525 largemouth bass (initial body weight: 11.48 ± 0.43 g) were equally divided into 15 buckets (three buckets per feed treatment) for a 67-day feeding experiment. The results showed that the addition of 1.5 and 2.0 mg/kg of selenium yeast significantly reduced weight gain rate, and specific growth rate (P < 0.05), disorganized the structure of liver cells and exacerbated the vacuolation of liver tissue in largemouth bass. The activities of glutathione peroxidase and superoxide dismutase in serum were significantly increased by 1.0 mg/kg selenium yeast (P < 0.05). After selenium yeast was supplemented, villus length and villus width were significantly increased (P < 0.05), NO contents and respiratory burst activities in hematocyte were not significantly changed (P > 0.05). With the addition of 1.5 mg/kg of selenium yeast, the relative expression of growth hormone 1, insulin-like growth factor-1, superoxide dismutase, catalase, Kelch-like ECH-associating protein 1, and nuclear factor erythroid 2 related factor 2 in hematocyte was significantly increased (P < 0.05). The significantly lowest percentage of apoptotic hematocyte was also seen in the 1.5 mg/kg selenium yeast addition group (P < 0.05). Overall, the supplementation of 1.5 mg/kg selenium yeast was not conducive to the growth and liver development of largemouth bass, but it could improve the antioxidant capacity of hematocyte and inhibit the apoptosis. It provided a theoretical basis for the enhancement of immune capacity during the farming of largemouth bass.

Potential Symbiotic Effects of β-1,3 Glucan, and Fructooligosaccharides on the Growth Performance, Immune Response, Redox Status, and Resistance of Pacific White Shrimp, Litopenaeus vannamei to Fusarium solani Infection

The potential effects of dietary supplementation with β-1,3 glucan and fructooligosaccharides (β-1,3 GF) on antioxidant activities, immunological response, and growth performance of Pacific white shrimp (Litopenaeus vannamei) was investigated. Four diets (iso-energetic and iso-nitrogenous) with different levels of β-1,3 GF (0, 0.5, 1.0, and 1.5 g kg−1) were fed to healthy shrimp juveniles weighing 3 ± 0.5 g for 75 days. Shrimps were randomly distributed into 12 net enclosures at a density of 30 shrimp/net, and the experiment was performed in triplicate. The results revealed that long-term supplementation with 1.5 g kg−1 β-1,3 GF significantly improved shrimp weight gain, feed conversion ratio, and digestive enzyme profiles compared to the control diet group. However, there were no substantial variations in the contents of moisture, crude protein, total lipids, and ash in the muscles of shrimp fed on different diets. Surprisingly, all antioxidants (superoxide dismutase, catalase, glutathione peroxidase) and immune biomarkers (lysozyme, total hemocyte count, phenol oxidase, and respiratory burst) activities were significantly elevated with increasing levels of β-1,3 GF in the shrimp diet, and the highest values were recorded in the 1.5 g kg−1 diet groups. Challenge test results revealed that F. solani could cause a high mortality rate (86.7%) in a group fed a normal basal diet within 14 days at a dose of 5 × 104 conidia mL−1. Surprisingly, all dietary treated groups with different doses of β-1,3 GF showed high resistance against F. solani, represented by lower cumulative mortality rates (20–43.3%) compared to the control group. Moreover, most of the infected shrimp showed a typical black to brown gill lesion similar to that observed in the natural infection, where an identical fungus was successfully re-isolated from infected gills and muscles. Overall, this study recommends an appropriate incorporation level of β-1,3 GF that could enhance growth performance and improve the antioxidant activities, non-specific immunity, and disease resistance of L. vannamei, with an optimal level of 1.5 g kg−1.

Nano-copper Enhances Gene Regulation of Non-specific Immunity and Antioxidative Status of Fish Reared Under Multiple Stresses.

Arsenic pollution, water temperature, and pH are the major concern for aquaculture. Moreover, the aim of the present investigation was to delineate the role of nano-copper (Cu-NPs) in the mitigation of arsenic toxicity, high temperature (34°C) and low pH (6.5) stress on Pangasianodon hypophthalmus. Four isonitrogenous and isocaloric experimental diets of Cu-NPs at 0, 1.0, 1.5 and 2.0mgkg-1 were formulated and prepared. Arsenic pollution, low pH and high temperature stress significantly reduced the anti-oxidative status (super oxide dismutase, catalase, glutathione peroxidase and glutathione-s-transferase), lipid peroxidation, total anti-oxidative capacity and lipid profiling (cholesterol, total lipid, phospholipid, very low-density lipoprotein and triglyceride). Further, the supplementation of Cu-NPs at 1.5 and 1.0mgkg-1 diets noticeably improve the anti-oxidant status and capacity. The stressors groups (As + pH + T, As + T and As) significantly reduced fish immunity viz. albumin, globulin, total protein, albumin globulin ratio (A:G ratio), myeloperoxidase, respiratory burst activities, tumor necrosis factor, total immunoglobulin, and interleukin. Whereas supplementation of Cu-NPs at 1.5 and 1.0mgkg-1 diets improved the immunity of the fish reared under multiple stresses (As + pH + T). Tail DNA %, DNA damage-inducible protein (DDIP) and inducible nitric oxide (iNOS) synthase gene expression were significantly enhanced with exposure to arsenic, low pH and high temperature but supplementation of Cu-NPs protects the tissues against DNA damage and improved the gene expression of iNOS and DDIP. Cu-NPs at 1.5 and 1.0mgkg-1 diets significantly enhanced the body weight gain %, protein efficiency ratio, specific growth rate, daily growth index, relative feed intake and reduced the feed conversion ratio. Whereas, the growth-related gene expression such as myostatin (MYST), somatostatin (SMT) was downregulated by Cu supplementation and upregulated the gene expression of growth hormone regulator 1 and β (GHR1 and GHR β) and growth hormone (GH) gene in fish. Dietary Cu-NPs supplementation protects the fish against bacterial infection and enhances arsenic detoxification in different tissues. The present investigation revealed that supplementation of Cu-NPs at 1.5 and 1.0mgkg-1 diet has the potential to mitigate multiple stress (As + pH + T) in fish.

Response of hepatic biochemical parameters and neurotoxicity to carbamazepine and ibuprofen in Oreochromis mossambicus.

Globally, the prevalence and pollution of pharmaceutical drugs in aquatic environments have been steadily increasing. This study sought to evaluate the effects of 14days of exposure to environmental-relevant doses (ibuprofen 0.5, 5, and 50µg/L, and carbamazepine 0.005, 1, and 10µg/L) of the nonsteroidal anti-inflammatory drugs ibuprofen and carbamazepine in the freshwater fish Oreochromis mossambicus. The results showed a significant (P < 0.05) decrease in O. mossambicus superoxide dismutase, catalase, biotransformation enzymes, glutathione-s-transferase, glutathione peroxidase, oxidative stress lipid peroxidation, protein carbonyl activity, cellular damage metallothionine, reduced glutathione, immunological activities, and respiratory burst activity. Consequently, the acquired data revealed that O. mossambicus treated with ibuprofen and carbamazepine shows more significant alterations in metabolic depression, biochemical parameters, and oxidative stress. In addition, increased neurotoxic effects were observed in ibuprofen and carbamazepine treated O. mossambicus.

Immunostimulatory effect of mannan-oligosaccharides supplementation diet in milkfish (Chanos chanos)

The current study was designed to examine the impacts of dietary mannan-oligosaccharides (MOS) on growth, hemato-biochemical changes, digestive-antioxidant enzyme activity, immune response, and disease resistance of milkfish (Chanos chanos) fed diets contained MOS i.e. 1g, 2g, and 3g MOS. The growth parameters were significantly influence in milkfish fed all MOS diets, whereas the feed conversion ratio (FCR) and protein efficiency ratio (PER) were significantly influence with 2g or 3g MOS diets. The total protein (TP), globulin (GB), and glucose (GLU) levels, amylase, protease, liver enzymes were found significantly high in fish fed 2g or 3g MOS diets; but, lipase, trypsin, and alkaline phosphatase (ALP) enzymes were increased significantly at 3g MOS diet. All MOS inclusion levels were significantly increased total and Lactobacillus intestinal microflora population. The oxidative enzymes activity as superoxide desmutase (SOD) and catalyze (CAT) were progressively increased with all MOS supplementation diet, but the glutathione peroxidase (GPx) and lactate dehydrogenase (LDH) content were found significantly high in fish fed 2g or 3g MOS diets. Similarly, the reduced glutathione (GSH) and glutathione reductase (GR) contents were observed significantly high level in fish fed 3g MOS diet. The phagocytic (PC) and lysozyme (LYZ) activities were found gradually increase in fish fed increasing level of MOS diets, while the respiratory burst (RB) and malondialdehyde (MDA) activities were seen significant in fish fed 2g and 3g MOS diets. The current research work confirmed that C. chanos fed diets contained 3g kg−1 MOS recorded better growth performance, digestive-antioxidant, immune response, and disease resistance.

Sodium Alginate from &lt;I&gt;Padina australis&lt;/I&gt; Modulates Innate Immune and Immune Gene Expression in Red Tilapia (&lt;I&gt;Oreochromis&lt;/I&gt; sp.)

Highlight Research Development of immunostimulant from natural sources is a promising approach to improve the immune status of fish Dietary administration of sodium alginate from Padina australis at 2.0 g/kg was an effective immunostimulant for red tilapia Red tilapia fed diet with sodium alginate significantly increased innate immune responses of TLC, PA, respiratory burst, SOD, TPP, lysozyme, and ACH50 activity Red tilapia fed diet with sodium alginate significantly modulated immune genes expression of GPx, TNF-α, IFN-γ, IL-10, TGF-β1, and Hepcidin Abstract The study of immunostimulant derived from natural sources has received considerable attention in recent years. Indonesian coasts have various sources of immunostimulant, especially brown algae, which are rich in alginate. This research investigated innate immune response and immune genes in red tilapia (Oreochromis sp.) after being fed diets supplemented with sodium alginates (SA) from a brown alga, Padina australis, originating from Awur Bay, Jepara, Indonesia. This work provided comprehensive information regarding the interaction of cellular-humoral immunity and immune gene expression. Red tilapia was fed with SA-containing diet at doses of 0, 2, 3 and 4 g/kg for 12 days, then blood samples were collected on days 0, 3, 6, 9, and 12 to evaluate the total leucocyte count (TLC), phagocytic activity/index (PA/PI), respiratory burst, superoxide dismutase (SOD), lysozyme, alternative complement (ACH50) activity, total plasma protein (TPP), and immune genes expression (blood sample on day 6). The immunological parameters improved in response to the administered doses and period. Dietary SA enhanced total leucocyte count, phagocytic activity, respiratory burst, superoxide dismutase, lysozyme, and ACH50 activity, whereas the phagocytic index and total plasma protein were not significantly different. Furthermore, the mRNA levels of antioxidant-related gene (GPx), pro-inflammatory cytokines (TNF-α and IFN-γ, except for IL-1β), and antimicrobial peptides (Hepcidin) were upregulated. Meanwhile, anti-inflammatory cytokines (IL-10 and TGF-β1) were downregulated. Supplementation SA diet at 2.0 g/kg as the lowest dose revealed the most effective effects on innate immune and immune genes expression. Dietary SA was a potential immunostimulant in red tilapia aquaculture.

Immunomodulation, Fish Health and Resistance to Staphylococcus aureus of Nile Tilapia (Oreochromis niloticus) Fed Diet Supplemented with Zinc Oxide Nanoparticles and Zinc Acetate.

Recently some metal-based nanoparticles have gained serious attention from aquaculture and the fish feed industry as feed supplements. Oral supplementation of zinc oxide nanoparticles (ZnO-NPs) in fish feed, replacing Zn acetate (conventionally used zinc), is suggested as a cost-effective and efficient approach. Our study assessed the response of Nile tilapia,Oreochromis niloticus,fingerlings after its diet supplemented with chemically synthesized ZnO-NPs and zinc acetate under controlled conditions. ZnO-NPs were chemically synthesized and characterized. Tilapia fingerlings with an average body weight of 09.12 ± 1.23g were randomly distributed into five groups. An 8-week trial was set with control and four experimental groups. Basal diet (D1) was used as control, whereas D2, D3 and D4 comprising 20, 40, and 60 mgkg-1 ZnO-NPs supplementation were experimental diets. Additionally, D5 was composed of a basal diet supplemented with 40 mgkg-1 of conventionally used zinc acetate. Significant improvement (P < 0.05) was found in nanoparticles and Zn acetate supplemented groups as compared to control, while the 40 mgkg-1 Zn-NPs supplemented diet (D3) showed best performance in terms of health parameters, oxidative status and disease resistance. Antioxidant profiling was based on catalase, superoxide dismutase, glutathione's transferase, and malondialdehyde; hematology included Hb, WBCs, RBCs, HCT MCV, MCH and MCHC; immunological parameters comprised IgM, lysozyme activity, phagocytic activity, respiratory burst activity, cholesterol, aspartate aminotransferase, alanine aminotransferase, glucose content, and total serum proteins. We report that the D3 (40 mgkg-1 ZnO-NPs supplementation) significantly (P < 0.05) improved health-related parameters as compared to the other groups. Moreover, D3 also showed significantly decreased mortality percentage when challenged byStaphylococcus aureus, while the Zn acetate supplemented diet group showed better results as compared to control. Overall results suggest the basal diet supplemented with 40 mgkg-1 ZnO-NP for enhanced health parameters, oxidative status, immune response, and disease resistance. Hence, 40mgkg-1 ZnO-NP can be recommended to formulate the practical diet of fish to boost health improvement, immunomodulation, and resistance to bacterial disease.

Effects of dietary β-glucan on growth rate, antioxidant status, immune response, and resistance against Aeromonas hydrophila in genetic improvement of farmed tilapia (GIFT, Oreochromis niloticus)

This study was conducted to assess the influences of dietary β-glucan on growth rate, antioxidant status, immunity response, and resistance against Aeromonas hydrophila of genetically improved farmed tilapia (GIFT, Oreochromis niloticus). Four isonitrogenous and isoenergetic diets were formulated to contain 0%, 0.2%, 0.4%, and 0.8% β-glucan, respectively. Fish with similar size (average body weight, 3.43 ± 0.02 g) were randomly distributed into 12 tanks with 40 juveniles per tank and hand-fed to apparent satiation twice daily for 10 weeks. The fish were then challenged by A. hydrophila and their cumulative survival rate was recorded for the next week. No obvious influences on growth rate and feed utilization of GIFT were induced by dietary β-glucan addition (P > 0.05). However, dietary 0.4% β-glucan inclusion increased the serum high-density lipoprotein cholesterol level but decreased the triglyceride, total cholesterol and low-density lipoprotein cholesterol levels as well as alanine aminotransferase and aspartate aminotransferase activities (P < 0.05). Similarly, the increased intestinal superoxide dismutase (SOD), peroxidase and total antioxidant capacity (TAC) activities as well as the decreased intestinal malondialdehyde (MDA) and reactive oxygen species contents were also observed in GIFT fed diet with 0.4% β-glucan (P < 0.05). Dietary addition of 0.8% β-glucan increased the plasma SOD, glutathione peroxidase (GPx), catalase, and TAC activities, but reduced the plasma MDA content (P < 0.05). Dietary β-glucan inclusion regardless of levels improved the hepatic SOD, glutathione reductase, catalase, peroxidase, TAC, and acid phosphatase activities, but depressed the hepatic nitric oxide and MDA contents (P < 0.05). Plasma lysozyme activity was improved by the inclusion of 0.4% and 0.8% β-glucan (P < 0.05); plasma immunoglobulin M, complement 3 and complement 4 contents were improved by dietary β-glucan inclusion (P < 0.05); respiratory burst activity of macrophages was improved by the addition of 0.2% β-glucan (P < 0.05). Additionally, dietary β-glucan supplementation reduced the mortality rate and increased the relative percentage survival of GIFT challenged by A. hydrophila (P < 0.05). These results indicated that suitable β-glucan inclusion (0.4–0.8%) could promote the antioxidant status, immune response and disease resistance of GIFT, and β-glucan can be used as a feed additive for GIFT.

Microalgae with immunomodulatory effects on fish

AbstractIn recent decades, microalgal biotechnology has become one of the most valuable sources of functional feed additives. Many microalgae, such as Chlorella sp., Tetraselmis sp., Navicula sp., Phaeodactylum tricornutum, Porphyridium cruentum, Nannochloropsis gaditana, Dunaliella salina, Lobosphaera sp. and Schizochytrium sp., have been considered valuable in fish species as they enhance immunostimulant abilities, resistance against infectious diseases and tolerance to environmental stress. For instance, immunological parameters (i.e., respiratory burst activity, phagocytic capacity and IgM production) and immune‐related gene expression (i.e., proinflammatory cytokines) in fish have been manly analysed, demonstrating modulation upon microalgal administration. Moreover, disease resistance to microbial pathogen challenges has also been evidenced in several studies. In this context, the present literature review aims to assemble and reveal published research about the immunostimulant properties of some microalgae and their compounds in the immune systems of farmed fish species, whether model or new, with a high potential for aquaculture. Overall, it remains unclear how microalgae or their bioactive compounds may induce or modulate signals in the immune systems of fish species; however, the inclusion of different microalgal species in fish feed has improved the immune response and increased survival to stress conditions and infectious challenges.

Elevation in Rearing Temperature within Optimum Range Mitigate Immunosuppressive and Metabolic Stress Effect of High or Low Dietary Protein Level in Labeo Rohita Fingerlings

In the present study, Labeo rohita fingerlings were maintained either at ambient water temperature (26ºC) for five weeks or exposed to 32ºC for one week then later maintained at 26ºC for four weeks. Fingerlings reared under different temperature regimes were fed with any of the four experimental diets containing 20, 30, 40 or 45% protein. Serum cortisol level was higher at 26ºC compared to 32ºC, and decrease with the increase in dietary protein level up to 40%. Fingerlings fed 30% and 40% protein recorded similar WBCS count and respiratory burst activity which was lower and higher respectively compared to 20% and 45% protein fed fingerlings. Correspondingly, lower WBCS count and higher NBT were recorded following exposure to higher temperature (32ºC) for one week compared to 26ºC exposure group. Significantly lower survival was recorded in groups fed with lowest (20%) and highest (45%) dietary protein level whereas fingerlings exposed to 32ºC for one week exhibited higher survival (%) compare to 26ºC. Present results indicate that both lower and higher level of dietary protein may cause metabolic stress to fingerlings, as might consequently lead to the depressed immunity and exposure of elevated temperature (32ºC) for one week mitigates this immunosuppressive effect.

Influences of Bacillus subtilis and fructooligosaccharide on growth performances, immune responses, and disease resistance of Nile tilapia, Oreochromis niloticus.

The present study investigated the effects of Bacillus subtilis and fructooligosaccharide (FOS) on growth performances, immunity improvement, and disease resistance of Nile tilapia (Oreochromis niloticus). The fish (24.5 ± 1.6 g) were fed a basal diet (G1), diets supplemented with 1 g/kg (G2), 3 g/kg (G3) and 5 g/kg (G4) of FOS as well as diets supplemented with 1 × 109 CFU/g (G5), 3 × 109 CFU/g (G6) and 5 × 109 CFU/g (G7) of B. subtilis for 56 days. After the feeding trial, the complement C3, IL-1β, TNF-α, IFN-γ, hsp70 gene expression in the liver was then analyzed by a quantitative Real-time PCR. Then, fish were infected with Streptococcus agalactiae, and the survival rate was recorded. The results showed that FOS and B. subtilis had no significant effect (P > 0.05) on growth performances and survival rate. Lysozyme activity was significantly greater in the G4, G5, G6, and G7 groups. Also, all fish fed FOS and B. subtilis showed significantly (P < 0.05) higher respiratory burst activity than other groups. The expressions of complement C3, IL-1β, TNF-α, IFN-γ, and hsp-70 in the liver were significantly higher for fish fed 5 g/kg of FOS as well as for fish that received any concentration level of B. subtilis (P < 0.05) used in the study. After the S. agalactiae challenge test, the survival rate of fish-fed diets supplemented with FOS and B. subtilis was slightly higher than for the control group. The results indicated that FOS and B. subtilis could stimulate immune responses and immune-related genes in tilapia. However, further investigation of other prebiotics or herbs in combination with B. subtilis is encouraged at molecular levels and screening for beneficial metabolites that may increasingly improve digestive enzymes, growth performances, and health benefits in tilapia. In addition, on-farm experiments are needed.

Effect of Oral Immunostimulation of Humpback Grouper (Cromileptes altivelis) in Increasing Non-specific Immunity to Fight Disease (Review)

Cromileptes altivelis humpback grouper is a commodity with high economic value. Humpback grouper is Indonesia's main export commodity. The high value of sales and production of humpback grouper is constrained by the many disease attacks in the humpback grouper aquaculture industry. An alternative solution that has been continuously researched for several years to tackle humpback grouper disease is the use of immunostimulants. The use of immunostimulants orally can increase non-specific immunity of humpback grouper to ward off disease attacks and is environmentally friendly. Immunostimulants such as Lactococcus lactis HNL12, Lactococcus lactis HNL12 combined with Schizochytrium limacinum (algae flour), Peptidoglycan, Phycocyanin (Spirulina platensis), Bacterium, Vitamin C, Chromium Yeast, Commercial Yeast can enhance the non-specific immune system in humpback grouper (Cromileptes altivelis). These immunostimulants increase non-specific immunity such as Respiratory burst Activity (RB), superoxide dismutase (SOD), alkaline phosphatase (AKP), lysozyme (LZM), Phagocytosis Activity, Phagocytosis Index, Total leukocytes, hemoglobin, hematocrit, phagocytic activity, lysozyme activity, serum protein, albumin and globulin, total erythrocytes, haemoglobin, hematocrit. These immunostimulants can also prevent attacks of Vibriosis disease caused by the bacteria Vibrio harveyi, Vibrio algynoliticus and Vibrio parahaemolyticus as well as Viral Necrosis Virus (VNN) disease by increasing the survival rate compared to fish that are not given immunostimulants.Keywords: cromileptes altivelis, immunostimulant, non-specific immunity, fish disease

Respiratory Burst Activities of Peripheral Blood Neutrophils in Leukaemic Subjects in Northern Nigeria

Background: Polymorphonuclear neutrophils (PMN) are the only leucocytes that are competent to produce large amounts of reactive oxygen species (ROS) to kill phagocytized bacteria. In leukaemia, we hypothesized this bactericidal function might be hampered. Aim and Objectives: The goal of this study was to assess respiratory burst activity of PMN in leukaemia compared with non-leukaemic control subjects. We assessed the respiratory burst function of PMN as an effective defence against pathogens in leukaemia and control subjects. Materials and Methods: Peripheral blood samples were collected from leukaemia and control subjects (30 in each case) in lithium heparin anticoagulant containers. Our study cases were 30 leukaemia belonging to different subtypes (AML=12, CML=12, ALL=4 and CLL=2), 22 males and 8 females age range from 2.5–63 years (M±SEM, 28.0±3.4 years); the controls were 26 males and 4 females, age range from 17–53 years (M±SEM, 31.1±1.5 years), respectively. The respiratory burst activity was assessed using the nitroblue tetrazolium (NBT) dye reduction test on stimulated and un-stimulated PMN in leukaemia and control groups. Results: The cells purity in leukaemia was &gt;99% using Turk's solution and cells viability was &gt;95% by Trypan blue dye exclusion test. The respiratory burst activity of PMNs showed a statistically significant increase (P&lt;0.05) in controls compared with leukaemia subtypes. Similarly, comparison within leukaemia subtypes indicates a statistically significant increase (P&lt;0.05) RBA in CML compared with AML, ALL and CLL, respectively. Conclusion: the respiratory burst activity of PMNs in leukaemia is variable with enhanced activity in CML subjects, while depressed in AML, ALL and CLL subtypes; suggesting impaired bactericidal capacities of PMNs in these diseases.

Probiotic Debaryomyces hansenii CBS 8339 yeast enhanced immune responses in mice.

This study aimed to examine the effect of Debaryomyces hansenii CBS 8339 on innate immune responses in mice. Thirty BALB/c mice were randomly treated with phosphate buffered saline (PBS) (control) and two D. hansenii (Dh) doses: Dh 10ˆ6 CFU (colony forming units) and Dh 10ˆ8 CFU daily for 15days. Spleen, blood, and gut samples were taken on days7 and 15. Mouse splenocytes were isolated and challenged with Escherichia coli. Immunological assays and immune-related gene expressions were performed. Serum was obtained from blood for total IgA and IgG antibody titer determination. Gut samples were taken for yeast colonization assessment. Phagocytosis, respiratory burst activity, and nitric oxide production in mice were mainly enhanced (p < 0.05) upon 7days of D. hansenii intake at a concentration of 10ˆ8 CFU before and after bacterial challenge. Moreover, oral D. hansenii in mice upregulated (p < 0.05) gene expression of pro-inflammatory cytokines (INF-γ, IL-6 and IL-1β) before or after E. coli challenge on day7 but downregulated (p < 0.05) on day15. Furthermore, total serum IgG and IgA titers were higher (p < 0.05) in Dh 10ˆ8 CFU at days7 and 15, and only at day7, respectively, than that in the other dose and control groups. Finally, D. hansenii was detected in the gut of mice that received the treatments, suggesting that yeast survived gastrointestinal transit. Altogether, a short period (7days) of D. hansenii CBS 8339 oral delivery improved immune innate response on mice.

Ammonia toxicity in Nile tilapia: Potential role of dietary baicalin on biochemical profile, antioxidant status and inflammatory gene expression

The privileges of dietary baicalin intervention as a prophylaxis were assessed in Nile tilapia (Oreochromis niloticus) pre and post-acute ammonia challenge. The experimental trial lasted for 4 weeks, the first group served as a control (fed a basal diet), while, the second group received diet supplemented with baicalin (0.8 g/kg diet). The third group was fed a basal diet and exposed to acute ammonia toxicity in the last week. Meanwhile, the fourth group fed diet supplemented with baicalin (0.8 g/kg diet), then exposed to unionized ammonia challenge (5 mg/L) in the last week. Dietary baicalin notably augmented immune indicators (lysozyme, respiratory burst activity, phagocytic activity assay, and immunoglobulin (Ig) and tissue antioxidant biomarkers (total antioxidant capacity (TAC), glutathione (GSH), superoxide dismutase (SOD), and catalase (CAT)). Meanwhile, it decreased malondialdehyde (MDA), hepato-renal indicators (aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea, uric acid, creatinine), and stress indicators including glucose, cholesterol, triglycerides, and cortisol. Ammonia exposure significantly elevated stress parameters and MDA, but lessened SOD, GSH, CAT, and TAC levels in liver, gills, and kidneys tissue. Dietary baicalin and ammonia exposure induced interaction impacts on urea, creatinine, cortisol, glucose, lysozyme, alanine aminotransferase (ALT), aspartate aminotransferase (AST). Prior to ammonia exposure, baicalin administration clearly decreased urea. Fish fed enriched diet with - baicalin showed no changes in stress biomarkers before ammonia exposure, meanwhile reduced it after the ammonia challenge. Dietary baicalin alleviated fluctuations induced by ammonia in all above-mentioned parameters. And alleviated the altered gene expression for Hsp70, Nf-κB, Myd88, traf6, Nrf2, Ho-1, Il-1β, Il-8, TNF-α, and TLR-4 induced by ammonia toxicity. Taken together, it has been recommended to use baicalin for 3 weeks to mitigate ammonia-induced-oxidative damage, immune impairment, stress condition, hepato-renal failure, antioxidant and inflammatory gene expression in O. niloticus.

Efficacy of Different Routes of Formalin-Killed Vaccine Administration on Immunity and Disease Resistance of Nile Tilapia (Oreochromis niloticus) Challenged with Streptococcus agalactiae

Vaccines prepared from formalin-killed Streptococcus agalactiae were administered to Nile tilapia (Oreochromis niloticus) via three different routes: immersion in a water-based vaccine, injection with an oil-based vaccine, and as a water-based oral vaccine. All vaccination treatments increased lysozyme and peroxidase activity in skin mucus of Nile tilapia by 1.2- to 1.5-fold compared to their activities in unvaccinated control fish. Likewise, alternative complement, phagocytosis, and respiratory burst activities in the blood serum of the vaccinated fish were 1.2- to 1.5-times higher than in the unvaccinated fish. In addition, the expression transcripts of interleukin-1 (IL-1), interleukin-8 (IL-8), and lipopolysaccharide-binding protein (LBP) were 2.3- to 2.9-fold higher in the vaccinated fish compared to those in the unvaccinated control. The unvaccinated fish challenged with Streptococcus agalactiae had a survival rate of 25% compared to a survival rate of 78–85% for the vaccinated fish. The differences between the unvaccinated and vaccinated fish were all statistically significant, but there was no significant difference in any of the indicators of immunity between the three vaccinated groups. Collectively, these results confirm that vaccination with formalin-killed Streptococcus agalactiae significantly improved the resistance of Nile tilapia to infection by the pathogen. Overall, the efficacy of oral administration of the vaccine was comparable to that of vaccine administered via injection, indicating that oral vaccination is a viable cost-effective alternative to administering vaccines by injection.

Pyrogallol downregulates the expression of virulence-associated proteins in Acinetobacter baumannii and showing anti-infection activity by improving non-specific immune response in zebrafish model

Acinetobacter baumannii, a virulent uropathogen with widespread antibiotic resistance, has arisen as a critical scientific challenge, necessitating the development of innovative therapeutic agents. This is the first study reveal the proteomic changes in A. baumannii upon pyrogallol treatment for understanding the mechanisms using nano-LC-MS/MS-based quantitative proteomics and qPCR analysis. The obtained results found that pyrogallol treatment dramatically downregulated the expression level of several key proteins such as GroEL, DnaK, ClpB, SodB, KatE, Bap, CsuA/B, PgaA, PgaC, BfmR, OmpA, and SecA in A. baumannii, which are involved in chaperone-mediated oxidative stress responses, antioxidant defence system, biofilm formation, virulence enzyme production, bacterial adhesion, capsule formation, and antibiotic resistance. Accordingly, the pyrogallol dramatically enhanced the lifespan of A. baumannii-infected zebrafish by inhibiting bacterial colonization, demonstrating the anti-infective potential of pyrogallol against A. baumannii. Further, the histopathological results also demonstrated the disease protection efficacy of pyrogallol against the pathognomonic sign of A. baumannii infection. In addition, the pyrogallol treatment effectively improved the immune parameters such as serum myeloperoxidase activity, leukocyte respiratory burst activity, and serum lysozyme activity in zebrafish against A. baumannii infection. Based on the results, the present study strongly proposes pyrogallol as a promising therapeutic agent for treating A. baumannii infection.

Effect of dietary celery (Apium graveolens) on the growth performance, immune responses, and bacterial resistance against Vibrio anguillarum of European seabass (Dicentrarchus labrax).

In this study, we evaluated to reveal the effects of aqueous methanolic extract of celery (Apium graveolens) on the growth performance, immune responses, and resistance against Vibrio anguillarum in European seabass (Dicentrarchus labrax). For this purpose, twenty fish (initial mean weight of 4.80 ± 0.06g) were placed into twelve tanks (400 L) in triplicate and fish were fed with control (C) and three different levels (0.01, 0.05, and 0.1g/kg) of A. graveolens (AG) extract-containing diets (AG0.01, AG0.05, and AG0.1) for 30days. Blood and tissue (kidney, spleen, and intestine) samples were taken from the fish every 10days during the study to determine the immune responses of the fish. Respiratory burst activity (RBA) was significantly decreased in the AG0.1 group compared to all other groups on the 10th day of the study (P < 0.05). Significance was noticed in the RBA of fish in all AG groups compared to the C group (P < 0.05) on the 30th day of the experiment Lysozyme activity (LYS) was raised on the 10th day of the study in all celery groups compared to the C group (P < 0.05). No differences in the myeloperoxidase activity (MPO) were observed among the experimental groups (P > 0.05). The final mean weight (FMW) was not affected in any experimental groups (P > 0.05). However, in the AG0.05 group, the specific growth rate (SGR) increased, and the feed conversion ratio (FCR) decreased compared to other groups (P < 0.05). IL-1β in the kidney was highly elevated in the AG0.01 group on the 20th day of the study (P < 0.05). Similar results were observed on IL-6, IL-8, and TNF-α expression in the kidney (P < 0.05). Anti-inflammatory responses (IL-10 and TGF-β) also increased in all experimental groups and tissues compared to the C group (P < 0.05). COX-2 was upregulated on the 20th day of the study in all tissues (P < 0.05). At the end of the feeding trial, the survival rate of the AG0.1 group in fish infected with Vibrio anguillarum infection was higher than the C group. Dietary celery extract did not affect growth performance directly but increased innate immune responses and a high survival rate. Overall, compared to the control group, the growth, immunity, and resistance of European seabass fed with a diet containing 0.05g/kg celery aqueous methanolic extract has been improved, and this could be used as an immunostimulant feed additive.

Effects of a Dietary Multi-Strain Probiotic and Vaccination with a Live Anticoccidial Vaccine on Growth Performance and Haematological, Biochemical and Redox Status Indicators of Broiler Chickens.

A total of 256 male Ross 308 chickens were assigned to four treatments in a 2 × 2 factorial design with two levels of the anticoccidial vaccine (ACV) Livacox T (none or 1 × dose) with or without dietary supplementation with the probiotic Protexin® (P). The growth performance parameters for the test periods (1-21, 22-42, and 1-42 d) and oocyst per gram (OPG) at weekly intervals were analysed. Blood samples were collected at 16 post-vaccination (pv) days to measure selected haematological, biochemical, redox, and immunological parameters. ACV administration worsened the performance parameters of the chickens for 1-21 d pv, while supplementation with P reduced this negative effect with a significant improvement in 1-21 d body weight gain and feed conversion ratio. ACV administration increased % phagocytic cells (%PC), phagocytic index (PI), respiratory burst activity, proportion of monocytes, and activities of aspartate aminotransferase (AST) and lactate dehydrogenase, while it decreased the catalase activity and concentration of malondialdehyde and peroxides. The dietary administration of P significantly increased counts of red blood cells and white blood cells and increased %PC and PI, while it decreased the heterophil proportion, heterophil/lymphocyte ratio (p = 0.059), and alanine aminotransferase and AST activities. The oocyst counts were comparable in all sampling periods, except on 14 d pv, as supplementation with P significantly decreased 14 d OPG, thus indicating a positive influence of P on immunity development. In conclusion, dietary supplementation with P led to improved performance, better immunity, and benefits in health status in broilers vaccinated with the ACV, without interfering with the circulating vaccine strains.