Antibacterial and antioxidant activity of clove oil against Streptococcus iniae infection in Nile tilapia (Oreochromis niloticus) and its effect on hepatic hepcidin expression.

This study was designed to evaluate the modulating effect dietary clove essential oil (CL) has on the antioxidant and immunological status of Nile tilapia following Streptococcus iniae (Si) infection. Fish were placed on either control or (1.5 and 3%) CL-supplemented diets for 4 weeks. After sampling, the remaining fish in the control group were divided into 2 groups: an unchallenged (negative control) and an Si-challenged positive control. On the other hand, the remaining fish in CL-supplemented groups were challenged with Si, and mortality was checked for two weeks before the final sampling. Serum immunological parameters, tissue antioxidants, and oxidative stress markers were determined. Moreover, hepatic hepcidin expression was also measured in different groups. The obtained results showed improvements in blood phagocytic, bactericidal, lysozyme, and respiratory burst activities in CL-supplemented fish before and after the Si challenge. Si-challenge caused a remarkable increase in tissue malondialdehyde (MDA) levels that was inhibited by CL supplementation. The activities of glutathione peroxidase (GPx) and superoxide dismutase (SOD) in tissues were significantly elevated in a dose-dependent manner in CL-supplemented groups in both pre- and post-challenge experiments; renal SOD did not show any differences. Hepatic nitric oxide (NO) level was significantly decreased in CL-supplemented fish in a dose-dependent manner. In the post-challenge experiment, nitrosative stress was apparent in the liver and kidney; however, CL supplementation was sufficient to reverse it. Interestingly, a remarkable induction of the hepatic hepcidin expression was observed in all CL-supplemented groups in the pre-challenge experiment and Si-challenged fish, underscoring the role of CL as an antibacterial through inducing hepatic hepcidin expression to combat S. iniae infection. CL-supplementation was associated with lower mortality rates after Si-challenge, which was more pronounced in CL-3% supplemented fish. In conclusion, our results demonstrate that CL has a potent antioxidant role via increasing antioxidant enzymes' activities and antagonizing lipid peroxidation. Moreover, CL has an immune-stimulant effect by inducing the hepatic hepcidin expression and immunological markers in response to S. iniae infection.

Molecular identification of Trichodina compacta Van As and Basson, 1989 (Ciliophora: Peritrichia) from cultured Oreochromis niloticus in Egypt and its impact on immune responses and tissue pathology.

Trichodinids are peritrichous ciliated protozoa that affect both wild and cultured fishes. Several Trichodina species have low host specificity and are morphologically distinct, facilitating their identification based primarily on the presence of adhesive discs and the number of attached denticles. A trichodinid species named Trichodina compacta was first reported by Van As and Basson (1989) (Protozoa: Ciliophora: Peritrichia). However, in trichodinid infestations, morphological characteristics are insufficient for identifying the infesting species. Therefore, molecular and phylogenetic analyses are considered to be promising and useful tools for identifying the infesting species. This study aimed to achieve the molecular identification of a trichodinid infestation in Nile tilapia and to construct the phylogenetic relationships between the identified species and other peritrichous parasites. Moreover, we also aimed to study the pathological and immunological impacts of trichodinids on fry tissue to improve our understanding of the immune responses of teleost fish to trichodinae parasitic infestations and develop a better control method. Here, we used molecular techniques to identify the isolated trichodina species as T. compacta and demonstrated that Trichodina infestation in Nile tilapia is associated with remarkable immunogenic and inflammatory responses (increased il-1ß expression and decreased il-8 and tgf-ß expression). These findings improve our understanding of the responses of teleost fish to trichodinid parasite infestation and will be helpful for the development of novel control strategies that reverse the inflammatory and immunogenic alterations that occur in infested fish.

Antagonistic activity of dietary allicin against deltamethrin-induced oxidative damage in freshwater Nile tilapia; Oreochromis niloticus.

Allicin, the main biologically active component of garlic clove extracts, has been evaluated for its' efficacy in preventing deltamethrin-induced oxidative damage in Nile tilapia; Oreochromis niloticus. Fish were fed on 2 different doses of 0.5 g and 1 g of allicin/kg diet for 28 days. Serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), cholesterol, urea, uric acid, creatinine, total protein, albumin and globulin were estimated. Moreover, the level of malonaldehyde (MDA) was analyzed as a lipid peroxidation marker. In addition, reduced glutathione (GSH), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT) were analyzed as antioxidant biomarkers in liver, kidney and gills. Results show that deltamethrin subacute intoxication (1.46 µg/L for 28 days) increased serum AST, ALT, ALP, cholesterol, urea, uric acid, creatinine and tissue MDA. At the same time, serum total protein and albumin as well as tissue level of GSH, GSH-Px, SOD and CAT were reduced. Allicin supplemented diets enhanced all the altered serum biochemical parameters as well as tissues' lipid peroxidation and antioxidant biomarkers in a dose-dependent manner. The results suggest that feeding allicin can ameliorate deltamethrin-induced oxidative stress and might have some therapeutic properties to protect Nile tilapia on subacute deltamethrin toxicity.

Molecular evidence for the existence of two distinct IL-8 lineages of teleost CXC-chemokines.

Interleukin-8 (IL-8) is a CXC-type chemokine with a chemotactic activity mainly on neutrophils and plays a key role in promoting inflammation. In teleosts, several CXC-chemokines have been cloned and characterized as being IL-8-like. Phylogenetic data however indicate that the reported teleost IL-8-like chemokines are substantially remote from mammalian IL-8, forming a fish-specific clade of IL-8-like chemokines distinct from that of tetrapod IL-8. In the present study, a novel IL-8-like chemokine, designated CaIL-8, has been found in the expressed sequence tags of carp gills and identified as an orthologue of mammalian IL-8. The CaIL-8 transcript encodes 99 amino acids containing a typical CXC motif but lacks an ELR motif, as in most teleost IL-8-like chemokines. Phylogenetic tree constructed by the maximum likelihood method suggests a closer relationship of CaIL-8 with mammalian IL-8 than with other teleost CXC-chemokines reported to be IL-8-like. In a normal unstimulated carp, CaIL-8 mRNA was detected by RT-PCR only in gills, kidney, spleen, heart and peripheral blood leukocytes, in contrast to a previously reported carp IL-8-like chemokine CXCa, which shows ubiquitous basal expression. The results, taken together, are strongly indicative of the presence of two major IL-8-like lineages of CXC-chemokines in teleost.

Protective role of dietary Spirulina platensis against diazinon-induced Oxidative damage in Nile tilapia; Oreochromis niloticus.

The current study was performed to investigate the ameliorating effect of dietary supplementation of 0.5 and 1% Spiurolina platensis (SP) diet against the sub-acute toxicity of diazinon (DZN) 0.28mg/L in Nile tilapia. At the end of experiment after 28days, hepatic and renal damage markers (aspartate transaminase, alanine transaminase, alkaline phosphatase, urea, uric acid and creatinine), serum biochemical parameters (total proteins, albumin, cholesterol and glucose) and tissue antioxidant status (superoxide dismutase, catalase, glutathione peroxidase, reduced glutathione and malondialdehyde) were detesrmined. The results of the current study revealed significant improvement in hepatic and renal damage markers after SP supplementation in fish exposed to DZN toxicity. Moreover, SP improved serum biochemical markers through increasing serum albumin and globulins with a significant decrease in serum glucose and cholesterol. In addition, liver, kidneys and gills antioxidant status showed a significant improvement after SP supplemented to fish exposed to DZN where a significant increase in tissue antioxidant activity were observed with a significant decline in lipid peroxidation levels. It can be concluded that, SP supplementation attenuated the toxic effect of DZN toxicity in Nile tilapia through improving liver and kidney functions with a significant enhancement of tissue antioxidant status.

Functional analysis of membrane-bound complement regulatory protein on T-cell immune response in ginbuna crucian carp.

Complements have long been considered to be a pivotal component in innate immunity. Recent researches, however, highlight novel roles of complements in T-cell-mediated adaptive immunity. Membrane-bound complement regulatory protein CD46, a costimulatory protein for T cells, is a key molecule for T-cell immunomodulation. Teleost CD46-like molecule, termed Tecrem, has been newly identified in common carp and shown to function as a complement regulator. However, it remains unclear whether Tecrem is involved in T-cell immune response. We investigated Tecrem function related to T-cell responses in ginbuna crucian carp. Ginbuna Tecrem (gTecrem) proteins were detected by immunoprecipitation using anti-common carp Tecrem monoclonal antibody (mAb) and were ubiquitously expressed on blood cells including CD8α(+) and CD4(+) lymphocytes. gTecrem expression on leucocyte surface was enhanced after stimulation with the T-cell mitogen, phytohaemagglutinin (PHA). Coculture with the anti-Tecrem mAb significantly inhibited the proliferative activity of PHA-stimulated peripheral blood lymphocytes, suggesting that cross-linking of Tecrems on T-cells interferes with a signal transduction pathway for T-cell activation. These findings indicate that Tecrem may act as a T-cell moderator and imply that the complement system in teleost, as well as mammals, plays an important role for linking adaptive and innate immunity.

Pharmacodynamic interaction of Spirulina platensis and deltamethrin in freshwater fish Nile tilapia, Oreochromis niloticus: impact on lipid peroxidation and oxidative stress.

Spirulina platensis (SP) is one of the most commonly used dietary supplements in human and many animal species, including fish. Recently, it has gained more attention in fish not only for its growth-promoting and immunomodulatory effects but also for its antioxidant potential. The present study was conducted to investigate the protective role of two different dietary levels of SP on freshwater Nile tilapia; Oreochromis niloticus exposed to subacute deltamethrin (DLM) intoxication. Spirulina was supplemented at levels of 0.5 and 1 % in the diet along with DLM at a concentration of 1.46 µg/l for 28 days. Serum biochemical parameters, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total protein, albumin, cholesterol, urea, uric acid and creatinine, were estimated. In addition, the level of malondialdehyde (MDA) was analysed as a lipid peroxidation marker. Reduced glutathione (GSH) content and glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT) activities were analysed as antioxidant biomarkers in liver, kidney and gills. The results revealed that DLM intoxication increased serum AST, ALT, ALP, cholesterol, urea, uric acid, creatinine and tissue MDA, while decreased serum total protein and albumin as well as tissue GSH level and GSH-Px, SOD and CAT activities. SP supplementation at the two tested levels enhanced all altered serum biochemical parameters as well as tissue lipid peroxidation and antioxidant biomarkers. Therefore, it could be concluded that SP administration could minimize DLM-induced toxic effects by its free radical scavenging and potent antioxidant activity.