GLOBAL JOURNAL OF MEDICAL SCIENCES VOL. 6, NO. 1, 2006: 1 5 COPYRIGHT (C) BACHUDO SCIENCE CO. LTO. PRINTER INNIGERIA. ISSN 1596-2911 RELATIONSHIP BETWEEN ACID AND ALKALINE PHOSPHATASE ACTIVITIES IN PARASITIC INFECTIONS C. E. J. UDIONG, U. S. EBONG, (Received 21 February, 2006; Revision Accepted 5 April, 2006) TOH ABSTRACT Serum acid and Alkaline Phosphatase levels of 80 subjects diagnosed of helminthic or protozoal infections were assayed (excluding pregnant women and children) while 64 non-infected age and sex-matched subjects served as control. Infected subjects had a significantly higher serum alkaline phosphatase activity (133.9 + 46.6 wl) than the control subjects (87.9 + 15.9 yl). But there was no significant difference between the total acid phosphatase values (7.45 + 1.76 yl) and prostatic acid phosphatase activity (1.04 + 0.31 WL) of infected subjects and total acid phosphatase values (6.60 + 1.84 iL) and prostatic acid phosphatase values (0.90 + 0.29 iL) of the control subjects. Females were more infected than the males. Infected females also had a significantly higher total acid phosphatase activity than males (P<0.5). There was no significant difference between the activities of prostatic acid phosphatase of male infected subjects and those for females subjects (P > 0.05). There was no significant difference between the acid and alkaline phosphatase activities of prostatic acid phosphatase of male infected subjects and those for females subjects (P > 0.05). There was no significant difference between the acid and alkaline phosphatase activities in helminthic and protozoal infection (P > 0.05) and also those infected with Entamoeba histolytica and Plasmodium felciparum had significantly higher alkaline phosphatase activities (P < 0.05) than other parasites. This work has established a high alkaline phosphatase activity among subjects with parasitic infections especially in amoebasis and falciparum malaria KEYWORDS: Acid and Alkaline Phosphatases, helminthic and protozoan infections, Relationship. INTRODUCTION ‘A high concentzation of helminthic parasites leads to anaemia, malnutrition, raised serum alkaline phosphatase and different forms of morbidities (WHO, 1981), These occur mostly in helminthic infections “affecting the liver such a8 hydatid disease, fascioliasis, opisthorchiasis and hepatic schistosomiasis (Mansour, 1982, Mackenjee, ef al, 1984). Serum alkaline phosphatase is also elevated in hepatobiliary disease (Mccomb, 1979) and bone disease associated with increased osteoblastic activity (Singer and Wallach 1991 » Helminthic infection is characterized by Increased eosinophils in peripheral blood, the eosinophilia being the host response to large parasité infections that cannot be phagocytosed. Extracellular kiling ot such parasites evolves to cope with the situation by increase in eosinophil output a reaction that is mediated by complement C3B. The C3B-parasite receptor triggers the release of lysosomes with ‘concomitant liberation of acid hydrolases which include ‘acid phosphatase (Roitt, 1988). Such events may lead to raised activity of acid phosphatase in the plasma of Alkaline phosphatase released is ‘Acid and alkaline phosphatases are enzymes. which are found in the liver, bone, intestine, kidney, placenta, prostate and erythrocytes (Obi and llor, 2002). Infection of these organs/cells by agents such as virus ‘and parasites is an obvious cause of cell membrane rupture and enzyme release into the plasma (Nakamura et al., 1988). In tropical Aftica where large parasitic infections are rampant, the esonophil defence response may provide. additional enzymes to host making interpretation of hast acid and alkaline phosphatase results difficult In a study on pathogenic free living amoeba, Warhust, (1985) reported a raised serum alkaline phosphatase due to the fact that enzymes leak out from damaged cells. In another study on severe falciparum malaria involving the liver, there was hepatic dysfunction resulting in inctéased serum bilirubin with increase in serum alkaline phosphate and serum transaminase concentrations (Bryceson, 1983). This study was designed to assess the influence of parasitic infections on plasma and'serum Alkaline and ‘Acid Phosphatase levels in human subjects. associated with similar response to complement. fl University of Calabar U-$. Ebong, School of Medical laboratory Science, University of Cala alt “Teaching Hospital, Calabar, Nigeria P.C. Inyang-etoh, Department of Medicel Microbiology & Parasitology. University af Calabar, C 9, Nigeria2 ‘Subjects and Methods One hundred and forty-four subjects attending the University of Calabar Teaching Hospital and General hospital, all in Calabar Municipality, Cross River State’ of Nigeria were examined. Eighty of them were found to be positive for parasitic infection after their stool and blood were examined for stool and malaria parasites respectively. The remaining 64 non-infected subjects served as control ‘Assay of acid and alkaline phosphatase was done using the test kit supplied by Randox diagnostic laboratories Ltd (United Kingdom) Stool samples were examined macroscopically for adult worms, blood stains, mucus and consistency according to WHO, (1991). Microscopic examination of the stool was done according to the method described by Siddiqui, (1981) while the Brine floatation concentration method was done according to the method described by Cheesbrough, (1998) For malaria parasite detection, thick and thin blood films were made on each blood sample (Laveran, 1980). Both films were placed on a flat surface of bench to air dry. There after the thin films were fixed in absolute methanol for 5 seconds and allowed to dry. Both the thick and thin fms were then stained in freshly prepared 2% Giemsa stain for 30 minutes (Payne, 1988). At the end of the staining, the films were removed C.E, J. UDIONG, U. S. EBONG, and P. C. INYANG-ETOH from the stain, rinsed in buffered water of PH 7.2 and stood vertically to dry. Both the stained thick and thin films were examined microscopically using the X 100 objective lens with oil immersion. The thin blood films were used for speciation of Plasmodium while thick films were used for detection and quantification of malaria parasitemia. Calculation of malaria parasite density was made by counting the parasite per white blood cells, in thick biood films and parasite density determined by ‘multiplying the figure by 8000 which is an average white blood cell count per microlitre (ul) of blood (Shute, 1986). Data obtained were analysed, using the t-test while the ‘relationship between G-6-PD and malaria density were compared using the Pearson correlation analysis. All analysis were done using the Microsoft Excel analytical tool of XP windows of IBM computer system RESULTS Table 1 shows @ comparism of biochemical parameters between infected and control subjects studied. The mean alkaline phosphatase level was significantly higher in helminthic infection than the control subjects. There was no significant difference between the acid phosphatase activity in infected and control subjects, TABLE 1: COMPARISM OF BIOCHEMICAL PARAMETER STUDIED BETWEEN INFECTED AND CONTROL ‘SUBJECTS Parameter Infected | subjects | (n= 80) | Controi ‘subjects (n= 64) ~~ | Calculated Tp Inference tvalue Alkaline phosphatase wh Total acid | 7.45+ 1.76 phosphatase (ul) | Protatic acid 193.92 466 660+ 1.84 1.044031 0.90 +029 phosphatase (ul Non-prostatic | 7.1521.31 6.11 41.75 | acid phosphatase (uw Table 2 compares the alkaline and acid phosphatase in male and female infected subjects. The toial acid phosphatase activity was significantly higher in infected males than females. But there was no significant 87.9415.9 seca P< 0.05 Significant | 0.724 P>005 Not significant 0.093 P> 0.05 Not significant | 1.793 P<0.05 Significant difference between the alkaline phosphatase activity of male and female subjects examined. There was also no significant difference between the prostatic acid phosphatase and non-prostatic acid phosphatase activities of male and female subjects examined.RELATIONSHIP BETWEEN ACID AND ALKALINE PHOSPHATASE ACTIVITIES IN PARASITIC INFECTIONS. TABLE 2: COMPARISM OF ALKALINE AND ACID PHOSPHATASES IN INFECTED MALES AND FEMALES Parameter | Infected males | Infected te _| females Alkaline j35.3 + 46.9 133.3 + 47.3 phosphatase (wil) Total acid | 8.31 +142 TAZEVTS phosphatase (wi) Prostatic acid | 1.13+0.22 1012034 phosphatase (wil) Non-prostatic 7.15 + 1.31 6.11414.75 aad phosphatase wu) | alculated tvaiue [p oft [P 2.017 Pp 1.106 Pp 1.793 TABLE 3: COMPARISM OF ACID AND ALKALINE PHOSPHATASES ACTIVITIES BETWEEN HELMINTHIC AND PROTOZOAN INFECTIONS ] inference 30.05 | Not significant ~ < 0.06 | Significant > 0,05 | Not significa, P > 0.05 | Not significant 3 Parameter ‘Alkaline phosphatase (ju!) Total acid phosphatase (/) Prostatic acid phosphatase (1/}) Non-prostatic acid phosphatase (ji!) TABLE 4: MEAN LEVEL OF ALKALINE PHOSPHATASE ACCORDING TO PARASITE DETECTED PARASITES — Lirichivra 7.854 1.42 4.014031 6.46 = 1.34 level __ OTs SD for +ve) Entamoeba | 2094.1 histolytica Plasmodium | 147 + 49.6 falciparum Ascaris 197.1 +48.1 lumbricoides Hookworm — | 126.8 + 45.5, Trichuris 1875+ 74.2 Helminthic ‘| Protozoal infection infection 130.3 + 464 149.92 49.6 7.40420 1.04030 6.37 41.94 Alkaline Phosphatase | Alkaline Phosphatase level (Ke 8D for n 120.7 + 36.8 119.34 40.2 192.0 + 46.5 116.3341 132.6 + 45.9 ~ | Calculated tvalue 1408 0.323 ont 0.186 _ | Calculated value jon +e) P > 0.05 | Not significant P>0.08 | Not significant P>0.05 | Not significant P Inference Significant Not significant Not significant Not significant lene | Sionificant — —]4 ‘Comparism of acid and alkaline phosphatase activiies between helminthic and protozoan infection is shown on table 3. There was no significant difference between the acid and alkaline phosphatase activities in helminthic and protozoan infections. Table 4 shows the mean level of alkaline phosphatase according to each parasite detected. The mean alkaline phosphatase activities of those infected with Entamoeba histolytica and Plasmodium falciparum was significantly higher than those of other parasites DISCUSSION This study attempts to assess the levels of acid ‘and alkaline phosphatases in helminthic and protozoa parasite infected patients. There was 10 significant difference in the activities of acid Phosphatase in infected and control subjects. NO significant difference was observed between the value for alkaline phosphatase in male and female infection showing that these infections are not gender bias. There was a significant difference between the total acid phosphatase activity in males and in females, the difference being contributed by the prostate gland of male subjects since the gland is absent in females and this can be compared to the result of Obi and lori (2002). The presence of parasitic infections does not appear to affect prostatic acid phosphatase activity but causes increase in the activity of alkaline phosphatase and non-prostatic acid phosphatase which is responsible for the raised total acid phosphatase activity. Those infected with E. histolytica had raised alkaline phosphatase levels the result which is similar to the work of Warhurst (1985). The higher prevalence of Ascaris lumbricoides compared to hookworm can be attributed to the usual large number of eggs produced by the parasite as reported by Ukoli, (1990) and their ability to resist desiccation. Ascaris lumbricoides eggs ate said to be Viable for years especially in faecally contaminated environment (Hallet al, 1982) Female subjects recorded a higher prevalence Of infection than the male subjects probably because females engage in more farming actvties than the males in this setting especially in areas where human waste serves as manure. Multiple infections with more than one parasite species were recorded by all gender. This is a regular feature among infected population with Ascaris lumbricoides and Plasmodium falciparum jon occurring concurrently with other parasites infections (Booth and Bundy, 1995; Adeyeba and Essiet, 2001). This study has shown that infected subject have significantly high alkaline phosphataes activities and total (non-prostatic) acid phosphatase activities. It has also established that those infected with Entamoeba histolytica and Plasmodium falciparum have significantly high” alkaline phosphatase activities. it has also confirmed that heavy burden of helminthic and protozoan infections can cause raised alkaline and acid phosphatase activities in the host thus complicating the interpretation of phosphatase results of such subjects, C.E, J. UDIONG, U.S. EBONG, and P. C. INYANG-ETOH, REFERENCES: ‘Adeyeba, O. A. and Essiet, U., 2001. Prevalence of Helminth and Protozoal infection among religious sect that walk barefooted in leeyin, Nigeria. The Nigerian Journal of Parasitology 22 (18 2) 85-94. Booth, M. and Bundy, D. A. P., 1995. Estimating the number of multiple-species of geohelminthic infections in human communities. Parasitology , 3: 645-653. Bryceson, A, 1983. Maiatia and splenomegaly. Transactions of the Royal society of Tropical Medicine and Hygiene 77, 6, 828 Cheesbrough, M., 1998. District Laboratory practice in Tropical Countries (Part 1), University press, ‘Cambridge, United Kingdom, 191-235. Hall, A., Anwar, K. S, Tomkins, A. Rahman, L.. 1999. ‘The distribution of Ascaris lumbricoides in human hosts: a Study of 1765 people in Bangladesh. Transaction of the Royal Society of Tropical Medicine and Hygiene 93: 503-610 Lavcran, A., 1980. Note sur un noveau parasite trouve dans le sang du plussiers maladies atteints de fievre palustre. Bull acad Natl. Med. Paris. 9: 12236, Mackenjee, M. K. R., Coovadia, H. M., and Chutte, C. H. J. 1984, Clinical recognition of mild hepatic Schistosomiasis in an endemic area. Transaction of the Royal Society of Tropical Medicine and Hygiene 78: 13-45. Mansour, M. M., 1982. Serum enzymes tests in hepatospleenic Schistosomiasis. Transaction of the Royei Society of Tropical Medicine and Hygiene. Page 76 McComb, R. 8, Bowers, G. N. Jr; Posen, S., 1979. Alkaline’ Phosphatase. New York, Plenum Press. Nakamura, |, Nakamura, K., and Stinson, R. A., 1988 Release of alkaline phosphatase from human ‘osteosarcoma cells by phosphatidyl inositol phospholipase C: Effect of tunicamycin. Arch. Biochem. Biophys 265: 190-196. ©, and lori 0. O., 2002. Effects of cadmium exposure on bone and kidney alkaline phosphatase activity abd acid phosphatase activity in testis and prostate gland in the male rat. Journal of applied science and environmental management, 6(1): 9-13 Obi, F.RELATIONSHIP BETWEEN ACID AND ALKALINE PHOSPHATASE ACTIVITIES IN PARASITIC INFECTIONS. 5 1888. Use and Limitations of Light microscopy for diagnosing malaria atthe primary health cate level. Bulletin of World Health Organization. 86(5) | 621-626. Plussiers maladies atteints de fievre palustre. Bull acad. Natl, Med, Paris. 9:12235 Roitt, |. M., 1988. The basis of immunology ~innate immunity in Essential immunology; 1-14 Oxford, Blackwell. Scientific Publications UK. ‘Shute, G. T., 1986. The microsopic diagnosis of malaria. In: Malaria principle and practice of malariology eds Wemederfer, WHO and McGregor L. ‘Churchill Livingstone Edinburg.pp. 78-814 Adiqui, M.A, 1981. The prevalence of human intestinal parasite in Albaha, Saudi Arat Annals of Tropical__ Medicine and Parasitology 75, 565-566. Singer, F. R and Wallach, S,, 1991. Pagets Disease of Bones Clinical Assessmnet, Recent and Future therapy. New -York, Elsevie. Ukoli, F. M. A., 1990. Introduction to Parasitology in Tropical Africa. Test Flow Ltd, Ibandan, Nigeria 105-108 Warhust, D. C., 1985, Pathogenic free living amoeba. Parasitology Today 1, 124-28 World Health Organization, 1981. Intestinal protozoan and helminthic infection. WHO Technical Report series. Genera. 66. World Health Organization, 1991. Basic laboratory method in Medical Parasitology. WHO Generation 10-24.