Sheep Red Research Articles

Suppressing Effect of Flavonoid Compounds on Lipids Photooxidation of Sheep Red Blood Cells and Oleic Acid Photooxidation

ABSTRACTPhotosensitizers and pigments in raw meat such as porphyrins, riboflavin, and myoglobin after incorporation with light beam prompt the generation of singlet oxygen (1O2) from triplet oxygen (3O2) and cause oxidative rancidity of meat products. In this study, the results of photooxidation reactions of sheep erythrocyte (red blood cell) model as a model rich in hemoglobin and phospholipids bilayer, and oleic acid model were obtained by 1H NMR spectroscopy, TBARS assay, and iodometric titration. In both models, the rate of lipid photooxidation in the presence of hydroalcoholic extracts of Turmeric (Curcuma longa L.) and Cumin (Cuminum cyminum L.) as natural antioxidants, Butyl hydroxytoluene (BHT) as a synthetic antioxidant, and sodium azide (NaN3) as a well‐known 1O2 scavenger were decreased in the order of NaN3 > Turmeric > Cumin > BHT. It was proven that during the photooxidation process, there is a direct association between the amount of flavonoid compounds and 1O2 scavenging.

Antimicrobial and Antiplasmodial Activities of Endophytic Fungi Associated with Psidium guajava

Infections due to antimicrobial-resistant microorganisms have become widespread in recent years. Thus, searching for novel antimicrobial agents to combat such pathogens has become crucial. The current study aimed to evaluate the antimicrobial, antiplasmodial, and immunomodulatory activities of the extracts of endophytic fungi isolated from Psidium guajava. Isolation, identification, fermentation, and extraction of the secondary metabolites of the fungal endophytes were carried out following standard procedures. The extracts were subjected to High-Performance Liquid Chromatography-Diode Array Detector (HPLC-DAD) analysis to detect their bioactive components. The Antimicrobial activity and Minimum Inhibitory Concentration of the fungal extracts were evaluated against pure cultures of Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Salmonella typhi, and Klebsiella pneumonia using Agar well diffusion and Agar dilution method respectively. The acute toxicity study (LD50) was carried out using Lorke’s method. The extracts were tested in vivo in mice for antiplasmodial activity against Plasmodium berghei and in vitro against Plasmodium falciparum using Peter and Reyley's curative test method and WHO standardized micro-test system with modification respectively. The immunomodulatory activity of the extracts was evaluated by cyclophosphamide-induced myelosuppression (hematological parameters). Active extracts were further subjected to Delayed-Type Hypersensitivity Response (DTHR) and Haemagglutination Inhibition Assay using Sheep Red Blood Cells as antigens. The result showed Alternaria sp. (PGL1, PGL2, PGL3), from P. guajava. The HPLC-DAD analysis revealed the presence of bioactive compounds previously reported to have antimicrobial, antiplasmodial, and immunomodulatory properties. The fungal extracts exhibited varying degrees of antimicrobial activity. The LD50 of the fungi extracts was>5000 mg/kg in mice. The extracts at 100 and 200 mg/kg body weight in mice showed varying degrees of antiplasmodial activity. Growth of P. berghei was significantly (p<0.001) inhibited, curative effect ranges from 59.09 – 100%. Schizont maturation of P. falciparum isolates was inhibited and the highest level of inhibition was observed at 1 mg/ml (p<0.05). The fungal extracts reversed the effect of cyclophosphamide-induced reduction in total white blood cell counts and % neutrophil. This study showed that the tested plant harbors species of endophytic fungi that contain numerous secondary metabolites. The endophytic fungi showed prophylactic, immunostimulatory, and antiplasmodial activities, which can be exploited to develop antimicrobial, antiplasmodial, and immunomodulatory agents.

Hypo-Osmotic Stress and Pore-Forming Toxins Adjust the Lipid Order in Sheep Red Blood Cell Membranes.

Lipid ordering in cell membranes has been increasingly recognized as an important factor in establishing and regulating a large variety of biological functions. Multiple investigations into lipid organization focused on assessing ordering from temperature-induced phase transitions, which are often well outside the physiological range. However, particular stresses elicited by environmental factors, such as hypo-osmotic stress or protein insertion into membranes, with respect to changes in lipid status and ordering at constant temperature are insufficiently described. To fill these gaps in our knowledge, we exploited the well-established ability of environmentally sensitive membrane probes to detect intramembrane changes at the molecular level. Our steady state fluorescence spectroscopy experiments focused on assessing changes in optical responses of Laurdan and diphenylhexatriene upon exposure of red blood cells to hypo-osmotic stress and pore-forming toxins at room temperature. We verified our utilized experimental systems by a direct comparison of the results with prior reports on artificial membranes and cholesterol-depleted membranes undergoing temperature changes. The significant changes observed in the lipid order after exposure to hypo-osmotic stress or pore-forming toxins resembled phase transitions of lipids in membranes, which we explained by considering the short-range interactions between membrane components and the hydrophobic mismatch between membrane thickness and inserted proteins. Our results suggest that measurements of optical responses from the membrane probes constitute an appropriate method for assessing the status of lipids and phase transitions in target membranes exposed to mechanical stresses or upon the insertion of transmembrane proteins.

Immunoprotective Effect of Cocos nucifera Oil on Sheep Red Blood Cell-Induced Immunocompromised Rats

The aim of this study is to investigate the immunoprotective effects of cold-pressed coconut oil in immunocompromised rat. Standard procedure was used to perform this research work with a modified method. The effect of immunomodulatory properties of coconut oil was evaluated after challenging the animals with 0.3 ml sheep red blood cells (SRBCs) intraperitoneally and further treated with graded doses (0.25, 0.5 and 1.0 ml/kg i.p) of the oil extract for 21 days. The hematological, CD4, CD8 biochemical and histopathological analysis were evaluated. Result obtained from this study showed a significant increase in hematological indexes (WBC: 11.70; LYM: 9.9 GRAN: 8.0 and HGB: 15.57) across the treated groups, but majorly at lowest dose (0.25 ml/kg). More so a significant increase in CD4 and CD8 Count specifically at 0.25 ml/kg (7.763 and 7.830). Anti-oxidant property of the oil extract at 0.25 ml/kg had a significant reduction in malondialdehyde with an increased in antioxidant enzymes (SOD, CAT and GPx) when compared with untreated control. The body weight of the animals in the treated groups showed a significant increase at 0.25 ml/kg of the oil extract when compared with the untreated control. Liver function test (ALT, AST and ALP) showed no significant increase in the treatment groups when compared with the controls. The histopathological result reveals a normal physiological structure of the heart, lungs, spleen, liver and kidney in lowest dose of the oil extract. In conclusion, this study validated the ethnomedicinal property of the plant.

The Antioxidant Modulating Properties of Justicia carnea Extract on Sheep Red Blood Cells Immunized Mice

Aims: Inflammatory cytokines as well as exogenously invading antigens among others are keenly involved in the generation of oxidative stress such as reactive oxygen species, reactive nitrogen species capable of causing related diseases. hence this study aimed at evaluating the In-vivo antioxidant modulatory properties of Justicia carnea aqueous leave extract on sheep red blood cells immunized mice.
 Study Design: The animals were grouped into six with five animals per group (Vehicle; distilled water 10 mL/Kg, Sheep Red Blood Cells 0.2 mL/Kg, Justicia carnea 125 mg/Kg, 250 mg/Kg and 500 mg/Kg, and Levamisole 7.5 mg/Kg )
 Place and Duration of Study: The animals used for this study were breed, feed and housed at ambient temperature and well-ventilated animal house of the Basic Medical Sciences in Delta State University, Abraka and Emma-maria laboratory Abraka. The study lasted for two (2) weeks.
 Methodology: All groups were immunized at day zero and day 5 with SRBC inducing immune response except for the VEH group that was not immunized. Treatment of the various groups with respective treatment agents commenced on day 3 to day 9. On day 10 the antioxidant enzymes (Determination of Superoxide Dismutase, Catalase, Myeloperoxidase, Malondialdehyde, Glutathione Peroxidase, Nitric oxide and Xanthine Oxidase) of the brain, liver, kidney and Spleen were carried out.
 Results: The findings in this study reveal the antioxidant enhancement by the plant Justicia carnea being significant when compared to the Vehicle and Sheep Red Blood Cells. Levamisole a standard immunostimulants drug also has significant antioxidant potentials.
 Conclusion: The study was able to showcase the antioxidants activities of the plant Justicia carnea. Pharmacological antioxidants may be developed from the plant, which might be helpful in immune related diseases.

Immunomodulatory Effects of the Somatic Antigens of Fasciola Hepatica and Teladorgasia Circumcincta in Mice Immunized with Sheep Red Blood Cells

Background & Objective: Infestation of humans with helminth parasites can reduce the severity of some auto-inflammatory symptoms in humans. This study was done to evaluate the impact of the somatic antigens extracted from Fasciola hepatica (as an indicator Trematode) ,and Teladorgasia circumcincta (as an indicator nematode) on the immune responses of NMRI mice challenged with sheep red blood cells (SRBCs). Materials & Methods: The mice in treatment groups were intraperitoneally immunized with 1×109 SRBCs twice with 14 days intervals. Concurrent with the immunization, the mice received the extract of each of the parasites (50,100. And 150 µg of protein) or placebo, throughout the study on a daily basis. The specific cellular immune responses and the anti-SRBC antibody titers were detected by footpad thickness and, microhemagglutination test, respectively. Splenocytes were also monitored for cytokine production, proliferation rate, and respiratory burst. Results: The extracts of F. hepatica and T. circumcincta had an opposite effect on the change of the Anti-sRBC antibody level. The extract of F. hepatica caused a significant decrease in the antibody level whereas, extract T. circumcincta did not show any significant changes in the anti-SRBC antibody. Both extracts caused a significant decrease in the level of delayed-type hypersensitivity. However, the extract of F. hepatica caused a more profound reduction in the severity of delayed-type hypersensitivity. The level of IFN-γ in the splenocytes of immunized mice receiving the F. hepatica showed a more pronounced decrease than the immunized mice receiving the extract of T. circumcincta extract. IL-10 levels were only increased in the immunized mice that received the extract of F. hepatica. Conclusion: The extract of F. hepatica may have an immunosuppressive property, while the extract of T. circumcincta may have immunomodulatory properties.

Effect of Media with Different Glycerol Concentrations on Sheep Red Blood Cells' Viability In Vitro.

Simple SummaryGlycerol is widely used as a feed supplement in ruminant nutrition. However, its administration at moderate and high doses results in an increase in plasma osmolality and in an alteration of red blood cell (RBC) indices. The present study aimed at further elucidating the effect of glycerol on RBCs’ functionality by evaluating the effect of different glycerol concentrations on RBC homeostasis under in vitro conditions. Obtained results showed that glycerol permeates the RBC membrane and leads to hemolysis when glycerol concentration exceeds 200 mg/dL. However, RBCs’ antioxidant defenses appear to protect cell membranes without causing an increase of oxidative stress markers. Moreover, no alteration in RBCs’ intracellular Ca2+ ion concentrations and metabolic activity were found. In conclusion, glycerol-based nutritional treatment should be designed in sheep to avoid exceeding 200 mg/dL glycerol circulating concentration to prevent RBC osmotic stress.The use of high doses of glycerol as a livestock feed supplement is followed by a rapid increase in plasma concentrations and consequently in plasma osmolality. Moreover, glycerol is a highly diffusible molecule that can readily permeate the red blood cell (RBC) membrane following a concentration gradient. A rise in glycerol plasma concentrations can thus alter RBC homeostasis. The present study aimed at investigating both glycerol osmotic effects on sheep RBCs and their oxidative response under in vitro conditions. Sheep blood samples were suspended in media supplemented with increasing glycerol concentrations (0, 25, 50, 100, 150, 200, 250, 300, 350, 400 mg/dL), which reflected those found in vivo in previous studies, and incubated at 37 °C for 4h. Thereafter, osmolality and hemolysis were determined in spent media, while cell extracts were used to assay intracellular concentration of glycerol, ATP, Ca2+ ions, oxidative stress markers and reactive oxygen species (ROS).The study confirmed that glycerol intracellular concentrations are directly related with its concentration in the incubation media, as well as hemolysis (p < 0.001) which increased significantly at glycerol concentrations higher form 200 mg/dL. ROS intracellular level increased at all glycerol concentration tested (p < 0.01) and total thiols decreased at the highest concentrations. However, RBCs proved to be able to cope by activating their antioxidant defense system. Superoxide dismutase activity indeed increased at the highest glycerol concentrations (p < 0.001), while total antioxidant capacity and malonyldialdehyde, a typical product of lipid peroxidation by ROS, did not show significant changes. Moreover, no alterations in intracellular Ca2+ ions and ATP concentrations were found. In conclusion, glycerol-induced hemolysis can be related to the induced osmotic stress. In sheep, nutritional treatments should be designed to avoid reaching glycerol circulating concentrations higher than 200 mg/dL.

The stimulation and inhibition of beta-2 adrenergic receptor on the inflammatory responses of ovary and immune system in the aged laying hens

BackgroundOvarian chronic inflammation has been known to incidence in the laying hen mainly via increasing laying frequency and microbial infection, especially during late stage of production period. This study was aimed to evaluate beta-2 adrenergic agonist (Beta-2 Adrenergic Agonist, BAA) Salmeterol and beta blocker (Beta Blocker, BB) Propranolol on the gene expression of the ovarian pro- and anti-inflammatory mediators, inflammatory responses of immune system, ovarian functions and, hormones in the laying hens on the late stage of production period. Forty-eight White Leghorn hens aged 92 weeks were used for 4 weeks to be supplemented by Salmeterol and Propranolol. Ovulation rate and follicular growth were determined based on laying frequency and ovarian visual evaluation, respectively; the mRNA expressions of follicular beta-2 adrenergic receptor (Beta-2 Adrenergic Receptor, β2ADR), cyclooxygenases (Cyclooxygenases, COX) 1 and 2, and cytokines were measured by real-time PCR. The plasma concentration of ovarian hormones, cellular, and humoral immune responses were measured via ELISA, heterophil to lymphocyte ratio (Heterophil to Lymphocyte ratio, H:L), and sheep red blood cell (Sheep Red Blood Cell, SRBC) test, respectively.ResultsAs compared to control, both of BAA Salmeterol and BB Propranolol resulted in a significant decrease in the mRNA expression of β2ADR, cyclooxygenases, and pro- and anti-inflammatory cytokines (P < 0.01). A significant elevation was observed in the ovulation rate (P < 0.05), plasma estradiol content on both treated groups (P < 0.05), and the content of progesterone and was just significantly (P < 0.05) increased in Salmeterol group. H:L was reduced in BAA group (P < 0.05), and immunoglobulin (Ig) M was elevated in both treated hens, when compared to control. The results indicated that Salmeterol significantly increases body weight (P < 0.05).ConclusionThe stimulation and inhibition of beta-2 adrenergic signaling could reduce ovarian inflammatory condition in addition to enhancing laying efficiency in the aged laying hens.

The Effect of Thaumatococcus danielli Leaf Extracts on Immunological and Oxidative Stress Markers in Rat

This work demonstrated the immunological and antioxidant activities of Thaumatococcus daniellii leaf extracts and their potential as the pharmaceutical factor for the treatment of some pathological disorders such as cancer, leucopenia, cells inflammation, dietary, atherosclerosis, neurodegenerative disease and other immunological and oxidative stress disorders.36 albino rats of average weight 140 ± 5 g were randomly grouped into 6 with six rats per group. The rats in group A (positive control) received only 0.1% DMSO. The rats in the negative control (group B) received pyrogallol (immunosuppressor) + 0.1% DMSO for 7 days and challenged with 0.1ml Sheep Red Blood Cell (SRBC). The test groups (C to F) also received pyrogallol + 0.1% DMSO for 7 days and were also challenged with SRBC for a period of 72hours after which they were treated with High and Low doses of either petroleum ether or water extracts of Thaumatococcus danielii for 21days. The assessment of immunomodulatory activity was carried out using Carbon Clearance Test. Haematological/ immunological parameters, Spleen and Plasma antioxidant parameters like Catalase (CAT), Superoxide dismutase (SOD), Glutathione Peroxidase (GPX) Activities, Lipid Peroxidation (LPO) and Malonaldehyde (MDA) concentrations were also determined. Result unveiled a significant (p&lt;0.05) increase in the phagocytic index of both high doses of water and petroleum ether extracts compared to the negative control group. There were no significant (p&lt;0.05) changes in the Heamatological parameters and plasma catalase, but there was a significant change in the white blood cell (WBC) count and also an improvement in the spleen and plasma antioxidant parameters. Further study is required to understand the mechanism of action of both extracts in order to exploit it in immunomodulation.

Production performance of sheep and goat breeds at a farm in a semi-arid region of Namibia.

This study retrospectively investigated the effect of breed and season on the lambing/kidding dynamics, growth performance, neonatal viability, and weaning dynamics of sheep (Damara, Dorper, and Swakara) and goats (Boer goat and Kalahari Red) at a farm in the Khomas Region of Namibia between 2004 and 2015. Litter size was dependent on breed (X2(12, N = 3388) = 796, p < 0.001), with twinning more frequent in Dorper sheep and Kalahari Red and Boer goats than in the Damara and Swakara sheep (p < 0.05), while triplets were more prevalent in the Dorper sheep and Kalahari Red goats (2.8% and 1.0%, respectively; p < 0.05). Distribution of birth weight categories was dependent on breed. There was a significant difference in the proportions of birth weight categories between breeds (X2(12, N = 3388) = 467, p < 0.001) whereby Dorper lambs were mostly born weighing below 3kg (2.6%, p < 0.05); Boer goat kids, Kalahari Red kids, and Damara lambs were mostly born weighing 3 to < 4kg (4.3%, 6.3% and 19.9%, respectively; p < 0.05); Swakara lambs were mostly born weighing 4 to < 5kg (12.2%, p < 0.05), and Swakara lambs were mostly born weighing ≥ 5kg (3.3% and 2.3%, respectively, p < 0.05). Weaning age categories were dependent on breed (X2(12, N = 3388) = 241, p < 0.001) whereby Dorper lambs were mostly weaned at below 3months of age (2.8%, p < 0.05); Damara lambs were mostly weaned at 3 to < 5months of age (12%, p < 0.05), and Boer goat kids were mostly weaned at ≥ 5months of age (0.9%, p < 0.05). Neonatal viability was dependent on breed (X2(8, N = 3388) = 49.2, p < 0.001) whereby Dorper lambs were more susceptible to abortions and neonatal deaths (0.6% and 1.5%, respectively; p < 0.05) than the rest of the breeds. Breed and lambing season interacted to produce effects on the birth weight of offspring although lambing season alone did not have a significant effect on Boer goat and Kalahari Red kids' birth weights.

The Study of extract ethanol of Pirdot (Saurauria Vulcani Korth.) as immunostimulant in Rats (Rattus norvegicus)

BACKGROUND: Ethanolic extract of Pirdot leaves (Saurauria vulcani Korth) (EES) has bioactive compound to decrease glucose and increase insulin levels.&#x0D; AIM: To know of effect of ethanolic extract of Pirdot leaves of immunoglobulin G and lysozyme activity in rats treated with Sheep Red Blood Cell (SRBC).&#x0D; METHODS: Experimental design used in this research was Completely Randomized Design (CRD). There were 24 white rats to be classified into four groups: a control group with water (G0), a group treated with 0.1 ml sheep red blood cell (SRBC) (G1), a group treated with 500 mg/kg ethanol extract pirdot leaves (EES) (G2), and a group treated with 500 mg/KgBW ethanol Extract pirdot leaves (EES) + 0.1 ml SRBC (G3). Immunostimulant activity was evaluated by analyzingthe levels of immunoglobulin G with ELISA method, lysozyme with ABX Micros 60 and histophatology kidney with Hematoxylin-Eosin stainning.&#x0D; RESULTS: The lysozyme activity in rats were treated with only SRBC () is higher and significantly different compared with a group of control (). Necrosis was not found in a group control () and rats treated EES 500 mg/kg ().There were no significant change in histophatology of kidney in rats treated EES 500 mg/kg. Disruption in bowman space with glomerulus was found in G1 (SRBC). Antigen induced disruption in G1 (SRBC) whereas giving extract ethanolic Pirdot leaves concurrently with antigen SRBC can protect kidney from disruption in G3.&#x0D; CONCLUSIONS: A significantly effect of immunoglobulin G, lysozyme activity and histophatology kidney in rats after given by ethanolic extract of pirdot leaves.

A New Method of Assessing Sheep Red Blood Cell Types from Their Morphology.

Simple SummaryHaematological tests are an important diagnostic tool for animal diseases. However, little is known beyond the standardly used haematological methods in sheep, especially those in which the detection of sheep red blood cells (RBCs) shape changes is crucial. Our goal is to obtain sheep RBC morphometric parameters as well as RBC subpopulations based on their morphometric parameters. Morphometric parameters of RBC size and shape were determined from stained blood smears using SFORM, a computer-assisted program for automated cell morphometric measurements. Based on their morphometric parameters, three RBC subpopulations were obtained using principal component and cluster analysis: the smallest and most elongated RBCs, the biggest and most rounded ones, and RBCs of average size and shape. When the values of RBC haematological parameters were higher or above the physiological range, a significantly higher proportion of both average size and shape RBCs, as well as the biggest and most rounded RBCs, was obtained. Since they were obtained from healthy animals, these results indicated the importance of determining morphometric parameters of RBCs and the proportion of each RBC subpopulation, which could serve as a basis for future possibilities in the diagnostic interpretation of haematological disorders in sheep, especially those for which the detection of shape changes in ovine RBCs is crucial.Data concerning the morphometric parameters of sheep red blood cells (RBCs) obtained using computer-assisted image analysis have not yet been investigated, and there are no data on any analyses of ovine RBC subpopulations based on their morphometric parameters. The aims of this study are to determine the values of RBC haematological and morphometric size and shape parameters, to form groups according to the obtained values of haematological parameters; to determine the differences in RBC morphometric parameters between the formed groups, and to determine RBC subpopulations and their respective proportions in the formed groups. Thirty-six blood samples were collected from the jugular vein of clinically healthy Lika pramenka sheep, aged between 2 and 5 years. Haematological parameters including haemoglobin (HGB), haematocrit (HTC), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), and RBC distribution width were analysed using a haematology analyser. Haematological parameters were categorized into two groups: those with lower values or values below the physiological range (Groups 1) and groups with higher values or values above the physiological range (Groups 2). Morphometric parameters of RBCs were determined from stained blood smears using SFORM, a computer-assisted program. Significantly higher values of RBC area, outline, convex, minimal and maximal radius, as well as length and breadth were established in Groups 2 compared to Groups 1 of HGB, HCT, MCV, MCH, and MCHC, respectively. Based on the morphometric parameters of RBCs, three RBC subpopulations were obtained using principal component and cluster analysis: ES 1—the smallest and most elongated RBCs, ES 2—the biggest and most rounded RBCs, and ES 3—average size and shape RBCs. Significantly higher proportions of ES 2 and ES 3 subpopulations, as well as a significantly lower proportion of ES 1 subpopulation, were established in Groups 2 compared to Groups 1 of HGB, HTC, MCV, and MCH, respectively. It can be concluded that ovine RBC subpopulations, based on their morphometric parameters, can be obtained by using computer-assisted image analysis of RBC morphometry and multivariate statistical methods, including principal component and cluster analysis. RBC morphometry, including classification into subpopulations, could serve as a basis for future possibilities in the diagnostic interpretation of anaemic syndromes in veterinary medicine, especially in normocytic, macrocytic, and microcytic anaemias in sheep.

Effects of Dietary Supplementation of L-Carnitine and Excess Lysine-Methionine on Growth Performance, Carcass Characteristics, and Immunity Markers of Broiler Chicken.

L-carnitine as well as lysine and methionine are amino acids of important nutritional and nutraceutical interest and are used in nutritional strategies as diet supplements to improve feed quality characteristics in animals and broiler chicken in particular. This study investigated the effect of different levels of L-carnitine and extra levels of lysine-methionine on growth performance, carcass characteristics, and some immune system markers. Two hundred seventy male Ross 308 broilers were a fed control diet (C) and eight different diets supplemented with an excess of amino acids. In the experimental diets, identified as D1, D2, D3, D4, D5, D6, D7, and D8, extra L-carnitine, lysine, and methionine were added in excess with respect to the American National Research Council (NRC) recommendations: L-carnitine equal to NRC (D1); control diet supplemented with lysine at 30% in excess of NRC, methionine at 30% in excess of NRC, and L-carnitine equal to NRC (D2); control diet supplemented with lysine equal to NRC, methionine equal to NRC, and L-carnitine at 15% in excess of NRC (D3); control diet supplemented control diet supplemented with lysine at 15% in excess of NRC, methionine at 15% in excess of NRC, and L-carnitine at 15% in excess of NRC (D4); control diet supplemented lysine at 30% in excess of NRC, methionine at 30% in excess of NRC, and L-carnitine at 15% in excess of NRC (D5); control diet supplemented with lysine equal to NRC recommendations, methionine equal to NRC recommendations, and L-carnitine at 75% in excess of NRC (D6); control diet supplemented with lysine at 15% in excess of NRC, methionine at 15% in excess of NRC, and L-carnitine at 75% in excess of NRC (D7); and control diet supplemented with lysine at 30% in excess of NRC, methionine at 30% in excess of NRC, and L-carnitine at 75% in excess of NRC (D8). During the starter and growth phases, feed intake was not affected by dietary treatment (p > 0.05). By contrast, body weight and FCR were both affected (p < 0.001) during the starter period. During the finisher phase, feed consumption was affected (p < 0.05) by dietary treatment. Feed intake of broilers fed on C, D3, D6, and D7 were statistically similar (p > 0.05) (1851.90, 1862.00, 1945.10, and 1872.80 g/pen/day, respectively) and were higher (p < 0.05) than 1564.40 g/pen/day (D5). With the exception of drumsticks, neck, back thoracic vertebrae, and proventriculus weights, economical carcass segments were not affected (p > 0.05) by the dietary supplementation of amino acids. Duodenum and ileum weights and lengths decreased with amino acid supplementation (p < 0.05). IgT and IgG titers against Sheep Red Blood Cells (SRBC) for both primary and secondary responses were not affected by dietary treatments (p > 0.05). Dietary amino acids supplementation did not affect IgM titer after the secondary challenge (p > 0.05) and had a significant effect (p < 0.05) on serum antibody titers in broilers vaccinated against Newcastle disease (NCD) and Gumboro ‘s disease at the 27th and 30th days, respectively.

The application of a halotolerant metalloprotease from marine bacterium Vibrio sp. LA-05 in liquid detergent formulations

The application of a halotolerant metalloprotease from marine bacterium Vibrio sp. LA-05 in liquid detergent formulations was investigated in detail. The washing simulation experiment revealed that blood stains were successfully removed from cotton cloths by washing them with three commercial liquid detergents (Liby, Bluemoon and Walch) supplemented with the crude protease at 15–45 °C for 30 min. However, macroscopic residues of stains were remained on the cloths treated with tap water, liquid detergent alone and liquid detergent supplemented with the commercial protease subtilisin A. The analyses of microstructure, chemical elements and tensile properties showed that the washing treatments with crude protease as a bio-additive almost did not affect the texture, elemental composition or tensile properties of cloths. Sheep blood agar plate and SDS-PAGE analyses indicated that its hydrolysis activity to blood proteins was not significantly influenced even in the presence of liquid detergents. The hydrolysis mechanism was further studied and results demonstrated that this protease could specifically hydrolyzed six proteins in Sheep Red Blood Cells (SRBCs) and four proteins in sheep plasma. These findings suggest that this crude halotolerant metalloprotease as a bio-additive may have considerable potentials in the application of liquid detergent formulations.

The Activity Test of Ethanol Extract Tea Parasite Herb (Scurrulla Artopurpurea) as an Immunostimulator on Wistar Strain Rat Sensititized with Sheep Red Blood Cell

Tea parasite (Scurrulla artropurpurea) is one of the hemiparasite plants suspected as an imustimulator agent. The aims of this study was to prove the activity of tea parasite herb extract as immunostimulator in Wistar strain rats that are sensitized by sheep red blood cell suspension (SRBC). Twenty eight Wistar rats were divided into 4 groups. The control group was given CMC 1%, the treatment group was given the tea parasite herb extract at doses of 750 mg, 1.5g and 3 g/kgBW/day, each group was sensitized by SRBC 10% as much 0.5 ml intraperitoneally on the 0th and 9th days and 0.1 ml intraplantar on 12th days. The total number of leukocyte and lymphocyte cells was performed using blood smear and analyzed by GLMRM. test. The macrophage cell activity and the capacity of the peritoneal fluid preparation on day 14 and analyzed by ANOVA test. The histopathological features are demonstrated after 48 hours of intraplantar injection, quantitatively perceived perivascular and periadnexal infiltrates. The results showed that there were average differences on leukocyte cell counts (F = 46.249) and total lymphocyte cells (F = 58.144) on each calculation (day), but there were no average differences between each treatment group. The highest activity of phagocytosis is 78% and continues to increase along with the additional dose of extract, without the increased capacity of macrophage phagocytosis. Histopathological features of slow-type IV reaction shows mild severity. It can be concluded that the active compound of the extract tea parasite herbare able to improve immune system.

Behavioral phenotype relates to physiological differences in immunological and stress responsiveness in reactive and proactive birds

It has now been demonstrated in many species that individuals display substantial variation in coping styles, generally separating into two major behavioral phenotypes that appear to be linked to the degree of physiological stress responsiveness. Laying hens are perfect examples of these dichotomous phenotypes; white laying hens are reactive, flighty, and exhibit large hormonal and behavioral responses to both acute and chronic stress, while brown laying hens are proactive, exploratory, and exhibit low hormonal and behavioral responses to stress. Given the linkages between stress physiology and many other body systems, we hypothesized that behavioral phenotype would correspond to additional physiological responses beyond the stress response, in this case, immunological responses. Because corticosterone is widely known to be immunosuppressive, we predicted that the reactive white hens would show more dampened immune responses than the proactive brown hens due to their exposure to higher levels of corticosterone throughout life. To assess immune function in white and brown hens, we compared febrile responses, corticosterone elevations, feed consumption, and egg production that occurred in response an injection of lipopolysaccharide (LPS) or saline, inflammatory responses to phytohemagglutinin (PHA) injection in the toe web, innate phagocytic activity in whole blood, and antibody responses to an injection of Sheep Red Blood Cells (SRBCs). Contrary to our predictions, white hens had significantly greater swelling of the toe web in response to PHA and showed a greater inhibition of feeding and reproductive output in response to LPS. These results indicated that reactive individuals are more reactive in both stress and immunological responsiveness.

Morphometric abnormalities in spleen and kidney of the progeny of mice fed American cranberry extract (Vaccinium macrocarpon) during pregnancy and lactation.

Although the observed differences between the control and experimental group were not large, caution is recommended in using cranberries and their extracts during pregnancy until more research will be done on this topic.

Immune-Challenged Fish Up-Regulate Their Metabolic Scope to Support Locomotion.

Energy-based trade-offs occur when investment in one fitness-related trait diverts energy away from other traits. The extent to which such trade-offs are shaped by limits on the rate of conversion of energy ingested in food (e.g. carbohydrates) into chemical energy (ATP) by oxidative metabolism rather than by the amount of food ingested in the first place is, however, unclear. Here we tested whether the ATP required for mounting an immune response will lead to a trade-off with ATP available for physical activity in mosquitofish (Gambusia holbrooki). To this end, we challenged fish either with lipopolysaccharide (LPS) from E. coli or with Sheep Red Blood Cells (SRBC), and measured oxygen consumption at rest and during swimming at maximum speed 24h, 48h and 7 days post-challenge in order to estimate metabolic rates. Relative to saline-injected controls, only LPS-injected fish showed a significantly greater resting metabolic rate two days post-challenge and significantly higher maximal metabolic rates two and seven days post-challenge. This resulted in a significantly greater metabolic scope two days post-challenge, with LPS-fish transiently overcompensating by increasing maximal ATP production more than would be required for swimming in the absence of an immune challenge. LPS-challenged fish therefore increased their production of ATP to compensate physiologically for the energetic requirements of immune functioning. This response would avoid ATP shortages and allow fish to engage in an aerobically-challenging activity (swimming) even when simultaneously mounting an immune response. Nevertheless, relative to controls, both LPS- and SRBC-fish displayed reduced body mass gain one week post-injection, and LPS-fish actually lost mass. The concomitant increase in metabolic scope and reduced body mass gain of LPS-challenged fish indicates that immune-associated trade-offs are not likely to be shaped by limited oxidative metabolic capacities, but may instead result from limitations in the acquisition, assimilation or efficient use of resources.

Phagosome Migration and Velocity Measured in Live Primary Human Macrophages Infected with HIV-1.

Macrophages are phagocytic cells that play a major role at the crossroads between innate and specific immunity. They can be infected by the human immunodeficiency virus (HIV)-1 and because of their resistance to its cytopathic effects they can be considered to be persistent viral reservoirs. In addition, HIV-infected macrophages exhibit defective functions that contribute to the development of opportunistic diseases. The exact mechanism by which HIV-1 impairs the phagocytic response of macrophages was unknown. We had previously shown that the uptake of various particulate material by macrophages was inhibited when they were infected with HIV-1. This inhibition was only partial and phagosomes did form within HIV-infected macrophages. Therefore, we focused on analyzing the fate of these phagosomes. Phagosome maturation is accompanied by migration of these compartments towards the cell center, where they fuse with lysosomes, generating phagolysosomes, responsible for degradation of the ingested material. We used IgG-opsonized Sheep Red Blood Cells as a target for phagocytosis. To measure the speed of centripetal movement of phagosomes in individual HIV-infected macrophages, we used a combination of bright field and fluorescence confocal microscopy. We established a method to calculate the distance of phagosomes towards the nucleus, and then to calculate the velocity of the phagosomes. HIV-infected cells were identified thanks to a GFP-expressing virus, but the method is applicable to non-infected cells or any type of infection or treatment.

Stat3 Is Important for Follicular Regulatory T Cell Differentiation.

The production of antibody is precisely controlled during the germinal center (GC) reaction. This process is dependent on the help from follicular T helper (Tfh) cells to germinal center (GC) B cells and is regulated by regulatory follicular T helper (Tfr) cells. How Tfr cells develop and how their suppressive activity functions are not well understood. Here, we found that Stat3 is indispensible for Tfr cell differentiation. After immunization with Sheep Red Blood Cells (SRBC), the loss of Tfr cells caused by deletion of Stat3 in Treg cells does not affect the size of Tfh or GC B cell population, but rather leads to strongly enhanced production of antigen-specific IgG1 and IgG2b. In Peyer’s patches (PPs) in the gut, we found that Stat3 expression in Treg cells is also required for Tfr cell formation to commensal organisms. However, loss of Tfr cells in the gut did not affect the numbers of Tfh cells and GC B cells, nor affect IgG1 or IgA switching by GC B cells. Overall, our study has uncovered unique roles of Stat3 in Tfr cell differentiation and the regulation of the antibody response.