Name: determining soil-transmitted helminth infection status and physical fitness of school-aged children
Uploaded: 2012-08-22T12:00:00
Description: Watch this Scientific Journal Video about Determining Soil-transmitted Helminth Infection Status and Physical Fitness of School-aged Children at JoVE.com

The authors are grateful to the communities in Yunnan province, People's Republic of China and Agboville, C&ocirc;te d'Ivoire, for their participation in the studies described above. We are indebted to the local field teams that have assisted in the testing and execution of the protocols. We are particularly grateful to the following colleagues from the Swiss Tropical and Public Health Institute: Yvette Endriss for providing the microscopic images, Hanspeter Marti and his team for laboratory support and Stefanie Krauth for her assistance during the video filming. Finally, we thank the two anonymous referees for a series of useful comments.

1. Kato-Katz Technique
<ol>
 <li>Place a standard Kato-Katz template with a hole for holding 41.7 milligrams (mg) of stool on a microscope slide.</li>
 <li>Scoop 2-3 grams (g) of a fresh fecal sample onto a piece of newspaper or aluminum foil, and press a piece of wire or plastic mesh on top to sieve it.</li>
 <li>Using a small plastic spatula, scrape the sieved material off the mesh and completely fill the hole in the Kato-Katz template with it. To remove excess fecal material, level the content of the hole with the s...

<ol>
	<li>Steinmann, P., Utzinger, J., Du, Z. W., Zhou, X. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiparasitism:+a+neglected+reality+on+global,+regional+and+local+scale.">Multiparasitism: a neglected reality on global, regional and local scale.</a> Advances in Parasitology. 73, 21-50 (2010).</li><li>Bethony, J., Brooker, S., Albonico, M., Geiger, S., Loukas, A., Diemert, D., Hotez, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+soil-transmitted+helminth+infections:+ascariasis,+trichuriasis,+and+hookworm.">The soil-transmitted helminth infections: ascariasis, trichuriasis, and hookworm.</a> Lancet. 367, 1521-1532 (2006).</li><li>Hotez, P. J., Molyneux, D. H., Fenwick, A., Kumaresan, J., Ehrlich Sachs, S., Sachs, J. D., Savioli, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Control+of+neglected+tropical+diseases.">Control of neglected tropical diseases.</a> New England Journal of Medicine. 357, 1018-1027 (2007).</li><li>Chan, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+global+burden+of+intestinal+nematode+infections-fifty+years+on.">The global burden of intestinal nematode infections-fifty years on.</a> Parasitology Today. 13, 438-443 (1997).</li><li>Olsen, A., van Lieshout, L., Marti, H., Polderman, T., Polman, K., Steinmann, P., Stothard, R., Thybo, S., Verweij, J. J., Magnussen, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strongyloidiases+-+the+most+neglected+of+the+neglected+tropical+diseases.">Strongyloidiases - the most neglected of the neglected tropical diseases.</a> Transactions of the Royal Society of Tropical Medicine and Hygiene. 103, 967-972 (2009).</li><li>Becker, S. L., Sieto, B., Silue, K. D., Adjossan, L., Kone, S., Hatz, C., Kern, W. V., N'Goran, E. K., Utzinger, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diagnosis,+clinical+features,+and+self-reported+morbidity+of+Strongyloides+stercoralis+and+hookworm+infection+in+a+co-endemic+setting.">Diagnosis, clinical features, and self-reported morbidity of Strongyloides stercoralis and hookworm infection in a co-endemic setting.</a> PLoS Neglected Tropical Diseases. 5, e1292 (2011).</li><li>World Health Organization. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Action+against+worms.">Action against worms.</a> WHO Newsletter. (11), (2008).</li><li>Montresor, A., Crompton, D. W. T., Hall, A., Bundy, D. A. P., Savioli, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Guidelines+for+the+evaluation+of+soil-transmitted+helminthiasis+and+schistosomiasis+at+community+level.">Guidelines for the evaluation of soil-transmitted helminthiasis and schistosomiasis at community level.</a> World Health Organization. , (1998).</li><li>Garc&#237;a, L. S., Bruckner, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Diagnostic medical parasitology. , (2001).</li><li>Brooker, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estimating+the+global+distribution+and+disease+burden+of+intestinal+nematode+infection:+adding+up+the+numbers+-+a+review.">Estimating the global distribution and disease burden of intestinal nematode infection: adding up the numbers - a review.</a> International Journal for Parasitology. 40, 1137-1144 (2010).</li><li>Stephenson, L. S., Latham, M. C., Kinoti, S. N., Kurz, K. M., Brigham, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improvements+in+physical+fitness+of+Kenyan+schoolboys+infected+with+hookworm,+Trichuris+trichiura+and+Ascaris+lumbricoides+following+a+single+dose+of+albendazole.">Improvements in physical fitness of Kenyan schoolboys infected with hookworm, Trichuris trichiura and Ascaris lumbricoides following a single dose of albendazole.</a> Transactions of the Royal Society of Tropical Medicine and Hygiene. 84, 277-282 (1990).</li><li>Stephenson, L. S., Latham, M. C., Adams, E. J., Kinoti, S. N., Pertet, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physical+fitness,+growth+and+appetite+of+Kenyan+school+boys+with+hookworm,+Trichuris+trichiura+and+Ascaris+lumbricoides+infections+are+improved+four+months+after+a+single+dose+of+albendazole.">Physical fitness, growth and appetite of Kenyan school boys with hookworm, Trichuris trichiura and Ascaris lumbricoides infections are improved four months after a single dose of albendazole.</a> Journal of Nutrition. 123, 1036-1046 (1993).</li><li>Bustinduy, A. L., Thomas, C. L., Fiutem, J. J., Parraga, I. M., Mungai, P. L., Muchiri, E. M., Mutuku, F., Kitron, U., King, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measuring+fitness+of+Kenyan+children+with+polyparasitic+infections+using+the+20-meter+shuttle+run+test+as+a+morbidity+metric.">Measuring fitness of Kenyan children with polyparasitic infections using the 20-meter shuttle run test as a morbidity metric.</a> PLoS Neglected Tropical Diseases. 5, e1213 (2011).</li><li>M&#252;ller, I., Coulibaly, J. T., F&#252;rst, T., Knopp, S., Hattendorf, J., Krauth, S. J., Stete, K., Righetti, A. A., Glinz, D., Yao, A. K., P&#252;hse, U., N'Goran, E. K., Utzinger, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+schistosomiasis+and+soil-transmitted+helminth+infections+on+physical+fitness+of+school+children+in+C&#244;te+d'Ivoire.">Effect of schistosomiasis and soil-transmitted helminth infections on physical fitness of school children in C&#244;te d'Ivoire.</a> PLoS Neglected Tropical Diseases. 5, e1239 (2011).</li><li>Yap, P., Du, Z. W., Chen, R., Zhang, L. P., Wu, F. W., Wang, J., Wang, X. Z., Zhou, H., Zhou, X. N., Utzinger, J., Steinmann, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Soil-transmitted+helminth+infections+and+physical+fitness+in+school-aged+Bulang+children+of+southwest+China:+results+from+a+cross-sectional+survey.">Soil-transmitted helminth infections and physical fitness in school-aged Bulang children of southwest China: results from a cross-sectional survey.</a> Parasites and Vectors. 5, 50 (2012).</li><li>L&#233;ger, L. A., Mercier, D., Gadoury, C., Lambert, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+multistage+20+metre+shuttle+run+test+for+aerobic+fitness.">The multistage 20 metre shuttle run test for aerobic fitness.</a> Journal of Sports Sciences. 6, 93-101 (1988).</li><li>Tomkinson, G. R., L&#233;ger, L. A., Olds, T. S., Cazorla, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Secular+trends+in+the+performance+of+children+and+adolescents+(1980-2000):+an+analysis+of+55+studies+of+the+20+m+shuttle+run+test+in+11+countries.">Secular trends in the performance of children and adolescents (1980-2000): an analysis of 55 studies of the 20 m shuttle run test in 11 countries.</a> Sports Medicine. 33, 285-300 (2003).</li><li>Mahon, A. D., Marjerrison, A. D., Lee, J. D., Woodruff, M. E., Hanna, L. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluating+the+prediction+of+maximal+heart+rate+in+children+and+adolescents.">Evaluating the prediction of maximal heart rate in children and adolescents.</a> Research Quarterly for Exercise and Sport. 81, 466-471 (2010).</li><li>Bogitsh, B. J., Carter, C. E., Oeltmann, T. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Human Parasitology. , (2011).</li><li>Steinmann, P., Zhou, X. N., Du, Z. W., Jiang, J. Y., Wang, L. B., Wang, X. Z., Li, L. H., Marti, H., Utzinger, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Occurence+of+Strongyloides+stercoralis+in+Yunnan+province,+China,+and+comparison+of+diagnostic+methods.">Occurence of Strongyloides stercoralis in Yunnan province, China, and comparison of diagnostic methods.</a> PLoS Neglected Tropical Diseases. 1, e75 (2007).</li><li>Martin, L. K., Beaver, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+Kato+thick-smear+technique+for+quantitative+diagnosis+of+helminth+infections.">Evaluation of Kato thick-smear technique for quantitative diagnosis of helminth infections.</a> American Journal of Tropical Medicine and Hygiene. 17, 382-391 (1968).</li><li>Dacombe, R. J., Crampin, A. C., Floyd, S., Randall, A., Ndhlovu, R., Bickle, Q., Fine, P. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time+delays+between+patient+and+laboratory+selectively+affect+accuracy+of+helminth+diagnosis.">Time delays between patient and laboratory selectively affect accuracy of helminth diagnosis.</a> Transactions of the Royal Society of Tropical Medicine and Hygiene. 101, 140-145 (2007).</li><li>Knopp, S., Mgeni, A. F., Khamis, I. S., Steinmann, P., Stothard, J. R., Rollinson, D., Marti, H., Utzinger, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diagnosis+of+soil-transmitted+helminths+in+the+era+of+preventive+chemotherapy:+effect+of+multiple+stool+sampling+and+use+of+different+diagnostic+techniques.">Diagnosis of soil-transmitted helminths in the era of preventive chemotherapy: effect of multiple stool sampling and use of different diagnostic techniques.</a> PloS Neglected Tropical Diseases. 2, e331 (2008).</li></ol>

The three protocols described in this paper have been tested and executed in the south-western part of Yunnan province, People's Republic of China among members of the Bulang ethnic minority21 and in different parts of Africa6,13,14.
There are several templates available for the Kato-Katz technique. Each distinct hole-size will deliver a distinct amount of fecal material. For the calculation of EPGs, it is therefore necessary to know the unit of the template, so as to mul...

Note: Materials in this list are generic and can be obtained from different sources.
<table border="1">
	<tbody>
		<tr>
			<td>Name of the reagent</td>
			<td>Company</td>
			<td>Catalogue number</td>
			<td>Comments </td>
		</tr>
		<tr>
			<td>Kato-Katz kit 
			[400 plastic templates with a hole of 6 millimeter (mm) (diameter) in a 1.5 mm thick template; 400 plastic spatula; a 20-m role of nylon screen of 80 mesh size; a 20-m role of hydrophilic cellophane, 34 &mu;m thick.]</td>
			<td>Vestergaard Frandsen</td>
			<td>&nbsp;</td>
			<td>Please note that the glycerin-malachite green solution is not included in this kit but can be bought from any chemical supplier.</td>
		</tr>
		<tr>
			<td>Glass funnel [about 8 centimeter (cm) wide and 6 cm deep]</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>The glass funnel for the Baermann technique should be able to hold about 60 mL of water. The size of the rubber hose and gauze is dependent on the type of glass funnel used.</td>
		</tr>
		<tr>
			<td>Biological microscope</td>
			<td>Olympus</td>
			<td>CX21LED/CX21</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>Shanghai Medical Instruments Group</td>
			<td>80 - 2T</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Pre-recorded signals</td>
			<td>Bitworks Design</td>
			<td>Team Bleep Test Version 1.3.1</td>
			<td>iPhone application</td>
		</tr>
		<tr>
			<td>Heart rate monitor</td>
			<td>POLAR</td>
			<td>FT1</td>
			<td>Watch and transmitter</td>
		</tr>
	</tbody>
</table>

determining soil-transmitted helminth infection status and physical fitness of school-aged children

Soil-transmitted helminth (STH) infections are common. Indeed, more than 1 billion people are affected, mainly in the developing world where poverty prevails and hygiene behavior, water supply, and sanitation are often deficient1,2. Ascaris lumbricoides, Trichuris trichiura, and the two hookworm species, Ancylostoma duodenale and Necator americanus, are the most prevalent STHs3. The estimated global burden due to hookworm disease, ascariasis, and trichuriasis is 22.1, 10.5, and 6.4 million disability-adjusted life years (DALYs), respectively4. Furthermore, an estimated 30-100 million people are infected with Strongyloides stercoralis, the most neglected STH species of global significance which arguably also causes a considerable public health impact5,6. Multiple-species infections (i.e., different STHs harbored in a single individual) are common, and infections have been linked to lowered productivity and thus economic outlook of developing countries1,3.
For the diagnosis of common STHs, the World Health Organization (WHO) recommends the Kato-Katz technique7,8, which is a relatively straightforward method for determining the prevalence and intensity of such infections. It facilitates the detection of parasite eggs that infected subjects pass in their feces.
With regard to the diagnosis of S.stercoralis, there is currently no simple and accurate tool available. The Baermann technique is the most widely employed method for its diagnosis. The principle behind the Baermann technique is that active S.stercoralis larvae migrate out of an illuminated fresh fecal sample as the larvae are phototactic9. It requires less sophisticated laboratory materials and is less time consuming than culture and immunological methods5.
Morbidities associated with STH infections range from acute but common symptoms, such as abdominal pain, diarrhea, and pruritus, to chronic symptoms, such as anemia, under- and malnutrition, and cognitive impairment10. Since the symptoms are generally unspecific and subtle, they often go unnoticed, are considered a normal condition by affected individuals, or are treated as symptoms of other diseases that might be more common in a given setting. Hence, it is conceivable that the true burden of STH infections is underestimated by assessment tools relying on self-declared signs and symptoms as is usually the case in population-based surveys.
In the late 1980s and early 1990s, Stephenson and colleagues highlighted the possibility of STH infections lowering the physical fitness of boys aged 6-12 years11,12. This line of scientific inquiry gained new momentum recently13,14,15. The 20-meter (m) shuttle run test was developed and validated by L&eacute;ger et al.16 and is used worldwide to measure the aerobic fitness of children17. The test is easy to standardize and can be performed wherever a 20-m long and flat running course and an audio source are available, making its use attractive in resource-constrained settings13. To facilitate and standardize attempts at assessing whether STH infections have an effect on the physical fitness of school-aged children, we present methodologies that diagnose STH infections or measure physical fitness that are simple to execute and yet, provide accurate and reproducible outcomes. This will help to generate new evidence regarding the health impact of STH infections.

Watch this Scientific Journal Video about Determining Soil-transmitted Helminth Infection Status and Physical Fitness of School-aged Children at JoVE.com

Determining Soil-transmitted Helminth Infection Status and Physical Fitness of School-aged Children

Chronic infection with soil-transmitted helminths (STHs) causes malabsorption, stunting, and wasting in the growing child. Hence, it is plausible that these infections also reduce the physical fitness of children. Here, we visualize two techniques for the diagnosis of STHs and the 20-meter shuttle run test for assessing children's physical fitness.

Soil-transmitted helminth (STH) infections are common. Indeed, more than 1 billion people are affected, mainly in the developing world where poverty ...

Research

JoVE Journal

Immunology and Infection

Determining the Reactivity and Titre of Serum using a Haemagglutination Assay

Haemagglutination is a specific form of agglutination and is used when antibodies bind to red blood cells, which act as a particulate antigen. Red blood ...

Determining the Phagocytic Activity of Clinical Antibody Samples

Antibody-driven phagocytosis is induced via the engagement of Fc receptors on professional phagocytes, and can contribute to both clearance as well as ...

Using Eggs from Schistosoma mansoni as an In vivo Model of Helminth-induced Lung Inflammation

Using Eggs from Schistosoma mansoni as an In vivo Model of Helminth-induced Lung Inflammation

Schistosoma parasites are blood flukes that infect an estimated 200 million people worldwide 1. In chronic infection with Schistosoma, the severe ...

Analysis of Pulmonary Dendritic Cell Maturation and Migration during Allergic Airway Inflammation

Dendritic cells (DCs) are the key players involved in initiation of adaptive immune response by activating antigen-specific T cells. DCs are present in ...

Fluorescence Microscopy Methods for Determining the Viability of Bacteria in Association with Mammalian Cells

Central to the field of bacterial pathogenesis is the ability to define if and how microbes survive after exposure to eukaryotic cells. Current protocols ...

Total Protein Extraction and 2-D Gel Electrophoresis Methods for Burkholderia Species

Total Protein Extraction and 2-D Gel Electrophoresis Methods for Burkholderia Species

The investigation of  the intracellular protein levels of bacterial species is of importance to  understanding the pathogenic mechanisms of diseases ...

Pairwise Growth Competition Assay for Determining the Replication Fitness of Human Immunodeficiency Viruses

In vitro fitness assays are essential tools for determining viral replication fitness for viruses such as HIV-1. Various measurements have been used to ...

Determining Immune System Suppression versus CNS Protection for Pharmacological Interventions in Autoimmune Demyelination

A major hallmark of the autoimmune demyelinating disease multiple sclerosis (MS) is immune cell infiltration into the brain and spinal cord resulting in ...

Visualizing the Effects of Sputum on Biofilm Development Using a Chambered Coverglass Model

Biofilms consist of groups of bacteria encased in a self-secreted matrix. They play an important role in industrial contamination as well as in the ...

High-throughput Parallel Sequencing to Measure Fitness of Leptospira interrogans Transposon Insertion Mutants During Golden Syrian Hamster Infection

High-throughput Parallel Sequencing to Measure Fitness of Leptospira interrogans Transposon Insertion Mutants During Golden Syrian Hamster Infection

In this manuscript, we describe a transposon sequencing (Tn-Seq) technique to identify and quantify Leptospira interrogans mutants altered in fitness ...