Archives of Clinical Microbiology

Keywords

Health assessment; School children; Flood affected areas; Ogbaru Local Government Anambra State

Introduction

Floods are the most common natural disaster in both developed and developing countries, and they are occasionally of devastating impact. Their impacts on health vary between populations for reasons relating to population vulnerability and type of flood event. There is potential for increased faecal-oral transmission of disease, especially in areas where the population does not have access to clean water and sanitation. Such diseases include, nonspecific diarrhoea [1], poliomyelitis, rotavirus, typhoid and paratyphoid, cholera, dysentery [2].

Intestinal parasites account for one of the major faecal orally transmitted diseases which are public health problem worldwide. It is estimated that 3.5 billion people are infected by intestinal parasites and 350 million fall ill due to these organisms [3]. At highest risk of morbidity are pre-school, school-age children and pregnant women [4]. Ascaris lumbricoides, Trichuris trichiura, the blood-feeding hookworms Ancylostoma duodenale and Necator americanus, which cause ankylostomiasis and Strongyloidies stercoralis is estimated to have infected 1.2 billion, 800 million, and 740 million people respectively worldwide [5]. Worldwide, 320 million school-age children are infected with A. lumbricoides [6]. The high incidence of intestinal parasite has been attributed to a number of factors from unhygienic habits, contaminated food and water to poor sanitary conditions [7].

It is an important cause of morbidity in school age children who harbor the highest intensity of worm infestation, living in conditions of poor sanitation, and their impact on morbidity and mortality is more severe in malnourished populations [8]. Also 68.2% prevalence rate of intestinal helminthes from stool samples of children 0-17 years was reported from Ibadan [9]. Ova of helminths can also be isolated from the underneath of fingernails [4] and on the surface of Nigerian currency [10].

Anaemia is a major health problem worldwide. According to the World Health Organisation two billion people suffer from anaemia in the world [11] and common among children as a result of nutrition deficiencies, parasitic infections and haemoglobinopathies [12]. Iron deficiency anaemia accounts for most of the anaemia that occurs due to parasitic infections. In the tropics, parasitic infections such as hookworm, schistosomiasis and malaria are predictors for anaemia [12-14]. The contribution of single parasitic infections on the risk of anaemia in schoolchildren is well known, with risk correlated with infection intensity [15].

With the recent flooding, the people of Ogbaru local government area of Anambra state are faced with so many challenges such as poor sanitary system; overflowing of toilets, poor drainage etc. These have affected their lives negatively. This study tends to find out the prevalence of intestinal parasite, malaria parasite and anaemia amongst the children in the environment given these recent challenges (Tables 1-4).

Protozoa helminths
School Location	No Examined	No of Infected	A lumbricoidies (%)	T tricuria (%)	Hookworm (%)	H.nana (%)	E.vermicularis (%)	Taenia (%)	E.histo/disp (%)	G.lamblia (%)	Total
ODEKPE,	64	36(56.25)	22(6.5)	4(1.17)	9(2.65)	1(0.29)	0	0	9(2.65)	9(2.65)	54(15.93)
OHITA	32	21(65.62)	7(2.10)	3(0.88)	1(0.29)	2(0.59)	0	5(1.47)	8(2.35)	8(2.35)	34(10.02)
OSSOMALA	64	38(59.38)	10(2.95)	4(1.17)	3(0.88)	4(1.17)	4(1.17)	4(1.17)	8(2.35)	15(4.42)	52(15.34)
AKILIOZIZO	64	33(51.56)	13(3.83)	0()	4(1.17)	0	3(0.88)	2(0.59)	9(2.65)	9(2.65)	40(11.79)
UMUNAKWO	32	15(46.88)	5(1.47)	1(0.29)	1(0.29)	0	1(0.29)	0	4(1.17)	4(1.17)	16(4.72)
OGBAKUBA	64	34(53.13)	12(3.54)	0()	1(0.29)	0	2(0.59)	6(1.76)	13(3.83)	7(2.10)	41(12.09)
OKPOKO	64	29(45.31)	8(2.35)	2(0.59)	2(0.59)	0	0	10(2.95)	9(2.65)	10(2.95)	41(12.09)
OCHUCHE	32	15(46.88)	5(1.47)	1(0.29)	1(0.29)	0	0	1(0.29)	2(0.59)	6(1.76)	16(4.72)
AMIYI	64	31(48.44)	17(5.01)	1(0.29)	3(0.88)	1(0.29)	1(0.29)	5(1.47)	13(3.83)	4(1.17)	45(13.27)
		252(52.50)	99(29.20)	16(4.72)	25(7.37)	8(2.35)	11(3.24)	33(9.73)	75(22.12)	72(21.24)
TOTAL	480										339(100.0)

Table 1: Prevalence of intestinal parasites from school children in ogbaru lga.

Intestinal parasites	Male 250(52.08)	Female 230(47.92)	Total 480
A lumbricoidies (%)	50(20.00)	49(21.30)	99
T tricuria (%)	6(2.40)	10(4.35)	16
Hookworm (%)	10(4.00)	15(6.52)	25
H.nana (%)	3(1.20)	8(3.48)	11
E.vermicularis(%)	4(1.60)	4(17.39)	8
Taenia (%)	15(6.00)	18(7.82)	33
E.histo/disp (%)	39(15.60)	36(15.65	75
G.lamblia(%)	36(14.4)	36(15.65)	72
Total	163(48.08)	176(51.29)	339

Table 2: Sex distribution of the intestinal parasites among the school children.

Variable	Frequency	Prevalence (%)	P value
Sex
Male	250	48.81	0.289
Female	230	51.19
No of people in the family
6-Apr	306	50.65
9-Jul	127	48.81	0
>9	47	74.47
Parent occupation
Farmer	296	58.12
Trader	35	51.42	0
Civil servant	71	33.8
Other	78	48.72
Source of water
Borehole	379	49.34	0
Others sources	101	64.35
Previous treatment
De-wormed	172	45.34	0.01
Not de-wormed	308	56.49
Refugee camp during flood
Stayed at camp	106	47.17	0
Off camp	374	54.01
Defecation site
Bush	128	49.21
Toilet	352	53.69	0
Behavioral pattern
Bite finger nails	314	50
Does not Bite finger nails	166	57.23	0.891
Walk bare foot	237	54.43
Does not walk bare foot	243	50.62	0.891
Geophilia
Yes	189	53.97	0
No	291	51.55

Table 3: Social and demographic factors of school children.

School location	No examined	No Infected (%)	Infected male (%)	Infected female (%)
Odekpe	64	56(87.5)	37(57.81)	19(29.68)
Ohita	32	29(90.63)	16(50.0)	13(40.63)
Ossomala	64	58(90.63)	26(40.63)	32(50.0)
Akiliozizo	64	59(92.18)	33(51.56)	26(40.63)
Umunankwo	32	29(90.63)	14(43.75)	15(46.88)
Ogbakuba	64	58(90.63)	30(46.88)	28(43.75)
Okpoko	64	48(75.0)	19(29.69)	29(45.31)
Ochuche	32	23(71.88)	14(43.75)	9(28.13)
Amiyi	64	55(85.94)	31(48.44)	24(37.5)
Grand Total	480	415(86.46)	220(45.83)	195(40.63)

Table 4: Prevalence of malaria parasite among school children from different towns visited.

Aims

This study is designated to determine the prevalence of malaria parasite, intestinal parasites and anaemia among primary school children in Ogbaru LGA, ascertain the potability of their drinking water and determine the prevalence of pathogenic fungi in their classrooms.

Materials and Methods

Study area

The study was done in Ogbaru Local Government Area in Anambra State south-eastern Nigeria.

Study population

The study population was primary school children from nine towns in Ogbaru LGA who were present during the study period. Not more than two primary schools from each community within Ogbaru LGA were selected with at least 32 children all within 6-17 years that fulfilled the inclusion criteria, in all 480 children.

Sampling technique

Pupils were selected randomly after volunteering based on age and sex. Age range of 6-17 from both sex consisted the number of children from primary [1-6].

Ethical considerations

Approval was sort from Anambra State Universal Basic Education Board (ASUBEB), The consent of Ogbaru Local Government Educational Secretary, school authorities, and ethical clearance from the Faculty of Health Science and Technology, Nnamdi Azikiwe University, Nnewi campus was also obtained. Verbal and written informed consent was sought from all eligible individuals and from their parents.

The investigation was carried out at the Department of Medical Laboratory Science, Nnamdi Azikiwe University Nnewi Campus (Tables 5-7).

Age range	No. of children tested (%)	Intestinal parasites (%)	Anaemia (%)	Malaria parasites (%)
6-9 (YEARS)	118(23.96)	62(52.54)	66(55.93)	106(89.83)
10-13 (YEARS)	209(43.96)	118(56.45)	119(57.77)	178(85.17)
> 13 (YEARS)	153(32.08)	75(49.02)	80(52.29)	131(85.62)
TOTAL	480(100.0)	252(52.50)	265(55.21)	415(86.46)

Table 5: The prevalence of malaria parasites, intestinal parasites and anaemia on school children in ogbaru lga by age range.

School location	No. of children	PCV (%)	HB(g/dl)
ODEKPE	64	38(59.38)	35(54.69)
OHITA	32	17(53.13)	15(46.88)
OSSOMALA	64	36(56.25)	32(50)
AKILIOZIZO	64	44(68.75)	34(53.13)
UMUNAKWO	32	21(65.63)	17(53.13)
OGBAKUBA	64	33(51.56)	31(48.44)
OKPOKO	64	43(67.19)	40(62.5)
OCHUCHE	32	22(68.75)	18(56.25)
AMIYI	64	55(85.94)	43(67.19)
GRAND TOTAL	480	309(64.38)	265(55.21)

Reference value HB-11-16g/dl  PVC 34-35%

Table 6: PCV and hb of children lower than the accepted reference value from various towns in ogbaru lga.

Malaria parasite	No of children with anaemia	Mean ± std	p-value	mean ± std	p-value
Level of parasitemia  (+)		hb (g/dl)		pcv (%)
Level of parasitemia  (++)	93	9.95 ± 0.76	0.156	30.03 ± 2.54	0.017
No parasite seen
Total	265	10.02 ± 0.73		30.50 ± 2.63

+= 1-10 Malaria parasite per 100 high power field.
++= 10-100 Malaria parasite per 100 high power field.

Table 7: Effects of malaria parasitaemia on haemoglobin concentration and packed cell volume.

Stool Sample Examination

Macroscopic examination

All specimen were examined to detect the presence of adult worms, or segment, the consistency, colour, presence of mucus and blood were also noted. Ethyl acetate concentration technique as described by Cheesbrough [16] was used to examine the stool for the presence of larvae, cysts or ova of parasites. Using a stick, about 1g from each of the faecal specimen was emulsified in 4 ml of 10% formol water. Another 3 ml of 10% formal water was added. The emulsified faeces were sieved into a beaker. The suspension was later transferred into a glass centrifuge tube and about 4 ml of ethyl acetate was added. The tube was stoppered and shaken vigorously for 1munite and then centrifuged at 3,000 rpm for 1munite. The layer of faecal debris was loosened from side of the tube using an applicator stick and the supernatant poured away. The deposit was re-suspended by tapping the bottom of the tube with finger. The deposit was transferred to a slide using a Pasteur pipette. The slide was covered with a cover slip and examined under x10 and x40 objectives of the microscope. The ova/larvae of parasites were identified with reference to Atlas of Parasitology [17].

Malaria parasite test

Thick and thin blood smears were prepared and stained with giemsa stain and examined according to the procedure described by Cheesbrough [16].

Haematological test

Estimation of Packed cell volume (PCV) using haematocrit packed cell volume capillary method.

Estimation of Haemoglobin Concentration by Cyanmethaemoglobin Method

Culture media

CHROMagar liquid ECC, KF Streptococcus Agar and Reinforced Clostridium agar were prepared according to manufacturers’ instruction.

Culture

KF Streptococcus agar was prepared based on formulation described by Kennor et al. [18]. The filtered sample on the membrane filter was transferred directly on the agar medium avoiding the formation of bubbles. The plates were inverted and incubated at 37°C for 48hours. After 48hours the red and pink colonies were counted and reported as faecal streptococci per 100ml. To count the spores of sulphite-reducing Clostridia, the volume (100 ml) of the water Sample was heated to 60 ± °C and the whole volume maintained at this temperature for 15 minutes. Cooked meat medium were prepared according to manufactures instruction and the samples were inoculated into the medium for enrichment and incubated for 24hours at 37°C. Prior to filtration, reinforced clostridium differential agar was prepared and allowed to solidify. The samples were filtered and the filter membrane placed on the Petri dishes containing the medium and incubated at 37°C in an anaerobic jar containing gas-pak for 24 hours [19].

After incubation, all black or grey colonies were counted an expressed as number of colonies per 100 ml.

The colonies were sub-cultured on blood agar and incubated anaerobically for 24 hours according to method described by Chessbrough [16]. The colonies showed marked haemolysis on blood agar after 24hours.

Gram stain as described by Chessbrough [16] and spore staining using Schaeffer-Fulton Method [20] was carried out to identify the organism. Gram-positive organisms-purple, Gram-negative organisms-pink. Organism was large gram positive rods.

Statistical Analysis

The data collected were analysed using the statistical package for the social sciences-SPSS software version 16.0, ANOVA, ttest and Frequency distribution. Level of significance was set at 95% confidence interval, p<0.05 was considered statistically significant.

Discussion

In this study, a high prevalence of intestinal parasites (52.50%) was obtained. The children were infected with at least one species of these parasite; Ascaris lumbricoides, Entamoeba histolytica/ dispar, Giardia Lamblia, Taenia spp, Hook worm, Trichuris trichura, Enterobius vermicularis Hymenolepis nana. This prevalence is comparable with the rate reported in similar studies in Nigeria by Banke et al. [21] 54.13%, Awolaju and Morenikeji [22] 48.4%. However it is above the rate of 25.8% recorded by Akingbade et al. [23] among children with diarrhea in Abeokuta, Ogun State, Odu et al. [24] recorded 15.7% prevalence among primary school children in Rivers State. The higher prevalence and highest occurring intestinal parasite reported for Ascaris lumbricoides (29.20%) in this study is lower than that of Akingbade et al. [23] 54.8%, Odu et al. [24] 57.4%, Omorodion et al. [25] 47.7% and 25% in Delta and Edo respectively. Hymenolepis nana (2.35%) was the least occurring intestinal parasite. This is in agreement with the report of Emmy-Egbe et al. [26] with prevalence of 2.6%. Intestinal parasitic infections are mainly acquired through faecal-oral route. High prevalence of intestinal parasite had been attributed to poor environmental conditions; shortage of good water supply, indiscriminate defecation and poor personal hygiene [26] which was also observed in this study and most of the children have never been dewormed. Entamoeba histolytica was the second most prevalent parasite amongst the children (22.1%). Banke et al. [21] observed a prevalence of 7.06%, Akingbade et al. [23] 19.4%. Infection of human by E. histolytica commonly results in amoebiasis [27]. E. histolytica is indicator organism of faecal contamination of food or water and are frequently present in unhygiencally prepared foods and can lead to outbreaks of ameobiasis. The higher prevalence and incidence of Entaemoeba histolytica and Giardia lamblia are a reflection of the poor environmental sanitation and poor personal hygiene and unclean habits practiced by these children. Ignorance and illiteracy of the parents can also be attributed to these infections due to their lack of information on the mode of transmission and symptoms of intestinal parasite infection. Survey in Nigeria, by Okonko et al. [27] indicated that intestinal parasitic infections such as amoebiasis are growing problem in the country.

Most of the pupils especially those from primary schools in Odekpe, okpoko and Ossomala had mixed infection of Ascaris lumbricoides, Entamoeba histolytica/dispar, Giardia Lamblia or Hookworm. Multiple parasitisms have also been reported by Chukwuma et al. [28].

Parents occupational prevalence showed that infection rate was highest among the children whose parents were farmers (58.12%), followed by the pupils whose parents where traders (51.42%) while the pupils whose parents where civil servants showed the least prevalence (33.80%) of infection with intestinal parasites. Difference in parents occupation prevalence was found to be statistically significant (p<0.05).

Afzal and Sultan [29] reported a malaria prevalence of 75% in a flood affected area of Muzaffargarh District, Southern Punjab, Pakistan. Children in the rural area are less protected and are more prone to mosquito bite [30]. Brown [31] stated that in hyperendemic areas, the malaria is mild and asymptomatic in older children. Age and nutritional status of the host might represent natural or acquired resistance and can play a role in the severity of the disease produced [30].

From this study, most of the children were anaemic, therefore anaemia is of public health importance in Ogbaru LGA especially among the school children. The rate (55.21%) recorded in this study is higher than the prevalence rate (38.6%) recorded in Edo state by Osazuwa and Oguntade [32]. The highest prevalence of anaemia (57.77%) was seen within age 10-13 followed by age 6-9 years (55.93%) and the least in age 14-17 years (52.29%).

Poor nutrition [32], parasitic infections [33] and other factors have also been noted to increase the incidence of anaemia.

The effect of malaria parasitaemia on anaemia using the mean and standard deviation of Hb and PCV of the children with malaria parasite showed no significant difference between malaria parasitaemia and Hb concentration (p>0.05), however there was a significant difference between malaria parasitaemia and PCV (p<0.05). Okafor et al. [34] and Cheesbrough [16] reported that anaemia as the commonest complication of malaria among 1-5 age groups. This study shows that anaemia found in some of the children could be due to other causes other than malaria.

Conclusion

The prevalence rate of intestinal parasite, malaria and anaemia in this study indicates that they are important public health problems in Ogbaru Local Government Area. There should be collective effort engineered towards prevention from infection and treatment of already established cases.

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