Biochemical and molecular investigation of thermal manipulation protocols during broiler embryogenesis and subsequent thermal challenge.

BACKGROUND: The aim of the current study was to evaluate the effect of different thermal manipulation (TM) protocols during embryogenesis on thermotolerance acquisition parameters during subsequent thermal challenge (TC) at posthatch day 28. A total of 1500 fertile chicken eggs were divided randomly into five treatments (300 eggs each): control was maintained at 37.8 °C and 56 % relative humidity (RH) whereas, TM1, TM2, TM3 and TM4 were subjected to 38.5, 39, 39.5 and 40 °C for 18 h and 65 % RH daily during embryonic days ED 12-18. Hatched chicks from each treatment group allocated randomly into two sub-treatment groups (thermo-neutral, naïve (TN) and thermal challenge (TC). At day 28 of age, chicks subjected to TC by adjusting room temperature to 42 °C for 6 h while naïve chicks kept under regular conditions (22 ± 1 °C and 50-60 % RH). Chick's response to TC evaluated by determination of plasma T3, T4, corticosterone, total proteins, albumin, selected enzymes and some electrolytes at the beginning (0 h) and after 1, 3 and 5 h of TC in TM and TN chicks. Furthermore, pectoral and thigh muscles mRNA expression of Atrogin-1, CK, avUCP, DIO3, DIO2 were evaluated in TC and TN sub-treatment groups. RESULTS: TM induced a significant reduction in free T3 and elevation in total proteins and albumin in plasma with significant down-regulation of Atrogin-1 and DIO2 and significant up-regulation of DIO3 mRNA expression in muscle of TM chicks compare to control. During TC at day 28, decrease in the concentrations of plasma free T3, total proteins and albumin with increase in T4 have been detected in control and TM chicks. TC induced up-regulation of Atrogin-1 and DIO3 with down-regulation of DIO2 gene expression in muscles of all TC chicks. CONCLUSION: The present study indicated that, TM improved thermotolerance acquisition by decreasing basal metabolic rate and muscle injury during thermal stress. Basal metabolic rate decreased via reduction of plasma T3 concentration with up and down regulation of expression of DIO3 and DIO2, respectively in muscles. Muscle injury protected by stimulation of protein biosynthesis and down-regulation of Atrogin-1 expression.

Curcumin regulates gene expression of insulin like growth factor, B-cell CLL/lymphoma 2 and antioxidant enzymes in streptozotocin induced diabetic rats.

BACKGROUND: The effects of curcumin on the activities and gene expression of antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), glutathione-S-transferase (G-ST), B-cell CLL/lymphoma 2 (Bcl-2) and insulin like growth factor-1 (IGF-1) in diabetic rats were studied. METHODS: Twenty four rats were assigned to three groups (8 rats for each). Rats of first group were non diabetic and rats of the second group were rendered diabetic by streptozotocin (STZ). Both groups received vehicle, corn oil only (5 ml/kg body weight) and served as negative and positive controls, respectively. Rats of the third group were rendered diabetic and received oral curcumin dissolved in corn oil at a dose of 15 mg/5 ml/kg body weight for 6 weeks. RESULTS: Diabetic rats showed significant increase of blood glucose, thiobarbituric acid reactive substances (TBARS) and activities of all antioxidant enzymes with significant reduction of reduced glutathione (GSH) compare to the control non diabetic group. Gene expression of Bcl2, SOD, CAT, GPX and GST was increased significantly in diabetic untreated rats compare to the control non diabetic group. The administration of curcumin to diabetic rats normalized significantly their blood sugar level and TBARS values and increased the activities of all antioxidant enzymes and GSH concentration. In addition, curcumin treated rats showed significant increase in gene expression of IGF-1, Bcl2, SOD and GST compare to non diabetic and diabetic untreated rats. CONCLUSION: Curcumin was antidiabetic therapy, induced hypoglycemia by up-regulation of IGF-1 gene and ameliorate the diabetes induced oxidative stress via increasing the availability of GSH, increasing the activities and gene expression of antioxidant enzymes and Bcl2. Further studies are required to investigate the actual mechanism of action of curcumin regarding the up regulation of gene expression of examined parameters.

Aqueous Ginger (Zingiber officinale) Extract Ameliorates the Harmful Effects of High-Dose Lornoxicam in Albino Male Rats.

Lornoxicam is a potent oxicam-class nonsteroidal anti-inflammatory drug (NSAID) with analgesic, anti-inflammatory, and antipyretic effects. Its impacts on many biological functions are not fully understood. We measured various biomarkers in male albino rats provided an oral aqueous ginger extract before IM administration of therapeutic and 2× the therapeutic doses of lornoxicam. The aqueous ginger plant extract was characterized by mass spectroscopy, and its effects were determined by examining free radical scavenging activity, blood parameters, renal and hepatic function, semen quality, proinflammatory cytokines, antioxidant markers, and histopathology. Rats administered lornoxicam had significantly higher liver and kidney function biomarker values, TNF-α, interleukin-6, and sperm abnormalities than the control rats. The overall erythrocyte count, packed cell volume, prostaglandin, and sperm counts were all considerably lower in the experimental animals. Histological changes were found in the liver, spleen, and testes of rats administered lornoxicam alone. In rats, pretreatment with ginger extract reduced the majority of the negative effects of conventional and high dosages of lornoxicam.

Enrichment of Broiler Chickens' Meat with Dietary Linseed Oil and Lysine Mixtures: Influence on Nutritional Value, Carcass Characteristics and Oxidative Stress Biomarkers.

This study aimed to evaluate the effect of four combinations of dietary linseed oil and lysine mixtures on performance, fatty and amino acid profiles, oxidative stress biomarkers, cell energy and meat quality parameters of broiler chickens. One hundred and sixty broiler chicks were allocated into four groups. Birds of groups 1-4 were fed diets containing optimum lysine and 2% of linseed oil, optimum lysine and 4% of linseed oil, high lysine and 2% of linseed oil, and high lysine and 4% of linseed oil, respectively, for a period of 35 days. High linseed oil or lysine levels did not affect the performance of the tested birds, but the high level of dietary linseed oil decreased the concentrations of muscles' saturated fatty acids (SFA). The highest values of ω-3 polyunsaturated fatty (ω-3 PUFA) and arachidonic acids with lowest levels of monounsaturated fatty (MUFA) were detected in the muscles of birds fed diets containing high linseed oils and/or lysine levels. High linseed oil or lysine levels provided the best essential amino acid profile and improved antioxidant components as well as cell energy, and tenderness and redness of the meat. Conclusively, high dietary lysine and linseed oil combinations improved the nutritional value, antioxidant status and cell energy of broiler chickens' meat.

Biosynthesized Iron Oxide Nanoparticles from Petroselinum crispum Leaf Extract Mitigate Lead-Acetate-Induced Anemia in Male Albino Rats: Hematological, Biochemical and Histopathological Features.

This study aimed to investigate the ameliorative effects of iron oxide nanoparticles (IONPs) prepared from leaf extract of Petroselinum crispum compared to those prepared using a chemical method in lead-acetate-induced anemic rats. Twenty rats were divided into four groups (five rats each). Throughout the experimental period (8 weeks), the rats in group 1 were not given any therapy. The rats in groups 2, 3 and 4 were given 400 ppm lead acetate orally for 2 weeks to make them anemic. Following that, these rats were either left untreated, given 27 ppm of chemical IONPs orally or given 27 ppm of natural IONPs orally for the remaining 6 weeks of the experiment. TEM analysis indicated that the chemically and naturally prepared IONPs had sizes of 6.22-9.7 and 64-68 nm, respectively. Serum ferritin and iron concentrations were reduced, whereas the total iron-binding capacity (TIBC), ALT, AST, urea and creatinine were significantly increased in the non-treated lead-acetate-induced anemic rats compared to those of the control. In addition, congestion, hemorrhage, necrosis, vacuolation and leukocytic infiltration in the kidneys, liver and spleen were observed in non-treated lead-acetate-induced anemic rats compared to the control. The effects of lead acetate were mitigated by IONPs, particularly the natural one. In conclusion, IONPs produced from Petroselinum crispum leaf extract can be used as an efficient and safe therapy in lead-acetate-induced anemic rats.

Impact of Dietary Betaine and Metabolizable Energy Levels on Profiles of Proteins and Lipids, Bioenergetics, Peroxidation and Quality of Meat in Japanese Quail.

Three different diets were formulated with three levels of metabolizable energy (ME) (optimum; 2900, restricted; 2800 and low; 2700 kcal ME/kg diet) without or with (0 and 0.15%) betaine supplementation in 2 × 3 factorial design to evaluate the effect of six experimental diets on performance, proteins and lipids profiles, bioenergetics, peroxidation and meat quality of Japanese quail. Therefore, 360 quails allocated into six groups in a 23-day experiment. Dietary betaine and ME levels did not affect the performance, meat energy indices (ATP and AMP) and malondialdehyde (MDA) levels of Japanese quail meat. Dietary betaine and/or ME levels induced significant changes in serum triacylglycerol (TAG), total cholesterols (TC), low-density lipoprotein cholesterol (LDL-c), very low-density lipoprotein cholesterol (VLDL-c), meat total lipids and cholesterol of Japanese quail. Optimum and restricted ME levels reduced total volatile basic nitrogen (TVBN) whereas dietary betaine increased ecosapentaenoic (EPA), docosahexaenoic acids (DHA) and glutamine concentrations in breast meat of Japanese quail. Dietary betaine and low energy diet improved cooking loss, thawing loss (ThL) and water holding capacity (WHC) in breast meat of Japanese quail. Conclusively, dietary betaine improved meat quality of Japanese quail fed diets containing either restricted or low ME by enrichments the meat with omega-3 fatty acids and reduction of lipids levels.

Influence of Dietary Garlic (Allium sativum) and/or Ascorbic Acid on Performance, Feed Utilization, Body Composition and Hemato-Biochemical Parameters of Juvenile Asian Sea Bass (Lates calcarifer).

The current study investigated effects of garlic (Allium sativum) and/or ascorbic acid on growth performance, feed utilization, biochemical body composition, and hemato-biochemical parameters of juvenile Asian sea bass. A total of 600 fish (43.14 ± 0.23 g body weight) were divided into four groups. Fish in the first group were fed basal diet and served as a control group. Fish in groups 2, 3 and 4 were fed a basal diet mixed with garlic (40 g/kg diet), ascorbic acid (1.5 g/kg diet), or garlic (20 g/kg diet)/ascorbic acid (0.75 g/kg diet) mixture, respectively, for 12 weeks. A significant (p < 0.05) increase was observed in growth performance, feed utilization, and chemical body composition in fish fed garlic alone in comparison with the control and other treated groups. All hematological indices, biochemical parameters, and survival rate were not changed significantly (p > 0.05) in all groups throughout the experimental period when compared with the control. Total cholesterol and feed conversion ratio were significantly (p < 0.05) decreased in fish fed garlic alone in comparison to the control and other treated groups. Conclusively, dietary supplementation of garlic alone (40 g/kg diet) was highly effective in improving most of the studied parameters in comparison with that of ascorbic acid alone or a mixture of garlic (20 g/kg diet) and ascorbic acid (0.75 g/kg diet).

Season's Effects on Some Clinical, Hematological Parameters and Blood Cortisol Level in Sedated Arabian Horses With Xylazine.

Influence of heat or cold stress in sedated animals is unclear and requires further investigations. The present study aimed to evaluate the season's effects on some clinical, hematological parameters and blood cortisol level in sedated Arabian horses with xylazine. Therefore, seven Arabian horses were used to investigate heart and respiratory rates, and capillary refill time and serum cortisol level were recorded before (0) and at 5, 15, 60, and 180 minutes postsedation. Heparinized venous samples were collected before (0) and 3 hours postsedation for analysis of hematological analysis. Arterial blood samples were collected before and 1 hour postsedation for arterial blood gases and electrolytes analysis. Repeated analysis of variance was performed (P < .05). Significant decreases have been observed in heart and respiratory rates at 5, 15, and 60 minutes postsedation in summer and only at 5 minutes postsedation in winter. Arterial oxygen pressure and arterial carbon dioxide pressure showed a significant decrease and increase, respectively at 1 hour postsedation in summer and winter. The serum cortisol levels were significantly higher in summer than in winter at 5, 15, and 60 minutes postsedation. In summer, the postsedation concentrations of cortisol did not change significantly than its values before sedation. However, in winter, the cortisol concentration decreased significantly at 5, 15, and 60 minutes postsedation compared with their value before sedation. The present study suggests that these season's effects on the sedated Arabian horses could take into consideration in xylazine-sedated Arabian horses.

Simultaneous multiresidue determination of metronidazole and spiramycin in fish muscle using high performance liquid chromatography with UV detection.

An efficient multiresidue method for the simultaneous determination of metronidazole (MET) and spiramycin (SPY) in tilapia fish muscle, based on high performance liquid chromatography with UV detection (HPLC-UV), has been developed. The drugs were extracted with 0.2% orthophosphoric acid-methanol (6:4), and the extracts were cleaned up on a solid phase extraction cartridge, C18 Sep-Pak light column. The LC separation was performed on a RP stainless-steel C-18 analytical column (150 mm x 4.6 mm, 5 microm) with a gradient elution system of 0.05 M phosphate buffer adjusted to pH 2.4-acetonitrile as the mobile phase at the flow rate of 1.0 ml min(-1). A wavelength programming was applied for the UV detection of the analytes. The method not only enabled the determination of the parent drugs, MET and SPY, but also permitted the determination of their metabolites, hydroxymetronidazole (HMET) and neospiramycin (NSPY). The calibration graphs for each drug were rectilinear in the range of 0.005-1.000 microg g(-1) for MET and HMET and 0.025-1.000 microg g(-1) for SPY and NSPY. With this method, the cited drugs with their metabolites were determined in fortified fish muscle tissues at levels of 0.025, 0.1 and 1.0 microg g(-1) with good accuracy and precision. LOD and LOQ obtained for each drug were as follows: 0.002 and 0.005 microg g(-1) for MET and HMET and 0.005 and 0.025 microg g(-1) for SPY and NSPY. Utilization of the method to successfully analyze tilapia fish muscle samples incurred with MET and SPY was described.

Serum biochemical profile and molecular detection of pathogens in semen of infertile male dromedary camels (Camelus dromedarius).

Detection of pathogens in the semen of camels has not been completely elucidated. Therefore, the current study aimed to determine the association of some economically important pathogens with infertility in 94 male infertile camels through molecular detection and estimation of selected biochemical parameters in serum of these animals compared with a control non infected fertile animals (n=40). PCR analysis of semen samples of infertile camels indicated that, four potential pathogens namely Mycoplasma spp., Leptospira spp., Brucella melitensis, and Bovine viral diarrhea virus (BVDV) were detected in 50 semen samples of infertile camels whereas, 44 semen samples of infertile camels were free of pathogens and all tested semen samples were negative for bovine herpes virus 1, Salmonella spp. and Trypanosoma evansi. Single and mixed infection was detected in 88% and 12% of the infected semen samples, respectively. Mycoplasma spp., Leptospira spp., Brucella and Bovine viral diarrhea virus infection represented 66%, 27.2%, 4.5% and 2.3% of the single infected semen samples. Mycoplasma spp.+Leptospira spp. and Mycoplasma spp.+Brucella spp. were detected in 83.3% and 16.7% of mixed infected semen samples, respectively. Testosterone concentration decreased significantly in infertile infected camels compare to both control and infertile non infected animals that remained comparable. The current findings reported the molecular detection of mixed infection in camel semen for the first time. Mycoplasma spp. is the most widely recognized microorganism in the present study and together with Leptospira spp., Brucella spp. and Bovine viral diarrhea virus, might be associated with infertility in dromedary camels.

Cancer cell death induced by nanomagnetolectin.

Magnetic nanoparticles represent a new paradigm for molecular targeting therapy in cancer. However, the transformative targeting potential of magnetic nanoparticles has been stymied by a key obstacle-safe delivery to specified target cells in vivo. As cancer cells grow under nutrient deprivation and hypoxic conditions and decorate cell surface with excessive sialoglycans, sialic acid binding lectins might be suitable for targeting cancer cells in vivo. Here we explore the potential of magnetic nanoparticles functionalized with wheat germ lectin (WGA) conjugate, so-called nanomagnetolectin, as apoptotic targetable agents for prostate cancer. In the presence of magnetic field (magnetofection) for 15min, 2.46nM nanomagnetolectin significantly promoted apoptosis (∼12-fold, p value <0.01) of prostate cancer cells (LNCaP, PC-3, DU-145) compared to normal prostate epithelial cells (PrEC, PNT2, PZ-HPV-7), when supplemented with 10mM sialic acid under nutrient deprived condition. Nanomagnetolectin targets cell-surface glycosylation, particularly sialic acid as nanomagnetolectin induced apoptosis of cancer cells largely diminished (only 2 to 2.5-fold) compared to normal cells. The efficacy of magnetofected nanomagnetolectin was demonstrated in orthotopically xenografted (DU-145) mice, where tumor was not only completely arrested, but also reduced significantly (p value <0.001). This was further corroborated in subcutaneous xenograft model, where nanomagnetolectin in the presence of magnetic field and photothermal heating at ∼42°C induced apoptosis of tumor by ∼4-fold compared to tumor section heated at ∼42°C, but without magnetic field. Taken all together, the study demonstrates, for the first time, the utility of nanomagnetolectin as a potential cancer therapeutic.

Thermal manipulation during broiler chicken embryogenesis: Effect on mRNA expressions of Hsp108, Hsp70, Hsp47 and Hsf-3 during subsequent post-hatch thermal challenge.

Effects of thermal manipulation during broiler chicken embryonic days 12-18 on body temperature (T(b)) and mRNA expressions of Hsp108, Hsp70, Hsp47 and Hsf-3 in muscle, heart and brain tissues during subsequent thermal challenge (TC) were investigated. Fertile chicken eggs were divided randomly into four groups (n=375): eggs in the control group were maintained at 37.8°C and 56% (RH). Eggs in TM1 group were subjected to TM at 39°C for 9h during ED 12-18. Eggs in the TM2 and TM3 groups were subjected to the same protocol of TM1 except for increasing the period of exposure to 12h and 18h, respectively. During TC (43°C for 6h) at days 10 and 28, T(b) of TM chicks was significantly lower compared to controls. Furthermore, significant changes in mRNA expressions of Hsp108, Hsp70 and Hsp47 in muscle, heart and brain tissues were observed.

Biochemical and histopathological study in rats intoxicated with carbontetrachloride and treated with camel milk.

The unique characters of camel's milk make it used extensively in the field of medicine as anti-microbial, anti-diabetic and hepatoprotective agent. The lack of studies demonstrating the protective effect of camel's milk against hepatotoxic compound was the main reason beyond the conduction of the current experiment which aimed to investigate the protective effects of camel's milk against carbontetrachloride (CCl4) induced hepatotoxicity. Therefore, 24 rats were fed on standard diet and divided into four groups. Rats of the first group and second groups were injected i/p with paraffin oil and received either tap water (control 1) or camel's milk (control 2), respectively. Rats of the third and fourth groups were injected i/p with CCl4 and received either tap water or camel's milk, respectively. At the end of the experiment (5 weeks), blood and liver samples were collected for biochemical and histopathological analysis. The present findings revealed that, CCl4 elevated serum enzyme activities of liver and some biochemical parameters, but these effects were prevented by the treatment of rats with camel milk. Histopathologically, a great amount of mononuclear cells infiltration, necrotic cells and few fibroblasts were observed in liver of CCl4 treated group. The present study concluded that camel milk treatment may play a protective role against CCl4-induced liver damages in rats. These protective effects were in the form of improving of liver enzyme activities, blood biochemical parameters and histological picture of liver of intoxicated rats. In the future, examination of the liver protective effect of camel milk against CCl4 in dose dependant manner could be investigated.

Investigation of selected biochemical indicators of Equine Rhabdomyolysis in Arabian horses: pro-inflammatory cytokines and oxidative stress markers.

A total of 30 horses were divided into two groups, one served as a control whereas other was rhabdomyolysis diseased horses. After blood collection, the resulted sera were used for estimation of the activities of creatin kinase (CK), aspartate transaminase (AST), lactate dehydrogenase (LDH), lactic acid, triacylglycerol (TAG), glucose, total protein, albumin, globulin, urea, creatinine, Triiodothyronine (T(3)), calcium, sodium, potassium, phosphorus, chloride, vitamin E, interleukin-6 (IL-6) and tumor necrosis-α (TNF-α). In addition, whole blood was used for determination of selenium, reduced glutathione (G-SH) and prostaglandin F2-α (PGF2α). The erythrocyte hemolysates were used for the determination of the activities of super oxide dismutase (SOD), catalase (CAT), total antioxidant capacity (TAC), nitric oxide (NO) and malondialdehyde (MDA). The present findings revealed a significant (p ≤ 0.05) increase in the values of CK, AST, LDH, glucose, lactate, TAG, urea, creatinine, phosphorus, MDA, TNF- α, IL6 and PGF2- α in diseased horses when compared with the control. Furthermore, the values of calcium, SOD, CAT, TAC, NO and GSH in diseased horses were significantly (p ≤ 0.05) lower than the control. The other examined parameters were not statistically significant. In conclusion, the examined pro-inflammatory cytokines were useful biomarkers for the diagnosis of Equine rhabdomyolysis (ER) in Arabian horses beside the old examined biomarkers. In the future, efforts should be made to confirm this in other breed. If this could be achieved, it would open up new perspectives in research fields dealing with ER.