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20 September 2024 | Story André Damons | Photo Supplied
zebrafish-blue-in-aquarium
Zebrafish blue in an aquarium.

A researcher from the University of the Free State (UFS) hopes to make living with epilepsy and other diseases of the central nervous system (CNS) easier by using South African plants extracts which may have anti-epileptic properties and testing them on zebrafish larvae.

Prof Anke Wilhelm, Associate Professor and Divisional Head of Organic Chemistry in the UFS Department of Chemistry, focuses her research on the isolation of active GABAergic compounds (substances that affect the brain’s GABA system, which helps control nervous system activity) by using a test that measures the movement of zebrafish larvae.

Even though obtaining regulatory approval for use as a treatment for epilepsy is a long and complex process, Prof Wilhelm hopes to contribute to the better pain management of people suffering from epilepsy and diseases of the CNS through an affordable alternative drug with less side effects.

The tests are done in a zebrafish bioassay (an analytical method to determine the potency of a substance by its effect on living animals) housed at the UFS’ Chemistry Department.

Why zebrafish larvae?

Prof Wilhelm, who is a National Research Foundation Y2-rated synthetic organic chemist, says zebrafish share about 70% of their genes with humans, and about 84% of human genes known to be associated with diseases have a counterpart in zebrafish. This makes them a valuable model for studying human biology and disease.

“Zebrafish are powerful tools for modelling a wide range of CNS diseases, contributing significantly to the understanding of disease mechanisms and the development of potential treatments,” she says. “Mood disorders, anxiety, insomnia, and attention deficit hyperactivity disorder (ADHD) are all diseases which may be studied through this bioassay.”

She explains that the zebrafish larvae are studied seven days after fertilisation in their bioassay. The larvae are incubated with the specific plant extract at a certain (non-toxic) concentration for three hours. Pentylenetetrazol (PTZ), a GABAA receptor antagonist that has been extensively used in rodent models for acute seizure and anxiety, is then administered to induce concentration-dependent seizures in the zebrafish larvae.

“GABA receptor antagonists are drugs that inhibit the action of gamma-aminobutyric acid, the chief inhibitory neurotransmitter in the mammalian central nervous system,” Prof Wilhelm says. “A specialised infrared camera is then used to track the movement of the larvae inside a chamber. The data is then converted into a graph which shows the movement of each larva over 30 minutes.

“If lowering of movement is observed at a specific concentration it means that the plant extract may have the potential to be used as an epileptic drug, since it has the ability to counteract the induced seizure in the larvae. This bioassay is extremely useful in drug discovery and toxicity screening of plant extracts.”

Zebrafish embryos, she says, develop quickly, with major organs forming within 36 hours of fertilisation. This rapid development allows researchers to observe the effects of experiments in a short period. The maintenance of a zebrafish model is less costly and labour-intensive than using a rodent model. “The use of zebrafish larvae allows for high-throughput screening due to their small size and transparency, which facilitates observation of CNS-related effects. Their genetic and physiological similarities to humans make them a valuable model for early-stage drug discovery.”

Potential uses

The next step in the research, according to Prof Wilhelm, is to identify a single compound from a natural source which may have potential anti-epileptic activity while causing less side effects than current drugs on the market. Researchers would then investigate the possibility of synthesising such a compound on a large scale, to eliminate the use of a natural resource and promote sustainability.

“Many plant extracts which I have screened show a synergistic effect in the zebrafish bioassay, meaning that the extract or the combination of compounds shows potential, but the isolated compounds are inactive. Even if a plant extract shows promise in preclinical and early clinical studies, obtaining regulatory approval for use as a treatment for epilepsy is a long and complex process.

“This includes demonstrating consistent efficacy, safety, and quality in large-scale clinical trials. One of the major challenges in using plant extracts is the lack of standardisation. The concentration of active compounds in plant extracts can vary depending on factors like the plant's growing conditions, harvest time, and extraction methods. This variability makes it difficult to ensure consistent efficacy and safety, therefore this is a time-consuming process.”

Green chemistry

After being approached by Dr Glen Taylor, Senior Director of the UFS Directorate Research Development (DRD), in 2017, regarding funding for Noldus Daniovision equipment, Prof Wilhelm received training from Prof Matthias Hamburger of the University of Basel in Switzerland on how to use such equipment. The larval zebrafish locomotive bioassay was established at the UFS Chemistry Department during 2017 and 2018 and now provides a third-stream income for the department, in conjunction with the Department of Genetics, where the adult zebrafish are housed.

Prof Wilhelm’s other research interests include green chemistry, food sustainability, and recycling. She is looking into green extraction techniques using non-conventional extraction methods to recover valuable bioactive compounds from agricultural and food residues. “Techniques like ultrasound, microwave-assisted extraction, and the use of deep eutectic solvents are becoming popular for their efficiency and alignment with circular economy principles.”

News Archive

Research into veld fires in grassland can now help with scientifically-grounded evidence
2015-04-10

While cattle and game farmers are rejoicing in the recent rains which large areas of the country received in the past growing season, an expert from the University of the Free State’s Department of Animal, Wildlife, and Grassland Sciences, says that much of the highly inflammable material now available could lead to large-scale veld fires this coming winter.

Prof Hennie Snyman, professor and  researcher in the Department of Animal, Wildlife, and Grassland Sciences, warns that cattle and game farmers should be aware, in good time, of this problem which is about to rear its head. He proposes that farmers must burn firebreaks as a precaution.

At present, Prof Snyman focuses his research on the impact of fire and burning on the functioning of the grassland ecosystem, especially in the drier grassland regions.

He says the impact of fire on the functioning of ecosystems in the ‘sour’ grassland areas of Southern Africa (which includes Kwazulu-Natal, Limpopo, Mpumalanga, the Eastern Cape, and the Harrismith environs) is already well established, but less information  is available for ‘sweet’ semi-arid grassland areas. According to Prof Snyman, there is no reason to burn grassland in this semi-arid area. Grazing by animals can be effectively used because of the high quality material without having to burn it off. In the sourer pasturage, fire may well form part of the functioning of the grassland ecosystem in view of the fact that a quality problem might develop after which the grass must rejuvenate by letting it burn.

Prof Snyman, who has already been busy with the research for ten years, says quantified data on the impact of fire on the soil and plants were not available previously for the semi-arid grassland areas. Fires start frequently because of lightning, carelessness, freak accidents, or damaged power lines, and farmers must be recompensed for this damage.

The shortage of proper research on the impact of fires on soil and plants has led to burnt areas not being withdrawn from grazing for long enough. The lack of information has also led to farmers, who have lost grazing to fires, not being compensated fairly or even being over-compensated.

“When above-and below-ground plant production, together with efficient water usage, is taken into account, burnt grassland requires at least two full growing seasons to recover completely.”       

Prof Snyman says farmers frequently make the mistake of allowing animals to graze on burnt grassland as soon as it begins to sprout, causing considerable damage to the plants.

“Plant roots are more sensitive to fire than the above-ground plant material. This is the reason why seasonal above-ground production losses from fire in the first growing season after the fire can amount to half of the unburnt veld. The ecosystem must first recover completely in order to be productive and sustainable again for the long term. The faster burnt veld is grazed again, the longer the ecosystem takes to recover completely, lengthening the problem with fodder shortages further.  

Prof Snyman feels that fire as a management tool in semi-arid grassland is questionable if there is no specific purpose for it, as it can increase ecological and financial risk management in the short term.

Prof Snyman says more research is needed to quantify the impact of runaway fires on both grassland plant productivity and soil properties in terms of different seasonal climatic variations.

“The current information may already serve as valuable guidelines regarding claims arising from unforeseen fires, which often amount to thousands of rand, and are sometimes based on unscientific evidence.”

Prof Snyman’s research findings have been used successfully as guidelines for compensation aspects in several court cases.

 

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