Magnoflorine from Berberis vulgaris Roots-Impact on Hippocampal Neurons in Mice after Short-Term Exposure.

In search of novel potential drug candidates that could be used as treatments or prophylactics for memory impairment, an aporphine alkaloid magnoflorine (MAG) isolated from the root of Berberis vulgaris was proven to exhibit beneficial anti-amnestic properties. Its effects on immunoreactivity to parvalbumin in the mouse hippocampus were assessed together with a study on its safety and concentration in the brain and plasma. For this purpose, four experimental groups were created: the MAG10 group-treated with 10 mg MAG/kg b.w. i.p., the MAG20 group-treated with 20 mg MAG/kg b.w. i.p., the MAG50 group-treated with 50 mg MAG/kg b.w. i.p., and a control group-injected with saline i.p. at a volume corresponding to their weight. Our results indicated that the hippocampal fields CA1-CA3 were characterized by an elevated number of parvalbumin-immunoreactive neurons (PV-IR) and nerve fibers in mice at the doses of 10 and 20 mg/kg b.w. (i.p.). No significant changes to the levels of IL-1ß, IL-6 or TNF-α were observed for the above two doses; however, the administration of 50 mg/kg b.w. i.p. caused a statistically significant elevation of IL-6, IL-1beta plasma levels and an insignificant raise in the TNF-alpha value. The HPLC-MS analysis showed that the alkaloid's content in the brain structures in the group treated with 50 mg/kg b.w. did not increase proportionally with the administered dose. The obtained results show that MAG is able to influence the immunoreactivity to PV-IR in hippocampal neurons and might act as a neuroprotective compound.

Peroxidation and unsaturation indices as potential biomarkers of multifarious zinc and copper micro-supplementation in Helix pomatia L.

The work is a continuation of two previous studies in which biomarker fatty acids (12 of 56 FA pools) were analysed in Helix pomatia L. after heterogeneous micro-supplementation of Zn and Cu (administered in five micro-doses in the form of salts and EDTA and lysine chelates). This time, peroxidation (PI) and unsaturation coefficients (UI) as biomarker were analysed. These indices were calculated based on the FA profile in the foot and hepatopancreas of snails. The correlation of frequently used oxidation status indicators of organisms (catalase - CAT, glutathione peroxidase - GPx, selenium-dependent peroxidase - se-GPx, superoxide dismutase - SOD, glutathione transferase - GST, glutathione reductase - GR, glutathione - GSH, carbonyl protein - CP, thiobarbituric acid reactive substances - TBARS) with the rarely used UI and PI ratios was analysed. It was found that the 12-week micro-exposure to Zn and Cu did not inhibit but rather stimulated antioxidative defence at a sufficient level to increase the values of peroxidation/unsaturation indices in comparison to the control groups. Induction of an opposite process to oxidation of fatty acids was demonstrated. Maximum activities and amounts of antioxidants as well as minima of protein and lipid decomposition were recorded in groups supplemented with 0.75mg/l Zn and 1.0mg/l Cu. The possibility of a direct use of fatty acids as well as peroxidation/unsaturation indices as sensitive and reproducible biomarkers of exposure and oxidative physiological status in snails was confirmed.

Comparative effect of different dietary inulin sources and probiotics on growth performance and blood characteristics in growing-finishing pigs.

The study was focused on assessment of the effect of an extract of long-chain inulin (LCI) and dried tubers of Jerusalem artichoke (JA) and a multispecies probiotic preparation as well as a combination thereof on growth performance and blood parameters of fattening pigs. In total, 144 pigs (initial body weight 30.0 ± 0.5 kg) were used in a 98-d experiment. The six dietary treatments consisted of the control diet (Con), diet Con supplemented with probiotics (ConP) and four diets supplemented with LCI or JA alone or with probiotics (diets LCIP and JAP). Throughout the fattening period, there was a beneficial effect of the probiotic supplementation to the inulin-containing diets and the average daily gain (ADG) was increased by supplementation of probiotics in combination with inulin sources (p < 0.05). At the end of the fattening period, ADG and feed conversion ratio (FCR) were higher after supplementation of LCI only (p < 0.05). Compared with group ConP, in groups LCI and JA, the ADG and FCR were improved (p < 0.05). Only in the first fattening stage, the addition of the prebiotics and/or probiotics had an impact on the level of white blood cells and some biochemical indices in pigs. In younger animals, probiotic or LCI supplementation increased the IgG level (p < 0.05). There was also an interaction between the probiotics and JA resulting in increased IgG and IgA concentrations (p < 0.05). In the finishing period, LCI addition increased the IgM level (p < 0.05), whereas JA addition increased IgG and IgM levels as well (p < 0.05). In conclusion, both dietary sources of inulin and probiotic supplementation can improve the fattening performance and health status of growing pigs.

Increased seizure susceptibility and other toxicity symptoms following acute sulforaphane treatment in mice.

Activation of Nrf2 with sulforaphane has recently gained attention as a new therapeutic approach in the treatment of many diseases, including epilepsy. As a plant-derived compound, sulforaphane is considered to be safe and well-tolerated. It is widely consumed, also by patients suffering from seizure and taking antiepileptic drugs, but no toxicity profile of sulforaphane exists. Since many natural remedies and dietary supplements may increase seizure risk and potentially interact with antiepileptic drugs, the aim of our study was to investigate the acute effects of sulforaphane on seizure thresholds and activity of some first- and second-generation antiepileptic drugs in mice. In addition, some preliminary toxicity profile of sulforaphane in mice after intraperitoneal injection was evaluated. The LD50 value of sulforaphane in mice was estimated at 212.67mg/kg, while the TD50 value - at 191.58mg/kg. In seizure tests, sulforaphane at the highest dose tested (200mg/kg) significantly decreased the thresholds for the onset of the first myoclonic twitch and generalized clonic seizure in the iv PTZ test as well as the threshold for the 6Hz-induced psychomotor seizure. At doses of 10-200mg/kg, sulforaphane did not affect the threshold for the iv PTZ-induced forelimb tonus or the threshold for maximal electroshock-induced hindlimb tonus. Interestingly, sulforaphane (at 100mg/kg) potentiated the anticonvulsant efficacy of carbamazepine in the maximal electroshock seizure test. This interaction could have been pharmacokinetic in nature, as sulforaphane increased concentrations of carbamazepine in both serum and brain tissue. The toxicity study showed that high doses of sulforaphane produced marked sedation (at 150-300mg/kg), hypothermia (at 150-300mg/kg), impairment of motor coordination (at 200-300mg/kg), decrease in skeletal muscle strength (at 250-300mg/kg), and deaths (at 200-300mg/kg). Moreover, blood analysis showed leucopenia in mice injected with sulforaphane at 200mg/kg. In conclusion, since sulforaphane was proconvulsant at a toxic dose, the safety profile and the risk-to-benefit ratio of sulforaphane usage in epileptic patients should be further evaluated.

Changes in fatty acid metabolism induced by varied micro-supplementation with zinc in snails Helix pomatia (Gastropoda Pulmonata).

We analyzed the changes in the profile of fatty acids (FA) in the foot tissues and hepatopancreas (HP) of snails Helix pomatia exposed to five microdoses of zinc (0.1, 0.25, 0.5, 0.75, or 1mg/l) administered in the form of a pure salt solution and in the form of EDTA and lysine chelates. Selection from a pool of 56 fatty acids analyzed in snail tissues yielded a set of 12 biomarker acids undergoing significant changes in contact with toxic substances. The selection criteria included the greatest percentage among the FA profile and their significant role in physiological processes. The proposed palette of acids of the biomarker FAs comprised C16:0; C18:0; C23:0; C18:1 n-9; C20:1 n-9; C18:2 n-6; C18:3 n-3; C20:2; C20:4 n-6; C20:5 n-3; C22:4 n-6; and C22:5 n-3, and saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), and polyunsaturated fatty acids (PUFAs), determined separately in the foot tissues and hepatopancreas. The significant (p=0.01) influence of the dose as well as the source of the zinc on its' concentration in the tissues and on changes in the fatty acid profiles. Among the three zinc forms administered to the snails, the highest bioaccumulation of zinc in both tissues was noted in the group receiving the Zn-EDTA chelate. The content of PUFAs increased as the supplementation with zinc increased up to 0.75mg/l, but at 1mg/l, the share of these FAs began to decrease. This trend was observed in both analyzed tissue types - foot and hepatopancreas. The dose of 1mg Zn/l might be considered as a threshold dose above which the saturation of FAs increases. The results proved that determination of FA profile in snails can be used in ecotoxicological research as a reliable test of the effect of trace doses of stressors. The micro-supplementation of the mollusks diet with zinc is an example of a non-routine approach to issues connected with both diet and toxicology.

N-3 Polyunsaturated Fatty Acids as a Nutritional Support of the Reproductive and Immune System of Cattle-A Review.

This paper focuses on the role of n-3 fatty acids as a nutrient crucial to the proper functioning of reproductive and immune systems in cattle. Emphasis was placed on the connection between maternal and offspring immunity. The summarized results confirm the importance and beneficial effect of n-3 family fatty acids on ruminant organisms. Meanwhile, dietary n-3 fatty acids supplementation, especially during the critical first week for dairy cows experiencing their peripartum period, in general, is expected to enhance reproductive performance, and the impact of its supplementation appears to be dependent on body condition scores of cows during the drying period, the severity of the negative energy balance, and the amount of fat in the basic feed ration. An unbalanced, insufficient, or excessive fatty acid supplementation of cows' diets in the early stages of pregnancy (during fetus development) may affect both the metabolic and nutritional programming of the offspring. The presence of the polyunsaturated fatty acids of the n-3 family in the calves' ration affects not only the performance of calves but also the immune response, antioxidant status, and overall metabolism of the future adult cow.

Barley, Triticale, or Rye? The Type of Grain Can Affect the Growth Performance and Meat Quality of Sustainable Raised Pigs.

The effect of high level of barley, triticale, or rye as base of complete mixtures for growing-finishing pigs on growth performance, carcass traits, meat quality, and fatty acid profile in meat and backfat was investigated. The 100-day experiment involved 72 pigs, assigned into three groups (24 animals each). Pigs of each group were kept in six pens (two gilts and two barrows per pen). Diets offered to pigs differed in the proportion of cereals as the leading ingredients in the mixture formulation: I-barley, II-triticale, III-rye. The results showed diversified grain influence on the production results and meat quality. Triticale- and barley-based diets ensured better weight gain and lower carcass fatness than rye (p ≤ 0.05). The basic nutrients digestibility of mixtures containing triticale was comparable to that containing barley and higher than that of rye (p ≤ 0.05). The meat and backfat of pigs receiving diet with triticale or barley was characterized by more favorable fatty acids profile in respect to the health-promoting indicators (atherogenicity and thrombogenicity indexes; hypocholesterolemic/hypercholesterolemic ratio). The cholesterol level in various tissues was the lowest in pigs fed with rye diet, and their meat characterized by better water holding capacity and more SFA. Higher fat saturation indicate better resistance to oxidation during storage and longer meat shelf life. It seems that the supplementation of triticale to diet may improve the growth efficiency of pigs and the health-promoting value of meat, while the supplementation of rye may be better for the production of traditional or long-matured meat products.

Effects of Supplementation with an Herbal Mixture on the Antioxidant Capacity of Milk.

The aim of this study was to assess the effect of the addition of a standardized herbal mixture to the feed ration for Holstein-Friesian cows on the antioxidant capacity of milk. The study was carried out on a farm specialized in breeding dairy cattle. The exact study involved 30 cows in lactation III, which were in the first phase of lactation at the beginning of the experiment (15 cows-control group; 15 cows-experimental group). The nutrition supplied to the cows was based on the TMR (total mixed ration) system, with roughage and concentrate fodder used as the basis of the feed ration. The addition of a standardized blend of dried herbs, i.e., oregano (Origanum vulgare), thyme (Thymus vulgaris), purple coneflower (Echinacea purpurea), and cinnamon bark (Cinnamomum zeylanicum), was the experimental factor. Powdered herbs were administered as a component of the concentrate fodder at the dose of 3% DM ration/day/head. Milk samples were collected four times during the experiment (term 0 after the colostrum period and then after lactation weeks 2, 4, and 6). The following parameters were determined in the milk: the basic chemical composition, i.e., the content of total protein, fat, lactose, and casein; somatic cell count; content of selected whey proteins (α-lactalbumin, ß-lactoglobulin, lactoferrin, BSA); and fat-soluble vitamins (A, D3, E). Additionally, the milk antioxidant capacity (ABTS, FRAP, DPPH) was determined and the degree of antioxidant protection (DAP) was calculated. It was shown that the milk from cows receiving the herbal blend-supplemented fodder had a higher content of casein, compared to the control group. The herbal supplementation contributed to a significant increase in the content of bioactive compounds, i.e., selected whey proteins (ß-lactoglobulin, lactoferrin) and lipophilic vitamins (A, E). The milk was also characterized by significantly higher antioxidant potential (regardless of the measurement method) and a higher degree of antioxidant protection (DAP).

The Effect of a Diet Supplemented with Organic Minerals and l-Carnitine on Egg Production and Chemical Composition and on Some Blood Traits of Pheasant Hens (Phasianus colchicus).

The study aimed to determine the effect of replacing 75% of inorganic calcium, iron, zinc, and copper salts with organic forms (glycine chelates of these elements) with or without the addition of l-carnitine on some reproductive traits and the blood lipid and mineral profile, as well as mineral and fatty acid profile of pheasant egg yolk. The study was performed on three groups of pheasant hens using glycine chelates with calcitriol (group II) or analogical treatment with the addition of l-carnitine at the level of 100 mg/kg of feed (group III) instead of Ca, Fe, Cu, and Zn salts (control). The replacement of inorganic forms with glycinates contributed to an increase in the number of laid eggs with a concomitant lower share of rejected eggs. The supplementation of organic forms of minerals improved mineral absorption and bioavailability in blood serum as well as in the egg yolk of experimental groups. Egg yolk fat was characterized by a higher proportion of polyunsaturated fatty acids and a favorable ratio of PUFA ω-3/ω-6. The proposed nutritional supplementation of the pheasant's diet might be a good strategy for increasing the nutritional reserves of poultry and improving their reproduction.

Can Bioactive Compounds in Beetroot/Carrot Juice Have a Neuroprotective Effect? Morphological Studies of Neurons Immunoreactive to Calretinin of the Rat Hippocampus after Exposure to Cadmium.

Cadmium ions (Cd2+) penetrate the blood-brain barrier and can, among other effects, influence intracellular calcium metabolism, leading to neurodegeneration. In the presented work, we estimated the effect of Cd2+ on the expression of calretinin in the neurons of the rat hippocampus and analyzed the reverse effect of freshly pressed beetroot/carrot juice in this context. In the 12-week lasting experiment, 32 8-week-old male Wistar rats were divided into four experimental groups (n = 8): the control group (C) received pure tap water; the Cd group (Cd)-received Cd2+ dissolved in tap water (5 mg Cd2+/kg b.w.); and two groups received beetroot/carrot juice: the BCJ group was administered only juice, and the Cd + BCJ group received juice with the addition of Cd2+ (5 mg Cd2+/kg b.w.). The exposition to low doses of Cd2+ caused a significant decrease in calretinin-immunoreactive (Cr-IR) neurons compared to the non-exposed groups. Moreover, the addition of Cd2+ to tap water reduced the numbers and length of Cr-IR nerve fibers. The negative effect of Cd2+ was significantly attenuated by the simultaneous supplementation of beetroot/carrot juice (Cd + BCJ). The study showed that the bioactive compounds in the beetroot/carrot juice can modulate Ca2+ levels in neurons, and thus, potentially act as a neuroprotective factor against neuronal damage.

Effect of heat treatments on the physicochemical and sensory properties of the longissimus thoracis muscle in unweaned Limousin calves.

The aim of the study was to assess the effect of methods of heat treatment on selected quality parameters of longissimus thoracis muscle of Limousin calves, subjected to grilling, steaming, and sous vide cooking. The type of heat treatment did not significantly affect shear force or water activity. Cooking loss in the grilled and steamed meat was significantly higher than in the sous vide. While the content of vitamin E was significantly lower in the cooked meat, it was retained to the greatest degree in the grilled meat and least in the steamed meat. The TBARS index significantly increased during all cooking methods with the highest level in steamed meat. The sensory analysis revealed a preference for the sous vide and grilled meat, while the steamed meat received the lowest scores. Although veal is commonly assumed to be highly nutritious and palatable, its quality can be significantly influenced by the type of heat treatment.

The Impact of Zn, Cu and Fe Chelates on the Fatty-Acid Profile and Dietary Value of Broiler-Chicken Thigh Meat.

This study aimed to compare the effect of Zn, Cu and Fe glycine chelates on the proximate composition, cholesterol levels, fatty-acid profile and dietary value of the thigh meat of broiler chickens. The experiment involved three hundred and fifty Ross 308 chickens divided into seven groups. The chickens were administered Zn, Cu and Fe glycine chelates in an amount corresponding to 50% of the requirement or 25% of the requirement for 42 days. It was found that the use of Zn, Cu and Fe glycine chelates did affect the fatty acid profile and dietary value of meat. A positive impact was most frequently (p < 0.05) noted in chickens receiving Zn chelate in an amount covering 50% of the requirement: the lowest levels of SFA and atherogenic and thrombogenic indices, the highest content of PUFA n-3 and PUFA/SFA ratios and hypocholesterolemic/hypercholesterolemic indices. Positive effects were more often recorded for chickens receiving Zn in an amount corresponding to 50% of the requirement. The results did not show that the use of Cu and Fe glycine chelates can reduce the dietary value of thigh meat in broiler chickens since, generally, the outcomes were not worse than those in the control group. It should be highlighted that due to ambiguous results, it is impossible to determine a dose of Cu and Fe glycine chelate which would be more efficient for broiler chickens. However, chickens receiving chelates in amounts corresponding to 25% of the requirement showed far better results.

Nutritional and Anti-Nutritional Factors in Vicia sativa L. Seeds and the Variability of Phenotypic and Morphological Characteristics of Some Vetch Accessions Cultivated in European Countries.

Agricultural research has traditionally focused on staple crops, while relatively little attention has been given to minor crops. Therefore, this study aimed to determine the nutrient contents and some anti-nutritional factors in the seeds of common vetch (Vicia sativa L.) and the variation of phenotypic and morphological traits in field studies of 44 European accessions, originating from Russia, Ukraine, Poland (east and east-central Europe), the Czech Republic, the Slovak Republic, Hungary (south-central Europe), and Germany (west-central Europe) and grown in the same soil-climate conditions. A three-year field study was conducted from 2010 to 2012. Accessions originating from west-central Europe flowered three days earlier than those from south-central Europe. They also had the lowest seed number per pod (5.9) but the highest thousand seed weight (58 g) (p < 0.05). Vetch lines coming from south-central Europe contained the highest level of crude protein in comparison with vetch seeds originating from west-central Europe (353 vs. 324 g kg-1, respectively) (p < 0.05), but the latter had the highest essential amino acid index value (75 vs. 71 in west-central Europe and south-central Europe, respectively) (p < 0.05). The highest protein level was noted in the seeds of Slovak origin (average 358 g kg-1), whereas the lowest protein level (324 g kg-1) was determined in the Russian and German lines. Vetch grain fat was rich in linoleic (53%) and linolenic (14%) acids. The best n-6/n-3 fatty acid ratio (4) and the highest α-linolenic acid level were exhibited by the Slovak and Polish accessions (p < 0.05). The seeds of vetch accessions from east, east-central, and south-central Europe contained higher levels of (p < 0.05) iron than those originating from west-central Europe. The concentration of tannins, trypsin inhibitors, and hydrogen cyanide reached on average 8, 3, and 81 mg kg-1, respectively. The highest hydrogen cyanide (HCN) levels was determined in the accessions of German and Russian origin (126 and 119 mg kg-1, respectively), and the lowest values were noted in the Slovak and Polish lines (50 and 67 mg HCN kg-1, respectively). Furthermore, the Polish accessions contained the lowest levels of tannins and trypsin inhibitors. Common vetch seeds may be valuable protein feed/food stuff, but their nutritional usefulness is limited by the presence of anti-nutritional factors, especially HCN, which is difficult to remove. Further selection in this direction may be postulated.

Peroxidation and unsaturation coefficients as biomarkers of environmental micro-exposure to molluscicides in Helix pomatia L.

This study was a follow-up of a previous study that investigated a set of selected fatty acids (FAs; 12 of 56 pools) in Helix pomatia L. as biomarkers of chemical stress induced by applying micro-doses of molluscicides. Here, the potential of rarely used peroxidation (PI) and unsaturation (UI) coefficients were tested as biomarkers. These indices were calculated based on the FA profiles of foot and hepatopancreas tissues of H. pomatia L. Snails were treated with three molluscicides: metaldehyde, methiocarb, and potassium chloride, in three doses each (5, 10, or 15 µl, 0.01% w/v concentration), for 12 weeks, under laboratory conditions. Correlations were evaluated between frequently used oxidation status indicators (catalase, glutathione peroxidase, selenium-dependent peroxidase, superoxide dismutase, glutathione transferase, glutathione reductase, glutathione, carbonyl protein, and thiobarbituric acid reactive substances, in the form of MDA) and UI and PI ratios. These results confirmed that fatty acids could be directly used as biomarkers of exposure and oxidative physiological status in snails. Moreover, the UI and PI, calculated based on FAs, clearly reflected the current oxidation status in snails. These indices changed with the application of micro-doses of molluscicides. In conclusion, these indices could serve as sensitive biomarkers of chemical stress in snails.

Dietary Intake and Content of Cu, Mn, Fe, and Zn in Selected Cereal Products Marketed in Poland.

This paper aims to verify whether cereal products are a good source of Cu, Mn, Fe, and Zn in the diets of Poles. The study material comprised of 445 cereal products including baked goods, breakfast cereals and groats, pasta, and rice. Products that required culinary treatment (pasta, groats, rice) were boiled in drinking quality water as recommended by the producer. The content of Cu, Mn, Fe, and Zn was determined by using the FAAS method. The average content of microelements in the analyzed products can be represented as Fe (17.9 mg kg-1 ± 10.3) > Zn (12.4 mg kg-1 ± 5.2) > Mn (9.6 mg kg-1 ± 6.7) > Cu (3.9 mg kg-1 ± 2.9). Considering the daily requirement of the analyzed minerals among adult Poles, it was determined that cereal products supply 58% RDA Cu, 61% AI Mn (men)-78% AI Mn (women), 19% RDA Fe (women)-34% RDA Fe (men), and 16% RDA Zn (men)-22% RDA Fe (women). Baked goods account for as much as about 90% of all Cu, Mn, Fe, and Zn consumed as cereal products in a daily diet. Based on the results, it can be concluded that cereal products are the main source of Cu, Mn, Fe, and Zn in the diets of Poles. In connection with low assimilability of minerals, they should not be considered the fundamental source of those microelements in the diets of Poles.

Estimation of weekly intake of cadmium and lead by consumption of commercial ready-to-feed infant foods.

The purpose of this survey was to estimate the safety of ready-to-eat infant foods in terms of Pb and Cd content. The studied samples were ready-to-eat infant products: dinners (n = 74), soups (n = 27) and desserts (n = 82) containing components of animal origin: meat and/or milk. Cd and Pb content was determined using a GF-AAS method. The analysed products contained 1.82-3.54 µg Pb and 1.32-1.50 µg Cd per kg. The content of Pb per kg of the product can be represented as dinners > soups > desserts, whereas the content of Cd was dinners > desserts > soups. The analysed ready-to-eat products could be regarded as safe, because they supply 12-month-old infants with Pb in an amount accounting for nearly 22% BMDL01 and Cd accounting for ca. 8.6% of the total weekly intake.

Selected fatty acids as biomarkers of exposure to microdoses of molluscicides in snails Helix pomatia (Gastropoda Pulmonata).

Two stages of selection from a pool of 56 fatty acids analyzed in Helix pomatia yielded a set of 12 biomarker acids undergoing significant changes in contact with three microdoses of toxic substances, i.e. three molluscicides containing metaldehyde, methiocarb, and potassium chloride (PC). The proposed palette of acids, including saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), and polyunsaturated fatty acids (PUFA), determined separately in the foot tissues and hepatopancreas of Gastropoda, can be used in ecotoxicological research as a reliable test of the effect of trace doses of stressors. The final set of the biomarker FA comprised C16:0; C18:0; C23:0; C18:1 n-9; C20:1 n-9; C18:2 n-6; C18:3 n-3; C20:2; C20:4 n-6; C20:5 n-3; C22:4 n-6; and C22:5 n-3. A clear physiological response manifested as changes in the content of fatty acids (FA) was observed in the snails even in the case of the lowest doses of the pollutants. All experimental factors analyzed, i.e., the dose (5, 10, or 15 µl 0.01% w/v concentration) and the type of preparation (metaldehyde, methiocarb or PC), had a significant (p ≤ 0.01) impact on the FA composition of the foot and hepatopancreas. Limitation of the analysis to a narrow pool of reactive FA meets the requirements of parameters of biomarkers of exposure and facilitates and accelerates visualization of the bioindicator organism's response to the presence of the stressor in the environment.