Materials and Method

2. Materials and Method

no public or commercial reliable laboratories with

2.1 Objective

skilled staff.

Theoretically, several sensitive lab analyses like The overall objective was to test the hypothesis that

locally available natal plum fruits could reduce IDA hematocrit, serum ferritin, Mean Corpuscular Volume through a dietary intervention with better (MCV), erythrocyte protoporphiryn and Hemoglobin

bioavailability than those of iron salts (HB) are ideal to secure an accurate diagnosis of IDA. supplementation and fortification practices. In this

However, as a result of inadequate facilities, the paper anemia and Iron Deficiency Anemia (IDA) are

research team measured HB only, thus under these interchangeable. If the hypothesis proves to be

assumptions, IDA and anemia due to dietary intake positive, consumption of the fruit (natal plum) could

are interchangeable throughout this paper. The HB

be encouraged to combat against iron deficiency cut-off for inclusion into the study was set at 8 g/dl anemia in developing countries, which import iron salt

rather than the standard 10 g/dL due to the general tablets from abroad with immense difficulties and

poor nutritional situation of the population in the costs. The second advantage of using natal plum

affected area and to the fact that our targeted would be to strengthen the vision of using locally

respondents were children and that IDA becomes available dietary inputs with presumably considerable

symptomatic around these HB concentrations [7, 8]. ease and acceptability. The third hypothetical

For the HB measuring device was the HemoCue advantage would be the introduction of an B-haemoglobin analyzer and accompanying environmental friendly scheme for planting more trees

replaceable microcuvettes with dried reagents thus reducing the production of pollutants produced

(HemoCue Ltd, Viking Court, Derbyshire UK). The by industry in manufacturing iron tablets. The fourth

blood samples were collected by pricking fingertips hypothetical advantage of this plant could be the use

with pen needles of 4 mm.

The Potentials of Locally Available Fruits Rich in Iron to Mitigate Iron

Deficiency Anemia in Least Developing Countries (LSD)

2.3 Respondents Selection and Sampling natal plum or Iron tablets for 15 days. During the first Procedures

week, the task of selecting new respondents, the Since there was no government or any other

follow up and at the same time giving out regimens authorities other than local chiefs due to the collapse

and doses to those already enrolled was a challenging of the government, a meeting was arranged between

job. The 15 days cumulative baseline totals of both the village chiefs, parents and the medical staff of the

experimental and control groups were n = 101 and n= implementing NGO to discuss and approve ethics of

100 respectively. Every effort was made to keep the study.

enrolment number constant but due to unpredictable Sample calculations were based on the village’s

daily respondents attendance, the number actually general child population of 1,471 (3,200 people of

fluctuated from sets as low as seven respondents to as which 46% were children between one and 14 years of

high as 20 respondents per day. Luckily, our resources age and a previous study in this community were more than enough at every one day should the unpublished local NGO DARYEEL report) with

respondents number should be swell unexpectedly–we population standard deviation (SD) of 18 units and a

planned for 30 respondents for each in experimental sample mean gain HB target of 4 g/dL. A power

and control groups.

calculation with 95% confidence, based on the mean Those whose anemia was due to malarial infections HB gain and SD determined in the previous study in

or hookworm infestations were excluded from the list this community, was done to determine sample size

on grounds of medical and ethical reasons. No other and was as follows:

selection criterion was practically applicable. = 1.96 * SD/ √n,

2.4 Randomization of Sample

√n = 1.96 * SD/ » (1.96 * 18*/4) ≈ 78 Where

and SD are the population age mean Parents, on behalf of their children, were happy to and standard deviations respectively, 1.96 = 95%

use pickup-your-luck system rather than computer confidence.

generated randomization selection. The respondents Therefore a sample size of 78 children was needed

were allocated with numbers that were written on for each of the control and experimental groups. An

small papers and twisted to hide the number and were extra 22 children were added to get a new sample size

rolled over several times to distribute evenly in an of 100 each for intervention and case groups to

empty medical jar. After that, any parents could begin address possible drop-outs.

the picking process from the now open container. Respondents were randomly selected from a pool of

Before the pickup the audience were told that the first anemic children patients that came every other day

draw would be for the fruit group, and the alternate and without appointments to the subject NGO health

second draw would be for the ferrous sulfate till the care centre. These children were aged from 5 to 12

last number of the day was drawn. years. It took a total of seven consecutive days to

For the experimental group (n = 101), about 400 identify and randomly choose those designed to use

grams fruits of ripe Natal plum, with bioavailability of natal plum fruits and those earmarked tas control

2 mg per 100 grams of the fruit [3], was given while group using ferrous sulfate tablets from the local

the control group (n = 100) received 30 mg of Ferrous NGOs. Each day of the seven days new group was

Sulfate tablets each day with bioavailability of 27% [7, chosen from the pool of anemic children while

9]. The adjustment was devised that both regiments previously selected group(s) continued to receive their

would deliver the same targeted amount of iron of 8 respective daily regimens/doses of either fruits of

mg delivery per day. For ethical reasons, after the

64 The Potentials of Locally Available Fruits Rich in Iron to Mitigate Iron

Deficiency Anemia in Least Developing Countries (LSD)

intervention period, all respondents (n = 190) the two means of the two groups were similar at 6.0 continued to receive 30 mg iron and vitamin C tablets

g/dL cut-point (P > 0.397, CI 95%) where unequal for another two months.

variances were assumed.

At baseline both treatment and control groups have

2.5 Blood Collection and Hemoglobin Analysis had similar low levels of HB (P > 0.397, at 95%

The blood sample was taken from fingertips using a confidence level). Though both groups seem to have recoil pen needle with replaceable needles. Before

gained HB level due to the intervention, the natal puncturing the fingertips, cotton swabs with natural

plum group had the highest gains (P < 0.008, at 95% alcohol cleansers were applied to avoid infection to

confidence level).

develop at the insertion point. At the final HB evaluations, both groups had The microcuvettes were supplied in tubs with

significant HB gains but when their respective final airtight lids designed to keep out moisture. The tubs

HB means were compared, using the independent were being kept closed and dated. The microcuvette

samples t-test, the fruit\experimental group gained was filled and there were no air bubbles within the

more than control group, 4.06 ± 1.59 compared to 2.8 chamber. Blood overflow outside microcuvette ± 0.99 to reach the targeted 8.0 g/dL cut-point windows was avoided. The loaded microcuvettes were

(P < 0.008, CL 95%), equal variances not assumed. tested within 10 minutes.