Ascorbic acid biosynthesis in hens producing strong and weak eggshells

1. An experiment was conducted with two strains of layers to ascertain whether the reduction in eggshell strength occurring at the end of the production cycle is the result of reduced ascorbic acid biosynthesis. 2. Hens producing strong and weak eggshells were identified within each strain and egg production, egg weight, per cent shell, shell surface density, plasma, adrenal and hepatic ascorbic acid and renal L-gulonolactone oxidase activity were measured. 3. The strains differed in ascorbic acid synthesis, as measured by L-gulonolactone oxidase activity, and tissue ascorbic acid concentration. 4. Comparison of results from birds producing eggs of similar weight but markedly different in shell strength detected neither a shell strength group x strain interaction nor an effect of shell strength group on plasma and hepatic ascorbic acid and activity of L-gulonolactone oxidase. 5. The results did not support the hypothesis that tissue ascorbate and ascorbic acid biosynthesis are reduced in old hens producing weak eggshells.     

Effect of chronic oxidative/corticosterone-induced stress on ascorbic acid metabolism and total antioxidant capacity in chickens (Gallus gallus domesticus)

The consequences of chronic corticosterone-induced stress (CCIS) on ascorbic acid (AsA) metabolism in chickens, an animal that syntheses the vitamin, are not known. This study was conducted to determine whether CCIS alters AsA synthesis, as measured by l-gulonolactone oxidase (GLO) activity, tissue AsA, lipid peroxides and tissue total antioxidant capacity (TAC). Stress was induced by dietary administration of corticosterone from 2 to 4 weeks of age and measurements were made at 0, 7 and 14 days post-treatment. Ascorbic acid synthesis was not influenced by CCIS but hepatic, cardiac, renal, bursal and duodenal AsA concentrations were significantly decreased and plasma TAC and uric acid concentrations were significantly elevated. Stress caused significant hepatomegaly and hepatic lipidosis but hepatic peroxides were not elevated despite the slight decrease in hepatic TAC. Tissue TAC varied in different organs. It was markedly elevated in the kidney, reduced by 49% in the spleen, and changes were not detected in the heart and duodenum even though AsA concentration was significantly decreased in all tissues. We conclude that CCIS caused a significant reduction in tissue AsA concentration but did not inhibit GLO activity. The change in AsA concentration was associated with increase, decrease or no change in TAC in tissues examined. The findings suggest that CCIS may alter AsA recycling, influx or turnover in different tissues of chickens.     

Ascorbic acid synthesis and transportation capacity in old laying hens and the effects of dietary supplementation with ascorbic acid

Background: Laying hens over 75 weeks of age commonly show great declines in immunity and production performance.It is unclear whether these declines can be relieved by supplementing with ascorbic acid(AA) in feed.Two trials were conducted to investigate the synthesis and metabolism of AA in layers of different ages and the effects of dietary supplemental AA on the performance and the immune and antioxidant statuses of 78 weeks old hens.Methods: In Exp.1,equal numbers(24 hens) of 35 weeks old(Young) and 75 weeks old(Old) layers were fed the same diet without AA supplementation for 4 weeks.In Exp.2,360 healthy 78 weeks old laying hens were randomly assigned to 4 treatments(basal diet supplemented with 0,0.25,0.5,or 1 g AA/kg diet) in an 8-week feeding trial.Results: The old hens tended to have decreased L-gulonolactone oxidase(GLO) synthase activity in the kidney and liver than that of the young hens(P = 0.07 and P = 0.05,respectively).Compared with the young hens,the old hens had lower hepatic antioxidant capacity allowing for the lower thioredoxin(TXN),thioredoxin reductase(TXNR) and cytochrome b5 reductase(CYB5 R) gene expression(P 0.05),whereas increased sodium-dependent vitamin C transporter(SVCT) 1 expression levels in the ileum and kidney and enhanced splenic and hepatic AA concentrations(P 0.05).Dietary supplementation with AA significantly decreased GLO enzyme activity but increased splenic AA concentration and anti-bovine serum albumin IgG levels(P 0.05) and tended to increase CD4+T lymphocyte numbers(P = 0.06) in serum.Supplementation of 0.25 g AA/kg diet significantly increased hepatic total antioxidant capacity(T-AOC,P 0.05) relative to the control group.Conclusions: Laying hens could synthesize AA in both the kidney and the liver,though the GLO enzyme activities were 100 times greater in kidneys than in livers.The old laying hens had greater absorption and reabsorption capacity and higher AA retention in some tissues that did the young hens.Dietary supplementation of AA can improve the health of old layers by enhancing immunity and antioxidant capacity.     

Plasma ascorbic acid concentrations in healthy dogs

Using a highly sensitive and selective analytical method and careful stability control, plasma concentrations of ascorbic acid were determined in German Shepherd Dogs, Labrador Retrievers and Siberian Huskies, a total 99 animals. Mean concentration was 35.9 micromol l(-1)(range 18.2-50.7), and no significant variation was observed neither between breeds nor between females and males. These and previous reported data on plasma ascorbic acid levels in dogs are discussed in the light of methodological aspects.     

Attenuation of ischemia-reperfusion injury by ascorbic acid in the canine renal transplantation

This study examined the effects of ascorbic acid on the attenuation of an ischemia-reperfusion (I/R) injury after a canine renal transplantation. Eight beagle dogs were subjected to a renal auto-transplantation followed by the administration of ascorbic acid (treatment group) and the same amount of vehicle (physiological saline, control group). Blood samples were collected from these dogs to perform the kidney function tests and the invasive blood pressure was measured in the renal artery at pre- and post- anastomosis. The antioxidant enzymes of level 72 h after the transplant were measured. The kidneys were taken for a histopathology evaluation at day 21. The kidney function tests showed a significant difference between the control and treatment group. The invasive blood pressure in the renal artery was similar in the groups. The activity of the antioxidant enzymes in the blood plasma was significant lower in the control group than in the treatment group. The histopathology findings revealed the treatment group to have less damage than the control group. The results of this study suggest that ascorbic acid alone might play a role in attenuating I/R injury and assist in the recovery of the renal function in a renal transplantation model.     

Factors affecting ascorbic acid biosynthesis in chickens: III. Effect of dietary fluoride on L-gulonolactone oxidase activity and tissue ascorbic acid (AsA) concentration

The inconsistent beneficial responses to dietary ascorbic acid (AsA) may be due to dietary factors that alter biosynthesis or tissue turnover of AsA. It has been suggested on the basis of altered tissue AsA that dietary fluoride is a determinant of biosynthesis in chickens. Fluoride may enter the food chain of poultry via industrial contamination, feed ingredients and drinking water. The goal of this study was to ascertain whether dietary fluoride at 300 mg/kg influences l-gulonolactone oxidase (GLO) activity in commercial meat-type chickens. The experimental diet was fed from day-old to 3 weeks and responses measured. Growth and feed conversion were not affected by fluoride in the diet. Dietary fluoride neither inhibited nor enhanced GLO activity nor did it increase or decrease AsA concentration in plasma, liver, kidney, adrenal gland and muscle (pectoralis major). Tissue AsA concentration in ascending order was adrenal > liver > kidney > pectoralis major > plasma. The results are consistent with that reported for the rat and calculations based on the results eliminate fluorine contamination for the inconsistent responses of immature chickens to dietary AsA.     

Pulmonary epithelial lining fluid and plasma ascorbic acid concentrations in horses affected by recurrent airway obstruction

OBJECTIVE: To determine the pulmonary epithelial lining fluid (ELF) concentrations and degree of oxidation of ascorbic acid in horses affected by recurrent airway obstruction (RAO) in the presence and absence of neutrophilic airway inflammation. ANIMALS: 6 RAO-affected horses and 8 healthy control horses. PROCEDURE: Nonenzymatic antioxidant concentrations were determined in RBC, plasma, and ELF samples of control horses and RAO-affected horses in the presence and absence of airway inflammation. RESULTS: ELF ascorbic acid concentration was decreased in RAO-affected horses with airway inflammation (median, 0.06 mmol/L; 25th and 75th percentiles, 0.0 and 0.4 mmol/L), compared with RAO-affected horses without airway inflammation (1.0 mmol/L; 0.7 and 1.5 mmol/L) and control horses (2.2 mmol/L; 1.4 and 2.2 mmol/L). Epithelial lining fluid ascorbic acid remained significantly lower in RAO-affected horses without airway inflammation than in control horses. Moreover, the ELF ascorbic acid redox ratio (ie, ratio of the concentrations of dehydroascorbate to total ascorbic acid) was higher in RAO-affected horses with airway inflammation (median, 0.85; 25th and 75th percentiles, 0.25 and 1.00), compared with RAO-affected horses without airway inflammation (0.04; 0.02 and 0.22). The number of neutrophils in bronchoalveolar lavage fluid was inversely related to the ELF ascorbic acid concentration (r = -0.81) and positively correlated with the ascorbic acid redox ratio (r = 0.65). CONCLUSIONS AND CLINICAL RELEVANCE: Neutrophilic inflammation in horses affected by RAO is associated with a reduction in the ELF ascorbic acid pool. Nutritional supplementation with ascorbic acid derivatives in horses affected by RAO is an area for further investigation.     

Factors affecting ascorbic acid (AsA) biosynthesis in chickens. II. Effect of dietary AsA and strain of chicken

The present study examined the effect of supplemental ascorbic acid (AsA) and ascertained if genotype is a determinant of biosynthesis and the response of strains to dietary AsA. Slow- (Ottawa Meat Control; OMC) and fast-growing (Peterson Enhanced x Hubbard; PEH) chicks were fed 1000 mg/kg AsA from 1 to 10 weeks of age. The activity of l-gulonolactone oxidase (GLO) was used to measure biosynthesis and estimated synthetic capacity (ESC) computed. Body weight was not affected by diets and relative kidney weight decreased with age. In 1 week, dietary AsA increased plasma AsA and inhibited GLO activity with a greater reduction in OMC birds. At 10 weeks, GLO activity was depressed almost uniformly in both strains. Strain by age and diet by age interactions were detected for GLO activity and ESC with significantly greater decline in PEH birds and birds fed supplemental AsA. The results demonstrated that dietary AsA inhibited biosynthesis in meat type chickens and the response at 10 weeks was not influenced by growth rate; and the age dependent decline in biosynthesis was more pronounced in the commercial PEH birds. The result suggests that such strains may be compromised in some situations. Research using multiple dietary levels of AsA, commercial strains, and defined stressors is warranted.     

Oral administration of ascorbic acid to horses

The effects of oral administration of high doses of ascorbic acid on plasma concentrations were investigated in both experimental Thoroughbred horses and those within racing stables. A single oral dose (20 g) did not result in any increase in plasma concentrations. However, daily administration of either 4.5 g or 20 g doses resulted in significant increases in plasma concentrations. Monthly variations in plasma ascorbate concentrations were found in both supplemented (20 g daily) and unsupplemented stables. It is concluded that oral supplementation with ascorbic acid is a satisfactory route to increase plasma and tissue concentrations.     

Bioavailability of ascorbic acid in horses

The bioavailability of ascorbic acid administered to thoroughbreds by intramuscular injection was investigated. For intramuscular injection two preparations were studied, and the percentage bioavailability up to 24 h of 10 g of ascorbic acid was 95% +/- 22 in four horses and 60% in two horses with preparations A and B, respectively. Bioavailability at 24 h in three horses injected subcutaneously with 10 g of preparation B was 82%. Intramuscular injection of both preparations was apparently well tolerated while subcutaneous injection of preparation B (pH 6.0) was associated with marked irritancy. In a cross-over trial in seven thoroughbreds the effect of 13 or 15 days of oral administration of crystalline ascorbic acid (20 g) or ascorbyl palmitate (47 g) on plasma ascorbic-acid concentrations was investigated. Marked differences occurred between individuals. There was a greater increase in plasma ascorbic-acid concentration with ascorbyl palmitate compared to ascorbic acid at 6 and 24 h following administration. In two horses there was no increase in plasma ascorbic acid at 6 h following either oral preparation. The finding of lowered plasma ascorbic-acid concentrations following a period of supplementation warrants further investigation to assess its significance.     

Effect of dietary vitamin A, ascorbic acid and their interaction on turkey bone mineralisation.

At early ages, added dietary ascorbic acid did not affect growth or femur mineralisation of poults. In poults after 8 and 12 weeks, femur mass was reduced by 3000 mg ascorbic acid per kg diet. 2. Plasma calcium and magnesium were not influenced differently by 1000, 4000 and 16,000 IU vitamin A/kg diet, but plasma phosphorus was decreased by the 16,000 level. 3. Vitamin A at 1000 or 16,000 IU/kg diet reduced growth in both sexes to 12 weeks of age compared with the NRC recommended 4000 IU/kg diet. In males, the growth retardation continued to 18 weeks. 4. Vitamin A at 16,000 IU/kg reduced femur development as measured by femur weight to body weight or to femur length ratios.     

Habitual diet and ascorbic acid status in Sudanese camels

There is suggestive evidence that a low status of ascorbic acid in camels enhances their risk for infectious diseases. This study was carried out to find clues as to the role of diet in affecting ascorbic acid status. In a crossover trial with feeding periods of 3 weeks each, six camels (Camelus dromedarius) were fed either a composite of their habitual diet or alfalfa. The simulated habitual diet contained grass (hummra), Acacia mellifera and Blepharis persica. The habitual diet significantly lowered ascorbic acid concentrations in plasma and leukocytes. It is concluded that camels kept on natural desert vegetation may not have optimal disease resistance due to a diet-induced low ascorbic acid status.     

Concentration of ascorbic acid in the blood of cows with subclinical mastitis

The aim of the investigations was evaluation of ascorbic acia concentration in the blood or cows in the subclinical form of mastitis. The research was conducted on 56 cows. The cows were divided into 4 groups: A, B, and C with subclinical form of mastitis caused by Staphylococcus aureus, Streptococcus agalactiae and Escherichia col, and D control. The ascorbic acid concentration in the serum of cows was measured by the hydrazine method. We observed a marked decrease in ascorbic acid concentration in the serum of cows from experimental groups A, B and C, respectively: 34.2, 35.9, and 39.4 micromol/dm3 when we compared them with control group D--69.8 micromol/dm3. In cows with subclinical form of inflammation of the mammary gland, a decreased potential of antioxidant protection in the blood was noticed, which manifested itself as a lower ascorbic acid concentration.     

Effect of supplementing the hen's diet with vitamin A on the accumulation of vitamins A and E,ascorbic acid and carotenoids in the egg yolk and in the embryonic liver

1. The effect of a range of supplementations of vitamin A to the laying hen on the concentration of vitamins A, E, ascorbic acid and carotenoids in the maternal liver, the egg yolk and the embryonic liver were investigated. 2. Four groups of 25 Rhode Island Red hens were fed on standard layer-breeder diets with concentrations of supplemented vitamin A ranging from 0 to 120 mug/g retinol equivalents from 28 weeks of age. After 3 months, the concentration of vitamin A in the maternal liver was found to be greatly enhanced in proportion to the increasing rates of supplementation with the vitamin. However, the concentration of vitamin E in the maternal liver was markedly reduced by high dietary contents of vitamin A. 3. The concentration of vitamin A in the yolk of the hens' eggs was markedly increased by the dietary supplementation. However, the concentration of both vitamin E and carotenoids in the yolks were significantly reduced by high dietary contents of vitamin A. 4. The concentration of vitamin A in the liver of the embryo and the day old chick was greatly increased by the high concentrations of maternal vitamin A provision. However, the concentration of vitamin E, carotenoids and ascorbic acid in the embryonic/neonatal liver were significantly reduced by high contents of vitamin A in the maternal diet. 5. The susceptibility of the embryonic/neonatal liver to lipid peroxidation was significantly increased as a result of high provisions of maternal vitamin A. 6. It is concluded that excessive provision of vitamin A to the laying hen results in an adverse effect on vitamin E, carotenoids and ascorbic acid in the embryonic/neonatal liver and can compromise the antioxidant status of the progeny.     

Content of beta-carotene, vitamin E and ascorbic acid in blood plasma of female calves, cattle, bulls, castrates and ox throughout the course of the year]

The concentration of beta-carotin, of vitamin E and of ascorbic acid in the plasma of female calves (beginning with an age of 4 weeks), of cows in the 2nd and 5th lactation, of bulls, of short scrotum bulls and of oxes in the course of the year was analysed. The concentration of beta-carotin in the plasma of calves after driving up to pasture increased slower than that of cows. After transition to stable feeding there was a significant decrease in the concentration after 4 weeks. During the period of suckling the concentration of vitamin E in the plasma of calves was low. After transition to pasture the concentration of vitamin E in the plasma of calves increased slower than in that of the cows. The concentration of ascorbic acid in the plasma during the winter was higher than during the summer. There was no influence of experimental cryptorchism or castration on the mentioned values.     

Ascorbic acid levels of aqueous humor of dogs after experimental phacoemulsification

Phacoemulsification has been successfully employed in humans and animals for lens extraction. This ultrasonic extracapsular surgical technique induces hydroxyl radical formation in the anterior chamber, which accumulates despite irrigation and aspiration. In this paper we determined the total antioxidant status of aqueous humor after phacoemulsification by measuring aqueous humor ascorbic acid levels. Mixed-breed dogs (n = 11; weighing about 10 kg) with normal eyes as determined by slit-lamp biomicroscopy, applanation tonometry, and indirect ophthalmoscopy had phacoemulsification performed in one eye with the other eye used as a control. Samples of aqueous humor were obtained by anterior chamber paracentesis before surgery and at days 1, 2, 3, 7, and 15 after surgery. Total aqueous humor antioxidant status was inferred from the capacity of aqueous humor to inhibit free radical generation by 2,2-azobis (2-amidopropane) chlorine. Ascorbic acid concentrations were measured by high-pressure liquid chromatography with UV detection. Protein content was determined with the biuret reagent. Statistical analysis was performed by anova followed by the paired t-test. Total antioxidant capacity was reduced from 48 to 27 min during the first 24 h with a gradual increase thereafter, remaining statistically lower than the control eye until 7 days postoperatively. Reduced levels of ascorbic acid followed this reduction in antioxidant capacity (from 211 to 99 microm after 24 h), remaining lower than the control eye until 15 days postoperatively. Protein concentration in aqueous humor increased from 0.62 mg/mL to 30.8 mg/mL 24 h after surgery, remaining statistically lower than the control eye until 15 days postoperatively. Paracentesis alone did not significantly alter the parameters measured. These results indicate that after phacoemulsification, the aqueous humor ascorbic acid levels and antioxidant defenses in aqueous humor are reduced, indirectly corroborating free radical production in the anterior chamber as a result of phacoemulsification. The inflammatory process consequent to the surgical procedure demonstrated by increased protein content in aqueous humor can also contribute to free radical production and ascorbic acid consumption.     

Effect of high environmental temperatures on ascorbic acid, sulfhydryl residue and oxidized lipid concentrations in plasma of dairy cows

Information on oxidative stress under hot conditions from the levels of cells to organs and the whole body has accumulated in the last decades. Although a hot climate decreased dairy performance, changes of oxidative stress markers under hot conditions have remained obscure. Therefore, the effect of high environmental temperature on ascorbic acid, sulfhydryl (SH) residue and oxidized lipids concentrations in plasma from a total of 128 dairy cows was investigated. The monthly average maximum day temperature varied from 9.2°C in January to 32°C in August of 2004 in this institute. High ambient temperatures increased the rectal temperature of dairy cows up to 39.3°C in August. One of the reducing equivalents in plasma, SH residue concentration, decreased in July compared with December (P < 0.05). Another antiradical molecule, ascorbic acid concentration in plasma, also decreased in July (P < 0.01). The oxidative stress index, thiobarbituric acid reactive substance (TBARS), which was produced from the oxidation of polyunsaturated fatty acids under oxidative conditions, increased in summer (P < 0.05). A significant positive relationship of SH residue and ascorbic acid concentrations in the hot season was observed (P < 0.01). A negative correlation between rectal temperatures and ascorbic acid concentrations in the hot season was obtained (P < 0.01). However, TBARS concentration varied independently of the SH residue and ascorbic acid concentration. These results suggest that the response of oxidative stress markers of SH residue, ascorbic acid and TBARS concentration to oxidative stress under hot conditions were not shown to be the same, and that oxidative stress in dairy cows in the hot season increased.     

The influence of different doses of α-tocopherol and ascorbic acid on selected oxidative stress parameters in in vitro culture of leukocytes isolated from transported calves

The objective of the study was to assess in vitro the influence of various doses of two different antioxidants, α-tocopherol and ascorbic acid, on protective mechanisms against ROS in white blood cells obtained from calves exposed to transport, and to compare these results with those obtained from non-transported animals.The concentrations of lipid peroxidation products in leukocytes and in the retained medium were assessed by determining the level of ThioBarbituric Acid Reactive Substances (TBARS). Total antioxidant status in the leukocytes and the medium were estimated using a ferric-reducing ability of plasma (FRAP) assay. Leukocyte viability was determined using the trypan blue reduction test.The study demonstrated that after bovine leukocytes (WBC) were incubated in vitro with α-tocopherol and ascorbic acid, peroxidation intensity decreased and total antioxidant capacity increased. The results of the study reveal that these antioxidants in concentrations over 0.1 mg/ml have a major impact on free radical activity on bovine white blood cells and on cell viability during transport of animals.Based on this study, we suggest that incubation of the leukocytes with antioxidants decreases the oxidative stress development, which can be helpful in protection of the immunological cells during bovine respiratory disease.     

Vitamin C concentrations in blood plasma,tissues and urine of camels (Camelus dromedarius) in Sudanese herds

There is suggestive evidence that a low status of ascorbic acid in ruminants is related with decreased disease resistance. In a first attempt to identify conditions in camels that could affect their health, an inventory was made of ascorbic acid (vitamin C) concentrations in plasma and tissues as related to breed, gender, sexual activity and season. A total of 3429 camels were studied and sub-samples were used for selected comparisons. The highest concentrations of ascorbic acid were found in adrenals (152 mg/100 g wet tissue) and the lowest in heart (8 mg/100 g), the levels being unrelated with season. Arabi camels had higher plasma concentrations of ascorbic acid (6.42 microg/ml) than did Anafi and Bishari camels, the latter breed showing the lowest concentrations (3.24 microg/ml). Female camels of the Anafi breed had higher concentrations urinary ascorbic acid than did their male counterparts. It is suggested that in camels the main elimination route of vitamin C is with urine. Female and male Arabi camels that were sexually active had 52 and 23% lower plasma ascorbic acid concentrations than did their sexually inactive counterparts. It is suggested that especially Bishari camels during the breeding season might be sensitive to disease.     

Effects of dietary supplemental L-carnitine and ascorbic acid on performance, carcass composition and plasma L-carnitine concentration of broiler chicks reared under different temperature.

The present study was initiated to determine whether dietary supplemental L-carnitine and ascorbic acid affect growth performance, carcass yield and composition, abdominal fat and plasma L-carnitine concentration of broiler chicks reared under normal and high temperature. During the experiment, two temperature regimes were employed in two experimental rooms, which were identical but different in environmental temperature. The regimes were thermoneutral (20-22 degrees C for 24 h) or recycling hot (34-36 degrees C for 8 h and 20-22 degrees C for 16 h). One-day-old broiler chicks (ROSS) were used in the experiment. A 2 x 2 x 2 factorial arrangement was employed with two levels (0 and 50 mg/kg) of supplemental L-carnitine and two levels (0 or 500 mg/kg) of supplemental ascorbic acid in drinking water under thermoneutral or high temperature regimes. Body weight gain was affected by high temperature. However, body weight gain was significantly improved in animals receiving supplemental L-carnitine, ascorbic acid or L-carnitine + ascorbic acid compared to animals receiving unsupplemented diet under high temperature. On the other hand, supplemental L-carnitine or L-carnitine + ascorbic acid reduced body weight gain under thermoneutral condition. Supplemental ascorbic acid significantly improved feed conversion efficiency, the improvement was relatively greater under high temperature. The L-carnitine content in the plasma was higher in the groups receiving supplemental L-carnitine and ascorbic acid under high temperature, while broilers fed supplemental L-carnitine and ascorbic acid had a decreased level of plasma L-carnitine concentration under normal temperature. It is concluded that dietary supplemental L-carnitine or L-carnitine + ascorbic acid may have positive effects on body weight gain, carcass weight under high temperature conditions.