Reference intervals for CTX and P1NP in a multi-ethnic Malaysian cohort.

BACKGROUND: Well defined reference intervals are central to the utility of serum C-terminal telopeptide of type 1 collagen (CTX) and N-terminal propeptide of type I procollagen (P1NP), designated as reference markers in osteoporosis, and useful for monitoring therapeutic response in that condition. This study reports the reference intervals for plasma CTX and serum P1NP in a multi-ethnic Malaysian population. METHODS: Ethnic Malay, Chinese or Indian subjects aged 45-90 years old were recruited from Selangor, Malaysia from June 2016 to August 2018. Subjects with known medical conditions (e.g., bone disorders, malnutrition, immobilisation, renal impairment, hormonal disorders) and medications (including regular calcium or vitamin D supplements) that may affect CTX and P1NP were excluded. Additionally, subjects with osteoporosis or fracture on imaging studies were excluded. The blood samples were collected between 8 a.m. and 9 a.m. in fasting state. The CTX and P1NP were measured on Roche e411 platform in batches. RESULTS: The 2.5th-97.5th percentiles reference intervals (and bootstrapped 90%CI) for plasma CTX in men (n = 91) were 132 (94-175) - 775 (667-990) ng/L; in post-menopausal women (n = 132) 152 (134-177) - 1025 (834-1293) ng/L. The serum P1NP reference intervals in men were 23.7 (19.1-26.4) - 83.9 (74.0-105.0) µg/L, and in post-menopausal women, 25.9 (19.5-29.3) - 142.1 (104.7-229.7) µg/L. CONCLUSION: The reference intervals for plasma CTX and serum PINP for older Malaysian men and post-menopausal women are somewhat different to other published studies from the region, emphasising the importance of establishing specific reference intervals for each population.

Effects of bacterial organic selenium, selenium yeast and sodium selenite on antioxidant enzymes activity, serum biochemical parameters, and selenium concentration in Lohman brown-classic hens.

This study compares the effects of sodium selenite, selenium yeast, and enriched bacterial organic selenium protein on antioxidant enzyme activity, serum biochemical profiles, and egg yolk, serum, and tissue selenium concentration in laying hens. In a 112-d experiment, 144 Lohman Brown Classic hens, 23-wks old were divided into four equal groups, each has six replicates. They were assigned to 4 treatments: 1) a basal diet (Con), 2) Con plus 0.3 mg/kg feed sodium selenite (SS); 3) Con plus 0.3 mg/kg feed Se-yeast (SY): 4) Con plus 0.3 mg/kg feed bacterial enriched organic Se protein (ADS18) from Stenotrophomonas maltophilia bacteria. On d 116, hens were euthanized (slaughtered) to obtain blood (serum), liver organ, and breast tissue to measure antioxidant enzyme activity, biochemical profiles, and selenium concentration. The results show that antioxidant enzyme activity of hens was increased when fed bacterial organic Se (ADS18), resulting in a significant (P < 0.05) increase in serum GSH-Px, SOD, and CAT activity compared to other treatment groups. However, ADS18 and SY supplementation increase (P < 0.05) hepatic TAC, GSH-Px, and CAT activity, unlike the SS and Con group. Similarly, dietary Se treatment reduced total cholesterol and serum triglycerides concentrations significantly (P < 0.05) compared to the Con group. At 16 and 18 weeks, selenium concentration in hen egg yolks supplemented with dietary Se was higher (P < 0.05) than in Con, with similar patterns in breast tissue and serum. Supplementation with bacterial organic Se (ADS18) improved antioxidant enzyme activity, decreased total serum cholesterol and serum lipids, and increased Se deposition in egg yolk, tissue, and serum. Hence, organic Se may be considered a viable source of Se in laying hens.

Effect of organic and inorganic dietary selenium supplementation on gene expression in oviduct tissues and Selenoproteins gene expression in Lohman Brown-classic laying hens.

BACKGROUND: The oviduct of a hen provides a conducive environment for egg formation, which needs a large amount of mineral elements from the blood via trans-epithelial permeability. Eggshell is the calcified layer on the outside of an egg that provides protection and is critical for egg quality. However, little is known about the genes or proteins involved in eggshell formation, and their relationship to dietary microminerals. We hypothesized that dietary selenium supplementation in chickens will influence genes involved in eggshell biomineralization, and improve laying hen antioxidant capacity. The objective of this research was to investigate how organic and inorganic dietary selenium supplementation affected mRNA expression of shell gland genes involved in eggshell biomineralization, and selenoproteins gene expression in Lohman Brown-Classic laying hens. RESULTS: Shell gland (Uterus) and liver tissue samples were collected from hens during the active growth phase of calcification (15-20 h post-ovulation) for RT-PCR analysis. In the oviduct (shell gland and magnum) and liver of laying hens, the relative expression of functional eggshell and hepatic selenoproteins genes was investigated. Results of qPCR confirmed the higher (p < 0.05) mRNA expression of OC-17 and OC-116 in shell gland of organic Se hen compared to inorganic and basal diet treatments. Similarly, dietary Se treatments affected the mRNA expression of OCX-32 and OCX-36 in the shell gland of laying hens. In the magnum, mRNA expression of OC-17 was significantly (p < 0.05) higher in hens fed-bacterial organic, while OC-116 mRNA expression was down-regulated in dietary Se supplemented groups compared to non-Se supplemented hens. Moreover, when compared to sodium selenite, only ADS18 bacterial Se showed significantly (p < 0.05) higher mRNA levels in GPX1, GPX4, DIO1, DIO2 and SELW1, while Se-yeast showed significantly (p < 0.05) higher mRNA levels in TXNRD1 than the non-Se group. CONCLUSIONS: Dietary Se supplementation especially that from a bacterial organic source, improved shell gland and hepatic selenoproteins gene expression in laying hens, indicating that it could be used as a viable alternative source of Se in laying hens. The findings could suggest that organic Se upregulation of shell gland genes and hepatic selenoproteins in laying hens is efficient.

Influence of bacterial organic selenium on blood parameters, immune response, selenium retention and intestinal morphology of broiler chickens.

BACKGROUND: Several studies indicated that dietary organic selenium (Se) usually absorbed better than an inorganic source, with high retention and bioavailability. Dietary Se as an antioxidant element affects the immune system and hematological status in animals. Therefore, the aim of this study was to evaluate the effect of dietary supplementation of bacterial selenium as an organic source on hematology, immunity response, selenium retention, and gut morphology in broiler chickens. RESULTS: The present results revealed that supplementation of inorganic Se was associated with the lowest level of RBC, HB, and PCV with significant difference than ADS18-Se. In the starter stage, both T2 and T5 were associated with the significantly highest IgG level compared to the basal diet, while all supplemented groups showed higher IgM levels compared to the control group. In the finisher phase, all Se supplemented groups showed significant (P ˂ 0.05) increases in IgG, IgA, and IgM levels compared to T1. Birds fed bacterial-Se showed high intestinal villus height and better Se retention more than sodium selenite. The organic selenium of ADS18 had a superior action in improving Se retention compared to ADS1 and ADS2 bacterial Se. CONCLUSIONS: Bacterial organic Se had a beneficial effect on the villus height of small intestine led to high Se absorption and retention. Thus, it caused a better effect of Se on hematological parameters and immunity response.

Effects of vitamin E, inorganic selenium, bacterial organic selenium, and their combinations on immunity response in broiler chickens.

BACKGROUND: Selenium (Se) and vitamin E (Vit E) can act synergistically and affect biological processes, mainly antioxidant and immunity. The use of excess dietary Vit E and Se in animals' feed could enhance immune response and induce disease resistance. Moreover, different Se sources may provide different alterations in the immune system. Accordingly, the aim of the current study was to assess the impact of dietary supplementation of Vit E, inorganic Se (sodium selenite, SS), bacterial organic Se of ADS18, and their different combinations on the plasma immunoglobulins, ceacum microbial population, and splenic cytokines gene expression in broiler chickens. RESULTS: Present results showed that, Se and Vit E synergistic effect was clear in plasma IgM level at day 42 and in splenic cytokines expression (TNF-α, IFN-γ, IL-2, IL-10). The combination of 0.3 mg/kg ADS18-Se with 100 mg/kg Vit E showed the highest IgM level compared to Vit E- SS complex. The combination of either SS or ADS18-Se with Vit E had no significant effect on IFN- γ and IL-10 compared to Vit E alone, while Vit E alone showed the significantly lowest TNF-α compared to the Se combinations. Supplementation of 100 mg/kg Vit E had no effect on microbial population except a slight reduction in Salmonella spp. The main effect of Se sources was that both sources increased the day 42 IgA and IgG level compared to NS group. ADS18-Se modulate the caecum microbial population via enhancing beneficial bacteria and suppressing the E-coli and Salmonella spp. while both Se and Vit E factors had no effect on lymphoid organ weights. CONCLUSIONS: The inclusion of 100 mg/kg Vit E with 0.3 mg/kg ADS18-Se, effectively could support the immune system through regulation of some cytokines expression and immunoglobulin levels more than using ADS18-Se alone, while no difference was observed between using SS alone or combined with Vit E.

The effect of dietary bacterial organic selenium on growth performance, antioxidant capacity, and Selenoproteins gene expression in broiler chickens.

BACKGROUND: Selenium (Se) is an essential trace mineral in broilers, which has several important roles in biological processes. Organic forms of Se are more efficient than inorganic forms and can be produced biologically via Se microbial reduction. Hence, the possibility of using Se-enriched bacteria as feed supplement may provide an interesting source of organic Se, and benefit broiler antioxidant system and other biological processes. The objective of this study was to examine the impacts of inorganic Se and different bacterial organic Se sources on the performance, serum and tissues Se status, antioxidant capacity, and liver mRNA expression of selenoproteins in broilers. RESULTS: Results indicated that different Se sources did not significantly (P ≤ 0.05) affect broiler growth performance. However, bacterial organic Se of T5 (basal diet +0.3 mg /kg feed ADS18 Se), T4 (basal diet +0.3 mg /kg feed ADS2 Se), and T3 (basal diet +0.3 mg /kg feed ADS1 Se) exhibited significantly (P ≤ 0.05) highest Se concentration in serum, liver, and kidney respectively. Dietary inorganic Se and bacterial organic Se were observed to significantly affect broiler serum ALT, AST, LDH activities and serum creatinine level. ADS18 supplemented Se of (Stenotrophomonas maltophilia) bacterial strain showed the highest GSH-Px activity with the lowest MDA content in serum, and the highest GSH-Px and catalase activity in the kidney, while bacterial Se of ADS2 (Klebsiella pneumoniae) resulted in a higher level of GSH-Px1 and catalase in liver. Moreover, our study showed that in comparison with sodium selenite, only ADS18 bacterial Se showed a significantly higher mRNA level in GSH-Px1, GSH-Px4, DIO1, and TXNDR1, while both ADS18 and ADS2 showed high level of mRNA of DIO2 compared to sodium selenite. CONCLUSIONS: The supplementation of bacterial organic Se in broiler chicken, improved tissue Se deposition, antioxidant status, and selenoproteins gene expression, and can be considered as an effective alternative source of Se in broiler chickens.

Influence of postbiotic RG14 and inulin combination on cecal microbiota, organic acid concentration, and cytokine expression in broiler chickens.

This study examined the effects of different combinations of inulin and postbiotics RG14 on growth performance, cecal microbiota, volatile fatty acids (VFA), and ileal cytokine expression in broiler chickens. Two-hundred-and sixteen, one-day-old chicks were allocated into 6 treatment groups, namely, a basal diet (negative control, NC), basal diet + neomycin and oxytetracycline (positive control, PC), T1 = basal diet + 0.15% postbiotic RG14 + 1.0% inulin, T2 = basal diet + 0.3% postbiotic RG14 + 1.0% inulin, T3 = basal diet + 0.45% postbiotic RG14 + 1.0% inulin, and T4 = basal diet + 0.6% postbiotic RG14 + 1.0% inulin, and fed for 6 weeks. The results showed that birds fed T1 and T3 diets had higher (P < 0.05) final body weight and total weight gain than NC and PC birds. A lower (P < 0.05) feed conversion ratio was observed in birds fed T1 and T2 compared with those fed the NC diet. Birds fed PC, T1, T2, and T3 had higher (P < 0.05) cecum total bacteria and Bifidobacteria compared to the NC birds. Diet had no effect on cecum Lactobacilli, Enterococcus and Salmonella. The NC birds had higher (P < 0.05) Enterobacteria and E. coli than other treatments. Concentration of acetic acid was higher in birds fed PC, T1, and T4 compared to the NC birds. However, the concentration of butyric acid, propionic acid, and total VFA did not differ (P > 0.05) among diets. The NC birds had higher (P < 0.05) expression of interferon (IFN) and Lipopolysaccharide-induced tumor necrosis factor-alpha factor (LITAF) gene compared with those fed other diets. The mRNA expression of interluken-6 (IL-6) was up-regulated in birds fed T3 and T4 compared to the NC birds. However, the expression of interluken-8 (IL-8) gene was not influenced by diet. Postbiotic and inulin combinations are potential replacements for antibiotic growth promoters in the poultry industry.

Individual non-essential amino acids fortification of a low-protein diet for broilers under the hot and humid tropical climate.

A study was conducted to investigate the effects of feeding low-protein diets fortified with individual non-essential amino acids (NEAA) on growth performance, serum metabolites (uric acid, UA; triglycerides, TG; total protein, TP; and albumin, Alb), organ weight, breast yield, and abdominal fat weight in broiler chicks raised under the hot and humid tropical climate. Eight isocaloric (3,017 kcal/kg) experimental diets were formulated and fed to male broiler chicks from d 1-21 as follows: 1) 22.2% crude protein (CP) (positive control; PC); 2) 16.2% CP + mixture essential amino acids (EAA) to meet or exceed the National Research Council (1994) recommendations (negative control; NC); 3) NC + glycine (Gly) to equal the total glycine + serine level in the PC; diets 4 through 7 were obtained by supplementing NC diet with individual glutamic acid, proline, alanine, or aspartic acid (Glu, Pro, Ala, or Asp, respectively); 8) NC + NEAA (Gly + Glu + Pro + Ala + Asp) to equal the total level of these NEAA in the PC. Fortifying NC diet with mixture NEAA resulted in a similar growth performance as PC. However, fortification of low-CP diet with individual NEAA failed to improve body weight (BW) (P < 0.0001), feed intake (FI) (P = 0.0001), and feed conversion ratio (FCR) (P = 0.0001). Serum uric acid (UA) was lower (P = 0.0356) in NC birds and NC diet supplemented with individual NEAA birds, whereas serum triglyceride (TG) (P = 0.007) and relative weight of abdominal fat (P = 0.001) were higher in these birds. In conclusion, no single NEAA fortification may compensate the depressed growth performance attributed to a low-CP diet. However, fortification with Gly may improve FCR. There is a possibility that broilers raised under the hot and humid climate require higher Gly fortification than the level used in this study.

Dietary putrescine effects on performance parameters, nutrient digestibility, intestinal morphology and tissue polyamine content of broilers fed low protein diet.

This study was carried out to investigate the effects of dietary putrescine (PUT) on broiler's response fed low crude protein (CP) diets. A total of 192 male day old chicks were fed with four dietary treatments including two levels of PUT (0 and 0.03%) and two levels of CP (normal and low) with factorial combinations. Weekly growth performance, nutrient digestibility and intestinal morphology (at the age of 21 days) and liver and intestinal tissue polyamines content were measured. As a result of this study lower dietary CP had a significant (P<0.05) lower body weight gain (BWG) and improved protein efficiency ratio (PER). PUT improved energy efficiency ratio (EER) significantly (P<0.05). Dry matter (DM) digestibility was decreased by lower dietary CP whereas 0.03% PUT significantly (P<0.05) increased it. Low CP caused significant (P<0.05) greater calcium digestibility, while this effect was not found when PUT was added. PUT had no effect on intestine villous height and crypt depth. Polyamine content of intestine and liver was influenced by the age of the birds, while PUT had no effects on them. In conclusion, dietary PUT has beneficial effects on EER in chicks fed CP-deficient diet, indicating possible involvement of PUT in energy metabolism. PUT supplementation did not moderate the reduced BWG of the chicks fed low protein. Intestinal and liver polyamine concentration was mainly affected by dietary CP and age of the birds rather than dietary PUT.

Dietary supplementation of Zingiber officinale and Zingiber zerumbet to heat-stressed broiler chickens and its effect on heat shock protein 70 expression, blood parameters and body temperature.

The present study was conducted to assess the effects of dietary supplementation of Zingiber officinale and Zingiber zerumbet and to heat-stressed broiler chickens on heat shock protein (HSP) 70 density, plasma corticosterone concentration (CORT), heterophil to lymphocyte ratio (HLR) and body temperature. Beginning from day 28, chicks were divided into five dietary groups: (i) basal diet (control), (ii) basal diet +1%Z. zerumbet powder (ZZ1%), (iii) basal diet +2%Z. zerumbet powder (ZZ2%), (iv) basal diet +1%Z. officinale powder (ZO1%) and (v) basal diet +2%Z. officinale powder (ZO2%). From day 35-42, heat stress was induced by exposing birds to 38±1°C and 80% RH for 2 h/day. Irrespective of diet, heat challenge elevated HSP70 expression, CORT and HLR on day 42. On day 42, following heat challenge, the ZZ1% birds showed lower body temperatures than those of control, ZO1% and ZO2%. Neither CORT nor HLR was significantly affected by diet. The ZO2% and ZZ2% diets enhanced HSP70 expression when compared to the control groups. We concluded that dietary supplementation of Z. officinale and Z. zerumbet powder may induce HSP70 reaction in broiler chickens exposed to heat stress.

Dietary methionine and n-6/n-3 polyunsaturated fatty acid ratio reduce adverse effects of infectious bursal disease in broilers.

This study was carried out to investigate the modulatory effects of dietary methionine and n-6/n-3 polyunsaturated fatty acids (PUFA) ratio on immune response and performance of infectious bursal disease (IBD)-challenged broiler chickens. In total, 350 one-day-old male broiler chicks were assigned to 1 of the 6 dietary treatment groups in a 3 × 2 factorial arrangement. There were 3 n-6/n-3 PUFA ratios (45, 5.5, and 1.5) and 2 levels of methionine (NRC recommendation and twice NRC recommendation). The results showed that birds fed with dietary n-6/n-3 PUFA ratio of 5.5 had higher BW, lower feed intake, and superior FCR than other groups. However, the highest antibody response was observed in birds with dietary n-6/n-3 PUFA ratio of 1.5. Lowering n-6/n-3 PUFA ratio reduced bursa lesion score equally in birds fed with n-6/n-3 PUFA ratio of 5.5 and 1.5. Supplementation of methionine by twice the recommendation also improved FCR and reduced feed intake and bursa lesion score. However, in this study, the optimum performance (as measured by BW, feed intake, and FCR) did not coincide with the optimum immune response (as measured by antibody titer). It seems that dietary n-3 PUFA modulates the broiler chicken performance and immune response in a dose-dependent but nonlinear manner. Therefore, it can be suggested that a balance of moderate level of dietary n-6/n-3 PUFA ratio (5.5) and methionine level (twice recommendation) might enhance immune response together with performance in IBD-challenged broiler chickens.

Promoting positive health behaviours--'tooth worm' phenomenon and its implications.

UNLABELLED: 'Tooth worm' is a traditional belief about the pathogen of dental caries (tooth decay). Nevertheless, in our previous study, parental 'tooth worm' belief was linked to a reduced caries risk of their children. OBJECTIVES: This study aimed to further characterize the impact of parental 'tooth worm' belief on their children's caries experience and its psychobehavioural mechanisms. BASIC RESEARCH DESIGN: analytic observational study. SETTING: Thirteen randomly selected kindergartens in Singapore. PARTICIPANTS: 1,782 preschoolers aged 3-6 years. METHODS: Each child received an oral examination and microbiological tests. Parents completed a self-administered questionnaire on their socio-demographic background, oral health knowledge/attitude and child's oral health habits. RESULTS: Multivariate analysis confirmed a reduced chance of 'high caries rate' (number of affected teeth > 2) among children whose parents held the 'tooth worm' belief (Odds Ratio = 0.41; 95% Confidence Interval = 0.19-0.89). With such perception among parents, children brushed their teeth more frequently (p = 0.042). Since no difference in oral hygiene was observed, the health benefit of the "tooth worm" perception may be acquired through the delivery of fluoride (an agent with proven anti-caries effect) during frequent toothbrushing episodes. CONCLUSIONS: This study revealed a 'tooth worm' phenomenon, indicating that parental 'tooth worm' belief is associated with early establishment of regular toothbrushing habit and reduction of dental caries in children. This phenomenon and its psychobehavioural mechanisms, enriching our understanding of oral health behaviours, have implications for effective health education.

Effects of feeding metabolite combinations produced by Lactobacillus plantarum on growth performance, faecal microbial population, small intestine villus height and faecal volatile fatty acids in broilers.

1. Four combinations of metabolites produced from strains of Lactobacillus plantarum were used to study the performance of broiler chickens. 2. A total of 432 male Ross broilers were raised from one-day-old to 42 d of age in deep litter pens (12 birds/pen). These birds were divided into 6 groups and fed on different diets: (i) standard maize-soybean-based diet (negative control); (ii) standard maize-soybean-based diet + Neomycin and Oxytetracycline (positive control); (iii) standard maize-soybean-based diet + 0.3% metabolite combination of Lactobacillus plantarum RS5, RI11, RG14 and RG11 strains (com3456); (iv) standard maize-soybean-based diet + 0.3% metabolite combination of L. plantarum TL1, RI11 and RG11 (Com246); (v) standard maize-soybean-based diet + 0.3% metabolite combination of L. plantarum TL1, RG14 and RG11 (Com256) and (vi) standard maize-soybean-based diet + 0.3% metabolite combination of L. plantarum TL1, RS5, RG14 and RG11 (Com2356). 3. Higher final body weight, weight gain, average daily gain and lower feed conversion ratio were found in all 4 treated groups. 4. The addition of a metabolite combination supplementation also increased faecal lactic acid bacteria population, small intestine villus height and faecal volatile fatty acids and faecal Enterobacteriaceae population.

Vermicomposting of cattle and goat manures by Eisenia foetida and their growth and reproduction performance.

Vermicomposting is commonly adopted for the treatment of livestock organic wastes. In the present study, two types of livestock manure were used for culturing of the earthworm, Eisenia foetida. Each treatment group consisted of six replicates and worm vermicasts were examined after 5 weeks. The concentrations of total C, P and K in goat manure vermicasts were higher than those in cattle manure vermicasts. Cattle vermicasts had a higher N content than goat vermicasts but the C:N ratio of fresh manure was higher than that of vermicasts for both materials. Earthworm biomass and reproductive performance, in terms of number of worms after 5 weeks of experiment, were higher in cattle manure than in goat manure. The cocoon production per worm in cattle manure was higher than in goat manure. However, the hatchability of cocoons was not affected by manure treatments. In conclusion, cattle manure provided a more nutritious and friendly environment to the earthworms than goat manure.

The effects of iron supplementation in preweaning piglets.

The trial was carried out at a commercial pig farm in Bukit Pelanduk, Negeri Sembilan, Malaysia. The objective of the study was to assess the efficacy of supplemental iron in drinking water and iron in paste form in comparison with the iron dextran injection. A total of 12 litters were used divided into three treatments : iron dextran injection (ID), Fedexx" - iron in paste form (IP) and Opti-iron" - iron in drinking water (IW). Hemoglobin level and growth performance parameters were monitored weekly over a period of 28 days. ID piglets had significantly higher (p < 0.05) body weight and weekly weight gain compared with IW piglets at 21 and 28 days of age whereas no significant different (p > 0.05) results were found between IP and IW piglets. Hemoglobin level from days 0 to 28 showed statistical difference (p > 0.05) between piglets in IP and IW groups. Mild anemia was found in IP piglets but not for ID and IW piglets compared with normal value (> 9 g/dL). Hemoglobin (Hb) levels were positively correlated (p < 0.05, r = .47) with body weight at 28 days of age. In conclusion, pigs supplemented with iron in paste form and drinking water had poorer growth performance than iron dextran injection piglets. Piglets given iron in paste form had mild anemia. It is advisable to give multiple doses of oral iron to piglets in order to prevent iron deficiency anemia. During the first 4 weeks of life, the body weights of the piglets increased by up to 5-fold. The dietary requirement for iron during this period is 7 mg per day but only 1 mg per day can be supplied by the sow's milk (Cunha, 1977; Miller and Ullrey, 1997; Roberts, 1998). The need to provide the piglets with an adequate amount of iron before weaning is therefore imperative because sow's milk alone will not meet the iron requirements of rapid growth and expanding blood volume (Hannan, 1971). Several different methods can be used to offer iron to preweaning piglets. The most common administration method for iron in piglets is through an intramuscular injection of iron dextran complex at 3 days of age. A single dose of 200 mg/ml iron-dextran is effective against iron deficiency anemia. However, the method is very stressful to the piglets. They will suffer more pain if a greater dosage of iron is given intramuscularly. Furthermore, poor iron injection techniques may cause considerable trauma to the muscles, staining of hams or create abscesses and lead to downgrading of the carcasses (Roberts, 1998). Since there are several drawbacks to the iron injection, alternative methods need to be considered in intensive farming i.e. supply iron orally (Miller and Ullrey, 1997). The oral administration of iron has two advantages: cost and the fact that absorption is regulated by the intestinal mucosae. However, oral administration requires multiple doses. This is because a single dose may not be sufficient to protect the piglets from iron deficiency anemia for the first 4 weeks post-partum. The iron bioavailability in oral iron depends greatly on iron status of animals (Amine et al., 1972; Susan and Wright, 1985). In addition, dietary factors such as amino acids and protein sources (Martinez et al., 1981), pectin content, phytate (Morris and Ellis, 1982) and the other minerals (Elvehjem and Hart, 1932; Hedges and Kornegay, 1973; Suttle and Mills, 1966) may also influence the bioavailability of iron. Oral iron can be given as paste or drinking water. Early administration of oral iron within the first few days of life will meet the iron needs of the suckling pig. However, it is critical to administer early before gut closure to large molecules (Harmon et al., 1974; Thoren-Tolling, 1975). This study was conducted to determine the efficacy of iron supplementation in drinking water or paste form in comparison with the injection of iron compounds in suckling pigs.

Heterophil to lymphocyte ratio and tonic immobility reactions to preslaughter handling in broiler chickens treated with ascorbic acid.

Stress and fear responses were evaluated in broiler chicks that were pretreated for 24 h with 0 ppm (control) or 1,200 ppm of L-ascorbic acid (AA) in their drinking water. The birds were subsequently subjected to either upright handling (UH) or inverted (IH) handling for about 45 s. Heterophil (H) counts, lymphocyte (L) counts, and H/L ratios (H/L) ratios were determined immediately (T0) and at 20 h (T20) following the handling treatment. The H/L ratios were similar for both groups at T0, whereas 20 h after the handling treatment, AA-supplemented birds had lower ratios than controls, resulting in a significant water treatment x time of blood sampling interaction. Inverted handling had negligible effect on H/L ratios but augmented tonic immobility (TI) durations as compared with UH. Irrespective of handling procedure, supplemental AA reduced underlying fearfulness, as measured by TI reaction. Neither water treatment nor handling method had significant effect on number of attempts to induce TI.

Specific interaction between tetrandrine and Quillaja saponins in promoting permeabilization of plasma membrane in human leukemic HL-60 cells.

Spontaneous Ni2+ entry (leak), measured as fluorescence quench in fura-2-loaded HL-60 cells at the excitation wavelength of 360 nm, was strongly inhibited by tetrandrine (TET, 100 microM), a Ca2+ antagonist of Chinese herbal origin. Exposure of the cells for 5 min to saponins from Quillaja saponaria (QS, 30 microg/ml), surfactants well known to permeabilize the plasma membrane by complexing with cholesterol, promoted Ni2+ entry without causing fura-2 leak-out. Unexpectedly, TET caused an immediate (within 2.5 min) augmentation of QS-promoted Ni2+ entry; and a 5-min treatment with both TET and QS resulted not only in an enhanced Ni2+ entry, but also a fura-2 leak-out. Ginseng saponins (100 microg/ml) alone or together with TET did not cause such a permeabilization. Permeabilization induced by 1-3 microM digitonin, another cholesterol-complexing glycoside, could not be enhanced by TET. TET did not affect permeabilization induced by Triton X-100 (0.01%), a detergent which non-specifically disrupts the hydrophobic interaction at the plasma membrane. TET also did not enhance Ni2+ entry triggered by ionomycin (0.35 microM) or SK&F 96365 (20 microM). Further, it did not augment Ni2+ entry when the plasma membrane fluidity was modulated by changes of temperature (27-47 degrees C) or treatment with 5% ethanol. This QS-promoted Ni2+ entry could not be amplified by other lipophilic Ca2+ antagonists, such as diltiazem (100 microM) and verapamil (100 microM). The results hence indicate that TET enhanced Ni2+ entry (or permeabilization) elicited by QS treatment, but not other perturbations of the plasma membrane. We suggest that pore formation at the plasma membrane, a consequence of QS-cholesterol interaction, can be specifically enhanced by TET. Also, a comparative study of the effects of TET and its very close analogues, hernandezine and berbamine, reveals that the methoxyl group at the R2 position of TET appears to be crucial in enhancing QS-promoted Ni2+ entry.

Dual effects of tetrandrine on cytosolic calcium in human leukaemic HL-60 cells: intracellular calcium release and calcium entry blockade.

1. Tetrandrine (TET, a Ca2+ antagonist of Chinese herbal origin) and thapsigargin (TSG, an endoplasmic reticulum Ca2+ pump inhibitor) concentration-dependently mobilized Ca2+ from intracellular stores of HL-60 cells, with EC50 values of 20 microM and 0.8 nM, respectively. After intracellular Ca2+ release by 30 nM TSG, there was no more discharge of Ca2+ by TET (100 microM), and vice versa. 2. Pretreatments with 100 nM rauwolscine (alpha 2-adrenoceptor antagonist), 100 nM prazosin (alpha 1-adrenoceptor antagonist), 10 nM phorbol myristate acetate (PMA, a protein kinase C activator) or 100 nM staurosporine (a protein kinase C inhibitor) had no effect on 100 microM TET-induced intracellular Ca2+ release. 3. After intracellular Ca2+ release by 30 nM TSG in Ca(2+)-free medium, readmission of Ca2+ caused a substantial and sustained extracellular Ca2+ entry. The latter was almost completely inhibited by 100 microM TET (IC50 of 20 microM) added just before Ca2+ readmission. In Ca(2+)-containing medium, 30 nM TSG caused a sustained phase of cytosolic Ca2+ elevation, which could be abolished by 100 microM TET. TET was also demonstrated to retard basal entry of extracellular Mn2+ and completely inhibit TSG-stimulated extracellular Mn2+ entry. 4. TSG-induced extracellular Ca2+ entry was insensitive to the L-type Ca2+ channel blocker, nifedipine (1 microM), but was completely inhibited by the non-selective Ca2+ channel blocker La3+ (300 microM). Depolarization with 100 mM KCl did not raise the cytosolic Ca2+ level. 5. These data suggest that (a) TET and TSG mobilized the same Ca2+ pool and TET-induced intracellular Ca2+ release was independent of protein kinase C activity and ox-adrenoceptor activation,and (b) TET blocked the voltage-insensitive Ca2+ entry pathway activated by TSG. These dual effects on HL-60 cells were also observed with hernandezine (HER), a TET-like compound and in another cell type, murine B lymphoma M12.4 cells.

Apotransferrin can elevate intracellular free calcium ion and stimulate mitogenesis in human leukemic HL60 cells.

In order to resolve the question whether or not transferrin could have a growth-promoting effect on cells independent of its action in iron transport, we investigated the effect of the iron-free form of transferrin, apotransferrin on cell activation and proliferation in the human leukemic HL60 cell line. Within a minute of its addition to HL60 cells, apotransferrin caused a rise in intracellular free calcium concentration ([Ca2+]i) in a dose-dependent manner and the higher the apotransferrin, the quicker it was to attain the calcium peak, showing the physiological characteristics of an agonist-induced [Ca2+]i elevation. The source of calcium appears to be extracellular since this signal could be abolished by nickel or when the reaction was carried out in calcium-free medium. Addition of apotransferrin in the serum-free medium could markedly promote DNA synthesis whereas addition of iron citrate could not. However, apotransferrin could not sustain cell proliferation and hypertrophism without other growth or nutritional factors. Antitransferrin receptor antibody inhibited the growth of HL60 cells cultured in serum-free medium supplemented with transferrin and insulin in a dose-dependent manner, whereas addition of ferric citrate could not reverse cell growth. Generation of the calcium signal probably reflects the initiation of the cell activation processes which could culminate into mitogenesis. Hence, our results suggest that apotransferrin, not iron, is bioactive in HL60 cells.

The interaction of resealed reticulocyte ghosts with Fe3+-transferrin-CO3(2-).

The present studies were designed to investigate the interaction of Fe3+transferrin-CO3(2-) with the transferrin receptors of the resealed reticulocyte ghost and to assess the degree to which the iron release reaction can be reconstituted in resealed ghosts supplemented with entrapped cytoplasmic components. Reticulocyte, but not erythrocyte, ghosts displayed an intact Fe3+transferrin-CO3(2-) binding capability. When ATP, NADH and ferritin were included during the resealing process, some iron release to the ghosts was observed.