TITLE: EFFECT OF MALARIA INFECTION ON PACKED CELL VOLUME AND HAEMOGLOBIN LEVELS AMONG

PATIENTS AT OJO DISTRICT OF LAGOS STATE NIGERIA.

IWUNZE VINCENT, NNEKA ELOTA, OKEY OWOLife – link Medical Diagnostic & labs Ltd

ABSTRACT Malaria is a worldwide migrating diseases associated with

anaemia especially in the tropical Africa. It causes high rate of mortality and morbidity among Sub-Saharan Africans. Anaemia is a Hematological disorder caused by a decrease in the production of Red blood cells, a born marrow failure or by an increase in the Red blood cells destruction. The study is to evaluate the effect of malaria infection on the Packed cell volume and haemoglobin levels of patients. Blood samples were collected in EDTA bottles from patients attending Life-link Medical Diagnostic and Laboratories Ltd and Patients from Ojo health Center, all in Ojo District of Lagos state Nigeria. Presence or absence of malaria identified by staining with Giemsa stain and examined under oil immersion , PCV and Hb were estimated using Hawskley Haematocrit machine and Microfield colorimeter respectively.

• A total of 250 patients were studied between June to October 2013, 180 (72%) malaria infected patients, 160 (89% Adult), 20 (11% children) and 70 (28%) of non-infected. The overall means for the Packed Cell volume and Haemoglobin showed a significant difference in malaria infected patients.

• The mean Hb, PCV in infected were lower than the non infected. Conclusively, there was a decrease in Packed cell volume and Haemoglobin which seems to be due to the excessive destruction of of Red blood cells by the malaria parasite.

BIOGRAPHY• IWUNZE VINCENT OKECHUKWU graduated from the

University of Calabar, Cross River state Nigeria in 1991. He specialized on Haematology and Blood Transfusion. Presently he is the MD/CEO of Life-link Medical Diagnostic and Laboratories Nigeria Ltd, one of the fast growing Diagnostic and Research Centers in the country. It is situated at the former capital of Nigeria, Lagos, the commercial hurb center of Africa. He has delivered many scientific papers on some conferences especially in Nigeria. He has also headed many scientific workshops and served in different Ad-Hoc committees. He is working with his team to contribute their quota to the profession of Haematology research.

INTRODUCTION• Malaria is a worldwide migrating health diseases especially in

many developing countries e.g Africa and Asia. It was first isolated in blood of a patient in 1881 by Alpores Laveren . Malaria is gotten if bitten by an infected anophelene mosquito, if one receives infected blood from someone during blood transfusion and from mother to child during pregnancy. The causative agent of malaria is Plasmodium which is made up of four species; Plasmodium Falciparium. Plasmodium Vivax, Plasmodium Ovale and Plasmodium Malariae. Plasmodium is a single celled parasite that multiplies in the red blood cells of human and mosquito. The two most common species in Africa are Plasmodium Vivax and Plasmodium Falciparium with P Falciparium more pathogenic with high morbidity and mortality rate in Sub-Saharan Africa. P. Falciparium modifies the surface of RBC causing it to become sticky. This nature makes the cells to stick to the walls of the vessels leading up to major organs

• like the brain. This build up is called “Sequestration” and results in reduced blood flow and oxygen deprivation in the organs hence causing anaemia and eventual death of the patient. Anaemia is a blood disorder and one of the fatal consequences of malaria infection in the whole world. The pathophysiology of anaemia caused by plasmodium falciparium is both complex and multifactorial and results in a condition which is a major cause of high mortality and morbidity in patients.

• According to World Health Organization 2013, 3.4 billion people ( half of world population) live in areas at risk of malaria transmission in 106 countries and territories. Report also had it that in 2012, malaria caused an estimated 207 million clinical episodes and 627,000 death yearly. An estimated 91% of death in 2010 were in the African region.

• MATERIALS AND METHOD • SAMPLE COLLECTION; Venous Blood samples were collected

in EDTA containers and test carried out the same day.• MATERIALS: Glass slide, Capillary tubes, Colorimeter,

Drabkin,s solution, Haematocrit Centrifuge and reader, Test tubes. Automatic pipettes

• METHOD• PACKED CELL VOLUME: It is the proportion of whole blood

occupied by the red cells expressed in percentage. It is used for the measurement of anaemia.

• PRINCIPLE: Anticoagulated blood in a glass capillary of specified length, bore size and wall –thickness is centrifuged in a microhaematocrit centrifuge at RCF 12000-15000 xg for 3-5 minutes to obtain a constant packing of the cells . The PCV value is read from the scale of a microhaematocrit reader.

HAEMOGLOBIN ESTIMATION: Measurement of haemoglobin concentration in a blood is a basic screen for anaemia and its severity or for polycythaemia.PRINCIPLE: Blood is diluted 1 in 201 in a solution containing potassium cyanide and potassium ferricyanide (Drabkins solution). The red cells are haemolyzed and the haemoglobin is oxidized by the ferricyanide to methaemoglobin. This is converted by cyanide to more stable haemoglobincyanide (HiCN). The absorbance of the solution is read in a Microfield colorimeter at 540nm wavelength. The absorbance obtained is compared with that of the reference standard on a haemoglobin chart prepared from a calibration graph.

• STATISTICAL ANALYSIS• The descriptive data were given as Mean ±

Standard Deviation SD. Independent student t-test was used to test and compare the significance of the difference between Mean values. The Pearson Chi Square test was used for analytical assessment and the differences were considered to be statistically significant when the P- values obtained was ≤ 0.05. Packed Cell volume is considered to the Hb in this evaluation.

ADULT MALE ADULT FEMALEGROUP PCV Hb PCV HbCONTROL 43.08±1.57 13.68±0.52 33.1±0.98 10.36±0.32POSITIVE for malaria

31.93±3.58 9.96±1.18 17.40±3.54 11.08±0.53

MALE CHILDREN FEMALE CHILDRENGROUP PCV Hb PCV HbCONTROL 21.74±3.88 6.57±1.29 31.93±3.59 9.96±1.18POSITIVE for malaria

17.4±3.54

4.91±1.13 43.08±1.57 13.68±0.52

Table 1; Showing PCV and Hb values with the mean and Standard Error of mean values of male and female adults.

Table 2; Showing PCV and Hb values with the mean and Standard Error of mean values of male and female children

Table 3; Showing comparism between GROUP 1&2, GROUP 3&4.

A STATISTICAL SIGNIFICANT DIFFERENCE WAS SEEN BETWEEN THE GROUPS. KEY; *P < 0.05 **P <0.01

GROUP MEAN STANDARD DEVIATION

STANDARD ERROR OF MEAN

SIG.- (2-TAILED)

GROUP 1-(control for male adult) AndGROUP 2-(malaria positive for male adult)

11.16 21.23 3.75 0.006*

GROUP 3-(control for female adult) AndGROUP 4-(malaria positive for female adult)

15.72 21.05 3.72 0.000*

TABLE 4: Showing comparism between GROUP 5&6, GROUP 7&8. GROUP MEAN STANDARD

DEVIATIONSTANDARD DEVIATION OF ERROR

SIG.-(2-TAILED)

GROUP 5-(control for male children) AndGROUP 6-(malaria positive for male children)

4,34 10.81 1.91 0.030*

GROUP 7-(control for female children)GROUP 8-(malaria positive for female children)

−8.84 17.54 3.10 0.008*

A STATISTICAL SIGNIFICANT DIFFERENCE WAS SEEN BETWEEN THE GROUPS. KEY; *P < 0.05

**P <0.01

• DISCUSSION

• The table above demonstrated that malaria caused more destruction of Red blood cells in male positive children in table 2 than the control male adult in table 1. It was also statistically significant (P<0.05) when the Haemoglobin values were compared. There was an insignificant difference between the control adult Female in group 3 and positive female children in group 4 with two-tailed P values less than 0.05. Packed cell volume values of control male children in group 5 and positive male children in group 6 had two-tailed P values less than 0.05 and by conventional criteria , this difference is considered to be very statistically significant. They also had Haemoglobin values which showed P values less than 0.05 thus was statistically significant. In group 7 , PCV for control female children and group 8 for positive female children , there is a significant difference with two-tailed P value of 0.05 and also had Haemoglobin values with P greater than 0.05 thus was statistically significant.

• The significant difference seen between the Packed cell volume values of the control groups and the positive groups is due to the increased destruction and reduced production of the Red blood cells.

CONCLUSION• The result in this research work showed that the P values of the

Packed cell volume and Haemoglobin values for control and positive malaria patients studied from Ojo district of Lagos Nigeria was statistically significant thus a difference was established between malaria positive and non malaria patients which were used as a control group. This study have proved other studies done which showed a decrease in PCV and Haemoglobin levels of malaria positive subjects. It has also established presence of critical anaemia in patients with malaria parasitaemia.

• Studies conducted in areas of stable, perennial malaria transmission in Tanzania, Cameroon and Kenya, showed that malarial infection correlates with decline in haemoglobin concentration of less than 7 gl/dl.

REFERENCES• (2008). Severe anemia in Malawian children, The New England Journal of Medicine

(358): 888-899.• Cox-Singh, Davis TM, Lee KS (2008). Plasmodium knowlesi malaria in humans is

widely distributed and potentially life threatening. Clinical Infectious Disease; (46):165

• Weatherall DJ & Abdalla S (1982), The Anaemia Of Plasmodium Falciparum Malaria, British Medical Bulletin, vol, 38, (2) pp 147-151

• Takem Ebako, Eric Achidi and Peter Ndumbe (2010); An update of malaria infections and anemia in adults in Beea, Cameroon. License BioMed Central Research Notes].

• Schellenburg, D, Schellenberg, and J.R., and Moshi, A. (2003); The silent burden of anaemia in Tanzanian children: a community – based study, Bull World Health Organ. (18): 581 – 90.

Monica Cheesbrough. District Laboratory Practice in Tropical Countries Part 2, Cambridge Second Editions. Published by Press Syndicate of the University of Cambridge, chp. 8, page 300-312. (2004).

• Phillips RE, Pasvol G (1992): Anaemia of Plasmodium falciparum malaria. Baillierers Clinical Haematology