Nigerian Journal of Paediatrics

6Prevalence of rotavirus among children under five years of age with diarrhea in Kaduna State Nigeria

Niger J Paediatr 2016; 43 (4): 264 – 268

ORIGINAL

Mohammed AA

Prevalence of rotavirus among

Aminu M

Ado SA

children under five years of age

Jatau ED

with diarrhea in Kaduna State,

Esona MD

Nigeria

DOI:http://dx.doi.org/10.4314/njp.v43i4.6

Accepted: 16th August 2016

Abstract : Rotavirus (RV) is a

(33.0%:63/111) compared to fe-

major etiological agent of acute

males (31.4%:66/210). The highest

Mohammed AA (

)

infantile gastroenteritis and is

burden was detected in children 25

Aminu M, Ado SA

associated with 20%-25% of diar-

-36 months of age (37.3%:22/59).

Jatau ED, Esona MD

Department of Applied Science,

rhea cases in infants. Nigeria con-

Highest prevalence was detected in

College of Science and Technology

tinues to be among the first five

children whose parents had pri-

Kaduna Polytechnic, Kaduna State,

countries with greatest number of

mary education (35.8%:19/53) and

Nigeria

RV disease associated deaths per

those whose parents were civil

Email: aisha2zra@yahoo.com

year. The objective was to deter-

servants (35.6%:36/101). There

mine some demographic factors

was no statistically significant as-

that might be associated with rota-

sociation between breast feeding

virus diarrhea among children in

and RV infection (P> 0.05). The

Kaduna State. From September

study has revealed that rotavirus

2013-August 2014, 401 diarrheic

remains an important cause of

stool samples were collected from

acute diarrhea in children under

children under 5 years of age in

five years in Kaduna State, Nige-

Kaduna State, Nigeria and ana-

ria. Hence the need to introduce

lyzed for RV antigen using

the vaccines into the childhood

ELISA. An overall RV prevalence

immunization program in the

of 32.2% (129/401) was obtained

country

with

the infection

occurring

throughout the study period. The

Keywords : Prevalence, Rotavirus,

infection was higher in males

Children, Kaduna State, Nigeria

Introduction

greatest number of RV disease associated deaths per

year. Estimates attribute up to 33,000 deaths annually to

RV disease in Nigerian children less than 5 years old.

7,9

Diarrheal disease kills 1.8 million children under five

years of age yearly. It is the second leading cause of

Prevalence of 11.0%-56% have been reported in Nige-

ria.

5,12,13,14, 15,16,17,18,19,20

death, and accounted for 9.9% of the 6.9 million deaths

in this age group in 2011. In Nigeria it is encountered

both in urban and rural areas. It is estimated that 1.3

The introduction of two effective RV vaccines (Rota

billion episodes and 4 million deaths occur each year in

Teq and Rotarix), licensed in 2006 and have been rec-

children under five years old with about 80% of deaths

ommended for use in all countries by WHO, particularly

occurring in the first two years of life.

in those countries with high diarrhea-related mortality in

children younger than 5 years. Substantial declines in

Rotavirus (RV) gastroenteritis is a mild to severe dis-

ease, with incubation period of about 1-2 days. The

morbidity and mortality attributable to RV and all-cause

symptoms often starts with fever, nausea, and vomiting,

diarrhea have been recorded in high-income and middle-

followed by abdominal cramps and frequent watery diar-

income countries that have introduced RV vaccines so

far. Therefore, there is the need to introduce RV vac-

rhea, which may last for 3-8 days. Infected children may

also have a cough and runny nose.

5,6

Rotavirus infection

cine into the National Immunization Program.

is more frequent in Africa, especially West Africa in-

cluding Nigeria in the cooler, drier winter months.

7,8

Rotavirus infection is not routinely diagnosed in most

Rotavirus is a major etiological agent for acute infantile

Nigerian hospitals probably due to the cost of its diagno-

gastroenteritis and is associated with 20%-25% of diar-

sis and because clinical spectrum of signs and symptoms

rhea cases in infants.

9,10

The number of deaths caused

are similar to other gastroenteritis. There is the need for

yearly by rotavirus has been estimated to be 453,000 in

regular detection of RV strains, because this information

children less than five years old annually worldwide.

is needed to interpret the results of vaccine studies and

Nigeria continues to be among the 10 countries with

epidemiologic surveillance.

265

Rotavirus surveillance has been going on in Nigeria

Specimen Preparation

since 2010 at the Institute of Child Health,

University of Nigeria Teaching Hospital, Enugu.

Exactly 1 ml of sample diluent was added to properly

The aim of the study was therefore to determine the

marked tube using a pipette. For solid stool, the sample

prevalence of Rotavirus associated diarrhea among chil-

was pressed into transfer pipette to the first mark. For

dren under 5 years with diarrhea in Kaduna state

liquid stool, samples were aspirated into transfer pipette

Nigeria.

to the first mark. Samples were re-suspended in 1 ml of

sample diluents. This makes a 10% fecal suspension.

Detection of Human Rotavirus

Materials and Methods

Study Area

Each 10% fecal suspension was screened for the pres-

ence of rotavirus antigens using commercially available

The study was carried out in Kaduna state, Nigeria. The

enzyme immunoassay (EIA) kit (Premier Rotaclone

state has a total number of 23 Local Government Areas

Meridian Bioscience, Inc. USA). All assays were per-

(LGAs) and three senatorial districts; that include south,

formed according to the manufacturers’ instructions.

north and central senatorial zones. Six of the LGAs were

The samples, microtiter wells and reagents were brought

selected by simple random sampling for this research.

to room temperature before the test was carried out.

These LGAs include Kachia and Kagarko (south); Chi-

Wells for samples and controls were inserted into the

kun and Giwa (central) and Soba and Sabon gari (north).

microtiter well holder. About 2 drops (100µl) each of

The health care facilities selected were Primary Health

diluted fecal sample, positive control and negative con-

Care Unit (PHCU) Ladduga for Kachia LGA, PHCU

trol (sample diluents) was added to the bottom of sepa-

Buruku for Chikun LGA, PHCU Maigana for Soba

rate wells. About 2 drops (100µl) of enzyme conjugate

LGA, General Hospital Kagarko for Kagarko LGA,

was added to each well, mixed by gently swirling on

PHCU Gangara for Giwa LGA and Major Abdullahi

tabletop and was incubated at room temperature for 60 ±

Memorial Hospital Sabon gari for Sabon gari LGA.

5 minutes. After incubation the liquid was poured out of

the wells into a discard vessel, and the microtiter well

Study Design

holder was tapped upside down vigorously against ab-

sorbent paper to ensure complete removal of liquid from

The research was a descriptive cross sectional study, and

the wells. All the wells were filled to overflow with

was conducted in a hospital or PHCU in each of the

washing buffer and the liquid was poured out. The mi-

LGA. Children under 5 years of age were studied. A

crotiter well holder was tapped upside down vigorously

diarrhea case was defined as a child passing 3 or more

against absorbent paper to ensure complete removal of

loose, liquid, watery stool in a 24 hour period.

liquid from the wells. The washing procedure was re-

peated two more times for a total of three washes.

Ethical Approval

About 2 drops (100µl) of substrate A solution was added

to each well, and 2 drops (100µl) of substrate B solution

Ethical clearance was obtained from the ethical commit-

was added to each well, and it was incubated for 10 min-

tee of Kaduna state Ministry of Health.

utes at room temperature. Visual determination was

made after 10 minutes incubation. Samples with blue

Sample Size

color greater than negative control are taken to be posi-

tive, while samples showing equal or less color than

A prevalence rate of 36.5% reported in a previous study

negative control are taken to be negative. Spectropho-

by Wada-Kura

was used to calculate the sample size

tometric determination was done by adding 2 drops

using the equation by Sarmukaddam and Garad.

The

(100µl) of stop solution (Sulphuric acid) to each well

calculated sample size was 356.15. However to have a

after the incubation, and the absorbance of each well

good representation of the target population and to in-

was read at 450nm against an air blank within 60 min-

crease the chances of having positive samples, a total of

utes. Specimens with absorbance units (A 450 ) greater

401 diarrheic stool samples were used for the study.

than 0.150 were considered positive, while Specimens

with absorbance equal to or less than 0.150 are consid-

Sample Collection

ered negative.

A total of 401 stool samples were collected from chil-

Analysis of Results

dren 0-5 years of age across the six selected LGAs.

About 5ml of fecal sample was scooped with a wooden

Data obtained from the questionnaire and the EIA was

spatula or decanted respectively into clean, labeled

analyzed using statistical package for the social sciences

screw capped tubes with the assistance of the laboratory

(SPSS) version 21. Chi-square and odds ratio was used

technologist. All samples were transported in ice box to

as test of association and risk respectively at 95% confi-

the Department of Microbiology, Faculty of Science,

dence interval with p ≤0.05 taken as statistically signifi-

Ahmadu Bello University, Zaria and stored frozen at -

cant.

20 C until analyzed.

266

Results

The result of the distribution of rotavirus with respect to

mode of feeding is shown in Table 3. There was no

Out of the 401 fecal samples screened for the presence

statistically significant difference between breast feeding

( χ =3.124,df=1,

of human rotavirus in children, 32.2% (129/401) were

and

the prevalence of rotavirus

positive for rotavirus antigens (Figure 1).

p=0.562).

The distribution of rotavirus infection in children with

respect to location is shown in Table 1. There was statis-

Fig 2: Monthly

tically significant difference between prevalence of rota-

Distribution of

virus infection and location ( χ =13.651, df=5 p=0.031).

Rotavirus Among

Children in Kaduna

Table 1: Distribution of Rotavirus among Children in Kaduna

State, Nigeria

Location (LGA) No examined No positive (%) No negative (%)

Chikun

21(36.2)

37(63.8)

Giwa

131

36(27.5)

95(72.5)

Kachia

25(61.0)

16(39.0)

Kagarko

14(22.6)

48(77.4)

Table 3: Prevalence of Rotavirus in Children with Respect to

Sabon gari

23(27.7)

60(72.3)

Breast Feeding and Introduction of Other Food

Soba

10(38.5)

16(61.5)

Mode of feeding Tested Positive Negative OR95% CI on ORp-value

Total

401

129(32.2)

272(67.8)

No (%) No (%)

Breast feeding

( χ =13.651, df=5, p= 0.031)

Yes 198

72(36.4) 126(63.6) 2.453 2. 500-2.732 0.562

203

57(28.1) 146(71.9)

Introduction of other food

Fig 1: Prevalence

Yes 328

100(30.5) 228(69.5) 3.1322.9 83-3.365 0.025

of Rotavirus among

29(39.7) 44(60.3)

Children in Kaduna

State, Nigeria

Discussion

The monthly distribution of rotavirus infection in chil-

This study shows that rotavirus remains an important

dren in parts of Kaduna State was determined as shown

cause of infantile diarrhea in children. Rotavirus antigen

in Figure 2. The highest prevalence was recorded in

was detected in children 0-5years in parts of Kaduna

March (75%:3/4), and least in July (14.0%:3/21)

state with a prevalence of 32.2% in this study. The result

( χ =30.661, df=11, p=0.002).

is similar to the 36.5% recorded in Kano Northern Nige-

ria

and 35.0% recorded in Lagos Southern Nigeria.

The result was analyzed according to some demographic

However the prevalence is higher than the 15.6% and

factors, and the result is shown in Table 2.There was no

previously reported in Zaria, the 11.0%

23.8%

statistically significant difference between sex, age and

and

13.8% reported in Jos, 18.0% in North-western Nige-

parents’ educational level and the presence of rotavirus

ria, 19.2% in Benin City and 56.0% in Enugu.

infection.

other African Countries, similar prevalence of 36.0%,

28.9% and 32.8% have been reported in Madagascar,

Table 2: Prevalence of Human Rotavirus in Relation to Demo-

Cote d’voire and Malawi respectively. In other parts

graphic Factors in Children in Parts of Kaduna State

Parameter Examined

Positive

Negative

p-value

of the world, prevalence of 31.0% and 34.4% have been

reported in Bangladesh and Italy

No (%)

Age group

(months)

The result however contradicts higher values of 57.4%

0-12

101

34(33.7)

67(66.3)

reported in Oman, 39.8% in Ghana, 45.5% in Ugan-

13-24

285

60(32.4)

125(67.6)

da and 61.0% in Indonesia. Lower values of 14.0%,

25-36

22(37.3)

37(62.7)

0.039

[33]

26.4% and 26.2%

37-48

290

9(31.1)

20(68.9)

have been recorded in Namibia,

40-60

4(14.8)

23(85.2)

South Africa and Tunisia respectively.

Sex

Male

111

63(33.0)

128(67.0)

This observed difference in prevalence could be due to

Female

210

66(31.4)

144(68.6)

1.002

the method used, time and season of sample collection,

Parents’ educational level

sample storage, geographical location or environmental

Informal

170

51(30.0)

119(70.0)

factors. The hospital-based WHO global networks for

Primary

(35.8)

34(64.2)

Secondary 95

33(34.7)

62(65.3)

0.635

surveillance of rotavirus diarrhea report estimated the

Tertiary

26(31.3)

57(68.7)

rotavirus rate to range from 39-52% in the African re-

gion.

Parents’ occupation

The 32.2% obtained in this study, though high,

Self-employed 288

90(31.3)

98(68.7)

did not fall in this WHO African region range probably

Civil servant 101

36(35.6)

65(64.4)

0.043

because the sample were collected mainly from primary

Unemployed 12

3(25.0)

9(75.0)

health care units and not the main hospitals where seri-

ous cases are referred or taken to.

267

Rotavirus was detected in children throughout the study

cant associated with the prevalence of rotavirus, this

agrees with the findings of Aminu et al. and Junaid et

period occurring with a slightly higher rate during the

al . Children whose parents had primary education had

dry season. This agrees with the observation of previous

studies

7,8,10,15,23,25,34

who found the circulation of rotavi-

the highest prevalence while those whose parents had

rus lower during the wet season with slightly increased

informal education had the least.

seasonal peaks during the cooler, dryer months from

October to March. The higher prevalence of rotavirus

Children whose parents were unemployed had the least

infection in the dry season may be attributed to geo-

prevalence and civil servants had highest prevalence of

graphical and environmental factors with low relative

rotavirus, there exist statistically significant association.

humidity being the most important environmental fac-

This observation, could be due to chance and not a cer-

tor.

tainty, and this implies that rotavirus infect children re-

Rotavirus was recorded in all age groups in this study.

gardless of parents demographic characteristics.

Although highest prevalence was recorded among chil-

Breast feeding was not associated with the prevalence of

dren in age group 25-36 months, there was statistically

rotavirus in children. This contradicts the findings of

Aminu et al.

significant difference between age and the prevalence of

who reported that breast feeding confer

rotavirus. This agrees with the findings of Wada-Kura

some protection against rotavirus infection. In this

who reported higher prevalence in children between 41-

study, children that were breast feed, and did not start

50 months in Kano and Coluchi et al . who recorded

any food had higher prevalence; which could be due to

higher prevalence in children within age group 24-35

the fact that at this age, children are more vulnerable to

months in Paraguay. The result however, contradict the

rotavirus infection. Breast feeding may only be protec-

report of Junaid et al . and Aminu et al .

in studies

tive if it is practiced with intensity and frequency that

conducted in Jos and Zaria respectively, who reported

allows continuous high level protection of the intestinal

mucosa rather than sporadic or low volume feeds.

that the detection of rotavirus infection is more in chil-

dren under the age of two years. The higher prevalence

recorded in children among this age group (25-36

months) in this study could be due to behavioral activi-

ties of children at this age, who tend to play outside with

Conclusion

possibly fecally contaminated materials. Least preva-

lence was recorded in children above 40 months of age.

The study has revealed that rotavirus remains an impor-

This could be due to the fact that older children tend to

tant cause of acute diarrhea in children under five years

become protected from severe form of rotavirus infec-

in Kaduna. There is the need for the introduction of the

tion as a result of protection acquired from multiple re-

licensed rotavirus vaccine in the study area as a means

infection.

of preventing rotavirus infection.

Sex was found not to be associated with rotavirus diar-

rhea in children, this agrees with previous findings con-

Limitations

ducted in Nigeria.

12,13,15,16,34

Even though in this study,

males had a slightly higher prevalence than females.

VP6 subgroup specificity could not be detected because

This difference could be due to chance because at this

of lack of monoclonal antibodies specific for VP6.

age there is no difference in life styles between the boy

and girl child. The result contradicts the finding of Jun-

aid et al . in Jos, who reported statistically significant

Conflict of interest: None

association between sex and rotavirus infection.

Funding: None

Educational level of parents was not statistically signifi-

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