Reproductive ability of queen bees under the influence of heavy metals and fatty acids of bee pollen

Bees feel the smell and taste of various fatty acids and amino acids of plant pollen [39].

The smell of amino acids and especially fatty acids, unlike pollen sterols, strongly attracts bees to honey plants [26, 39, 40]. At the same time, among the fatty acids of plant pollen, which attract the special attention of honey bees, there are monounsaturated and polyunsaturated fatty acids containing 16 or more carbon atoms in their composition [40]. At the same time, the fatty acids of plant pollen are much more valuable for the body of bees in terms of energy than amino acids [14, 22, 39, 40]. The attractive function of fatty acids in plant pollen is due to their relatively high volatility and specific smell [34], and the energetic function is due to the high ratio of hydrogen and carbon atoms to oxygen atoms in their composition [34, 40].

In addition, monounsaturated and polyunsaturated fatty acids of plant pollen provide antibacterial and antifungal protection of the body of honey bees, bee combs and beehive [14, 19, 25]. Non-esterified forms of fatty acids are particularly active in this regard [15, 38].

Unsaturated fatty acids (linolenic, linoleic, palmitoleic, oleic, and eicosaenic) of bee pollen have an antimicrobial effect due to their high ability to increase the surface activity of the tissues of microorganisms and thus, under normal osmotic pressure of the surrounding environment, strongly inhibit their vital activity [16, 19].

It was established that the content of linolenic, linoleic, palmitoleic, oleic and eicosaenic acids, which are an attractive and energetic material for bees and provide antimicrobial protection of their body, bee combs and hive, in bee honey obtained from beehives located in the foothills (21.09 g/kg of air-dry mass) and forest-steppe (18.19) zones of the Carpathian region, compared to bee honey taken from hives located in the mountain zone (24.32 g/kg of air-dry mass), is smaller (table. 5).

The main mass of polyunsaturated fatty acids of the omega-3 and omega-6 families in the composition of phospholipids is included in the structure of cellular and cytoplasmic membranes of body tissues and ensures their functional activity and ultimately the vital activity of bees [34]. At the same time, linolenic acid and its longer-chain and unsaturated fatty acids in the body of bees are the initiators of anti-inflammatory processes, and linoleic acid and its longer-chain and unsaturated derivatives are pro-inflammatory [34, 46]. Linolenic and linoleic acids and their longer- chain and unsaturated derivatives act on the body of bees through the corresponding anti-inflammatory and pro-inflammatory cytokines of a peptide nature [21, 36] and such anti-inflammatory and pro-inflammatory eicosanoids as various types of prostaglandins [19].

The effects of pro-inflammatory cytokines, which are abundant in the body of bees, can be characteristic of each tissue separately, are inhibited by certain types of anticytokines [19]. At the same time, there are very few anti-inflammatory cytokines in the body of bees, since they exert their effect on all tissues and do not have inhibitory endogenous anticytokines.

It should be emphasized that pro-inflammatory eicosanoids (certain groups of prostaglandins) in the body of bees are synthesized from polyunsaturated fatty acids of the rn-6 family (eicosatrienoic, eicosatetraenoic-arachidonic, docosatetraenoic) [19]. Data from the literature indicate that the most active pro-inflammatory eicosanoid in the body of bees, especially queen bees, is prostaglandin F2a, which is synthesized in their tissues from eicosatetraenoic-arachidonic acid and, in turn, from linoleic acid [19, 24]. Pro-inflammatory prostaglandin F2a promotes the growth and maturation of sperm and the growth and fertilization of oocytes in the reproductive tracts of drones and queen bees, respectively [24]. Thus through functionally active cell and cytoplasmic membranes and biologically active substances, in particular prostaglandins, unsaturated fatty acids, primarily polyunsaturated fatty acids of the omega-3 and omega-6 families, have the most complete and pronounced effect on the reproductive capacity of bees.

Table 5 shows that the largest amount of structural, biologically active and anti-inflammatory forms of linoleic and linolenic acids is contained in bee pollen of the mountain zone of the Carpathian region (19.74 g/kg of air-dry mass), a smaller amount is found in bee pollen in the foothill zone (17.21), even smaller in the forest-steppe zone (14.88 g/kg of air-dry mass).

A very low level of unsaturated fatty acids, in particular polyunsaturated fatty acids of the omega-3 and omega-6 families in plant pollen, which is the basis of bee pollen, can cause a decrease in the permeability of its structural components to water and water-soluble substances and thereby inhibit the intensity of metabolic processes in it [14, 31]. In turn, a low level of unsaturated fatty acids in bee pollen can contribute to a decrease in the functional activity of cellular and cytoplasmic membranes of the honey bee body and thus inhibit their vital activity [26, 31].

It was established that the total content of unsaturated fatty acids of the omega-3, omega-6, omega-7 and omega-9 families in the abdominal tissues of honey bees obtained from hives located in the foothills (17.29 g/kg of raw mass) and especially forest-steppe (16.36) zones of the Carpathian region, compared to the abdominal tissues of honey bees selected from hives located in the mountain zone (21.33 g/kg of raw mass), is smaller (Table 6). Data from the literature indicate that the relative level of unsaturated fatty acids in some tissues of honey bees and queen bees corresponds to their relative concentration in other tissues [34].

6. The content of unsaturated fatty acids of the omega-3, omega-6, omega-7, and omega-9 families in the abdominal tissues of honey bees in different natural zones of the Carpathian region, g/kg raw weight (M±m, n=3)

It can be seen from Table 6 that the largest amount of monounsaturated and polyunsaturated fatty acids, which can serve as energy and antimicrobial material, is contained in the abdominal tissues of honey bees of the mountain zone of the Carpathian region (21.33 g/kg of raw weight), a smaller amount is in the abdominal tissues bees in the foothill zone (17.29), even less in the forest-steppe zone (16.36 g/kg of raw weight).

It was recorded (Table 6) that the highest level of polyunsaturated fatty acids of the omega-3 and omega-6 families, which are part of the structure of cell and cytoplasmic membranes and are precursors of prostaglandins and initiators of anti-inflammatory and pro-inflammatory processes, respectively, is contained in the abdominal tissues of honey bees of the mountain zone of the Carpathian region (respectively 8.51 and 8.03 g/kg of raw mass), their level is lower in the tissues of the abdomen of honey bees in the foothills (6.81 and 6.47), and even lower in the foreststeppe zone (6.47 and 6.14 g/kg of raw weight).

It should be noted that in the abdominal tissues of honey bees of the mountain zone of the Carpathian region, compared to the foothills and forest-steppe zone, there is a 1.2-1.3 times higher content of the direct precursor of the highly active prostaglandin F2a - eicosatetraenic- arachidonic acid (Table 6). Prostaglandin F2a synthesized from this acid can maintain the reproductive capacity of drones and especially queen bees at a very high level [21].

Thus, in the forest-steppe natural zone of the Carpathian region, among other zones, due to a decrease in the content, the value of monounsaturated and polyunsaturated fatty acids of bee pollen for the organism of bees, bee combs and beehive decreases most intensively.

At the same time, the value of the mentioned fatty acids in the tissues of the abdomen of bees decreases.

The decrease in the content of monounsaturated and polyunsaturated fatty acids in bee pollen and in the tissues of honey bees of the foothills and especially the forest-steppe zones of the Carpathian region, compared to the bee pollen and tissues of honey bees in the mountain zone, is clearly related to their greater transition to the anionic form and the formation of salts. This is due primarily to the binding of fatty acids by heavy metal cations [13, 26].

High levels of heavy metals, including toxic ones, but low levels of unsaturated fatty acids, in particular polyunsaturated fatty acids of the omega-3 and omega-6 families, in bee pollen and abdominal tissues of honey bees obtained from hives located in foothill and especially foreststeppe zones of the Carpathian region, is a consequence of the urbanization and industrialization of the territory.

From the above follows that in the direction from the mountainous zone of the Carpathian region to the foothills and further to the forest-steppe zone, due to the high level of heavy metals, including toxic ones, the attractive, energetic, structural, biologically active, antimicrobial and antiinflammatory value of the unsaturated fatty acids of bee pollen decreases for the organism of bees, bee combs and beehive. In the direction mentioned above, the energetic, structural, biologically active, antimicrobial and antiinflammatory value of unsaturated fatty acids in the tissues of the abdomen of honey bees also decreases for the specified reason. Thus, a high level of Zinc, Copper, Plumbum, and especially Cadmium, but a low level of unsaturated fatty acids, in particular polyunsaturated, in bee honey and in the tissues of the abdomen of bees affects the reproductive capacity of queen bees kept in hives located in the mentioned areas of the Carpathian region.

Table 7 shows that the egg-laying intensity of queen bees kept in hives located in the foothills and especially in the forest-steppe zone of the Carpathian region, compared to queen bees in the mountain zone, in the second half of the spring period is lower by 1.12 and 1.17 times respectively.

A decrease in the egg-laying intensity of queen bees kept in hives located in the foothills and especially in the forest-steppe zone of the Carpathian region, compared to queen bees in the mountain zone, as in the second half of the spring period, due to a decrease in the number of worker bees, can in the future have an extremely negative effect on productive and reproductive features of the bee family. The decrease in the egg-laying intensity of bee queens is clearly an integrated indicator of the influence of the ecological state of the environment, since the latter also equally affects the reproductive capacity of drones, in particular, their quantitative and qualitative indicators of sperm production. Other scientists also point to the same negative impact of territories polluted with heavy metals on the reproductive characteristics of bees [17].

7. Egg-laying intensity of queen bees in the second half of the spring period in different natural zones of the Carpathian region, pieces of eggs (M±m, n=3)

Conclusions

In the direction from the mountain to the foothills and forest-steppe zone of the Carpathian region, the level of Zinc, Copper, Plumbum, and especially Cadmium increases in the topsoil, bee pollen, and abdominal tissues of honey bees.

In the mentioned direction, the coefficient of transition of Zinc from the arable layer of the soil to the bee colony decreases. At the same time, the assimilation of Cadmium by plants increases.

In the direction from the mountain to the foothills and forest-steppe zone of the Carpathian region, the level of unsaturated fatty acids of the omega-3, omega-6, omega-7, and omega-9 families in bee pollen and abdominal tissues of honey bees decreases.

High levels of heavy metals, but low levels of unsaturated fatty acids in bee pollen and abdominal tissues of honey bees obtained from hives located in the foothills and especially the forest-steppe zones of the Carpathian region are a consequence of the urbanization and industrialization of the territories.

Zinc, Copper, Plumbum and Cadmium affect the value of unsaturated fatty acids of the omega-3, omega-6, omega-7 and omega-9 families of bee pollen and affects the reproductive capacity of honey bees in the Carpathian region.

In particular, in the direction from the mountain to the foothills and further to the forest-steppe zone of the Carpathian region, due to the high concentration of Zinc, Cuprum, Plumbum, and especially Cadmium, both in the second half of the spring period and in the second half of the summer period, the value of unsaturated fatty acids of the omega-3, omega-6, omega-7 and omega-9 families in bee pollen decreases for intensive egg-laying of bee queens.

In the second half of the spring period, the egg-laying intensity of queen bees kept in hives located in the foothills and especially in the foreststeppe zones of the Carpathian region is 1.12 and 1.17 times lower, respectively, compared to queen bees in the mountain zone.

Bee pollen and abdominal tissues of honey bees can serve as a bioindicator of the ecological state of the environment by the content of heavy metals and unsaturated fatty acids. An integrated indicator such as the intensity of egg-laying by queen bees can also serve as a good bioindicator of the ecological state of the environment.

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