ANCYLOSTOMA DUODENALE SEPERATED FROM CONTAMINATED SOIL

SHAHIDA AZHAR ALI1, TANVEER AKHTAR

2, AZHAR MAQBOOL

3, AMMARA HUSSAN

4 &

SAMMAN IKRAM5

1,2,4&5Department of Zoology, University of the Punjab, Quaid-e-Azam Campus, Lahore, Pakistan

3Department of Parasitology, University of Veterinary and Animal Sciences, Lahore, Pakistan

ABSTRACT

Soil samples showed an overall prevalence 1.79 % of Ancylostoma duodenale‟s eggs in six slums of Lahore,

Punjab from Nov 2006 to Oct 2008. Significant prevalence was observed in Missrisha (0.89 %) followed by Bhutto Colony

(0.83 %) , Siddiquia Colony (0.72 %), Khairdin Park (0.53 %), Sheikhupura Road (0.39 %) whereas lowest in Scheme No

2 ( 0.22 %) respectively. Season wise high prevalence was observed in autumn (3.92 %), summer (1.88 %) and winter

(1.29 %) as compared to spring (0.50 %). Month wise highest prevalence (4.83 %) was observed in October followed by

3.67 % in September that gradually decreased to 0.83 % in December and declined to 0.50 % in March respectively.

Demographic data showed 33.47 % people used gurkies / pit, 27 % defecated in open fields, 23 % had very poor sewerage

system and 14.52 % availed satisfactory sewage conditions. Children who had habit of pica (10.50 %), nail biting

(12.51%), sucking thumb (11 %), did not wash hand before meal (12.10 %), after defecation (21.76 %), walked bare footed

(25.51 %) and used junk food (7.54 %) were found respectively.

KEYWORDS: Ancylostomiaisis, Prevalence, Slums, Gurkies, Defecation

Part of Ph.D thesis of first author

INTRODUCTION

Ancylostomiaisis was a widely spread parasitic disease in tropical and subtropical areas (Jiraanankul et al. (2012)

caused by A. duodenale exclusively man parasite, a soil transmitted helminth (STHs) (CDC, 2012), especially in areas with

poor sanitary conditions (Shaikh et al. (2009) ; Ziegelbaueret al. (2012 ). One quarter of the world population i.e. Africa,

Middle East, Eastern Europe and South East Asia (de Silva et al. (2003) were infected with hookworms (Hotez et al.,

2004). out of 122 million, 21 million children of age 1-5 years old were found infected with hookworm, (Kirwan et al.

(2009), 4-15 years of agein India (Norhayati et al. (2003), (52%) hookworm prevalence (Verle et al. (2003) was present in

school going children and decreased with age (Clements et al. (2010) and caused about 22.1 million disabilities annually

in the world (Kirwan et al. (2009). Larvae of A.duodenale survive for several weeks in dirt (WHO, 2011), could also reside

in dogs, cats etc and they acted as paratenic host till they find their way to final host (Brinksworth et al. (2000), it

constituted a significant risk for public health (Kollataj et al. (2012). Ancylostomiaisis was more common in 2-7 years old

children due to habit of pica (Al Mekhlafi et al. (2008) and those who did not wash their hands after defecation and before

meal (Speare et al. (2006) ; Mehmood et al. (2009) ; Khanum et al. (2010). A. duodenale„s eggs from soil could be

transferred onto vegetables (Maipanich et al. (2004) when children eat these contaminated raw vegetables (Ulukanligi et

al. (2001) they might have helminthiasis. In Argentina from city of Resistencia, from 5 public places , 475 soil samples,

17 from kindergarten sandpits and 124 from housing estates were collected, high prevalence of ancylostoma‟s eggs were

observed in 20 % contaminated public park playgrounds with a value of 100%, followed 19.4% found in housing estates

and 11.8% in kindergarten samples (Alonso et al. (2001). Increased urbanization and population density was directly

proportionate to STH infection (Brooker et al. (2006). 288 faecal samples were collected and observed in 2000-2006 from

International Journal of Zoology

and Research (IJZR)

ISSN 2278–8816

Vol.3, Issue 2, Jun 2013, 27-38

© TJPRC Pvt. Ltd.

28 Shahida Azhar Ali, Tanveer Akhtar, Azhar Maqbool, Ammara Hussan & Samman Ikram

contaminated parks; A. duodenale prevalence 20.47% (Rubel and Wisnivesky, 2010) was observed on sidewalks

surrounding the parks. Improvement in socioeconomic status (Rokni, (2009) ; Lin et al. (2010), personal hygiene /

behavior and inadequate sanitation reduced the rate of infection (Ziegelbauer et al. (2012).Many surveys had been

conducted in various parts of Pakistan (Shaikh et al. (2000) Larkana, (Chaudhry et al. (2004) Muzzafarabad, (Kamran et

al. (2005) Karachi and (Khan et al. (2012) in Swat respectively but researchers did not represented relationship among

socioeconomic status (SES), inadequate education, poor sanitation, personal hygiene, family cluster and lack of health care

respectively. Soil contact activity was the basic reason for STHs infection in children.

OBJECTIVE

The study was launched to observe the status of ancylostomiasis in the slums of Lahore to assess the degree of

prevalence (%) in soil samples to improve the health status of children. These studies would be helpful to control

ancylostomiaisis by launching deworming programme at national level with collaboration of veterinarians and health

department.

MATERIAL AND METHOD

Soil samples were collected from the six slums i.e. Bhutto Colony, Siddiquia Colony, Scheme No 2, Sheikhupura

Road, Khairdin Park and Missri Shah of Northern Lahore, Punjab, Pakistan from November 2006 to October 2008 to find

out the prevalence (%) ratio (Map. a and b ). Demographic data was also collected along with social behavior of local

people (Performa 1).

Soil Collection

A total of 25 soil samples from each locality were monthly collected .i.e. from the public parks, fields and

playgrounds where local people defecate i.e. near the filth depots and from the dunghills where stray / wild dogs wander.

Soil was kept in screw caped plastic vial clearly labeled with date and place of collection. These samples were examined

on the same day by Sodium hypochlorite recovery technique (Urquhart et al. (2001) to observe the presence of

helminth‟seggs. The samples not processed on the same day were kept at room temperature.

Sodium Hypochlorite Technique

In a test tube 2 g. of sieved soil sample was placed with 5 ml of 30% sodium hypochlorite (NaOCl) solution. The

tube was shaken intermittently. A few drops of prepared solution was placed on glass slide, placed cover glass and

examined microscopically at 40 magnification, to identify the helminth‟s ovas on the basis of morphology described by

Urquhart et al. (2001).

Statistical Analysis

Collected soil data was entered into Excel Data Sheet version “7” of the computer and transferred to SPSS

Version 15.5, Z-test was applied as it was suitable for analysis, level of significance at the 0.05, 0.01 and 0.001 was

considered as .i.e. *, ** and *** respectively (Steel and Torrie, 1980).The prevalence of the infection / disease was

recorded following the modified formula described as.

RESULTS

3600 soil samples were observed from the 6 slums i.e. Bhutto Colony, Siddiquia Colony, Scheme No 2,

Ancylostoma duodenale Seperated from Contaminated Soil 29

SheikhupuraRoad, Khairdin Park and Missrishah of Northern Lahore, Punjab, Pakistan from Nov 2006 to Oct 2008.

Demographic Status

Out of 3600, children of age 4-10 years old were found with the habit of pica / geophagia (10.50 %), 4-10 years

with nail biting (12.15 %), 5-10 years old sucked thumb (11 %), 3-5 years old did not wash hands before meal (12.10 %)

and 6-11 years old after ( 21.76 %) defecation, 5-13 years old walked bare footed (25.52 %) and 4-11 years old used junk

food (7.54 %) respectively. 35.47% peoples were using gurkies / pit, 27 % defecated in open fields, 23 % had very poor

sewage system and 14.52 % availed satisfactory sanitary system (Table 1; Figure 1a,1b ; Plate 1a, 1b.1c,1d,1e,1f ).

Overall Prevalence

An overall prevalence (%) of A.duodenale’s eggs in the soil was found 1.79 %in Lahore, Punjab from November

2006 to October 2008 (Table 2).

Area Wise Prevalence (%)

Area wise prevalence of geohelminth‟s eggs in soil of six slums of Lahore, Punjab province,Pakistan was

observed, it was highest in Missrishah(0.89 %) followed by Bhutto Colony (0.83 %) Siddiqia Colony (0.72 %) Khairdin

Park (0.53 %) and Sheikhupura Road (0.39 %) whereas lowest in Scheme No 2 (0.22 %) respectively (Table 2; Figure 1c).

Month Wise Prevalence (%)

After analyzing month wise data, highest prevalence was observed 4.83 % in October followed by 3 % in

September that gradually decreased to 0.83 % in December and declined to 0.50 % in March. When statistical analysis (Z-

test) was applied on month wise data, prevalence of the month of the March was the lowest, so it was compared with other

months. There was no significant difference among prevalence (%) of the month January, February, May and July.

Whereas November (P<0.001), August (P<0.001) and Septembers (P< 0.01) showed significant difference with respect to

March (Table2; Figure 1d).

Season Wise Prevalence (%)

Statistically prevalence was significantly higher in autumn (P<0.001), summer (P<0.01) and in winter (P<0.05) as

compared to spring i.e it was observed 3.92 % during autumn followed by 1.88 % in summer, 1.29 % in winter and lowest

0.50 % in spring respectively (Table 2; Figure 1e).

DISCUSSIONS

Soil samples determines the prevalence / infection ratio as eggs deposited in soil by pet / stay animals and man,

hibernate and transmit to person who come in direct contact with that contaminated soil, eggs might be transferred from

soil to vegetables, fruits or being transmitted by insects, animals and man supported by the reports of (WHO, 2011).

Therefore soil contamination constituted a significant zoonotic risk. Unawareness about the transmission and biology of

parasite and inattentively promote ancylostomiasis and these observations are justified with findings of Habluetzel et al.

(2003) and Ejima and Ajugun (2011). Soil acts as a transmitting medium for fertilized eggs of helminth to develop and

mature. Present study showed an overall prevalence (1.79 %) of A. duodenale‟s eggs in the soil less than 14.3 % observed

in China (Uneke et al. (2007), 24.1 % in Lahore, Pakistan (Ashraf et al. (2008) and 53.6 % in Nigeria (Chukwuma et al.

(2009).The conflict existed due to difference in climatic, socioeconomic conditions, personal hygiene, social behavior,

population diversity, density, geographical distribution of helminth. Another reason is increase in soil contamination by the

people who defecate in open fields, sidewalks and on dunghills, throw excreta of animals and man on dunghills, flies, dust

30 Shahida Azhar Ali, Tanveer Akhtar, Azhar Maqbool, Ammara Hussan & Samman Ikram

transmit eggs here and there and ultimately ingested by man and contaminate the topography and playgrounds. Another

reason might be the use of recovery techniques, size, volume of collected data, season of collection, examination and depth

of soil effected prevalence (%) in soil. Similarly it was observed that soil contamination is going to be increased due to the

frequent use of Bhal (the soil taken from the sewerage drain) in lawns to enhance fertility for better growth of plant after

every year. The residents of the posh areas also practiced to add river soil (Kaasuo) in foundations of their houses, in this

way helminth ova along with termite‟s egg were transmitted from one area to another unintentionally. Similarly soil from

other parts of the country was commonly brought by people for construction purposes in old and new housing societies,

indirectly transmitted helminth infection. The children come in contact with soil when play in their lawns or elder sat there,

transmitted ova or infected stage larvae with their shoes inside the houses premises unknowingly. It s therefore suggested

to disinfect the soil before use to avoid ancylostomiaisis and educate people about health care practices.From the present

study infested soil samples with helminth ova were observed from the lawns of the houses, backyards, compoundsand

parks consistent with findings described by Orhun et al. (2009), indicated that direct contact with fur of pets was not

necessary (Zewdu et al. (2011). The outcomes of the present study showed decrease in temperature reduced the rate of

prevalence 3.92 % in autumn, declined to 1.29 % in winter supported by findings in Nigeria (Mabaso et al. (2003) and

increase in age also decreased the prevalence ratio of ancylostomiasis due to the improvement in personal hygiene with

growing age. The common habits of walking barefoot and improper disposal of human faeces were the important features

for high prevalence of helminthes in Pakistan. Present study indicated helminthiaisis had association with SES factors such

as inadequate sewerage, sanitary and education facilities, poor personal hygiene, water supply, social behavior of persons

like nail biting, sucking thumbs, pica, walking bare footed, defecate in open fields, do not wash hands before meal and

after defecation and improper health care facilities supported by the observations of [Khan et al. (2004), Gunawardena et

al. (2005), Speare et al. (2006), Ferguson et al. (2007), Saeed et al. (2008), Shaikh et al. (2009), Mishra et al. (2005),

Muznebin et al. (2007), Al Mekhlafi et al. (2008), Mehmood et al. (2009), Khanum et al. (2010) and Ziegelbauer et al.

(2012)]. Improvement in sewage system, SES, personal hygiene and provision of hygiene and physiology education

minimizes the risk of ancylostomiasis. Similarly poverty played important role to enhance that gusting situation, people

hesitate to spend money on their health.

CONCLUSIONS

Children health was neglected since now and the vulnerable class was of 4-13 years old as

1- They played in parks or playground come in contact with contaminated soil was a risk factor for health.

2- Did not wash their hands before meal and after defecation.

3- Had the habit of nail biting and sucking thumbs too.

5- Always wandered bare footed.

Performa 1

Serial No: Date: _______Area Rural/Urban: _____Fecal Sample No: ______Soil Sample No.: ______

Male/Female: ______

Age: _____Name of Hospital/School: ______________ Name of Patient: ______________

Address: ______________________ Education: _____________B.P.S: _____Monthly income:

__________Occupation: _________

Ancylostoma duodenale Seperated from Contaminated Soil 31

Family Size: ______________ No. of Old: __________No. of Young: ______________ No. of Children:

_______

House constructed of Concrete/Mud: _______________No. of Rooms:_________Fecal

Refuse……Low/Moderate/High: _____________

Domestic Refuse…..Open/Bin: _________

Personal Hygiene: ___________Habit of Nail biting Yes / No , sucking thumb Yes / No, hand washing before meal

and after defecation Yes / No, walk bare-footed Yes / No, fond of street junk food Yes / No.

Well Nourished/Under Nourished/Malnutrition: ______________Drainage…… Open/Underground:

________________

Locality surrounded by Water Body/Fields: _______________Fecal dispersal via Sanitary/Unsanitary/No Facility:

___________

Water supply... Tap/Hand Pump/Well/Spring/River: ___________

APPENDICES

Table 1: Showing the Prevelance (%) Social Habits of Children Residing in Soil Contaminated Slums of Northern

Lahore from Nov 2006-Oct 2008

Factors Habit of

Pica Nail Biting

Suck

Thumb

Don’t

Wash

Hands

Before

Meal

Don’t

Wash

Hands

After

Defecation

Walk Bare

Footed

Use Junk

Food from

Street

Hawkers

Total no

of

children

observed n=

36

00

429 792 871 1567 1837 543 689

Percentage

(%) 10.50% 12.51% 11% 12.10% 21.76% 25.51% 7.54%

Prevalence

(%)±S.E

10.50±0.51

***

12.51±0.39

***

11.00±0.37

***

12.10±0.38

***

21.76±0.49

***

25.51±0.51

***

7.54±0.31**

*

Age (years) 10-Apr 10-Apr 5-Mar 8-May 11-Jun 13-May 11-Apr

Sewerage System

Gurkies (pit) Open

defecation Poor sanitation Satisfactory

1277 972 828 523

35.47% 27% 23% 14.52%

Table 2: Areawise, Monthwise and Seasonwise Prevalence (%) of Ancylosto maduodenalein Soil Samples in Six

Slums of Northern Lahore, Punjab, Pakistan from Nov2006-Oct 2008

Total n=3600

Total No. of Infected Soil Samples (Ancylosto maduodenale) in Soil

Fa

ct

ors

n=1800 Prevalenc

e

(%)±S.E

n=180

0 Prevalence

(%)±S.E

n=3600 Prevalence

(%)±S.E

(1st

Year)

(2nd

Year)

(1+2Ye

ar)

Slu

ms

Bhutto Colony

n=

60

0/Y

ear

/

Are

a

12 0.67±0.19

*

18 1.00±0.23** 30 0.83±0.15*

** Siddiquia Colony 10 0.56±0.18 16 0.89±0.22 26 0.72±0.14

Scheme No. 2 3 0.17±0.10 5 0.28±0.12 8 0.22±0.08

Sheikhupura Road 6 0.33±0.14 8 0.44±0.16 14 0.39±0.10

Khairdin Park 9 0.50±0.17 10 0.56±0.18 19 0.53±0.12

Missri Shah 11 0.61±0.18 21 1.17±0.25 32 0.89±0.16

32 Shahida Azhar Ali, Tanveer Akhtar, Azhar Maqbool, Ammara Hussan & Samman Ikram

Table 2: Contd.,

Total 3600 51 1.42±0.20

*

78 2.17±0.24* 129 1.79±0.16*

Tim

e (M

on

ths)

Nov2006&2007

n=

30

0 /

Mo

nth

/ Y

ear

2 0.67±0.47 4 1.33±0.66 6 1.00±0.41

Dec 2006&2007 3 1.00±0.57 2 0.67±0.47 5 0.83±0.37

Jan 2007&2008 5 1.67±0.74 5 1.67±0.74 10 1.67±0.52

Feb 2007&2008 4 1.33±0.66 6 2.00±0.81 10 1.67±0.52

Mar 2007&2008 2 0.67±0.47 1 0.33±0.33 3 0.50±0.29*

Apr 2007&2008 0 0.00±0.00 3 1.00±0.57 3 0.50±0.29*

May 2007&008 5 1.67±0.74 7 2.33±0.87 12 2.00±0.57

Jun 2007&2008 6 2.00±0.81 6 2.00±0.81 12 2.00±0.57

Jul 2007&2008 3 1.00±0.57 6 2.00±0.81 9 1.50±0.50

Aug 2007&008 5 1.67±0.74 7 2.33±0.87 12 2.00±0.57

Sep 2007&2008 6 2.00±0.81 12 4.00±1.13 18 3.00±0.70

Oct 2007&2008 10 3.33±1.04 19 6.33±1.41** 29 4.83±0.88

Total 3600 51 1.42±0.20

*

78 2.17±0.24* 129 3.58±0.31

Sea

son

s

Winter 1200 14 1.17±0.31 17 1.42±0.34 31 1.29±0.23*

Spring 600 2 0.33±0.24

*** 4

0.67±0.33**

* 6

0.50±0.20*

**

Summer 1200 19 1.58±0.36 26 2.17±0.42**

* 45

1.88±0.28*

**

Autumn 600 16 2.67±0.66 31 5.17±0.90 47 3.92±0.56

Overall Prevelance (%) n=1800*2=3600 / 1+2 Year 129 1.79±0.16*

*P<.05, **P<.01, ***P<.001 Z-Test

Figure 1: Prevalence (%) of Socioeconomic Status of Children (a), Sewerage System (b), Prevalence (%) of

Ancylostomaduodenalein Soil Samples Area Wise (c), Month Wise (d) and Season Wise (e) Nov 2006-Oct 2008

Northern Lahore. Z-test *=P<0.05, **=P<0.01, ***P<.001

Ancylostoma duodenale Seperated from Contaminated Soil 33

Plate 1: Showing the Poor Sewerage System with Uneduquate Supply of Water (a),Open Defecated Ares (b), Poor

Drain Syatem (C), Gurki (D), Poor Sanitation (e)and Contact of Man with Animals and Dunghills (f)

34 Shahida Azhar Ali, Tanveer Akhtar, Azhar Maqbool, Ammara Hussan & Samman Ikram

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