Parasitic Worm Infections and Anthelmintic Drug Testing
Parasitic worm infections or helminthiasis caused by parasitic worms such as roundworms, tapeworms, and hookworms are a health issue in many countries, especially in areas with poor sanitation conditions. Infections can lead to malnutrition and growth disorders, even death, in both humans and animals. To address this issue, the development of anthelmintic drugs has been carried out, which is very important for controlling the spread of these parasitic infections.
The development and search for new deworming drugs must be accompanied by effectiveness testing, which can generally be conducted through in-vivo tests on animals infected with parasites. However, this method has several limitations, including ethical concerns about animal use and the variability of the results obtained. Therefore, a method has now been developed in vitro using parasite cultures as a more controlled and ethical alternative.
Parasite Culture in Anthelmintic Drug Testing
Parasit culture is a method where parasites are maintained in laboratory conditions, either in artificial media or using host tissue. The cultured parasite is not in the form of individual cells but rather in specific stages of its life cycle, such as eggs, larvae, or adults. Some parasites that are commonly used for in-vitro testing include Haemonchus contortus (a worm in ruminant animals), Strongyloides stercoralis, and Ascaris suum. (cacing pada babi).
The success of parasite culture is greatly influenced by environmental conditions and nutrient availability, including growth media, temperature, pH, and oxygen supply. Parasites can be cultured in liquid or solid media that contain essential nutrients that support their growth. One important stage is to culture the larvae in a medium that is adjusted to observe the effects of deworming medication on various developmental phases of the parasite.
In general, the creation of parasite cultures begins using the egg or larval stages of the parasite. Once the parasite reaches a certain stage, such as the L3 larva or adult larva, the anthelmintic drug to be tested is introduced into the culture.
The tested deworming medication was added to the culture at various concentrations to determine its effective dosage. Usually, several test concentrations are used to measure how the drug affects the parasite. After the addition of the medication, observations were made to determine the effects of the drug in inhibiting the development of the parasite from one stage of its life cycle to the next, for example, from larva to adult. In addition, observations were also made on the mobility of the parasites to determine whether the medication caused paralysis or immobilization in the parasites. The number of dead parasites is also counted so that it can serve as an indicator of the effectiveness of the drug in killing parasites. The test results were further analyzed to determine the effective concentration of the drug (EC50), which is the minimum dose required to inhibit the growth or kill 50% of the parasites in culture.
The use of parasite cultures in testing anti-parasitic drugs has several advantages, including:
Time and Cost Efficiency
In vitro methods are faster and cheaper compared to in vivo tests. Researchers can test many compounds in a short amount of time.
Better Environmental Control
Parasitic cultures allow for complete control over environmental conditions such as pH, temperature, and nutrients, which in turn enables more consistent test results.
Not Using Test Animals
From an ethical research perspective, parasite cultures reduce or even eliminate the use of test animals, making it more aligned with ethical standards in biomedical research.
Flexibility
Parasitic culture also provides flexibility in the life cycle, allowing for testing at various stages of the parasite's life cycle, which offers a comprehensive overview of the drug's effectiveness.
To achieve accurate results, testing should be conducted by individuals who possess expertise and experience in the fields of parasitology and pharmacology. Well-trained researchers will be capable of designing appropriate experiments, controlling variables effectively, and analyzing data with suitable statistical methods. Thus, the research results can be reliable and provide valid information regarding the effectiveness of the tested anthelmintic drug.
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REFERENCE
Frahm, S., Anisuzzaman, A., Prodjinotho, U. F., Vejzagić, N., Verschoor, A., & Prazeres da Costa, C. (2019). A novel cell-free method to culture Schistosoma mansoni from cercariae to juvenile worm stages for in vitro drug testing. PLOS Neglected Tropical Diseases, 13(1), e0006590.
Jackson, G. J. (1962). The parasitic nematode, Neoaplectana glaseri, in axenic culture. II. Initial results with defined media. Experimental Parasitology, 12(1), 25-32.
Voronin, D., Tricoche, N., Jawahar, S., Shlossman, M., Bulman, C. A., Fischer, C., … & Lustigman, S. (2019). Development of a preliminary in vitro drug screening assay based on a newly established culturing system for pre-adult fifth-stage Onchocerca volvulus worms. PLoS neglected tropical diseases, 13(1), e0007108.
