PREVALENCE OF NEMATODE LARVAE FROM FECAL SAMPLES AMONG CHILDREN IN TERESA ST., STA.

MESA MANILA
Garcia, Carla Mae D.C, Gumban, John Daniel P., Ocasla, Donitalyn D, Penetrante, Gerson Kim O. Department of Biology, College of Science Polytechnic University of the Philippines Group 9 ABSTRACT:

INTRODUCTION Intestinal nematodes infect people, especially children and constitute a formidable public health status problem (Savoli, L. et al., 1992). These worm infections are very common and often minimized causing strongyloidiasis. The infected person, especially the children may suffer nutritional deficits, serious illness, cognitive impairments and occasionally death (Barnish, G. et al., 1990). The prevalence of infection with helminth parasites is approximately two billion people worldwide and 60 000 deaths annually (Walsh, JA. et al., 1990). In Philippines, it is well documented that a prevalence of 38% of hookworms done by the U.S Naval Medical Research Unit No. 2 (Basaca-Sevilla, C., 1984). Another local study on children living in residential institutions in Metro Manila yielded 7% prevalence of hookworm. Strongyloidiasis is caused by 2 species of intestinal nematodes Strongyloides. The most common and globally distributed human pathogen of

clinical importance is Strongyloides stercoralis. The other species, Strongyloides fuelleborni, is found sporadically in Africa and Papua New Guinea (Grove, DI., 1996; Liu, LX., et al., 1993; and Genta, RM. 1989). S. stercoralis was first reported in 1876 in the stools of French soldiers on duty in Vietnam who had severe diarrhea, and the disease the organism produces was known for many years as CochinChina diarrhea (Grove, DI. 1996). The elucidation of the complete life cycle occurred 50 years after the discovery of the worm. S. stercoralis has a complex life cycle in which parthenogenic females (i.e., capable of reproducing without males) embedded in the intestinal mucosa lay embryonated eggs that hatch internally (Grove, DI. 1996; Mansfield, LS. 1996). The resultant first-stage larvae (L1; rhabditiform larvae) are passed out in the feces and may develop directly into second (L2)-stage and third (L3; filariform larvae)-stage larvae may develop through 4 free-living larval stages to become freeliving adult males and females. The freeliving adults reproduce sexually to

produce L1, which also develop to L3. The L3 of either cycle can penetrate the skin of the human host, pass through the circulation to the lungs, enter the airways, be swallowed, and finally reach intestine, where they mature into adult egg-laying females. In autoinfection, larvae that have developed to the infective third stage within the gastrointestinal tract penetrate the intestinal mucosa and then migrate to the definitive site in the small intestine or to parenteral sites (e.g., lungs) (Grove, DI. 1996; Mansfield, LS. 1992). Some have argued that the pulmonary route is just one of the several possible pathways for the larvae to reach the duodenum (Scad, GA. et al., 1989). In any event, this ability to establish a cycle of repeated endogenous reinfection within the host invariably results in chronic infection that can last for several decades; the current recorded appears to be 65 years (Liu, LX. et al., 1993). In order for to determine the nematode larval stages from a fecal sample, the Baermann funnel technique can be used to isolate larvae and can be employed to diagnose the lungworm infections. It requires equipment to hold the fecal sample in water so that larvae can migrate out and be collected (Zajac et al., 2006). The principle of this method is very simple, the nematodes move out of the substrate toward the water, and sink to the bottom of the funnel stem due to gravity forces. This type of nematode extraction is suitable to extract mobile

nematode from substrates like soils and sediments, plant material and litter. The advantages and disadvantages of this technique: it is cheap and simple; extraction efficiency is quite good if your sample is small relative to the funnel diameter (with only a thin layer of substrate on the sieve), but decreases very rapidly when your sample gets bigger than that (Coleman et al., 2004). Strongyloides stercoralis is a round worm occurring widely in tropical and subtropical areas that cause Strongyloidiasis infection (WGO 2004). Its clinical relevance increases when one considers its opportunistic behavior in compromised patients, in whom infection is frequent and sometimes leads to chronic infections, which may be fatal due to secondary bacterial infections. Strongyloidiasis diagnosis is still made by detection of larvae in stool samples by microscopic observation of stool smears. (Chavarra et al., 2001). Determining the prevalence of nematode or hookworm (Strongyloides stercoralis) is the main goal of this study. Specifically, it intends to apply Baermann Funnel Technique for the extraction of nematode worm in fecal samples among children in Teresa St., Sta. Mesa Manila. The result of the present study would provide descriptions and data regarding to the Strongyloides stercoralis. Additionally, this study will produce significant and relevant information for future studies.

METHODOLOGY Sample Site The study will be conducted at Teresa St., Sta. Mesa Manila City, which is located at 140 35 59.85 N latitude and 1210 0 43.86 E latitude. The residential areas at Teresa St. with the ocular inspection, are characterized with good level of environmental sanitation, because the barangay officials make sure that the environment are clean and green. Also, there are ready schedule in collecting the garbage. Each house has latrines or toilets. But the sewage system is poor because a rain of about 30 minutes can cause flood. And also, street children playing and go around the area barefoot and eating without washing hands. Sampling Size A total of 30 children from Teresa St. is the total sample size of this study. It is a random sampling that whosoever children that the researchers meet or interviewed and kindly ask them for their fecal samples with an exchange of tokens for them (candies, mini chocolates, cookies and junk foods) and with the consent of their parents. Sample Collection Thirty (30) stool samples will be collected of children. A fecal container for collection and storage will be used. These containers will be labeled with unique identifiers including the name of the patient, sex, age, data and the consistency of the stool. The patient will place the

stool specimen directly into the container or on a piece of paper. Specimens will be placed inside ice chest for transport to the laboratory. Sample Processing Upon the arrival of stool samples in the laboratory, they will be classified according to their physical characteristics. The stool samples that contain blood will be examined first, followed by watery specimens. Baermann Funnel Method was employed in the extraction of nematode larval stages from fecal samples. A set-up of glass funnel with supporting clamp is needed in this method. Kindly place a wire gauze or nylon filter over the top of the funnel, followed by a pad consisting of two layers of gauze. Then, close the pinch clamp at the bottom of the tubing, and fill the funnel with lukewarm tap water (about 450C) until it just soaks the gauze padding. The fecal sample will be placed on gauze and cover with a piece of filter paper. Then allow the set-up to stand for 2 hours longer. Draw off 10 ml fluid into the beaker by releasing the pinch clamp and centrifuge the fluid for 2 minutes at 400 to 500 revolution per minute (rpm). Examine the sediment under microscope. Use low power magnification (LPO) but for motile larvae use high power magnification (HPO).

RESULTS AND DISCUSSION A total of 30 stool samples of children were collected from Teresa St., Sta. Mesa Manila. Stool sample were investigated for the presence of nematode worm using Baermann Funnel Method. Of a total of 30 stool samples, hookworm or nematodes was seen in none of the stool samples of children from Teresa St., Sta. Mesa Manila. Present study showed no detection of nematodes in the stool samples of the children. This finding was in agreement with no detection of hookworms among school children in elementary of two areas in Metro Manila (Belizario, V. et al., 2007). Also, a study that is still conducting at Bagong Silang, San Jose Navotas, at the detection of Strongyloides stercoralis using Baermann Funnel Method has a findings of no detection of hookworms (Garcia, C., Gumban, JD. 2012) and it was in agreement also in this study. Interviewing the children, it is reported that at their school, they attend, a deworming program that is given free at the public schools by the Manila City government and in the Philippines, a proposed model for school-based worm control involving mass treatment and health education has been well documented (Belizario, V. et al., 200). And this study suggests that the hookworm infection are controlled by periodic mass treatment in the form of chewable and attractive preparations of albendazole or mebendazole tablets given single dose,

two to three times a year (Belizario, V. et al., 2007). Also, the most important factor for sustaining hookworm control is the support of the childrens parents and families. And of course, the Teresa St. as a community in general, with the help of information, families should be tapped as important resources to provide for the cost of the drugs, to improve hygiene and sanitation, and to reinforce health education activities. The result from this study provides us a glimpse of nematode status in Teresa St. Sta. Mesa Manila. The children in the study benefit the most because knowledge about nematode will enable them to be treated and prevent possible infection. Also, Strongyloidiasis should be one of the problems given priority in all countries, to enhance quality of life, health and productivity of children. And through this study, the parents knowledge regarding nematodes would be increased and prevention could be done to avoid more serious damage to their childrens health.

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