Effects of ß-carotene-enriched dry carrots on ß-carotene status and colostral immunoglobulin in ß-carotene-deficient Japanese Black cows.

Data from 18 ß-carotene-deficient Japanese Black cows were collected to clarify the effects of feeding ß-carotene-enriched dry carrots on ß-carotene status and colostral immunoglobulin (Ig) in cows. Cows were assigned to control or carrot groups from 3 weeks before the expected calving date to parturition, and supplemental ß-carotene from dry carrots was 138 mg/day in the carrot group. Plasma ß-carotene concentrations in the control and carrot groups at parturition were 95 and 120 µg/dL, and feeding dry carrots slightly improved plasma ß-carotene at parturition. Feeding dry carrots increased colostral IgA concentrations in cows and tended to increase colostral IgG1 , but colostral IgM, IgG2 , ß-carotene and vitamin A were not affected by the treatment. Feeding dry carrots had no effects on plasma IgG1 , IgA and IgM concentrations in cows, but plasma IgG1 concentrations decreased rapidly from 3 weeks before the expected calving date to parturition. These results indicate that feeding ß-carotene-enriched dry carrots is effective to enhance colostral IgA and IgG1 concentrations in ß-carotene-deficient cows.

Relationships between immunoglobulin M and immunoglobulin G or A in colostrum of Japanese black multiparous cows.

This study was conducted to clarify the relationships among immunoglobulin (Ig)M, IgG, IgA, ß-carotene, vitamin A and α-tocopherol contents in colostrum of 24 Japanese Black multiparous cows in order to evaluate the role of IgM on colostral IgG and IgA production. Compared with colostral IgG, colostral IgM and IgA were very low but varied widely. There was positive correlation between colostral IgM and IgG, but colostral IgM was not related with colostral IgA. There was no relationship between colostral IgM and age of cows, although colostral IgG was increased with aging. There were positive correlations among colostral ß-carotene, vitamin A and α-tocopherol and these vitamins were positively correlated with colostral IgM and IgG. These results indicate that fat-soluble vitamins may affect colostral IgG and IgM in cows and colostral IgG increases with the increase of colostral IgM.

Relationships between immunoglobulin and fat-soluble vitamins in colostrum of Japanese Black multiparous cows.

Data from 19 Japanese Black multiparous cows were collected to clarify the relationships among immunoglobulin (Ig) G, IgA, ß-carotene, vitamin A and α-tocopherol contents in colostrum of cows in order to evaluate the role of fat-soluble vitamins on colostral IgG and IgA production. Mean colostral IgG was 141 mg/mL, ranging from 65 to 208 mg/mL, whereas mean colostral IgA was 8.7 mg/mL, ranging from 1.0 to 34.6 mg/mL. Colostral IgG increased with aging in multiparous cows. There were positive correlations between colostral IgG and colostral vitamin A or colostral α-tocopherol in cows, and the higher adjusted R(2) was obtained in the prediction model of colostral IgG from age and colostral vitamin A. Colostral vitamin A was positively correlated with colostral ß-carotene or colostral α-tocopherol in cows, but there were no relationships between colostral IgA and colostral IgG or colostral fat-soluble vitamins. These results indicate that fat-soluble vitamin contents in colostrum of cows may change in similar patterns and high colostral vitamin A is related with high colostral IgG.

Effects of astaxanthin-enriched yeast on mucosal IgA induction in the jejunum and ileum of weanling mice.

The present study was conducted to clarify the effects of astaxanthin-enriched yeast on the concentration of immunoglobulin A (IgA), the numbers of IgA antibody-secreting cells (ASC) and the messenger RNA (mRNA) expression of IgA C-region in the jejunum and ileum of weanling mice. Weanling mice were fed rodent feed or astaxanthin-enriched yeast-supplemented rodent feed for 7, 14 or 21 days. Supplemental astaxanthin-enriched yeast increased the numbers of IgA ASC in the jejunum and ileum after 7, 14 and 21 days of treatment. Supplemental astaxanthin-enriched yeast increased IgA concentrations in the jejunum after 21 days of treatment, but IgA concentrations in the ileum were not affected by the treatment. The mRNA expressions of IgA C-region in the jejunum after 14 and 21 days of treatment and the ileum after 14 days of treatment were enhanced by supplementation of astaxanthin-enriched yeast. These results indicate that supplementation of astaxanthin-enriched yeast is effective to enhance the numbers of IgA ASC in the jejunum and ileum and IgA concentrations in the ileum of weanling mice.

Effects of supplemental ß-carotene on mucosal IgA induction in the jejunum and ileum of mice after weaning.

An adequate immune system is required to prevent diarrhoea in neonates, and IgA provides protection against microbial antigens on mucosal surfaces. Although ß-carotene supplementation has been expected to enhance the retinoic acid (RA)-mediated immune response in neonates, the exact mechanism of the enhancement of mucosal IgA production in the small intestine by ß-carotene is still unclear. In the present study, we investigated the effect of supplemental ß-carotene on the concentrations of IgA, the numbers of IgA antibody-secreting cells (ASC) and the mRNA expressions of IgA C-region, CCL25, retinoid X receptor (RXR) α, retinoic acid receptor (RAR) α and RARγ in the jejunum and ileum of weanling mice. Weanling mice were fed rodent feed or 50 mg/kg ß-carotene-supplemented rodent feed for 7, 14 or 21 d. The concentrations of IgA and the numbers of IgA ASC in the jejunum and ileum of mice increased markedly with age, and supplemental ß-carotene increased the concentrations of IgA, the numbers of IgA ASC and the mRNA expressions of IgA C-region, CCL25 and RARγ in the jejunum after 14 and 21 d of treatment. Supplemental ß-carotene increased the numbers of IgA ASC in the ileum after 14 and 21 d of treatment, but the concentrations of IgA in the ileum were not affected by ß-carotene supplementation. The mRNA expressions of RXRα and RARα in the jejunum and those of RXRα and RARγ in the ileum after 21 d of treatment were enhanced by ß-carotene supplementation. These results indicate that ß-carotene supplementation in weanling mice is effective to enhance mucosal IgA induction in the jejunum or ileum and that the effects are mainly due to the RA-mediated immune response.

Effects of high potassium chloride supplementation on water intake and bodyweight gains in pregnant and lactating mice.

Thirty-one ICR pregnant mice were assigned to a control or a potassium chloride (KCl) diet group to clarify the effects of KCl supplementation on water intake, bodyweight gains and serum components in pregnant and lactating mice, and 5% KCl was supplemented in KCl diets from 6.5 days post coitus to 1 or 14 days after parturition. Feed intake was not affected by treatment, but supplemental KCl decreased bodyweight gains of lactating mice and their neonatal mice. Water intake and urine volume of KCl supplemented mice were significantly higher than those of control mice during pregnancy and supplemental KCl decreased serum urea N in pregnant mice. Supplemental KCl increased water intake drastically in lactating mice immediately after parturition and increased serum K at 14 days after parturition. Histological alteration using hematoxylin-eosin was not found in the kidney of each mouse at 1 or 14 days after parturition. These results indicate that high KCl supplementation accelerates water intake in lactating mice and prevents bodyweight gains of maternal and neonatal mice during lactation.

Effects of coumestrol administration to maternal mice during pregnancy and lactation on immunoblogulin A-secreting cells in mammary glands.

Mortality and morbidity of neonates continue to be major problems in humans and animals, and immunoblogulin A (IgA) provides protection against microbial antigens at mucosal surfaces. The present study was conducted to clarify the effects of coumestrol administration to maternal mice during pregnancy and lactation on IgA antibody-secreting cells (ASC) in mammary glands in lactating mice. From 6.5 to 16.5 days post coitus and 1 to 13 days post partum (dpp), maternal mice were administered coumestrol at 200 µg/kg body weight/day. Coumestrol administration increased the number of IgA ASC and the messenger RNA expression of IgA C-region and vascular cell adhesion molecule-1 in mammary glands of maternal mice at 14 dpp, but coumestrol administration had no effect on the number of IgA ASC in the ileum. Coumestrol administration increased serum IgA concentration in maternal mice at 14 dpp, but IgA concentrations in serum, stomach contents, intestine and feces of neonatal mice were not affected by treatment. These results imply that coumestrol administration to maternal mice during pregnancy and lactation is effective in increasing the numbers of IgA ASC in mammary glands during lactation owing to the activated messenger RNA expressions of IgA C-region and vascular cell adhesion molecule-1 in mammary gland.

Effects of coumestrol administration to maternal mice during pregnancy and lactation on renal Ca metabolism in neonatal mice.

The present study was conducted to clarify the effects of coumestrol administration to maternal mice during pregnancy and lactation on serum Ca and Ca metabolism in their neonatal mice. From 6.5 to 16.5 days post coitus and from 3 to 10 days after parturition, maternal mice were administered at 200 µg/kg body weight/day of coumestrol. Coumestrol administration did not affect weight gains, serum Ca and the expression of vitamin D receptor (VDR) protein in the kidney of neonatal mice, but weight gains of maternal mice were decreased by coumestrol administration. Coumestrol administration increased the messenger RNA (mRNA) expressions of epithelial Ca channels 1 (ECaC1) and VDR in the kidney of neonatal mice, and also increased the mRNA expressions of ECaC2 in the kidney and small intestine of male neonatal mice. The mRNA expressions of ECaC1, ECaC2, calbindin-D(9k) (CaBP-9k) and estrogen receptor (ER)α in the kidney and VDR in the small intestine of male neonatal mice were higher than those of female mice. Thus, coumestrol administration to maternal mice during pregnancy and lactation may affect renal Ca metabolism in neonatal mice, especially male neonatal mice via maternal milk.

Effects of coumestrol administration to pregnant and lactating mice on intestinal alkaline phosphatase activity.

The present study was conducted to clarify the effects of coumestrol administration on Ca metabolism during pregnancy and in lactating mice. From 6.5 to 16.5 days post coitus (dpc), pregnant mice were administered coumestrol at 200 µg/kg body weight/day. The duodenum, jejunum and blood samples were obtained at 17.5 dpc or 10 days after parturition (dap). Coumestrol administration decreased alkaline phosphatase (ALP) activity and mRNA expression of IAP and estrogen responsive genes, c-fos and vascular endothelial growth factor (VEGF), in the duodenum and jejunum of pre-delivery mice. In lactating mice, the ALP activity and mRNA expression of IAP were not changed, although coumestrol administration decreased mRNA expression of c-fos in the duodeum and VEGF in the jejunum. Coumestrol did not affect serum Ca and the expression of vitamin D receptor protein in the duodenum and jejunum. Thus, coumestrol administration during pregnancy may decrease the mRNA expression of IAP and the ALP activity in the intestine of the pre-delivery mice through ERα, but coumestrol had little effect on intestinal ALP activity at 10 days after parturition.

Supplemental ß-carotene increases IgA-secreting cells in mammary gland and IgA transfer from milk to neonatal mice.

Mortality of neonates continues to be a major problem in humans and animals. IgA provides protection against microbial antigens at mucosal surfaces. Although ß-carotene supplementation has been expected to enhance retinoic acid-mediated immune response in neonates, the exact mechanism by which ß-carotene enhances IgA production is still unclear. We investigated the effect of supplemental ß-carotene for maternal mice during pregnancy and lactation on IgA antibody-secreting cells (ASC) in mammary gland and guts and on IgA transfer from milk to neonatal mice. Pregnant mice were fed untreated or 50 mg/kg ß-carotene-supplemented diets from 6·5 d postcoitus (dpc) to 14 d postpartum (dpp). Supplemental ß-carotene increased the numbers of IgA ASC in mammary gland (P < 0·05) and ileum (P < 0·001), and also mRNA expression of IgA C-region in ileum (P < 0·05) of maternal mice at 14 dpp, but few IgA ASC were detected in mammary gland at 17·5 dpc. IgA concentration in stomach contents, which represents milk IgA level, was significantly higher (P < 0·01) in neonatal mice born to ß-carotene-supplemented mothers at 7 and 14 dpp, and IgA concentration in serum, stomach contents and faeces increased (P < 0·001) drastically with age. These results suggest that ß-carotene supplementation for maternal mice during pregnancy and lactation is useful for enhancing IgA transfer from maternal milk to neonates owing to the increase in IgA ASC in mammary gland and ileum during lactation.

Effects of high potassium chloride supplementation on water intake, urine volume and nitrogen balance in mice.

Sixteen ICR male mice were assigned to a control diet group or a KCl diet group in metabolic cages to clarify the effects of KCl supplementation on water intake, urine volume and N balance, and 5% of KCl was supplemented in KCl diets for 4 or 8 weeks. Bodyweight of KCl supplemented mice was significantly higher than that of control mice from 24 to 28 days after treatment. Feed intake, water intake and urine volume of KCl supplemented mice were significantly higher than those of control mice, and the increased water intake and urine volume in KCl supplemented mice were 4.49 and 4.15 g, respectively. Urinary N, K and Cl excretion were significantly higher in KCl supplemented mice. Although N retention was not significantly different between control and KCl supplemented mice, N retention in KCl supplemented mice tended to be lower. Serum creatinine concentration at 8 weeks after treatment was lower in KCl supplemented mice. Histological alteration using hematoxylin-eosin and Sirius red staining was not found in the kidney of each mouse at 4 and 8 weeks after treatment. These results suggest that high KCl supplementation increases water intake, urine volume and urinary N excretion in mice.

Coumestrol decreases intestinal alkaline phosphatase activity in post-delivery mice but does not affect vitamin D receptor and calcium channels in post-delivery and neonatal mice.

In this study, we investigated the effects of administration of coumestrol during pregnancy on calcium (Ca) metabolism in post-delivery maternal and neonatal mice. From 6.5 to 16.5 days post coitus (dpc), pregnant females were administered daily doses of coumestrol (200 microg/kg body weight/day). One day after parturition, blood samples and the kidneys, liver, jejunum and duodenum were obtained from each of maternal mouse, and blood samples and the kidneys and liver were obtained from neonatal mice. Coumestrol did not have any significant effect on the Ca and inorganic phosphorus concentrations in the sera of the maternal and neonatal mice. No notable effects of coumestrol were observed in relation to Vitamin D receptor expression in the maternal and neonatal mice by immunohistochemical analysis. Coumestrol did not affect the Vitamin D receptor and epithelial calcium channel and 2 mRNA levels in any of the organs investigated. Enzyme histochemical analysis showed that coumestrol decreased intestinal alkaline phosphatase activity in the maternal jejunum and duodenum. In the duodenum, coumestrol decreased expression of intestinal alkaline phosphatase, c-fos and vascular endothelial growth factor at the mRNA level. However, we did not observe any significant effects of coumestrol on the expression of these genes. In conclusion, coumestrol decreased intestinal alkaline phosphatase activity in the small intestines of maternal mice at the level used in the present study, and the mechanisms underlying this effect are different for the jejunum and duodenum.