Identification of a Hemolysis Threshold That Increases Plasma and Serum Zinc Concentration.

Background: Plasma or serum zinc concentration (PZC or SZC) is the primary measure of zinc status, but accurate sampling requires controlling for hemolysis to prevent leakage of zinc from erythrocytes. It is not established how much hemolysis can occur without changing PZC/SZC concentrations.Objective: This study determines a guideline for the level of hemolysis that can significantly elevate PZC/SZC.Methods: The effect of hemolysis on PZC/SZC was estimated by using standard hematologic variables and mineral content. The calculated hemolysis threshold was then compared with results from an in vitro study and a population survey. Hemolysis was assessed by hemoglobin and iron concentrations, direct spectrophotometry, and visual assessment of the plasma or serum. Zinc and iron concentrations were determined by inductively coupled plasma spectrometry.Results: A 5% increase in PZC/SZC was calculated to result from the lysis of 1.15% of the erythrocytes in whole blood, corresponding to ∼1 g hemoglobin/L added into the plasma or serum. Similarly, the addition of simulated hemolysate to control plasma in vitro caused a 5% increase in PZC when hemoglobin concentrations reached 1.18 ± 0.10 g/L. In addition, serum samples from a population nutritional survey were scored for hemolysis and analyzed for changes in SZC; samples with hemolysis in the range of 1-2.5 g hemoglobin/L showed an estimated increase in SZC of 6% compared with nonhemolyzed samples. Each approach indicated that a 5% increase in PZC/SZC occurs at ∼1 g hemoglobin/L in plasma or serum. This concentration of hemoglobin can be readily identified directly by chemical hemoglobin assays or indirectly by direct spectrophotometry or matching to a color scale.Conclusions: A threshold of 1 g hemoglobin/L is recommended for PZC/SZC measurements to avoid increases in zinc caused by hemolysis. The use of this threshold may improve zinc assessment for monitoring zinc status and nutritional interventions.

Biomarkers of Nutrition for Development (BOND)-Zinc Review.

Zinc is required for multiple metabolic processes as a structural, regulatory, or catalytic ion. Cellular, tissue, and whole-body zinc homeostasis is tightly controlled to sustain metabolic functions over a wide range of zinc intakes, making it difficult to assess zinc insufficiency or excess. The BOND (Biomarkers of Nutrition for Development) Zinc Expert Panel recommends 3 measurements for estimating zinc status: dietary zinc intake, plasma zinc concentration (PZC), and height-for-age of growing infants and children. The amount of dietary zinc potentially available for absorption, which requires an estimate of dietary zinc and phytate, can be used to identify individuals and populations at risk of zinc deficiency. PZCs respond to severe dietary zinc restriction and to zinc supplementation; they also change with shifts in whole-body zinc balance and clinical signs of zinc deficiency. PZC cutoffs are available to identify individuals and populations at risk of zinc deficiency. However, there are limitations in using the PZC to assess zinc status. PZCs respond less to additional zinc provided in food than to a supplement administered between meals, there is considerable interindividual variability in PZCs with changes in dietary zinc, and PZCs are influenced by recent meal consumption, the time of day, inflammation, and certain drugs and hormones. Insufficient data are available on hair, urinary, nail, and blood cell zinc responses to changes in dietary zinc to recommend these biomarkers for assessing zinc status. Of the potential functional indicators of zinc, growth is the only one that is recommended. Because pharmacologic zinc doses are unlikely to enhance growth, a growth response to supplemental zinc is interpreted as indicating pre-existing zinc deficiency. Other functional indicators reviewed but not recommended for assessing zinc nutrition in clinical or field settings because of insufficient information are the activity or amounts of zinc-dependent enzymes and proteins and biomarkers of oxidative stress, inflammation, or DNA damage.

Meat supplementation increases arm muscle area in Kenyan schoolchildren.

The present study examines the effect of animal-source-food (ASF) intake on arm muscle area growth as part of a larger study examining causal links between ASF intake, growth rate, physical activity, cognitive function and micronutrient status in Kenyan schoolchildren. This randomised, controlled feeding intervention study was designed with three isoenergetic feeding interventions of meat, milk, and plain traditional vegetable stew (githeri), and a control group receiving no snack. A total of twelve elementary schools were randomly assigned to interventions, with three schools per group, and two cohorts of 518 and 392 schoolchildren were enrolled 1 year apart. Children in each cohort were given feedings at school and studied for three school terms per year over 2 years, a total of 9 months per year: cohort I from 1998 to 2000 and cohort II from 1999 to 2001. Food intake was assessed by 24 h recall every 1-2 months and biochemical analysis for micronutrient status conducted annually (in cohort I only). Anthropometric measurements included height, weight, triceps skinfold (TSF) and mid-upper-arm circumference (MUAC). Mid-upper-arm muscle area (MAMA) and mid-upper-arm fat area (MAFA) were calculated. The two cohorts were combined for analyses. The meat group showed the steepest rates of gain in MUAC and MAMA over time, and the milk group showed the next largest significant MUAC and MAMA gain compared with the plain githeri and control groups (P< 0.05). The meat group showed the least increase in TSF and MAFA of all groups. These findings have implications for increasing micronutrient intake and lean body mass in primary schoolchildren consuming vegetarian diets.

Titers of antibody to common pathogens: relation to food-based interventions in rural Kenyan schoolchildren.

BACKGROUND: Undernutrition is widely perceived to affect the development of an effective immune system. OBJECTIVE: We used a mini-analysis system to quantitate antibody titers and evaluate the sera of 200 Kenyan schoolchildren for antibodies to Helicobacter pylori [isotypes of immunoglobulins A (IgA), G (IgG), and M (IgM)], hepatitis A virus, rotavirus, tetanus toxoid (IgG), and a panel of recombinant malarial antigens (MSP1(19), MSP2, Ag512, MSP4, and MSP5). DESIGN: Children participated in a school-based feeding intervention with meat, milk, or nonanimal-source foods or in a nonintervention control group. Microvolumes (200 mL) of sera were analyzed at baseline and after 1 y. RESULTS: Nearly all children had elevated titers of antibody to H. pylori, hepatitis A virus, rotavirus, and malaria at the outset, despite a high prevalence of apparent biochemical micronutrient deficiencies and stunting, but many had titers of tetanus toxoid IgG antibodies below the protective concentration. Children with low hemoglobin had a greater proportion of elevated H. pylori IgM antibody titers at baseline, which suggests that current infection with H. pylori may be associated with anemia. Compared with the control subjects, only the group eating meat had a significant increase in H. pylori IgM antibodies during the intervention (P = 0.019). No other group comparisons with the control subjects were statistically significant. The additional finding that the sera of some children showed inadequate tetanus-protective antibodies, despite immunization, suggests that the vaccination program was suboptimal. CONCLUSIONS: A large battery of immune assays can be performed on microvolumes of sera. Furthermore, despite evidence of malnutrition, children do develop significant antibody-mediated responses to common pathogens.

Low plasma vitamin B-12 in Kenyan school children is highly prevalent and improved by supplemental animal source foods.

The high prevalence of vitamin B-12 deficiency in many regions of the world is becoming recognized as a widespread public health problem, but it is not known to what extent this deficiency results from a low intake of the vitamin or from its malabsorption from food. In rural Kenya, where a previous study identified a high prevalence of inadequate vitamin B-12 intakes, this study examined whether plasma vitamin B-12 concentrations were associated with dietary sources of the vitamin at baseline and could be increased by supplementation with animal source foods (ASF). The 4 experimental groups in 503 school children were: 1) control (no food provided); 2) githeri (a maize and bean staple with added oil); 3) githeri + meat (githeri + minced beef); or 4) githeri + milk (githeri + milk). Feedings were isocaloric. Dietary data were collected at baseline, and biochemical data at baseline and after 1 and 2 y of feeding. Baseline plasma vitamin B-12 concentration was 193.6 +/- 105.3 pmol/L and correlated with % energy from ASF (r = 0.308, P < 0.001). The odds ratio for low plasma vitamin B-12 (<148 pmol/L), which occurred in 40% of children, was 6.28 [95% CI: 3.07-12.82] for the lowest vs. highest ASF intake tertile (P < 0.001). Feeding ASF (meat or milk) greatly reduced the prevalence of low plasma vitamin B-12 (P < 0.001). The high prevalence of low plasma vitamin B-12 concentrations in these children is predicted by a low intake of ASF, and supplemental ASF improves vitamin B-12 status.

Kenyan school children have multiple micronutrient deficiencies, but increased plasma vitamin B-12 is the only detectable micronutrient response to meat or milk supplementation.

Animal source foods (ASF) can provide micronutrients in greater amounts and more bioavailable forms compared to plant source foods, but their intake is low in many poor populations. However, the impact of ASF on micronutrient status of undernourished populations has not been assessed. Supplemental meat (60-85 g/d), milk (200-250 mL/d) or energy (isocaloric with the meat and milk, 240-300 kcal/d) were randomly assigned to 555 undernourished school children aged 5-14 y in a rural malaria-endemic area of Kenya, at one school meal daily for one school year. Blood and stool samples were collected at baseline and after 1 y to assess stool parasites, malaria, hemoglobin, serum or plasma C-reactive protein, ferritin, iron, zinc, copper, vitamin B-12, folate and retinol, and erythrocyte riboflavin. At baseline, there was a high prevalence of micronutrient deficiencies (iron, zinc, vitamins A and B-12 and riboflavin), yet plasma ferritin was low in few children, and none had low serum copper. At the end of the year of supplementation, plasma vitamin B-12 concentrations were significantly increased in children fed the Meat or Milk meal; prevalence of severe plus moderate deficiency fell from 80.7% at baseline to 64.1% in the Meat group and from 71.6 to 45.1% in the Milk group, respectively. No significant improvement was observed in the status of other micronutrients compared to the Energy and Control groups, although malaria and other infections may have obscured effects. Supplementation with small amounts of meat or milk reduced the high prevalence of vitamin B-12 deficiency in these children.