Relationship between blood Lead status and anemia in Ugandan children with malaria infection
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RESEARCH ARTICLE
Open Access
Relationship between blood Lead status and anemia in Ugandan children with malaria infection Ambrose Mukisa, Denis Kasozi, Claire Aguttu, Peter C. Vuzi and Joseph Kyambadde*
Abstract Background: In Uganda, childhood anemia remains a health challenge and is associated with malaria infection as well as iron deficiency. Iron deficiency is intertwined with nutritional status, age and other comorbidities including helminths and Lead toxicity. Environmental Lead levels accounts for one’s blood Lead (BL) levels. Blood Lead competitively blocks iron absorption, inhibits hemoglobin (Hb) biosynthesis and elevates free erythrocyte protoporphyrin (FEP) levels. Lead toxicity’s contribution towards anemia pathogenesis, especially during malaria infection has not been studied. Concomitant exposure to both malaria infection and Lead pollution, exacerbates the anemia status. This study therefore aimed at expounding the anemia status of these Ugandan children aged under 5years who are exposed to both malaria infection and environmental Lead pollution. Methods: Briefly, venous blood samples from 198 children were microscopically assayed for malaria parasite density (PD), and hemoglobin (Hb) concentrations using the cyanmethemoglobin method, while BL and FEP levels were determined by the standard atomic absorption spectrophotometric and fluorometric methods respectively. Results: One hundred and fifty-one (76.3%) of the children analyzed had moderate anemia (Hb 5 g/dL) with Means of BLL=8.6 µg/dL, Hb =7.5 g/dL, FEP/Hb =8.3 µg/g and PD =3.21×103 parasites / µL, while eight (4%) were severely anemic ( 10 g Hb/dL (WHO cut off reference), mean parasite density = 1.7 × 103 parasites/ µL, mean BLL < 2 µg/ dL, Mean FEP/Hb = 7.4 ± 2.9 µg /g. The details of the distribution are shown in Table 1 below
The FEP was measured following a method described by [29] using a fluorospectrophotometer set at 405 nm excitation 610 nm emissions. The porphyrins were extracted by adding 20 µl aliquots of whole blood to a solution containing 100 µL of 10% ammonium sulfate and 5% celite and vortex mixed for 10sec. 400 µl of 95% ethanol was then added and vortex mixed for more 20 seconds. This was followed by addition of 600 µl of acetone and further vortex mixing for 20 seconds. All the samples were put on an ice bath for 20 min, vortex mixed for 20 sec. and centrifuged at 4 °C for 10 min. After which the supernatants were harvested into small borosilicate tubes and aliquots of 300 µl mixed with 300 µl of a solution containing propylene glycol and 1.5 N HCI, (4:1), let to stand for 20 min before reading at 405 nm excitation and 610 emissions. The FEP blood concentration was calculated using the following formulae; FEP ðμg=dL BloodÞ ¼ FEP μg=100 ml extract Fz Cs 2:7 100 ¼ Fs 1:1 0:2 Where Fz is the sample fluorescence, Cs concentration of the standard, Fs is the fluorescence of the standard, 2.7 is the final volume of HCl phase, 100 is the conversion factor to 100 ml of extract, 1.1 conversion factor for protoporphyrin m
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