FEDERAL STATE BUDGETARY INSTITUTION OF SCIENCE

Download Original PDF

2

REPORT

UNDER THE AGREEMENT ON SCIENTIFIC COOPERATION BETWEEN FSBIS PAVLOV INSTITUTE OF PHYSIOLOGY OF RAS AND AIRES HUMAN GENOME RESEARCH FOUNDATION "Study of high-frequency electromagnetic radiation impact and Aires resonators influence on behavior, genetic and epigenetic processes in cells of central and peripheral organs (models o Rattus norvegicus rat and Apis mellifera L. honey bee)".

STAGE FIVE (October 2018 - May 2019): Study of effect non-ionizing electromagnetic radiations and Aires Defender Pro resonators have on cognitive habits of rat lines with different excitation of nervous system.

Continuous improvement of communication systems and development of information

technologies result in an increased intensity of the effect anthropogenic electromagnetic radiation

sources (EMS) have on biological objects. Technology-caused electromagnetic fields are more

intensive and have higher radiation frequencies. Both a decreased depth of penetration and

increased energy influencing factor thereof characterize the impact they have on human beings.

Insufficient data on this EMR effect on animals and humans makes studies of possible

mechanisms of microbiological effect (MBE) forming, evaluation of aftereffect thereof on

humans, and development of possible methods and means of electromagnetic protection,

especially important (Lai, 2005).

The systems, protecting from this type of radiation, are different way developed, e.g., by

decreasing intensity and changing different parameters of EMR sources. Aires Foundation has

developed resonators that influence the living systems responses to non-ionizing EMR and have

a protective effect (Zhabrev at al., 2005; Jasitis et al., 2018).

The mechanisms of the effect the nervous system functional state has on congenital

elements of behaviour repertoire and cognitive body abilities, when exposed to EMR, and also

on the efficient operation of devices meant for body protection from harmful effect thereof, have

not been exploited to any great extent.

As shown earlier, the character of orientative-trying reaction of animals (“open field”

test) to reduced external electric and magnetic fields, to router EMR depends on inherited

produced excitation of the nervous system and centers around different components of

behaviour. At that, the effect of Aires Defender Pro resonators was tested on the line of rats with

low excitability threshold (LET) only and resulted in the animals changed behaviour, increased

activity thereof when placed in a new environment. Specific behaviour components of the

animals’ response to the effect of resonators were also revealed, showing their selective positive

impact on separate elements of behaviour.

Aires Defender Pro resonators effect under the same conditions using the line of rats with

high excitability threshold (HET) was evaluated herein.

3

As shown earlier, animals exposure in a Faraday cage without any additional influences

and combined with the router effect, resulted in problems to memorize the conditioned passive

avoidance reflex in Vistar male rats. The resonators partially prevented a negative influence on

the rats exposed in a Faraday cage in 24 hours after training. This protective effect reduced in 7

days.

It is important to evaluate the influence of the researched effects on changes in the

training and memory processes, using the same behaviour test in rats with contrasting

excitability of the nervous system.

The goal and objectives of stage five:

1) study of Aires Defender Pro resonators effect when exposed to standard WiFi router

EMR with a reduced external magnetic field and without any additional restrictions

regarding behaviour of low threshold excitability male rats in the open field test (final

fragment of stage 4) (section 1 hereafter)

2) study of WiFi router and Aires Defender Pro resonators electromagnetic radiation effect

on memorising the conditioned passive avoidance reflex (CPAR) in Vistar male rats and

in rat lines with contrasting excitability of the nervous system (HET and LET) ( section 2

hereafter).

MATERIALS AND METHODS

5 month old male rats of the standard line Vistar and HET and LET lines selected in the

Higher Nervous Activity Genetics Laboratory (Vaydo, 2000, Vaydo et al., 2018) and included in

the biocollection of FSBIS Pavlov Institute of Physiology (No GZ 0134-2018-0003) were

studied. 6-8 male rat groups were kept in standard cages on a standard diet.

Section 1.

In order to create a decayed external magnetic field (DMF hereafter), use was made of a

shielded chamber made from a non-magnetic material (cardboard) coated with several layers of

amphorous soft magnetic material AMAG-172, thereby the Earth’s magnetic field’s induction

was 40-fold reduced inside the chamber (from 48 mcTl to 1.2 mcTl) (Kuznetsov et al., 2006). A

simulation chamber (without magnetic field decay, NDMF hereafter) was made from cardboard,

had no shielding coating and was covered with black polyethylene. Both chambers were cylinder

shaped 60 cm in diameter and 140 cm long, open from one end and plugged from the other so

that a cage with rats easily fitted therein.

Use was made of a WiFi router (LinkSys E1200-EE/RU wireless router) with the

following technical parameters: carrier frequency of wireless communication: 2.4 Hz, number

and type of antennas: 2 internal antennas, standard antenna(s) gain ratio, dBi: 4 dBi.

4

Aires Defender Pro resonators were used in the experiments (Zhabrev et al., 2005;

Jasaitis at al., 2018). Like previously, 6 resonators were used to evaluate the effect thereof when

exposing rats to router EMR. The resonators were placed on top of the cage with animals

(Experiment 1) or in the center point of each edge of the Faraday cage (Experiment 2).

In order to study the effect of resonators exposed to router EMR under the decayed

external magnetic field (DMF) or without any additional restricting effects (NDMG), “a home”

cage with animals was inserted in the shielding chamber with the router located on a tray in the

central point on top of the cage upper cover jointly with resonators (group

DMF+router+resonators). The experimental groups were exposed for 12 hours (10:00 p.m. –

10:00 a.m.). Control groups were the groups of rats placed in the simulation chamber for the

same period with the router and resonators (group NDMF+ router + resonators).

The animals behaviour in the “open field” test were evaluated in an hour after the animals

exposition in the chambers was over.

The intact control groups of animals (Control 1 hereafter) were those in the vivarium and

not exposed to radiation.

The used “open field” test was a circular apparatus 160 cm in diameter walled to a height

of 35 cm. The circle bottom was divided into squares 20 cm in a side. 500 W lamp was fixed

over the circle center at a height of 60 cm, which mirror reflector provided bottom level

illumination of 2000 lux in the center and 1500 lux at the borders. The apparatus was placed in a

darkened room. In testing, a rat’s behaviour was observed for 5 minutes after the animal’s

placement in the central square. After testing, each animal was wiped with 35-40% alcohol

solution and then with dry gauze pads. The following behaviour parameters were recorded:

Latent periods from the first movement (c)

Horizontal locomotion (number of crossed squares)

Vertical locomotion (getting on hind legs, a number of rearing responses)

Emotionality – number of defecatory boluses.

Grooming (number of acts)

Freezing (number of acts)

Turns left (number of acts)

Turns right (number of acts)

Spinning (number of acts)

The “open field” apparatus is used to assay rodents’ orientative-trying reaction and

emotional behaviour when moved to a new environment; it enables complex evaluation of

natural controlled behaviour and changes thereof when exposed to various effects (Amiksheeva

et al., 2003): to measure the level of emotionality and locomotion activity (horizontal – number

5

of crossed squares, and vertical – rearing respoinses, showing orientative-trying activity), as well

as anxiodepressive component intensity (activity in the central most open and illuminated field

sector, number of turns and spins), level of fear by freezing reaction, stereotype behaviour

(grooming) (Kaluev 1998, Buresh et al., 1991). See our Report 2018 for this method detailed

description (Stage 4).

Section 2.

WiFi router electromagnetic radiation effect (rats were exposed for 4 days, 6 hours a day,

for total of 24 hours) on memorizing the conditioned passive avoidance reflex (CPAR) was re-

evaluated in the Vistar rats under the same conditions in the Faraday cage with Aires Defender

Pro resonators.

In order to study WiFi router electromagnetic radiation effect on memorizing the

conditioned passive avoidance reflex (CPAR), a standard cage with HET and LET rats and Aires

Defender Pro resonators in the Faraday cage was exposed to the router radiation for 24 hours. As

before (Stages 1-3), the resonators were placed in the central point of each edge of the Faraday

chamber. The each line intact animals (Control 1) and those in the Faraday cage not exposed to

any additional effects (Control 2) were used as the control ones.

One hour after learning CPAR, the rats of the relevant groups, apart from intact control 1,

were placed in the given experimental conditions. Memory consolidation. i.e. the process of

transferring new learning from short- to long-term memory storage, was tested through checking

CPAR retaining soon after the animals exposure in the Faraday cages with the router, router and

resonators, was over.

The passive avoidance reflex was conditioned by delivering a single adverse stimulus and

using a chamber divided into a lit compartment and a dark compartment, with a gate between the

two. As known, normally rats spent most time in the dark compartment due to instinctual drive

thereof for a dark and confined space, i.e. a hole (hole exploratory behaviour). This method is

based on rats acquiring the conditioned passive avoidance of the dark compartment in response

to an unconditioned stimulus in the form of an electric shock. A rat was placed in the center of

the lit compartment facing away from the dark compartment and was given 2 minutes to explore

the compartments, during which the rat was capable of finding a hole to the dark compartment

and enter it. Being in the dark compartment, the rat was delivered 1mA electric shock within 1

min. This was the final stage of the conditioning. If a rat failed to enter the dark compartment

within 2 minutes, it was excluded from further experiments. Recorded was a per cent of rats that

did not enter the dark compartment.

6

Statistical processing

Average values and medians were calculated for the results tabulation purpose. The

figures show medians. The statistical significance of differences between the groups was

determined by Mann-Whitney test, ANOVA, as well as by the significance of differences

between samples (Plokhinsky, 1970). Use was made of Statgraphics Centurion XV11 and

Statistica 6.0 software.

RESULTS AND DISCUSSION

Section 1.

During the last reported period (Stage 4) the open field test helped to study both the effect

of the cylinder shaped chamber, shielding the external magnetic field, and the non-shielding

simulation thereof, as well as the effect of a standard WiFi router EMR on the behaviour of HET

and LET rats, however the effect of resonators was tested in the HET line only. This year the

effect the resonators have on the rats exposed to WiFi router external magnetic field and without

it was tested in highly excitable LET rats. For convenience of data comparison and discussion,

total results obtained in LET rats are given below.

Fig. 1. Latent period of the LET male rats first reaction in the open field test. Designations: * -

differences between LET line and Control 1 are significant (P<0.05), Control 1 – intact rats,

DMF – decayed magnetic field (shielding chamber), NDMF – non-decayed magnetic field

(simulation chamber).

Latent period

DMF+router+resonators

NDMF+router+resonators

7

Fig.2. LET male rats horizontal locomotor activity (number of crossed squares) in the open field

test. Designations: # - differences from other LET line groups are significant (P<0.05), other

designations are similar to those in Fig. 1.

Fig.3. LET male rats vertical locomotor activity (number of rearing responses) in the open field

test. Designations are similar to those in Fig. 1.

Horizontal locomotor activity

Number of crossed squares

DMF+router+resonators

NDMF+router+resonators

DMF+router+resonators

NDMF+router+resonators

Number of rearing responses

Vertical locomotor activity

8

Fig.4. LET male rats emotionality (number of fecal boluses) in the open field test.

Designations: * - differences from Control 1 and DMF+router+resonators and

NDMF+router+resonators of the LET line are significant (P<0.05), other designations are similar

to those in Fig. 1.

Fig.5. LET male rats grooming acts in the open field test.

Designations: # - differences of LET line from Control 1 are significant (P<0.05), other

designations are similar to those in Fig. 1.

Emotionality

Number of fecal boluses

DMF+router+resonators

NDMF+router+resonators

DMF+router+resonators

NDMF+router+resonators

Grooming

Number of grooming acts

9

Fig.6. LET and HET male rats freezing acts in the open field test.

Designations: # - differences from LET rats of NDMF group are significant (P<0.05), & -

differences from LET rats of DMF and NDMF groups are significant (P<0.05); $ - difference

from LET rats of the relevant DMF+router and NDMG+router are significant (P<0.05). Other

designations are similar to those in Fig. 1.

Fig.7. LET male rats turns to the left in the open field test.

Freezing

Number of acts

DMF+router+resonators

NDMF+router+resonators

Turns to the left

Number

DMF+router+resonators

NDMF+router+resonators

10

Designations: # - differences from NDMF group are significant (P<0.05), & - differences from

Control 1 and NDMF groups are significant (P<0.05). Other designations are similar to those in

Fig. 1.

Fig.8. LET male rats turns to the right in the open field test.

Designations: * - differences from Control 1 and NDMF groups are significant (P<0.05), # -

differences from NDMF group are significant (P<0.05); Other designations are similar to those

in Fig. 1.

Turns to the right

Number

DMF+router+resonators

NDMF+router+resonators

11

Fig.9. LET male rats number of spins in the open field test.

Designations: * - differences from other groups are significant (P<0.05). Other designations are

similar to those in Fig. 1.

Compared to the router adverse impact, the effect of resonators causes restoration of the

following highly excitable LET line rats behaviour indicators: 1) horizontal locomotor activity

(HLA): under the decayed magnetic field, the router causes HLA decrease, while the exposure to

resonators restores HLA to the control levels (Fig.2); 2) freezing: independently on DMF

existence or absence thereof, the router intensifies freezing responses, while the resonators

decrease the number freezing acts and time thereof in both experimental groups (DMF and

NDMF) (Fig.6); 3) spins: a number of spins is increased by the router exposure, while the

resonators decrease it to the control level (Fig.9). However, it should be noted that the combined

resonators-router effect independently on DMF existence or absence thereof, resulted in an

increased number of rats spins (Figs. 7,8). It is hard to say why. No router and resonators caused

changes in the first reaction latent period, vertical locomotor activity, emotionality, and

grooming behaviour indicators were revealed (Figs. 1, 3, 4, 5).

The obtained results as a whole show a router caused possible growth of fear and anxiety

in the highly excitable LET line rats and decrease thereof when the resonators are used as an

additional factor of influence. It should be noted that the low excitable HET line rats exposed to

EMR of the router and resonators demonstrated the suppressed fear reaction, but increased

general locomotor and exploratory activity, which was considered as possible adaptive reactions

of the animals under the new conditions (for more details see Report 2018, Stage 4).

Spins

Number of acts

DMF+router+resonators

NDMF+router+resonators

12

Section 2.

The experiments in evaluating the router (4 days x 6 hours) and resonators effect on

CPAR forming and retaining aimed at confirming the earlier obtained results were repeated

using the Vistar rats.

Fig. 10. CPAR retaining after exposure to EMR UHF of the router and resonators in the Vistar

rats. Designations: *- differences from all other groups are significant (P<0.05). C1 – intact

control, C2 – Faraday cage, R- router, R+Rst -routers +resonators.

The effects demonstrated in Stage 3 Report were confirmed (Fig. 10). Router’s EMR

UHF caused 2-fold suppression of CPAR retaining in the Vistar rats compared to the intact

control and 3-fold suppression compared to the active control, i.e. the Faraday cage, while

introducing the resonators into the experiments restored the conditioned reflex retaining to the

control values.

No differences in retaining the reflex by HET line rats of Control 1, Control 2, router,

router+resonators groups were determined (Fig.11). The exposure of the highly excitable LET

line rats in the Faraday cage and without any additional effects, as well as exposed to the router

significantly affected memory consolidation, resulting in a considerably worsened CPAR

retaining ability (Fig. 11). Appearance of more than 2-fold interline differences in CPAR

retaining efficiency when exposed to the router should be noted (Fig.11). It means that the highly

excitable LET line rats are found to be more sensitive to EMR UHF. Use of the resonators had a

positive effect on these rats CPAR retaining (Fig.11).

C1 C2 R R+Rst

% of CR retainers

% of CR non-retainers

13

Fig. 11. CPAR retaining after exposure of two lines of rats (LET, HET) within 24 hours in the

Faraday cage without any additional effects (C2), in the Faraday cage with the router (R), in the

Faraday cage with the router and resonators R+Rnt). The intact control rats – C1. Significantly

different values are designated on the diagram with the lines (P<0.05).

The obtained results made it possible to arrive at the conclusion that the disturbed

memory consolidation in learning CPAR due to the decayed electric field in the Faraday cage

and combined with the router EMR additional effect is apparent in rats with the inherited high

excitability of the nervous system (LET line) and downplays once exposed to the resonators

effect. The used experimental effects did not cause any statistically important changes in the

ability to retain CPAR in rats with the low excitable nervous system (HET line) both between the

animals and compared with the intact control group.

The effect of a variable but not static electric field (35 kV/m) on the ability to learn and

spatial memory is shown in mice (Di at al., 2019). The effect high frequency electromagnetic

fields have on the adult male rats memory is demonstrated by the social discrimination tests

(Schneider at al., 2014). It is shown that ultra-high frequency EMRs cause metabolic

reprogramming of the cerebral mitochondria in the Vistar rats thereby increasing a speed of

forming superoxide radicals and nitrogen oxide that may initiate development of neuregenerative

diseases and cancer (Burlaka et al., 2016).

As was earlier revealed (Stage 2), the highly excitable LET rats’ chromosomes are more

vulnerable to the damaging effect of high frequency EMRs compared to the HET line. High

HET LET HET LET HET LET HET LET

C1 C2 R R+Rst

% of CR non-retainers

% of CR retainers

14

excitability of the nervous system determines a greater apparency of mitotic disturbances

reduction in presence of Aires Defender Pro resonators when exposed to standard WiFi router

UHF radiator. In studying destabilisation of EMR UHF exposed bone marrow cells genome, it

was demonstrated that the efficiency of the resonators’ protective effect may depend on the

animal nervous system functional state (Dyuzhikova et al., 2019).

The mechanisms of the effects EMR and magnetic fields have on the body are presently

under active studies with concomitant multiple discussions, regarding possible ways of effect

thereof on the body, displayed magnetobiological impacts and consequences thereof (Karthick et

al., 2017; Pall, 2016; Terzi et al., 2016, etc). A group of American and Japanese researchers,

using EEG method, has recently found a human ability to feel magnetic field changes (cerebral

activity changes in the alpha-range with a magnetic field differently oriented relative to a tested

person in the Faraday cage) (Wang et al., 2019), nevertheless, the mechanisms of

magnetoreception in humans are not clear yet.

The molecular concept (Bukachenko, 2014) based on the importance of a radical ion pair

as a magnetic fields receiver and source of magnetic effects is most substantiated among the

hypotheses under discussion. Non-paired electrons therein are the carriers of spin magnetism and

interact with constant and variable magnetic fields. Involvement of these pairs in the enzymic

synthesis of ATF (adenosine triphosphate), being the main energy carrier of live systems, and

replicative synthesis of DNA with polymerases has been shown.

Overall, the findings of this paper confirm an adverse effect the external electric and

magnetic fields and UHF range EMR have on the rats’ behaviour and memory and demonstrate

Aires Defender Pro resonators’ positive effect on restoration of a number of disturbed elements

of rats’ innate behaviour and cognitive abilities when exposed to non-ionising electromagnetic

radiations. They also show the necessity to consider typological features of the nervous system in

developing the means of protection against an adverse effect of EMR and correction of

magnetobiological effects produced by various radiation sources. This approach is important for

understanding the causes of an individual’s variable sensitivity to EMR and determining the

ways of correcting pathogenic processes in humans on this basis, which is a necessary link of the

evidence based medicine.

15

BIBLIOGRAPHY A.V. Amitsheeva. Functional observation: modern methods and equipment// Annals of Vavilov Society of Geneticists and Selectionists. 2009.-V.13.№3. P.529-542. Y. Buresh, O.Bereshova, D.P. Huston. Brain and behaviour research methods and main

experiments. M: Higher School. 1991. 399 p. A.L. Buchachenko. Megneto-dependant molecular and chemical processes in biochemistry, genetics, and medicine// Chemistry Successes, 2014, 83(1): 1-12. A.I. Vaydo. Physiologico-genetic analysis of laboratory rodents nervous system excitability and behaviour. Doctor’s of Biology thesis. SPb. 2000. 197 p. A.I.Vaydo, N.V.Shiryava, M.B.Pavlova, A.S. Levina, D.A.-A. Khlebaeva, O.A., Lyubashina, N.A. Dyuzhikova. Selected lines of rats with high and low excitability threshold: model to study desadaptive nervous system excitability dependent states// Laboratory animals for researches. 2018. №3: P.12-23. N.A. Dyuzhikova, A.I.Vaydo, E.V. Daev, S.V. Surma, B.F. Schegolev, A.V. Kopyltsov, I.N.Serov. The effect UHF range electromagnetic radiation has on destabilisation of bone marrow cells genome in the lines of rats with contrasting excitability of the nervous system //Environmental genetics. №2. 2019. https://doi.org/10.17816/ecogen%25v%25i%25p Zhabrev V.A., Lukyanov G.N., Margolin V.I., Potekhin M.S., Tupik V.A., Serov I.N., Soshnikov I.P., Study of Cu fractal structures, produced by ion magnetron deposition method// Composite materials structures 2005, No 4 A.V. Kaluev. Stress. Anxiety. Behaviour. Kiev.:1998, 98 p. P.A. Kuznetsov, B.V.Farmakovsky, A.I.Askinazi, T.V.Peskov, S.B.Bibikov, E.I.Kulikovsky, L.V.Orlova. Composite materials to protect from electromagnetic radiation. Patent №2324989. 2006. N.A. Plokhinsky. Biometry. 1970, 2nd edition - M.: MSU Publishing House, - 367 p. Burlaka A.P., Druzhyna M.O., Vovk A.V., Lukin S.M. Disordered redox metabolism of brain cells in rats exposed to low doses of ionizing radiation or UHF electromagnetic radiation.//Exp Oncol. 2016;38(4):238-241.

Di G., Kim H., Xu Y., Kim J., Gu X. A comparative study on influences of static electric field and power frequency electric field on cognition in mice. Environ Toxicol. Pharmacol. 2019 66:91-95.

Jasaitis D., Vasiliauskienė V., Miškinis P., Damauskaitė J., Jukna A., Kopyltsov A., Lukyanov G., Korshunov K., Serov I. Investigation of the Circle Fractal Structure Interaction with Gigahertz Frequency Electromagnetic Waves. Proc. “The 12th Int. Sci. Conf. Intelligent Technologies in Logistics and Mechatronics Systems (ITELMS’2018)”, Lithuania, Panevėžys, 2018: 81-87. P. 81-87.

16

Karthick T., Sengottuvelu S., Sherief H., Duraisami A review: biological effects of magnetic fields on rodents// Sch J. App. Med. Sci., 2017; 5(4E): 1569-1580

Lai N. Biological effects of radiofrequency electromagnetic fields//Encyclopedia of Biomaterials and Biomedical Engineering DOI: 10.1081/E-EBBE-120041846 Copyright # 2005 by Taylor & Francis.

Pall M.L. Microwave frequency electromagnetic fields (EMFs) produce widespread neuropsychiatric effects including depression.// J Chem Neuroanat. 2016;75(Pt B):43-51.

Terzi M., Ozberk B., Deniz O.G., Kaplan S. The role of electromagnetic fields in neurological disorders.// J Chem Neuroanat. 2016 ;75(Pt B):77-84.

Wang S.H., Hilburn I.A., Wu D-A., Mizuhara Y., Cousté C.P., Abrahams J.M.H., Sam Bernstein S.E., Matani A., Shimojo S., Kirschvink J.L. Transduction of the Geomagnetic Field as Evidenced from Alpha-band Activity in the Human Brain // eNeuro. 2019;6(2). pii: ENEURO.0483-18.2019. doi: 10.1523/eneuro.0483-18.2019

Schneider J., Stangassinger M. Nonthermal effects of lifelong high-frequency electromagnetic field exposure on social memory performance in rats.// Behav. Neurosci. 2014 ,128(5):633-7.

LIST OF PUBLICATIONS FOR THE REPORTED PERIOD:

1. N.V. Shirayeva, A.I. Vaydo, N.A. Dyuzhikova, B.F. Schegolev, S.V.Surma, I.N. Serov.

Impact of high-frequency electromagnetic radiation and resonators on behavior of rats with different excitability of the nervous system.//In scientific papers of VIII International Congress “Weak and super weak fields and radiations in biology and medicine”, September 10-14, 2018. Saint Petersburg, V.8, p. 160-161 (poster-report and abstracts)

2. N.V. Shirayeva, A.I. Vaydo, I.N. Serov. Impact of UHF electromagnetic radiation and

resonators on the memory consolidation in learning the conditioned passive avoidance reflex by the rats lines with contrasting excitability of the nervous system// All Russian “Integrative Physiology” Conference with foreign participants devoted to the 170th birth anniversary of I.P.Pavlov, September 24-26, 2019 (poster-report and abstracts)

3. N.A. Dyuzhikova, A.I.Vaydo, E.V. Daev, S.V. Surma, B.F. Schegolev, A.V. Kopyltsov,

I.N.Serov. The effect UHF range electromagnetic radiation has on destabilisation of bone marrow cells genome in the lines of rats with contrasting excitability of the nervous system //Environmental genetics. №2. 2019. https://doi.org/10.17816/ecogen%25v%25i%25p