Working on ants as models, we examined the side effects of six largely used products, separately published our findings, and here summarize them for the readers’ convenience. The herbicides, the active substance of which is pelargonic acid, must be used only for not cultivated area since they impact the live of the soil fauna. Imodium, the active substance of which is Loperamide, used to stop diarrhea, should be used during short time periods, monitoring the patients as for their food consumption, locomotion, social relationships, cognition and dependence on the drug use. Only small amount of the myorelaxant Epsipram, the active substance of which is Tetrazepam, should be used, monitoring the patients as for their activity, locomotion, social relationships, stress, cognition, memory and dependence on the drug consumption. The antiemetic Motilium, the active substance of which is Domperidone, should be used after having made an adequate clinical examination of the patients, take then care of them monitoring them as for their health since side effects may occur. The dietary supplement Cognizing, the active substance of which is citicoline, should not be used, being inefficient and causing stress, agitation, nervousness and strong dependence on its consumption. The dietary supplement PQQ is efficient and has no adverse effect; it could be used, but at low doses, since, being novel, some not yet observed side effects may occur.

Keywords: cognizing, domperidone, herbicide, imodium, loperamide, motilium, PQQ, tetrazepam

ang deg, angular degrees; ang deg/cm, angular degrees per centimeter; mm/s, millimeter per second; n°, number; n^os, numbers

Working on ants as models, we have until now studied the effects of 56 products or situations used by humans. Though having duly separately published our successive works, we moreover published up to now seven summaries of these studies for the readers’ convenience.^1–7 The last six products we examined are not included in these summaries. This is why we here briefly report our findings on these six last studied products not yet related in a summary. These six products are, in the order of their studies and of their present brief report, an herbicide still authorized, easily available and largely used; three drugs largely consumed by humans their use being still debated; two dietary supplements rather new and more and more consumed by humans. Before relating our results, we here below give available information on each of these six products, in the order we studied them and briefly here report our findings.

Pollution due to herbicides is far less examined than that resulting from the humans’ use of pesticides, insecticides and drugs. However, since they prevent plants living, herbicides are likely to affect the health of contaminated animals. There exist many kinds of herbicides, with different chemical structures, affecting different biological functions of the plants, differently acting. Their use leads to a decreased of weeds, and of birds.^8,9 They pollute natural water,^10,11 and cause illness in humans.^12–17 The most dangerous ones are no longer authorized in several countries. Those, the active substance of which is pelargonic acid, are still authorized. However, even if pelargonic acid is a natural substance present in nature, it may also have several adverse not yet known effects. We thus aimed, to study on ants as models, the effects of a largely used and easily available herbicide of this kind, Herbatak. Pelargonic acid attacks the cellulose wall of the plants which finally died. It affects humans’ skin and eyes. Noting else is known as for its potential physiological and ethological adverse effects, and we thus intended to at least partly fill this gap.

Imodium is an easily available drug allowing efficiently treating diarrhea, acting on the nervous system innerving the colon.^18–20 Its active substance is Loperamide which has a chemical structure similar to that of opiate receptors antagonists. It does not cross the blood-brain barrier, has no analgesic effect,²¹ and according to some authors has no significant adverse effect.²² However, side unwanted effects are reported by other researchers,^23–26 as well as in the notice for use joined to the Imodium packages (e.g., headache, dizziness, drowsiness). Estimating that not enough information can easily be found by practitioners and patients about such a largely used and easily available drug, we decided to study, on ants as models, the potential ethological and physiological adverse effects of Imodium.

The benzodiazepine Tetrazepam is the active substance of drugs (e.g., Epsipam) allowing decreasing painful muscular contractions.²⁷ It is efficient, has only few adverse effects,^28–30 but more trials should be conducted for having an obvious idea about its safety.^31,32 A bibliographical study also concludes that more clinical trials are required. ³³ In the notice for use joined to Epsipam packages, several adverse effects are reported, such as memory and behavioral impairments, aggressiveness, skin allergy, dependence. Consequently, to get the bottom of it, we aimed to examine, on ants as models, the potential impact of Epsipam consumption on several biological traits.

Domperidone is the active substance of the antiemetic drug Motilium. It is very efficient in numerous cases of health problems, among others, for children, adults, patients under chemotherapy or pulmonology treatment, women in menopause, all this with few adverse effects.^34–42 However, Motilium induces some side effects such as headache, anaphylactic shock, anxiousness, dizziness, drowsiness, asthenia, akathisia. In some European lands, it is no longer authorized for children. To have an opinion about this decision, and because nothing is reported as for the impact of Motilium on several important ethological and physiological traits, we intended to examine on ants as models such potential impacts of this drug.

Several dietary supplements contain essentially citicoline (cytidine-diphosphate-choline) and for instance a small amount of PQQ (e.g., Cognizing). Citicoline could ameliorate the brain functioning,⁴³ and help patients recovering after having had an ischemic stroke or other neurological disorders,^44–47 but these properties are not unanimously affirmed. Nevertheless, citicoline has a neuroprotective effect,⁴⁸ and ameliorates the memory in case of deficiencies,⁴⁹ thanks to its increase of choline in the cell membrane.^50–55 It is also beneficial to persons suffering from glaucoma,⁵⁶ and is very useful for caring of elderly persons.⁵⁷ All this is again related in three reviews.^58–60  Due to such very probable beneficial effects of citicoline, dietary supplements containing essentially this substance were created and sold to humans for improving their brain functioning. Because the wanted effect of such dietary supplements might not occur, and because only very few side effects due to their consumption are reported, we decided to study on ants as models the impact of Cognizing on several physiological and ethological traits, including memory, sensory perception, social relationships.

PQQ (pyrroloquinoline-quinone) is a co-enzyme present, among others, in the inner membrane of the mitochondria, and it valuably improves the functioning of this cellular element. Drugs and dietary supplement having PQQ as active component have been created and their beneficial impact on, among others, the mitochondria have been many times proved, e.g.^61–64 PQQ was also found to be beneficial to rats having had a cerebral artery occlusion.⁶⁷ It also helps caring of persons suffering from diabetes.⁶⁵ However, even if its mode of action becomes more and more elucidated over time, not all its beneficial effects are until now known, and nothing is not yet known about its potential side unwanted effects. We thus opted to study on ants as models the potential physiological and ethological wanted and unwanted effects this dietary supplement may present.

Why using ants, which species we used, what we know on it, and which traits, potentially affected by the examined products, we considered is explained in each of our works. We here only briefly recall the answers to these questions. The basic biological processes are similar in all animal species (genetics, nervous influx, muscles contraction, sensory perception, conditioning acquisition). They are thus, at least in a first time, studied on vertebrates and invertebrates, the latter being often preferred for their small size, easy maintenance in a laboratory, and short reproductive cycle. Hymenoptera (e.g., bees) are largely used, and ants can thus be used. They can be the more so because their maintenance is very easy, cheap, even if several colonies containing hundreds of individuals are required. We work on the species Myrmica sabuleti Meirnert, 1861, the biology of which we know rather well, having investigated, among others, on their visual perception, navigation, recruitment,⁶⁶ ontogenesis of some of their skills, self-recognition in a mirror, distance and size effects,⁶⁹  Weber’s law,⁶⁸ numerosity abilities and related topics (e.g., ^69–71). This ant species represents thus, at last for our research, a comfortable biological model.

There are explained in each of our works, those relative to the six here briefly reported ones,^72–77 as well as in all the previous 56 ones. We thus here only cite these methods and very briefly explain them.

Collection and maintenance of ants

The ants were collected in autumn and spring 2021 and 2022, in Belgium, in Ardenne and in Condroz, from abandoned quarries, under stones and in grass. The ant colonies contained about 500 to 1,000 workers, 1 to 3 queens and brood. In the laboratory, they were kept in 1 to 3 glass tubes half filled with water, a cotton plug separating the water from the place devoted to the ants’ nesting. The nest tubes of each colony were set in a tray the borders of which having been covered with talc. These trays served as foraging areas: food was there delivered (larvae of Tenebrio molitor three times per week; sugar water delivered in small plugged tubes), and experiments were performed in these trays. The lighting varied between 330 and 110 lux, the humidity always equaled 80%, the temperature constantly equaled 20°.

Products provided to the ants

The herbicide, at the concentration advised for humans’ use, was spilled on the bottom of the trays around the food and the nest entrance. The other products were provided to the ants at the adequate concentration (10 times more than that advised for humans) in their usual sugar water given in their usual sugar water tubes. The cotton plugs of these tubes were refreshed, and their entire contents were renewed as necessary. Control experiments were firstly conducted on ants normally maintained. Then, the product, the effects of which must be studied, was provided to the ants, and the test experiments started 12 to 24 later.

Assessment of ants’ traits potentially affected by the delivered products the effects of which must be examined

The ants’ food consumption and activity were assessed by separately counting several times per day, during six days, the ants being on their meat, at their sugar water tube entrance, and active at any part of their territory, by establishing the daily means of these counting, and finally the mean of these six daily means. The ants’ linear speed, angular speed, and orientation were assessed by recording ants’ trajectories and analyzing them thanks to adequate software, then establishing the median and the quartiles of the recorded values. The ants’ audacity was evaluated by counting those coming onto an unknown apparatus, and establishing the mean and extremes of the recorded numbers. The ants’ tactile perception was quantified by assessing their linear and angular speeds while they walked on a rough substrate (if perceiving the uncomfortable character of such a substrate, the ants walk more slowly and sinuously as usual), and by establishing the median and quartiles of the obtained values. The ants’ brood caring was quantified by removing larvae from the nest, then counting those among ten ones not yet re-entered over time. The ants’ social relationships were quantified by counting, in the course of dyadic encountering, the number of times an ant of the pair did nothing (level 0), contact the opponent with its antennae (level 1), opened its mandibles (level 2), gripped the opponent (level 3), tried to sting or stung the opponent (level 4). The results of ten experiments were correspondingly added, and a variable ‘numbers of levels 2 + 3 + 4 / levels 0 + 1’ was each time calculated. The ants’ state of stress and cognition was evaluated by counting those escaped over time from an enclosure, and correspondingly adding the results of several experiments. The ants’ cognition was evaluated by counting those having and having not be able to cross a twists and turns path over time, and correspondingly adding the numbers obtained during several experiments. The ants’ learning and memory was appreciated through their ability to acquire conditioning, then their loss of it after the cue removal, and by adding the results of two such experiments. The cue was a blue or green or yellow hollow cube set near the food. The ants’ adaptation as well as habituation to the side as well as the wanted effects of a product were assessed by quantifying, several days after having lived with this product, one or a few traits affected by the product, and comparing the results with those obtained soon after the ants lived with the product. The decrease of the effect of a product was studied by replacing at a given time the product solution by the ants’ usual sugar water, and quantifying over time one or two traits affected by the product until the obtained values equaled those corresponding to ants normally maintained. The mathematical function of such a decrease was tried to be defined.

Remark

The here given table is new. The photos are never published ones: taking several photos of each experiment, we could select not yet published ones.

The herbicide Herbatak

The herbicides are a kind of products, used by humans, which pollute the environment and are not studied to a large expend as are insecticides and drugs. The most toxic ones are no longer authorized. Those the active substance of which is pelargonic acid can still be used. We studied the side effects of one of them, largely used and easily available: herbatak®. On ants, this product decreased the food consumption, orientation, audacity, tactile perception, social relationships (Figure 1), cognition, learning (Table 1) and memory. No adaptation occurred to these side effects. The effect of Herbatak® vanished in 24 hours after its used was stopped. On the basis of these results, it can be deduced that the fauna of the areas treated with such kind of herbicides is impacted, and has its health even its life imperiled. In other words, though duly weeded, such treated areas have their fauna affected and may even be deprived of their fauna which is essential for efficiently cultivating. We thus advise to use herbicides the active substance of which is pelargonic acid only for not cultivated areas e.g., (sidewalks, driveways, parking lots) and never for areas on which plants are cultivated (e.g., gardens, areas with cultivated fruits and vegetables). We also advise to do so avoiding polluting the environment and the natural water, as well as taking care of the users’ health (e.g., their eyes and skin).

Figure 1 For each product, a photo of one of their side effects, except for PQQ, the photo being that of a wanted effect. Of course, each product had many other not illustrated effects reported in the text. Herbatak: decreased the ants’ brood caring; Imodium: decreased the ants’ ability to escape from an enclosure, so their cognition; Epsipam: induced some aggressiveness towards congeners, impacting thus the social relationships; Motilium: leaded to dependence on its consumption (the tube with a red dot contained the drug); Cognizing: did not affect the ants’ conditioning acquisition; the ant duly responded to the learned green cube; PGG: increased the number of ants coming onto an unknown apparatus, and thus their activity and audacity.

Imodium

Imodium is an easily available drug which allows efficiently treating diarrhea. Its active substance is loperamide, an opiate derivative. Few adverse effects of it are commonly reported, but not easily available bibliography informs that it has some important side effects and can lead to abuse. We thus examined on ants as models the potential side effects of Imodium, and found that this drug affected the food consumption (Table 1), locomotion, audacity, social relationships, cognition (Figure 1), learning and memory. No adaptation occurred to these side effects, and strong dependence developed. The effect of Imodium vanished in about 13 hours after weaning. On the basis of its efficiency and of the adverse effects we found, we advise to treat patients suffering from diarrhea thanks to Imodium, but to do so during the shortest possible time period and to monitor them as for their food intake, locomotion, social relationships, cognitive abilities and dependence of this drug consumption. Concerning the last advice, it must be noted that Loperamide consumption is increasing in the United States since high dose of this substance can lead to euphoric effects.⁷⁸ Consumers should thus be monitored as for their health and their potential increase of Loperamide consumption.

Tetrazepam (Epsipam)

The myorelaxant drugs, the active substance of which is Tetrazepam (e.g., Epsipam), are no longer authorized in some European lands, but are still largely used in other countries. In order to have a personal opinion about these two different directives, we studied on ants as models, the ethological and physiological potential side effects of Epsipam. Effectively, this drug impacted the ants’ food consumption, activity, orientation, audacity (Table 1), sensory perception, social relationships (Figure 1), state of stress, cognition, learning and memory.  Adaptation to such side effects occurred, and ants developed dependence on Epsipam consumption. On the basis of these results, we concluded that the European withdrawing is valid. However, drugs containing Tetrazepam are more efficient than the other kinds of myorelaxants. Therefore, we propose to treat patients with such efficient drugs, but using the smallest possible dose during the shortest possible time period, and to monitor the patients as for the occurrence of the side effects we revealed in ants, e.g., impairments of activity, locomotion, social relationships, stress, cognitive abilities, memory and dependence on the drug consumption.

The antiemetic Motilium

Motilium, the active substance of which is domperidone, is an efficient largely used antiemetic drug, which is nowadays no longer authorized in France. Several of its side effects are divulgated, but nothing is mentioned about important potential physiological and ethological impacts. We thus studied such impacts on ants as models, and found that Motilium affected these insects’ food consumption, activity (Table 1), locomotion, orientation, audacity, sensory perception, social relationships, state of stress, cognition, learning and memory. No adaptation to these side effects occurred, and ants developed dependence on Motilium consumption (Figure 1), the effect of which vanished in about 12 hours after weaning. Clinicals analysis now exist to predict if patients will strongly suffer from side effects of Motilium.⁴² Since this drug is really very efficient, we propose to still use it, but after having made the required medical exam, then, to treat the patients with the smallest possible dose and during the shortest possible time period, and to simultaneously monitor them as for the occurrence of the here reported side effects, such as a decrease of activity, some locomotor function disfunctioning, social relationships impairments, increase of stress, dependence on the drug consumption.

Cognizing

Cognizing is a dietary supplement, the active substance of which is citicoline, a drug allowing caring of persons suffering from cerebral disorders. It is thus used for potentially improving the brain functioning of human adults. Dietary supplements are not always effectively efficient, are often expensive, and may present some side effects. We thus studied on ants as models the potential wanted and side effects of Cognizing. We discovered that this dietary supplement caused stress, agitation, and nervousness, and consequently impacted the ants’ food intake, locomotion (Table 1), orientation, audacity, and social relationships. It did not affect the ants’ sensory perception, cognitive abilities, and memory (Figure 1). It may slightly improve the ants’ memory. No adaptation to the side effects occurred, as well as no habituation to the wanted effect occurred. The ants developed a strong dependence on Cognizing consumption, and the effect of this dietary supplement rapidly vanished after weaning in a total of only 11 hours. To conclude, Cognizing is not well appropriated for rapidly improving the brain functioning; it only slightly ameliorates the memorization. Also, it presents several side effects, essentially stress, nervousness, and a strong dependence on its consumption. We do not recommend it. Many expensive dietary supplements are inadequately efficient and cause several adverse effects. Humans should pay attention while using them, and stop to use them if health problems occur. On the other hand, citicoline is an efficient drug allowing patients recovering after having had or while having some cerebral damage.

PQQ

PQQ is a recently used drug as well as a dietary supplement in order to improve humans’ brain functioning and activity. Its wanted effects are rather well known, but its potential adverse ones are not yet so. This is why we studied such potential ethological and physiological side effects of PQQ using ants as biological models. It is the first time, all along our study of until now 56 products used by humans, that we found so few slight adverse effects and obvious beneficial ones. Indeed, PQQ favorably affected the ants’ activity, audacity (Figure 1), cognitive abilities, learning and memory. It did not impact or very slightly affected all the other examined biological traits. In fact, PQQ increased the ants’ sugar water consumption (Table 1) and sinuosity of movement. In addition, the ants did not develop dependence on this dietary supplement consumption, the effect of which regularly, linearly decreased after weaning, and vanished in a total of 33 hours. PQQ is thus not only a drug which could help caring of patients (e.g., suffering from cerebral artery occlusion or Alzheimer’s disease, requiring neuroprotectant or improvement of their mitochondria functioning), but is also a promising dietary supplement allowing improving humans’ (in good health) activity and brain functioning. However, all the effects of PQQ are not yet well known. Therefore, as also reported in the notice for use joined to the product package, we recommend to consume it with moderation ant to pay attention to any health problems occurrence.

For each examined product, the results of the different conducted experiments were in agreement with each other, and were also in agreement with the observations made by practitioners, pharmacists, and other researchers. In addition, each time, we could precise previously made observations (e.g., on the decrease of the effect of the product after its consumption was stopped), and add novel or not divulgated ones (e.g., impact on food intake, social relationships, stress, cognitive abilities). For each examined product, we could finally define how using it safely, at low cost. All these novel information should be known by practitioners, pharmacists and patients. As many times previously observed (e.g.⁷⁹), a dependence on a product developed when the effect of the product rapidly decreased after weaning. In general, for each of the six here studied products, as well as for the previously studied ones and for any other similar products, attention should be paid concerning its use, efficiency and safety, because, even if meticulous, honest and statistically significant experimental researches have been made, drugs, dietary supplements (above all) and several other products are produced with the aim to earn money, and are sold under well-organized marketing. Concerning the medicinal analysis of drugs, dietary supplements, pesticides and similar products, before experimenting on humans and even mammals, studies on ants could advantageously be made. Indeed, ants appeared to be excellent biological models, capable to reveal easily, rapidly, at low cost many physiological and ethological impacts of the product, though, of course, these studies are only the first step of the entire examination of the product.

For each examined product, on the basis of all our observations, we could define a procedure for use we here below summarize.

1. Herbatak (pelargonic acid): use as an herbicide; only for not cultivated area, paying attention to not contaminate other areas and natural water, as well as to the users’ eyes and skin.
2. Imodium (Loperamide): use to stop diarrhea; during the shortest possible time period, and paying attention to the users’ food intake, locomotion, social relationships, cognitive abilities, and above all dependence on its consumption.
3. Epsipam (Tetrazepam): use as a myorelaxant; at the smallest possible dose, during the shortest possible time period, paying attention to the users’ activity, locomotion social relationships, stress, cognition, memory and dependence on this drug use.
4. Motilium (Domperidome): use as an antiemetic drug; after having made the required medicinal analysis allowing knowing the users’ potential reactions to the product, then treating them shortly with small amount, and paying attention to their health, side effect being susceptible to occur.
5. Cognizing (citicoline): use as a dietary supplement for “improving humans’ brain functioning”; poorly efficient and induces stress, agitation, nervousness, strong dependence; to be avoided, not to use.
6. PQQ: use as a dietary supplement for improving humans’ activity and brain functioning; no adverse e ffect, efficient, recommended (but expensive).

We are very grateful to R. Cammaerts and D. Cammaerts who helped us for the writing, the bibliography and the statistical analysis of the present summarized six works.

We affirm having not conflict of interest as for the use of any of the here examined products. We work on ants and receive not money for making our research. We maintain the ants under the best possible conditions and collect on field the smallest possible number of colonies.

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