Samah R. Khalil

Efficacy of Spirulina platensis diet supplements on disease resistance and immune-related gene expression in Cyprinus carpio L. exposed to herbicide atrazine

Abstract The present study evaluated the immunotoxicological effects of the herbicide atrazine (ATZ) at sub‐lethal concentrations and the potential ameliorative influence of Spirulina platensis (SP) over a sub‐chronic exposure period on Cyprinus carpio L., also known as common carp. Common carp was sampled after a 40‐days exposure to ATZ (428 &mgr;g/L) and SP (1%), individually or in combination to assess the non‐specific immune response, changes in mRNA expression of immune‐related genes [lysozyme (LYZ), immunoglobulin M (IgM), and complement component 3 (C3)] in the spleen, and inflammatory cytokines (interleukins IL–1ß and IL–10) in the head kidney using real‐time PCR. Additionally, disease resistance to Aeromonas sobria was evaluated. The results revealed that ATZ exposure caused a significant decline in most of the hematological variables, lymphocyte viability, and lysozyme and bactericidal activity. Moreover, ATZ increased the susceptibility to disease, reflected by a significantly lower post‐challenge survival rate of the carp. ATZ may induce dysregulated expression of immune‐related genes leading to downregulation of mRNA levels of IgM and LYZ in the spleen. However, expression of C3 remained unaffected. Of the cytokine‐related genes examined, IL–1B was up‐regulated in the head kidney. In contrast, the expression of IL–10 gene was down‐regulated in the ATZ‐exposed group. The SP supplementation resulted in a significant improvement in most indices; however, these values did not match with that of the controls. These results may conclude that ATZ affects both innate and adaptive immune responses through the negative transcriptional effect on genes involved in immunity and also due to the inflammation of the immune organs. In addition, dietary supplements with SP could be useful for modulation of the immunity in response to ATZ exposure, thereby presenting a promising feed additive for carps in aquaculture. HighlightedsExposure to atrazine (ATZ) for 40 days exhibited immunotoxic effects in common carp (Cyprinus carpio).ATZ altered hematological variables and increased disease susceptibility.ATZ had a more pronounced immunosuppressive effect at the genomic level.Dietary Spirulina platensis (SP) (1%) could be useful for maintaining the altered immunity in response to ATZ exposure.

Apitoxin protects rat pups brain from propionic acid-induced oxidative stress: The expression pattern of Bcl-2 and Caspase-3 apoptotic genes.

The primary aim of this study was to determine the potential modulatory role of the apitoxin (bee venom; BV) against propionic acid (PPA)-induced neurotoxicity. The biochemical responses to PPA exposure in rat pups were assayed, including changes in the antioxidant barrier systems and lipid peroxidation and protein oxidation biomarkers in the brain tissue. DNA damage was measured by single-cell gel electrophoresis and differences in Bcl-2 and Caspase-3 mRNA expression were assessed using real-time PCR. Changes in amygdala complex ultrastructure were visually assessed using electron microscopy. Sixty rat pups were assigned into six groups: a control group, a PPA-treated group, a BV-treated group, a protective co-treated group, a therapeutic co-treated group, and a protective/therapeutic co-treated group. The results indicate that PPA induced a pronounced increase (64.6%) in malondialdehyde (MDA), and in DNA damage (73.3%) with three-fold increase in protein carbonyl concentration. A significant reduction was observed in the enzyme activities of superoxide dismutase (SOD) (48.7%) and catalase (CAT) (74.8%) and reduced glutathione (GSH) level (52.6%). BV significantly neutralized the PPA-induced oxidative stress effects, especially in the BV protective/therapeutic co-treated group. In this group, GSH levels were restored to 64.5%, and MDA, protein carbonyl levels and tail moment % were diminished by 69.5, 21.1 and 18.8% relative to the control, respectively. Furthermore, while PPA induced significant apoptotic neural cell death, BV markedly inhibited apoptosis by promoting Bcl-2 expression and blocking Caspase-3 expression. BV markedly restored the normal ultrastructural morphology of the amygdala complex neurons. These results conclusively demonstrate that BV administration provides both protective and therapeutic effects in response to the PPA-induced deleterious effects, including oxidative stress, DNA damage, and neuronal death in the brains of rat pups.

Antidotal impact of extra virgin olive oil against genotoxicity, cytotoxicity and immunotoxicity induced by hexavalent chromium in rat

Abstract An in vivo study was carried out to verify whether extra-virgin olive oil (EVOO) has the potential to modulate alterations resulted from exposure to hexavalent chromium (CrVI) as potassium dichromate in rats. For this purpose, CrVI was injected intraperitoneally (i.p.) at a dose of 0.4 mg/kg bw/day, EVOO was given orally at a dose of 300 μl daily either a lone or co-treated with CrVI at the same doses, routes and duration (26 days). At the end of the experiment, blood and spleen samples were collected. Genotoxicity, cytotoxicity and immunotoxicity biomarkers induced by CrVI were evaluated. Also, histopathological and immunohistochemical investigations of spleen tissue were conducted. A significant increase in genotoxicity and cytotoxicity biomarkers (micronucleus frequency, 8-hydroxy-2-deoxyguanosine level and lactate dehydrogenase activity) were recorded in CrVI treated rats. In addition, the immunotoxicity biomarkers showed a significant decrease in phagocytic%, stimulated nitric oxide production and decrease in the serum lysozyme activity. Histopathological and immunohistochemical studies support the cytotoxicity study. Oral administration of EVOO can ameliorate those effects but not restored to control level. Thus, authors recommend that regular consumption of this oil in the diet provides a constant supply of potential antioxidants that could reduce these alterations.

Combined Cytogenotoxic Effects of Bee Venom and Bleomycin on Rat Lymphocytes: An In Vitro Study

This study was carried out to determine the cytotoxic and genotoxic effects of bee venom (BV) and/or the chemotherapeutic agent bleomycin (BLM) on healthy isolated rat lymphocytes utilizing morphometric and molecular techniques. Using the Ficoll-Histopaque density gradient centrifugation technique, lymphocytes were isolated, divided into groups, and subjected to BV and/or BLM at incubation medium concentrations of 10 or 20 μg/mL respectively for 24 and 72 hrs. An MTT assay and fluorescent microscopy examinations were used to assess the cytotoxic effects. To determine the predominant type of BV and/or BLM-induced cell death, LDH release assay was employed beside quantitative expression analyses of the apoptosis-related genes (Caspase-3 and Bcl-2). The genotoxic effects of the tested compounds were evaluated via DNA fragmentation assay. The results of these assays demonstrated that BV potentiates BLM-induced cytotoxicity through increased LDH release and diminished cell viability. Nevertheless, BV significantly inhibited the BLM-induced DNA damage. The results verify that BV significantly attenuates the genotoxic effects of BLM on noncancerous isolated rat lymphocytes but does not diminish BLM cytotoxicity.

Melamine and/or formaldehyde exposures affect steroidogenesis via alteration of StAR protein and testosterone synthetic enzyme expression in male mice

The reproductive effects of melamine and formaldehyde, either alone or in combination, on mature male Swiss mice were investigated. The animals were orally administered melamine (50mg/kg/day), formaldehyde (25mg/kg/day), a mixture of melamine and formaldehyde, or a vehicle control for 65 consecutive days. As a result, the deterioration of sperm characteristics and inhibition of testicular enzyme activity were observed in the melamine- and formaldehyde-exposed groups. In addition, testosterone and luteinizing hormone levels were significantly reduced in the melamine but not in the formaldehyde-exposed group, which correlated with down-regulation of transcription levels of steroidogenic-related genes. Histopathologically, both compounds caused lesions in the testes. However, the co-exposure reduced the induced alterations in spermatogenesis, steroidogenesis, and testicular architecture that were obviously observed in the melamine-exposed group. Consequently, we demonstrated that melamine exhibited more pronounced reproductive impact in comparison with formaldehyde. In addition, formaldehyde was able to substantially temper the melamine -induced reproductive toxic effect.

Effect of dietary supplementation with Spirulina platensis on Atrazine-induced oxidative stress- mediated hepatic damage and inflammation in the common carp (Cyprinus carpio L.)

The present study evaluated the potential modulatory effect(s) of dietary supplementation with Spirulina platensis (SP) on Atrazine (ATZ)-induced oxidative stress and inflammation in common carp (Cyprinus carpio L.). Common carp was exposed to ATZ (428μg/L) and SP (1%), either alone or in combination, for 40 days. Subsequently, the treatment groups were evaluated for ATZ-induced oxidative stress-mediated hepatic damage and the potential antioxidant effect(s) of SP supplementation. The results indicated that ATZ exposure led to a significant increase in the oxidative stress as suggested by the increased levels of lipid and DNA oxidative damage markers and the significant decline of antioxidant status biomarkers. Further, a real-time PCR analysis of the liver tissues revealed that the ATZ exposure resulted in the significant modulation of the mRNA expression of cytokines involved in the inflammatory response pathway in the liver, such as Interleukin (IL)-1ß and IL-10. The expression of IL-1ß mRNA was up-regulated while that of IL-10 mRNA was down-regulated. The group subjected to supplementation with SP exhibited a significant decrease in ATZ-induced oxidative stress-mediated hepatotoxic and inflammatory responses; however, these did not attain the levels of the control group. Owing to its ability for protecting against ATZ-induced oxidative stress-mediated hepatic damage in carps, SP could be a potentially effective and promising candidate as a feed additive for carps in aquaculture.

Possible role of Arthrospira platensis in reversing oxidative stress-mediated liver damage in rats exposed to lead

Environmental pollutants, particularly metallic elements, mobilized and released into the environment, eventually accumulate in the food chain and thus pose a serious threat to human and animal health. In the present study, the role of Arthrospira (Spirulina platensis; SP) as a protector against oxidative stress-mediated liver damage induced by an exposure to lead acetate (LA; as a metallic pollutant) was assessed. To achieve this aim, rats were orally administered with 300 mg/kg bw SP for 15 days, before and concurrently with an intraperitoneal injection of 50 mg/kg bw LA (6 injections throughout 15 days). As a result, co-administration of SP with LA reduced the amount of lead that accumulated in both blood and liver tissue of the exposed rats and minimized the increased levels of lipid peroxidation, protein oxidation, DNA oxidative damage, and liver enzyme endpoints. In addition, because of SP administration, the levels of depleted biomarkers of antioxidant status and total antioxidant capacity in LA-exposed rats improved. Moreover, SP protected the liver tissue against the changes caused by LA exposure and also decreased the reactivity of HSP70 in the cytoplasm of hepatocytes. Collectively, our data suggest that SP has a potential use as a food supplement in the regions highly polluted with heavy metals such as lead.

Behavioral response and gene expression changes in fipronil-administered male Japanese quail (Coturnix japonica) ☆

Fipronil is an important member of the phenylpyrazole group of insecticides and is widely used for various crops and vegetables to control insects, thereby exposing birds, animals, and humans to fipronil. Currently, there is limited information on the effects of fipronil exposure in Japanese quail. Therefore, our aim was to assess the reproductive toxicological effects of fipronil in the Japanese quail in a 15-day gavage study and then its recovery over a period of 60 days. Fipronil-administration led to significant losses in both feed intake and body weight. Whereas, the gonadosomatic index was not affected, and histological changes observed in the testes were reversible, particularly by day 45 and day 60 of recovery. Cloacal gland atrophy, reduced foam quantity and a reduction in fertility, sexual and aggressive behaviors, and serum testosterone with elevated estradiol (E2) hormone levels were also observed. All these changes gradually reversed during various recovery periods. Further, alterations in hepatic vitellogenin (Vtg) and estrogen receptor α (ERα) gene expression, assessed by quantitative polymerase chain reaction, were also observed. Specifically, ERα1 was induced after fipronil administration, while the Vtg transcript was elevated during both exposure and recovery periods. Our results showed that fipronil exposure has a profound negative influence on reproductive traits in the male Japanese quail and exhibits an estrogenic activity that can raise the incidence of infertility in males. Nevertheless, most of the changes could be reversed after a recovery period of 30-45 days.

The role of apitoxin in alleviating propionic acid-induced neurobehavioral impairments in rat pups: The expression pattern of Reelin gene

The efficacy of apitoxin (bee venom; BV) in ameliorating propionic acid (PPA) -induced neurobehavioral impacts was studied. Sixty rat pups were enrolled in a split litter design to six groups: a control group, a PPA-treated group, a BV-treated group, a BV/PPA protective group, a PPA/BV therapeutic group, and a BV/PPA/BV protective and therapeutic group. Exploratory, social, locomotor, and repetitive/stereotype-like activities were assessed and prosocial, empathy, and acquired behavior were evaluated. Levels of neurotransmitter including serotonin, dopamine, and gamma-aminobutyric acid (GABA) were determined and a quantitative analysis of Reelin gene expression was performed. PPA treatment induced several behavioral alterations, as reduced exploratory activity and social behaviors, increased repetitive/stereotypic behaviors, and hyperactivity. In addition, a marked decline of neurotransmitters and down-regulation of Reelin mRNA expression were observed. BV exhibited high efficiency in ameliorating the PPA-induced neurobehavioral alterations, particularly when applied both before and after PPA administration. Overall, the results implied that BV has merit as a candidate therapeutic treatment to alleviate PPA-induced neurobehavioral disorders.

Neurotransmitters and neuronal apoptotic cell death of chronically aluminum intoxicated Nile catfish (Clarias gariepinus) in response to ascorbic acid supplementation

Few studies have been carried out to assess the neurotoxic effect of aluminum (Al) on the aquatic creatures. This study aims to evaluate the neurotoxic effects of long term Al exposure on the Nile catfish (Clarias gariepinus) and the potential ameliorative influence of ascorbic acid (ASA) over a 180 days exposure period. Forty eight Nile catfish were divided into four groups: control group, placed in clean water, ASA exposed group (5mg/l), AlCl3 received group (28.96 μg/l; 1/20 LC50), and group received AlCl3 concomitantly with ASA. Brain tissue was examined by using flow cytometry to monitor the apoptotic cell population, HPLC analysis for the quantitative estimation of brain monoamine neurotransmitters [serotonin (5-HT), dopamine (DA), norepinephrine (NE)]. The amino acid neurotransmitters [serum taurine, glycine, aspartate and glutamine and brain gamma aminobutyric acid (GABA)] levels were assessed, plus changes in brain tissue structure using light microscopy. The concentration of Al in both brain tissue and serum was determined by using atomic absorption spectrophotometery. The Al content in serum and brain tissue were both elevated and Al exposure induced an increase in the number of apoptotic cells, a marked reduction of the monoamine and amino acids neurotransmitters levels and changes in tissue morphology. ASA supplementation partially abolished the effects of AL on the reduced neurotransmitter, the degree of apoptosis and restored the morphological changes to the brain. Overall, our results indicate that, ASA is a promising neuroprotective agent against for Al-induced neurotoxicity in the Nile catfish.