Camel milk or silymarin could improve the negative effects that experimentally produced by aflatoxin B1 on rat's male reproductive system.

BACKGROUND: Camel milk and silymarin have many different beneficial effects on several animal species. Meanwhile, Aflatoxins are mycotoxins with extraordinary potency that pose major health risks to several animal species. Additionally, it has been documented that aflatoxins harm the reproductive systems of a variety of domestic animals. The present design aimed to investigate the impact of aflatoxin B1 (AFB1) on rat body weight and reproductive organs and the ameliorative effects of camel milk and silymarin through measured serum testosterone, testes pathology, and gene expression of tumor necrosis factor (TNF-α), luteinizing hormone receptor (LHR), and steroidogenic acute regulatory protein (StAR) in the testes. A total of sixty mature male Wister white rats, each weighing an average of 83.67 ± 0.21 g, were used. There were six groups created from the rats. Each division had ten rats. The groups were the control (without any treatment), CM (1 ml of camel milk/kg body weight orally), S (20 mg silymarin/kg b. wt. suspension, orally), A (1.4 mg aflatoxin/kg diet), ACM (aflatoxin plus camel milk), and AS (aflatoxin plus silymarin). RESULTS: The results indicated the positive effects of camel milk and silymarin on growth, reproductive organs, and gene expression of TNF-α, LHR, and StAR with normal testicular architecture. Also, the negative effect of AFB1 on the rat's body weight and reproductive organs, as indicated by low body weight and testosterone concentration, was confirmed by the results of histopathology and gene expression. However, these negative effects were ameliorated by the ingestion of camel milk and silymarin. CONCLUSION: In conclusion, camel milk and silymarin could mitigate the negative effect of AFB1 on rat body weight and reproductive organs.

Innate immune response to double-stranded RNA in American heritage chicken breeds.

Backyard poultry flocks that employ heritage breeds of chicken play a crucial role in the maintenance of poultry pathogens of economic and zoonotic importance. This study examined innate immunity to viral pathogens in heritage chicken breeds using a model of viral double-stranded RNA (dsRNA). Following intraperitoneal injection of high molecular weight (HMW) -poly(I:C)/Lyovec into 4-wk-old chicks, we evaluated gene expression in peripheral blood mononuclear cells (PBMCs) and splenocytes. There was a significant difference across breeds in the expression of IL-4, IL-12p40, IFNγ, and B-cell activating factor (BAFF) in the spleen. In PBMCs, a significant difference in IFN-α expression was seen across breeds. Approximately 57% of IFN-α transcripts in PBMCs was explained by levels of expression of MDA5 transcripts. Using flow cytometry, we showed that only monocytes/macrophages (KUL01+ cells) expressed the scavenger receptor CD163. Regression analysis showed that 42% of fold change in CD163 expression on PBMCs was explained by breed (P < 0.0004). In general, breeds that responded to HMW-poly(I:C) by showing higher upregulation of IFNγ, IL-1ß, and IL-12p40 transcripts in the spleen, and higher IFNα transcripts in peripheral blood, expressed less CD163 on blood monocytes. These findings suggest a genetic basis for the response of chickens to double-stranded RNA. Surface expression of the scavenger receptor CD163 in PBMCs following injection of high molecular weight poly(I:C) may be a rapid method to select chickens for breeding based on innate immune response to viral dsRNA.

Length-Dependent Modulation of B Cell Activating Factor Transcripts in Chicken Macrophage by Viral Double-Stranded RNA.

Viral double-stranded RNA (dsRNA) interacts with Retinoic-acid-inducible-gene-1 (RIG-1)-like receptors (RLRs) to induce type 1 interferons. Melanoma-derived-antigen-5 (MDA-5), an RLR, but not RIG-1, is found in chickens. Ducks express both RIG-1 and MDA-5, a possible cause of differences in susceptibility to influenza virus infection between chickens and ducks. Using the HD11 chicken macrophage cell line and an RT2 Profiler PCR-array system, we showed that high-molecular-weight poly(I:C), HMW-poly(I:C), upregulates CCL4, interferon-gamma, interleukin-1, and interleukin-6 mRNA transcripts. HMW-poly(I:C), an in vitro surrogate of long dsRNA species, also induces the upregulation of IL-12B and B cell activating factor (BAFF). Conversely, low-molecular-weight poly(I:C), LMW-poly(I:C) did not induce a distinct cytokine expression pattern. Nonetheless, co-transfection of LMW and HMW-poly(I:C) significantly reduced the upregulation of IL12B and BAFF by HMW-poly(I:C). These findings support previous studies that found no expression of RIG-1, a receptor for short dsRNA species, in chicken cells. Surprisingly, however, our data suggested that in the absence of RIG-1 in chicken macrophages, short dsRNA species may inhibit macrophage-mediated B cell development and survival by modulating the expression of BAFF without significantly reducing type 1 interferon response.

Ameliorative effects of camel milk and silymarin upon aflatoxin B1 induced hepatic injury in rats.

Aflatoxin B1 (AFB1) poses a major risk to both human and animal health because it contaminates food, feed, and grains. These dangerous effects can be mitigated using natural components. The purpose of this study was to examine the ameliorative effects of camel milk and silymarin supplementation upon aflatoxin B1 induced hepatic injury in rats. This improvement was assessed by measuring leukocytic and deferential counts, serum biochemical parameters, and gene expression of Tumor Necrosis Factor (TNF-α), antioxidant gene (NAD(P)H quinone oxidoreductase 1 (NQO1)), and base excision repair genes (APE1 and OGG1) in the liver tissue, in addition to liver histopathology. Sixty mature males Wister white rats were used to perform the present study; the rats were distributed in six groups (ten rats/group). The control group (without any treatment) received saline by gavage. The camel milk group received 1 ml of camel milk/kg body weight. The silymarin group received 1 ml of silymarin suspension solution at a dose of 20 mg of silymarin/kg of b.wt. The aflatoxin group received an aflatoxin-contaminated diet at a dose of 1.4 mg of aflatoxin /kg of diet and received saline. The camel milk + aflatoxin group received the same previous oral doses of camel milk and an aflatoxin-contaminated diet at the same time. The silymarin + aflatoxin group received the same previous doses of silymarin orally and an aflatoxin-contaminated diet at the same time. The obtained data indicated the deleterious effect of aflatoxin B1 on the leukocytic count, activity of AST and ALT, serum proteins, ferritin, alpha-fetoprotein, carcinoembryonic antigen, liver pathology, and the expression of the studied genes. However, these deleterious effects were mitigated by camel milk and silymarin supplementation. Thus, we could conclude that the ingestion of camel milk and silymarin mitigated the negative effects of AFB1 on the hematology, activity of AST and ALT, serum proteins, ferritin, alpha-fetoprotein, carcinoembryonic antigen, liver pathology, and gene expression in the rat model.

The synergetic effect of Bacillus species and Yucca shidigera extract on water quality, histopathology, antioxidant, and innate immunity in response to acute ammonia exposure in Nile tilapia.

Acute ammonia toxicity suppresses the immune function and enhances the inflammatory pathways in Nile tilapia. The aim of this study was to compare the effect of Bacillus strains probiotic mixture (BS) or Yucca shidigera liquid extract (YSE) alone or their combination in water treatment and in reliving toxicity of an acute ammonia exposure in Nile tilapia through the assessment of fish immune response, inflammatory pathway, oxidative stress response with respect to the histopathological changes, gene expression, enzymes levels and phagocytosis. Five groups were used; the 1st and 2nd groups fed the basal diet; the 3rd group fed basal diet with BS in water, 4th group fed basal diet and supplemented with YSE in water and 5th group received a combination of BS and YSE. After two weeks of treatments, the 2nd, 3rd, 4th, and the 5th groups were exposed to acute ammonia challenge for 72 h. Fish exposed to ammonia displayed significant decreases in RBCs, Hb, PCV, WBCs, phagocytic activity (PA) and index (PI), lysozyme activities and serum antioxidant enzymes (glutathione peroxidase (GPX) and catalase (CAT)). Also, a significant increase in Malondialdehyde (MDA), degenerative changes in the gills, hepatopancrease and spleen associated with an elevated un-ionized ammonia level. A significant restoration of the hematological parameters was observed with the use of BS, YSE or their combination. Additionally, they improved the innate immunity, antioxidant responses, and histopathological changes. At transcriptomic level, ammonia toxicity significantly lowered the mRNA transcription levels of Nuclear erythroid 2-related factor 2 (Nrf2), quinone oxidoreductase 1 (NQO-1), Heme oxygenase 1 (HO-1) and Heat shock proteins (HSP70). While nuclear factor kappa ß (NFкß), Tumor necrosis factor α (TNF-α), Interleukin 1ß (IL-1ß), and Interleukin 8 (IL8), transcription levels were increased. Interestingly, BS and YSE and their combination significantly increased the expression of these genes with the highest levels reported with BS and YSE combination. We observed that, the most pronounced restoration of some important inflammatory and immune related genes close to the control level was observed when BS-YSE mix was used. Furthermore, a restored water pH, and a maintained ammonia level to the control level were observed in this group. Otherwise, equal effects for the three treatments were observed on the assessed parameters. We recommend the used of BS-YSE mix for water ammonia treatment and relieving ammonia toxicity in fish.

Intra- and inter-breed variation in immune response to acute and sub-chronic Salmonella infection and commercial immune-stimulant in two-layer breeds.

Salmonella is one of the most hazardous diseases in poultry farms. Markedly, the application of active immunostimulants is illustrated as potential protective agents against infection in poultry farms. Thus, this work aimed to explore inter- and intra-breed variation in response to acute and subchronic Salmonella enteritidis infection in two-layer breeds (one commercial [Hy-line strain] and another native [Fayoumi breed]). Besides exploring the possible protective effect of a commercial immune modulator (STIMULAN) on the two breeds during the acute infection. The ELISA antibody titer in sub-chronic infections and the expression analysis of some selected genes (IL-1ß, LITAF, TGF-ß, HSP90 and HSP70) are used as the clinical signs for acute infections to assess the possible protective role of a commercial immunomodulator (STIMULAN). Five groups were used during the acute experiment: G1-control; G2a-susceptible; G2b-resistant birds, G3-which received STIMULAN and G4-which received the infection + STIMULAN. The groups with sub-chronic infections include G1 (control), G2 (high antibody titer) and G3 (low antibody titer). The gene expressions among the susceptible birds during acute infection of both breeds are nearly similar. They only differ in the expression of HSP90 in the Fayoumi breed. However, the resistant birds vary in their gene expression profile. The effect of STIMULAN as a feed additive in non-infected birds was an up-regulation of LITAF, TGF-ß, HSP90 in Fayoumi. Moreover, a powerful stimulatory role was observed when both breeds were infected. Both breeds were asymptomatic during the sub-chronic infection. Although, the increased expression of inflammatory-related genes in the Hy-line was considered as an indication of infection persistence. Fayoumi is capable of immune clearance for this infection. Thus, the Fayoumi breed is more resistant to acute Salmonella infection. HSP90 plays a vital role in its resistance. We recommend the use of STIMULAN as an immunomodulator during Salmonella infection.

Comparative Expression Analysis of Inflammatory and Immune-related Genes in Cattle During Acute Infection with Foot-and-mouth Disease Virus in Egypt.

INTRODUCTION: Foot-and-mouth disease is a highly infectious viral disease affecting all cloven-footed domestic animals. The three foot-and-mouth disease virus (FMDV) serotypes A, O and SAT2 are at present the greatest threat to susceptible animals in Egypt. The aim of the present study was, for the host factors associated with different FMDV infections in cattle during the acute phase, to compare these factors' influence on the expression of the IL-10, TLR-2, TNF-α, CXCL10, CD48, NFATC4 and IFNG inflammatory and immune-related genes. MATERIALS AND METHODS: Vesicular fluid and epithelium samples were obtained from at least three infected cattle on the same affected farm during three different FMDV outbreaks and were used for serotyping of the virus and for expression analysis of host genes. A two-step RT-PCR was used for diagnosis of the virus with primers specific for each serotype. RESULTS: In quantitative PCR analysis, the expression patterns of TLR-2 and IFNG were prominent, while NFATC4 expression was absent in all FMDV-infected cattle. The highest expression of CD48 was associated with increased expression of other inflammatory and immune-related genes (IL-10, TLR-2, TNF-α and IFNG), which may be an indication of rapid virus clearance. CONCLUSION: The use of vesicular fluid and epithelium for investigation of viral and immune-related gene expression levels in acute FMDV infection is possible. Host-dependent variation in the expression of the studied genes was observed in different FMDV serotype outbreaks.

Meta-analysis of Genetic Diversity of the VP1 Gene Among the Circulating O, A, and SAT2 Serotypes and Vaccine Strains of FMD Virus in Egypt.

INTRODUCTION: Three strains of the FMD virus (A, O, and SAT 2) were recognised as causes of the FMD circulating in Egypt. The aims of this study were to trace the FMDV isolates from outbreaks in Egypt to understand their epidemiology and evolution and to understand the situation of the vaccine strains compared with the circulating serotypes. MATERIAL AND METHODS: A meta-analysis was carried out by using the data available for FMD outbreaks in Egypt from GenBank and the World Reference Laboratory for Foot-and-Mouth Disease (WRLFMD); a comparison was done with both data sets for the three serotypes. MEGA-X was used for the evolution analysis, through constructions of phylogenetic trees for all sequences recorded in GenBank for each serotype in different Egyptian outbreaks in different years and also within the same year. Additionally, nucleotide substitution rate, molecular clock, and mean evolutionary rates were estimated for the three serotypes to understand and compare their evolution. RESULTS: Absence of some records of certain serotype outbreaks from the WRLFMD database was noted as were subsequent missing appropriate vaccine programmes. Genetic variation was recorded among the virus isolates within the same years and also the vaccine strain was associated with up to 26 amino acid substitutions. The evolution rate of the SAT2 strain was the highest of the circulating strains. SAT2 had high amino acid substitution per year at an important immunogenic site (130-170), serotype A had less, and serotype O the least. CONCLUSION: The need for different strategies for vaccine serotype selection is indicated.

The impacts of seasonal variation on the immune status of Nile tilapia larvae and their response to different immunostimulants feed additives.

Few data are available on the thermal tolerance of Nile tilapia fish larvae in relation to their immune status and survival. The aims of this work were to evaluate the immune status of one day old Nile tilapia (Oreochromis niloticus) larval stage collected at the beginning (March), middle (August) and at the end (October) of hatching season through morphometric assessment of the larvae parameters including yolk sac diameter, body length and width as well as the expression of some immune-related genes (rag, sacs and tlr), inflammatory (il1b and il8) and stress related genes (hsp27, hsp70). Also, to compare the effect of three different immunostimulants (ß-glucan, Vitamin C, and methionine/lysine amino acids mix) on the expression of the studied genes at two variant temperatures (23 ± 1 °C and 30 ± 1 °C) in experimental study for 21 days. The immune status of Nile tilapia is affected by thermal fluctuation throughout the hatching season reflected by altered yolk sac size, length, and expression of the immune and stress related genes of the larvae, the best performances was observed at the beginning of the hatching season (March). High temperature (30 °C) suppress immune and stress responses throughout downregulation of all the genes under study, mask any effects for the immunostimulants, increased mortality in fish larvae suggesting narrow thermal tolerance range for the larvae compared with the adult fish. We recommend the use of amino acid mix as immunostimulant for Nile tilapia larvae, it reduces the mortality percentage and improve cellular response. Also, the use of ß-glucan should be prohibited during this developmental stage of larvae, it induced the highest mortality percentage.

Partial ameliorative effect of Moringa leaf ethanolic extract on the reproductive toxicity and the expression of steroidogenic genes induced by subchronic cadmium in male rats.

The impact of Moringa oleifera leaf ethanol extract (MOLEE) was assessed on the expression of the steroidogenic genes (steroidogenic acute regulatory protein (StAR) and cytochrome P450c17 subfamily a (CYP17a) and luteinizing hormone receptor (LHR) gene) as well as on the cadmium chloride (CdCl2)-induced reproductive toxicity for 56 days in male rats. Four groups were used: control, Moringa-treated (MOLEE), CdCl2-treated, and CdCl2 + MOLEE groups. The reproductive toxicity of CdCl2 was confirmed; it caused a significant decrease in the accessory sex organ weights, testosterone level, testicular GST level, elevated MDA level (lipid peroxidation indicator), and histopathological alterations in seminiferous tubules, prostate, seminal vesicles, and epididymis as well as sperm characteristics. It also induced downregulation in the expression of StAR and CYP17a genes without change in the expression LHR gene. Eleven active compounds were detected in the GC-MS analysis of MOLEE; six of them have antioxidant properties, and five new compounds presented variable activities. MOLEE alone induced a stimulatory effect on the expression of steroidogenic and LHR genes. It restored the weight of reproductive organs to the control level; however, the recovery in sperm count, motility, abnormalities, percentage of alive sperm, testosterone, and MDA level are still comparable with the control level. Similar findings were also reported at the histological structure of the testes, epididymis, and accessory sex glands. Complete recovery of the GST enzyme activity was observed. Additionally, a restoration in the expression level of the steroidogenic genes was also reported. Our results indicated that the concurrent administration of MOLEE with CdCl2 can partially mitigate its harmful effects on male fertility.

Optimum salinity for Nile tilapia (Oreochromis niloticus) growth and mRNA transcripts of ion-regulation, inflammatory, stress- and immune-related genes.

We aim to study the optimum salinity concentration for Nile tilapia, through the assessment of its growth performance and the expression of its related genes (Gh and Igf-1), as well as its salinity adaptation and immune status through the assessment of the gene expression of ion-regulation genes (Na+/K+-ATPase α-1a and α-1b), stress-related genes (GST, HSP27, and HSP70), inflammatory-related genes (IL1, IL8, CC, and CXC chemokine), and immune-related genes (IgMH TLR7, MHC, and MX) at the osmoregulatory organs (gills, liver, and kidney). Based on the least mortality percentage and the physical appearance of the fish, three salt concentrations (6, 16, and 20 ppt) were chosen following a 6-month preliminary study using serial salt concentrations ranged from 6 to 36 ppt, which were obtained by rearing the fish in gradual elevated pond salinity through daily addition of 0.5 ppt saline water. The fish size was 10.2-12 cm and weight was 25.5-26.15 g. No significant differences in the fish weight gain were observed among the studied groups. The group reared at 16-ppt salt showed better performance than that of 20 ppt, as they have lower morality % and higher expression of ion-regulated gene (Na+/K+-ATPase α1-b), stress-related genes (GST, HSP27, and HSP70) of the gills and also GST, inflammatory-related genes (IL-1ß and IL8), and TLR in the liver tissue. Higher expression of kidney-immune-related genes at 20-ppt salt may indicate that higher salinity predispose to fish infection and increased mortality. We concluded that 16-ppt salinity concentration is suitable for rearing O. niloticus as the fish are more adaptive to salinity condition without changes in their growth rate. Also, we indicate the use of immune stimulant feed additive to overcome the immune suppressive effect of hyper-salinity. Additionally, the survival of some fish at higher salinity concentrations (30-34 ppt) increase the chance for selection for salinity resistance in the Nile tilapia.

Characterization of type IV antifreeze gene in Nile tilapia (Oreochromis niloticus) and influence of cold and hot weather on its expression and some immune-related genes.

The aim of this work is to study the effect of the thermal stress of ambient temperature during winter and summer on the expression of type IV antifreeze gene (ANF IV) in different tissues of Nile tilapia (Oreochromis niloticus) as well as some immune-related genes. At first, genomic ANF IV gene was characterized from one fish; 124 amino acids were identified with 92.7% similarity with that on the gene bank. Expression of ANF IV and immune-related genes were done twice, once at the end of December (winter sample, temperature 14 °C) and the other at August (summer sample, temperature 36 °C). Assessment of ANF IV gene expression in different organs of fish was done; splenic mRNA was used for assessment of immune-related gene transcripts (CXCl2 chemokine, cc-chemokine, INF-3A, and MHC IIß). Winter expression analysis of AFP IV in O. niloticus revealed significant upregulation of mRNA transcript levels in the intestine, gills, skin, spleen, liver, and brain with 324.03-, 170.06-, 107.63-, 97.61-, 94.35-, and 27.85-folds, respectively. Furthermore, upregulation in the gene was observed in some organs during summer: in the liver, gills, skin, intestine, and brain with lower levels compared with winter. The level of expression of immune-related genes in winter is significantly higher than summer in all assessed genes. Cc-chemokine gene expression was the most affected in both winter and summer. Variable expression profile of ANF IV in different organs and in different seasons together with its amino acid similarity of N-terminal and C-terminal with apolipoprotein (lipid binder) and form of high-density lipoprotein (HDL) suggests a different role for this protein which may be related to lipid metabolism.

Modulatory effect of different doses of ß-1,3/1,6-glucan on the expression of antioxidant, inflammatory, stress and immune-related genes of Oreochromis niloticus challenged with Streptococcus iniae.

ß-glucans are widely-known immunostimulants that are profusely used in aquaculture industry. The present study was conducted to evaluate the effect of different in-feed doses of ß-1,3/1,6-glucans on the expression of antioxidant and stress-related genes (GST, HSP-70, Vtg), inflammation related genes (Il-8, TNFα, CXC-chemokine and CAS) and adaptive immune-related genes (MHC-IIß, TLR-7, IgM-H, and Mx) of Oreochromis niloticus challenged and non-challenged with Streptococcus iniae. Six experimental groups were established: non-challenged control (non-supplemented diet), challenged control (non-supplemented diet), non-challenged supplemented with 0.1% ß-glucan, challenged supplemented with 0.1% ß-glucan, non-challenged supplemented with 0.2% ß-glucan and challenged supplemented with 0.2% ß-glucan. Fish were fed with ß-glucan for 21 days prior challenge and then sampled after 1, 3 and 7 days post-challenge. In non-challenged group, variable effects of the two doses of ß-Glucans on the expression of the studied genes were observed; 0.1% induced higher expression of HSP70, CXC chemokine, MHC-IIß and MX genes. Meanwhile, 0.2% induced better effect on the expression of Vtg, TNF-α, CAS and IgM-H, and almost equal effects of both doses on GST and IL8. However, with the challenged group, 0.2% ß-Glucans showed better effect than 0.1% at day one post challenge through significant up-regulation of GST, HSP, IL8, TNF-α, CXC, and MHC-IIß, meanwhile, the effect of 0.1% was only on the expression of HSP70, MHC-IIß, and TLR7 at day 3 post challenge. No stimulatory role for both doses of ß-Glucans on the expression of almost all genes at day 7 post-challenge. We conclude that both doses of ß-glucan can modulate the antioxidant, inflammation, stress and immune-related genes in Nile tilapia, moreover, 0.2% ß-Glucans showed better protective effect with Streptococcus iniae challange.

Modulatory Effect of Monochromatic Blue Light on Heat Stress Response in Commercial Broilers.

In a novel approach, monochromatic blue light was used to investigate its modulatory effect on heat stress biomarkers in two commercial broiler strains (Ross 308 and Cobb 500). At 21 days old, birds were divided into four groups including one group housed in white light, a second group exposed to blue light, a 3rd group exposed to white light + heat stress, and a 4th group exposed to blue light + heat stress. Heat treatment at 33°C lasted for five h for four successive days. Exposure to blue light during heat stress reduced MDA concentration and enhanced SOD and CAT enzyme activities as well as modulated their gene expression. Blue light also reduced the degenerative changes that occurred in the liver tissue as a result of heat stress. It regulated, though variably, liver HSP70, HSP90, HSF1, and HSF3 gene expression among Ross and Cobb chickens. Moreover, the Cobb strain showed better performance than Ross manifested by a significant reduction of rectal temperature in the case of H + B. Furthermore, a significant linear relationship was found between the lowered rectal temperature and the expression of all HSP genes. Generally, the performance of both strains by most assessed parameters under heat stress is improved when using blue light.

Assessment of complex water pollution with heavy metals and Pyrethroid pesticides on transcript levels of metallothionein and immune related genes.

Alteration of immunological function of an aquatic organism can be used as an indicator for evaluating the direct effect of exposure to pollutants. The aim of this work is to assess the impact of complex water pollution with special reference to Pyrethroid pesticides and heavy metals on mRNA transcript levels of Metallothionine and some immune related genes of Nile tilapia (Oreochromas Niloticus). Residues of six heavy metals and six Pyrethroid were assessed in water as well as fish tissues at three different sites of Lake Burullus, located at Northern Egypt. Variations of water physicochemical properties associated with different levels of heavy metals at the three different sections were recorded. Tissue residues of Fe, Mn and Zn, Cu, Ni exceed water levels in contrast to elevated water level of Pb. All assessed Pyrethroids are detected in fish tissue samples with higher concentration (3-42 folds) than that found in water samples especially Cypermethrin. Significant down-regulation of expression levels of metallothionein (MT) at the three sections of the lake was observed. The expression of immune related genes (IgM) and inflammatory cytokines (TNF, IL.8 and IL.1) were affected. IgM and TNF were significantly down-regulated at eastern and western section of the lake; meanwhile the expression of IL8 is down regulated at the three sections of the lack. IL1 was significantly up-regulated at eastern and middle sections. We conclude that, variable gene expression of MT and immune-related genes at the three sections of the lack impose different response to complex water pollution in relation to variable aquatic environment.

Vitamin C Modulates the Immunotoxic Effect of 17α-Methyltestosterone in Nile Tilapia.

The synthetic androgen 17α-methyltestosterone (MT) is profusely used and practically needed in the production of all-male Nile tilapia fry; however, such androgenic hormones badly disrupt the immune system. This study aimed to alleviate or counteract the immunotoxic effect of MT using vitamin C (ascorbic acid or vit C). Our results show that the highest phagocytic activity (PA), phagocytic index (PI), and lysozyme activity were detected in the vit C group and the MT plus vit C group. Furthermore, PA and PI were significantly suppressed, but lysozyme activity was stronger in the MT group than in the control. No differences were detected in the differential leukocyte count among the studied groups. Moreover, vit C obviously reduced the upregulated expression level of the innate immune-related genes, interleukin 1ß (il1ß), interleukin 8 (il8), tumor necrosis factor α (tnfα), CC-chemokine, Toll-like receptor 7 (tlr7), immunoglobulin M (IgM) heavy chain, and cellular apoptosis susceptibility (cas) induced by MT, excluding tnfα in the liver and CC-chemokine and tlr7 in the kidney. The micronucleus frequency was found to significantly improve in the vit C plus MT group in comparison to that in the MT group. Normal histoarchitecture of the liver, kidney, and spleen was observed in all the groups, except for the frequently observed melanomacrophage centers in the spleen and kidney of the fish that were treated with vit C and vit C plus MT. More importantly, our findings demonstrate that the upregulation of immune-related genes is not necessarily a sign of a stimulated or enhanced immune system.

Impaired antioxidant gene expression by pesticide residues and its relation with other cellular biomarkers in Nile Tilapia (Oreochromis niloticus) from Lake Burullus.

Organochlorines and Organophosphorus are the most commonly used pesticides. These pesticides constitute a considerable contaminating threat due to their excessive agricultural usage which in turn contaminates the aquatic system through agricultural drainage. The aim of this study was to evaluate water and tissue residues of both pesticides in O. niloticus obtained from three different sections in Lake Burullus, Egypt. Assessment of relative change in mRNA levels of GST and Vtg (oxidative stress indicator) was done and its relation with other cellular biomarkers including apoptosis, which is assessed by Cellular apoptosis susceptibility transcript level (CAS), comet assay and micronucleus assays (genotoxicity indicator). Pesticide residue levels in water are fluctuating. In fish tissues, most residues were higher than those found in water and were associated with down regulation of hepatic GST gene and Vtg expression. CAS gene involved in apoptosis, its transcript is down regulated in middle and western sections of the lake with higher pesticide residues. Different degrees of DNA damages in O. niloticus' liver cells were demonstrated by comet assay. Significant increase in the micronucleated cells in the three sections of the lake was observed; the western section fish showed the highest number. Persistent exposures of fish to pesticide caused impairment of antioxidant gene expression. This negatively affects apoptosis associated with damaging DNA and chromosome fragments.

The extent to which immunity, apoptosis and detoxification gene expression interact with 17 alpha-methyltestosterone.

Innate immunity is the first line of defence against invasion by foreign pathogens. One widely used synthetic androgen for the production of all-male fish, particularly commercially valuable Nile tilapia, Oreochromis niloticus, is 17 alpha-methyltestosterone (MT). The present study investigates the effect of MT on innate immunity, cellular apoptosis and detoxification and the mortality rate, during and after the feeding of fry with 0-, 40-and 60-mg MT/kg. Expression analysis was completed on interleukin 1 beta (il1ß), interleukin 8 (il8), tumour necrosis factor alpha (tnfα), CXC2- and CC-chemokines, interferon (ifn), myxovirus resistance (mx), toll-like receptor 7 (tlr7), immunoglobulin M heavy chain (IgM heavy chain), vitellogenin (vtg), cellular apoptosis susceptibility (cas) and glutathione S-transferase α1 (gstα1). Expression analysis revealed that MT had a significant impact on these genes, and this impact varied from induction to repression during and after the treatment. Linear regression analysis showed a significant association between the majority of the tested gene transcript levels and mortality rates on the 7th and 21st days of hormonal treatment and 2 weeks following hormonal cessation. The results are thoroughly discussed in this article. This is the first report concerning the hazardous effect of MT on a series of genes involved in immunity, apoptosis and detoxification in the Nile tilapia fry.

Vitamin C modulates cadmium-induced hepatic antioxidants' gene transcripts and toxicopathic changes in Nile tilapia, Oreochromis niloticus.

Cadmium (Cd) is one of the naturally occurring heavy metals having adverse effects, while vitamin C (L-ascorbic acid) is an essential micronutrient for fish, which can attenuate tissue damage owing to its chain-breaking antioxidant and free radical scavenger properties. The adult Nile tilapia fish were exposed to Cd at 5 mg/l with and without vitamin C (500 mg/kg diet) for 45 days in addition to negative and positive controls fed with the basal diet and basal diet supplemented with vitamin C, respectively. Hepatic relative mRNA expression of genes involved in antioxidant function, metallothionein (MT), glutathione S-transferase (GST-α1), and glutathione peroxidase (GPx1), was assessed using real-time reverse transcription polymerase chain reaction (RT-PCR). Hepatic architecture was also histopathologically examined. Tilapia exposed to Cd exhibited upregulated antioxidants' gene transcript levels, GST-⍺1, GPx1, and MT by 6.10-, 4.60-, and 4.29-fold, respectively. Histopathologically, Cd caused severe hepatic changes of multifocal hepatocellular and pancreatic acinar necrosis, and lytic hepatocytes infiltrated with eosinophilic granular cells. Co-treatment of Cd-exposed fish with vitamin C overexpressed antioxidant enzyme-related genes, GST-⍺1 (16.26-fold) and GPx1 (18.68-fold), and maintained the expression of MT gene close to control (1.07-fold), averting the toxicopathic lesions induced by Cd. These results suggested that vitamin C has the potential to protect Nile tilapia from Cd hepatotoxicity via sustaining hepatic antioxidants' genes transcripts and normal histoarchitecture.

Association of a novel SNP in exon 10 of the IGF2 gene with growth traits in Egyptian water buffalo (Bubalus bubalis).

Insulin-like growth factor 2 (IGF2) plays an important role in muscle growth and it might be used as a marker for the growth traits selection strategies in farm animals. The objectives of this study were to detect polymorphisms in exon 10 of IGF2 and to determine associations between these polymorphisms and growth traits in Egyptian water buffalo. PCR-single-strand conformation polymorphism (SSCP) and DNA sequencing methods were used to detect any prospective polymorphism. A novel single nucleotide polymorphism (SNP), C287A, was detected. It was a non-synonymous mutation and led to replacement of glutamine (Q) amino acid (aa) by histidine (H) aa. Three different SSCP patterns were observed: AA, AC, and CC, with frequencies of 0.540, 0.325, and 0.135, respectively. Association analyses revealed that the AA individuals had a higher average daily gain (ADG) than other individuals (CC and AC) from birth to 9 months of age. We conclude that the AA genotype in C287A SNP in the exon 10 of the IGF2 gene is associated with the ADG during the age from birth to 9 months and could be used as a potential genetic marker for selection of growth traits in Egyptian buffalo.