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Understanding drug penetration, distribution, and metabolization is fundamental for understanding drug efficacy. This also accounts for parasites during antiparasitic treatment. Recently, we established matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) in blood flukes and liver flukes. This label-free technique is capable of visualizing the molecular distribution of endogenous and exogenous molecules, such as drug compounds. Here, we conducted atmospheric-pressure scanning microprobe MALDI MSI (AP-SMALDI MSI) of tissue sections of adult Fasciola hepatica that have been treated in vitro with 100 µM of triclabendazole (TCBZ), the drug of choice for treatment of fasciolosis, and its main metabolite triclabendazole sulfoxide (TCBZ-SO). Measurements covered an m/z mass range of 250–1,000 and provided a high spatial resolution using a pixel size of 10 µm. To support the interpretation of drug distribution, we first identified endogenous lipids that mark characteristic tissues such as the gastrodermis, the tegument, and the parenchyma. The obtained results suggested an early tegumental route of TCBZ uptake within 20 min, followed by spreading throughout the parasite after 4 h, and an even distribution in most tissues after 12 h. This coincided with a strong reduction of parasite vitality. TCBZ-SO treatment demonstrated the accumulation of this metabolite in the same tissues as the parent drug compound. These data demonstrate the auspicious potential of MALDI MSI to visualize uptake and distribution patterns of drugs or drug-candidate compounds in parasites, which might contribute to preclinical drug discovery in liver fluke research and beyond.

Schistosomiasis caused by different species of schistosome parasites is one of the most debilitating helminthic diseases of humans worldwide. For decades, chemotherapy is the main method of controlling schistosomiasis. However, the fear of drug resistance has motivated the search for alternatives. It has been demonstrated that the ABL kinase inhibitor imatinib affected the development and survival of Schistosoma mansoni in vitro; however, there is still lack of information on whether imatinib also affects other schistosome species such as Schistosoma japonicum. In the present study, the anti-schistosomal potency of imatinib on adult S. japonicum was investigated in vitro, and the results showed that imatinib had a significant impact on various physiological processes of S. japonicum adult worms. Besides its negative effects on worm motility, pairing stability, and gonad development, imatinib caused pathological changes in the gastrodermis as well as the death of the parasite. Our findings suggest that imatinib is an intriguing candidate for further development as an option to fight S. japonicum.

Cercariae of Schistosoma mansoni approach towards their host's skin with a chemokinesis. They respond to human skin extracts by inserting shifts between backward and forward swimming and therewith increase the rate of change of direction (RCD). We identified the host attractants by fractionating human skin surface extracts, exposing the fractions and chemicals to the cercariae and recording cercarial swimming by video tracking. The cercariae responded specifically to three skin surface compounds by increasing the frequency of shifts in swimming direction. (1) Free fatty acids. The effectiveness was limited to chain lengths between 10 and 14 carbon atoms in saturated chains and increased by the number of double bonds. (2) Free L: -arginine at concentrations as low as 100 nM. (3) Small peptides with terminally located L: -arginine at concentrations as low as 50 microM. All chemokinesis stimuli are also involved in subsequent behavioural phases of host invasion: attachment and directed creeping on the skin surface, penetration and navigation within host tissues. The cercariae responded during swimming only to these small molecular water soluble cues and not to cues to which they respond in other behavioural phases of host invasion, but which are unsuitable as chemoattractants due to their chemical properties or distribution within the skin layers. This characterises the chemokinetic responses as adequate adaptation to advance transmission success.

Dermatitis caused by penetrating bird schistosome cercariae is an emerging global public health problem. Infections may be prevented by the use of topical formulations that inhibit cercarial skin penetration. We evaluated nine water resistant formulations by exposing treated arms of volunteers to Trichobilharzia szidati cercariae. Six formulations protected from cercarial invasion. However, after immersion of the treated skin in water (2 x 20 min), only two formulations offered full protection: (1) Safe Sea, a cream protecting against jelly fish, (2) niclosamide in water resistant sun protecting cream formulations at concentrations as low as 0.05%. In an in vitro system Safe Sea and a 0.1% niclosamide formulation caused a high damage rate in T. szidati (92% and 99% after 5 min; only niclosamide with lethal effect) but not in Schistosoma mansoni (1% and 72%; both formulations with lethal effect). However, a 1% niclosamide formulation damaged S. mansoni sufficiently (100% after 5 min) and might offer full penetration protection.

We studied the skin invasion of Schistosoma mansoni cercariae by placing gamma-irradiated and nonirradiated cercariae onto the living human skin and timing the behavior of 53 individuals. The skin invasion of S. mansoni was less efficient compared to the bird schistosome Trichobilharzia szidati. S. mansoni cercariae crept longer on the skin after attachment until they started penetration movements (median of 43 s [range of 15 s-6.58 min]; T. szidati, median of 8 s [range of 0-80 s]). Subsequent to this longer exploratory phase, 74% penetrated into wrinkles (T. szidati 84%), 22% into the smooth skin surface (T. szidati 0%), and 4% into hair follicles (T. szidati 16%). The S. mansoni cercariae needed, on average, 6.58 min (range of 1.57-13.13 min) for full entry, while T. szidati needed 4.0 min (range of 1.38-13.34 min); the fastest S. mansoni cercaria entered the skin within 94 s, while T. szidati entered within 83 s. Sixty percent of the S. mansoni cercariae had the tails still attached when the bodies disappeared in the skin whereas all T. szidati cercariae shed their tails within 0-105 s after the onset of penetration movements. The faster invasion of T. szidati may result from the more sophisticated host-finding mechanisms of this species. Regarding S. mansoni, cercarial dermatitis, as immediate skin response, developed after a sensitization period of 19 days.