Prospects for the use of Arthropoda in Kazakhstan



The manuscript describes some prospect for practical application of Arthropoda group in Kazakhstan. In this manuscript were considered and given bright examples of medical, pharmacological and commercial significances of arthropods. This manuscript describes potential of Kazakhstan for practical usage of arthropods. Chitin and chitosan production techniques from arthropods, especially insects and application of obtained chitosan in production of chitosan nanoparticles, that are very useful in medical research for treatment of several disease and botany were considered. Also, this work considers structure and potential of Kazakhstan for application of silk fibroins, that may be used in medicine. We also reported about natural red dyes production and potential of Kazakhstan for Artemia sp. production. Information was taken from databases such as Scopus and Google Scholar. Presented data is based on books, scientific journals and articles, presented in references.

Keywords : Arthropoda, Kazakhstan, silk, nanoparticles, nanocomposites, natural dyes, aquaculture.

1. Introduction

Phylum Arthropoda consists of the most diverse and the most complex organisms. About 84 % of all known animal species are represented by this taxon [1]. This group possess high adaptation. As a result, members of this phylum are present in all types of habitats. Arthropods play crucial toles in ecosystems and human’s life. These organisms serve as a source of food for human and as a major parts of food chains, providing vertebrate animals with nutrients. Also, people used them in religious rituals and nowadays they are mainly used as vectors of diseases and pests in agriculture [2], [3].

Kazakhstan is the largest country in Central Asia and the 9 ^th country in the world by the land territory. Kazakhstan possesses different ecological regions such as semiarid steppes, forested steppes, warm moderate deserts, cold semideserts and mountains. This large territory is occupied by various organisms. According to the USAID and The Ministry of ecology and natural resources of the Republic of Kazakhstan, Kazakhstan is a home for 50 000 species of insects [4, 5] and hosts 176 species of spiders [6]. Alot of works related to fauna of centipedes, different taxa of insects and spiders in Kazakhstan are being published every year [7, 8, 25–29]. It is important to evaluate potential of Kazakhstan in utilization of different groups of arthropods.

2. Aims

The aim of this paper is to determine and describe the potential of utilization of Kazakhstani arthropods in biotechnology for solving practical tasks in medicine, ecology and pharmacology.

3. Natural red dyes

Nowadays synthetic aniline dyes are widely used in various fields of life such as perfume, food industry, textile. Before 18 ^th century people used cochineal as a source of natural dyes. It’s suggested that several world-wide companies started to replace synthetic dyes with more eco-friendly dyes from cochineal as hazardous impact of synthetic dyes became obvious [9].

Application of synthetic aniline dyes is hazardous for human health and environment. Waheed et al., in their review article showed the impact of synthetic aniline dyes and intermediate compounds in their synthesis. Metabolism of such dyes occurs in liver and then metabolites are transported by blood. These metabolites may form complexes with proteins or get to kidneys where in acidic values of pH they can trigger carcinogen DNA and harm normal DNA [10].

Serrano et al. proposed new methodology of analyzing cochineal and red dyes made of them using HPLC. Also, abovementioned authors analyzed 50 species of cochineal and by comparing database identified that the most productive insect is American cochineal [11].

Jashenko mentioned the historical application of cochineal and also found the half of all known cochineal species on the territory of Kazakhstan. According to this work author mentioned that according to Ibn-Sina carmine released from cochineal has an antimicrobial effect and used in medical procedures. In his work Jashenko suggests the application of carmine bugs in perfume and textile industries as a replacement for synthetic dyes. Author determined South-East Kazakhstan as a favorable territory to start commercial growth of cochineal population. Especially, he considers that P.sophorae, P. polonica and P.violaceae may be the main source of red dye, as they have the highest concentration of carmine acid. Also, we may use P. lappulae, P. turaiginiensis and P. ketmeniensis as an alternative resource since they show high populations. [9].

4. Potential for production of Artemia sp. in Kazakhstan

Artemia sp. or brine shrimps are very important crustaceans, inhabiting saline waters. These shrimps are consumed by some birds (such as flamingos, grebes, avocets), water boatmen, fishes and play significant role in some ecosystems of dry lakes as a primary consumer. These organisms basically feed on green algae, using their filter-legs [12].

According to Dhont and Sorgeloos, the main production of Artemia sp. cyst belongs to The Great Salt Lake, Utah, US. Since 1930s cysts were adopted as a natural diet for fish larvae. Also, authors mention that Artemia sp. can control the number of green algae in lakes. Green algae influence on the production of solar salt because we can use algal blooms as an indicator of solar activity in that lake. Moreover, metabolites and decomposing animals serve as nutrient sources for the growth of Halobacteriumsp . in the crystallization ponds. High levels of these red halophilic bacteria lead to a decrease in dissolved organic matter and enhance heat absorption, thus speeding up the process of evaporation.

But the unpredictable variability in cyst production from the Great Salt Lake has prompted producers to seek out alternative locations, including Lake Urmia in Iran, Aibi Lake in China, Bolshoye Yarovoye in Siberia, Kara Bogaz Gol in Turkmenistan, and various lakes in Kazakhstan [13].

Talking about Kazakhstan’s lakes, Sharapova et al. in their work mention the importance of Artemia sp. not only for aquaculture, but also for poultry farming, as nauplii of Artemia sp. are very rich in amino acids, hormones, carotenoids, vitamins and suit for production of special feeds [14].

Sharapova et al. found that the lake debris of the Northern Caspi, and lakes like Bura, Kalatuz, Borli possess favorable conditions for Artemia sp. cultivation. These lakes and lake debris have optimal salinity not more than 300 g/l, absence of predators and competitors [15]. Also, they found that hydro chemical regime of lakes Ray № 1, № 2, № 3 and Tuzkol is suitable for Artemia sp . cultivation. Authors determined that Ray lakes have different productivity periods, since they’re inhabited by different generations of crustaceans. Authors detected significant growth of biomass in Tuzkol lake and suggest it for Artemia cyst production. Although Tuzkol have very low biomass level during summertime, the rapid growth of crustaceans is observed through the vegetation period. So, it was determined that in October, when lake reaches its highest salinity, population reaches the highest biomass productivity [14]. Mazhibayeva et al. determined another natural habitat with suitable hydro-chemical regime for Artemia sp. with more diverse biomass in Karabassky floods (usually called Zhantelikol lakes) during summer-autumn. [16]

As a result, Kazakhstan also can play significant role in Artemia sp. production.

5. Chitosan nanoparticles

Chitosan nanoparticles are a promising organic nanoparticle that have very high practical significance. They can act as a container with particular drugs, transfer a medical substance to a specific location in organism and release transported chemical.

First, let’s define what are chitosan and chitin. Chitin is a linear polysaccharide made up N-acetyl-2-amino-2-deoxy-D-glucopyranose monomers, that are bonded by 1–4 glycosidic bonds. In living things chitin is synthesized in special organelles called chitosomes and achieved by enzyme chitin synthetase. Chitin isn’t soluble in water.

Chitosan is a deacylated form of chitin. Thus, it is made up β-D-glucosamines. Chitosan is more soluble that chitin. NH ₂ -group of chitosan has pKa 6.3–6.5 and this makes possible to use chitosan in various ways. Because under that point chitosan is presented by well-soluble cation.

To obtain chitosan we use a great range of organisms like algae, arthropods, molluscs, some worms, and bacteria. To get chitosan from chitin we use 3 different methods [17]:

1. Chemical method . This method is based on serial treatment of raw material with acids and alkalis. The process of purification of chitin-containing raw material from proteins is achieved by alkalis like NaOH. After that, the stage demineralization begins by using highly concentrated HCl. Next step is to treat demineralized chitin with H ₂ O ₂ to clear all pigments. Then, chitin is deacetylated by heating it with highly concentrated alkali. Obtained chitosan is briefly washed with water and methanol. There is alternative way when we firstly demineralize and then remove proteins. Chitosan derived this way is better that the chitosan made by previous steps.
2. Biotechnological method . Chemical method has a lower price, though creates a lot of waste products. Biotechnological method means that we use certain enzymes to remove proteins, products of lactic and acetic acid fermentation are used in demineralization. Enzymes that possibly used in this method are pancreatin and some proteinases. This is very expensive method, and it doesn’t provide high level of purity from proteins.
3. Electrochemical method . This method provides us with chitin with high level of purity. This technology means that we procced deproteination, demineralization and discolouration stages in form of water-salt suspension in electrolyzers under the influence of an electromagnetic field, a flow of ions H ⁺ and OH ^- that are formed as a products of water electrolysis [16].

According to A. A. Vetoshkin insects have a low concertation of, minerals in their cuticle, so we should use other techniques to obtain chitosan. This process includes 4 steps [18]:

1. The water extraction of melanin with 10 % suspension of chitin-containing raw material in temperature of 80 ^◦ for 1 hour.
2. Filtrated and solidified sediment is deprotonated by 10 % solution of NaOH in temperature of 45–55 ^◦ for 2 hours to get chitin-melanin complex. Then this complex is washed with distilled water.
3. Bleaching stage. Chitin-melanin complex is treated with 3 % solution of H ₂ O ₂ under 45–55 ^◦ for 1 hour. Solidified sediment is discoloured chitin-melanin complex.
4. Deacetylation. This process is achieved by heating chitin-melanin complex with 50 % solution of NaOH under the temperature 125–130 ^◦ for 1–1.5 hours. Then, this suspension is cooled down to 50 ^◦ C and filtrated to get solid sediment- chitosan-melanin complex.

A review of Mohan et al. shows different steps and assays of chitosan extraction for different orders and species of insects [19]. The collective of scientists present comprehensive table where they evaluate steps of chemical extraction such as deproteinization, demineralization, decoloration, deacetylation.

Islam et al. in their review on application of chitosan in biomedicine describe various fields of medical science, where chitosan plays significant role. Authors describe artificial kidney membranes, wound healing materials, bone, liver, skin repairing material, artificial tendons, anticoagulation drugs, drug delivery systems and many more [20].

Abere and colleagues suggest following approaches to make chitosan nanoparticles [21]:

1. Ionotropic gelation,
2. Complex coacervation
3. Coprecipitation
4. Microemulsion
5. Emulsification solvent diffusion

This collective of scientists believe that chitosan may be used as an antimicrobial substance and drug delivery agent. Chitosan is cationic molecule and is very sensitive for pH. If we increase the level of NH ₃ ⁺ groups, we can enhance the activity of chitosan nanoparticles. When chitosan enters the nucleus, it inhibits RNA transcription. Also, chitosan interferes the work of signaling molecules like HAQ. They showed the new type of antibiotics. That is chitosan nanoparticle that contains antimicrobial substance. It is easy to put antibiotics into the nanoparticle as chitosan is chelatious substance.

Apart from that function Abere and colleagues describe several applications of chitosan nanoparticles in medicine like gene therapy for treatment of some neurological disorders (medulloblastoma, glioblastoma, sclerosis). Chitosan is cationic molecule, so it easy bonds with anionic molecule of nucleic acids. For instance, PEGylation process can make a gene delivery system. Amaral et al. showed that PEGylated chitosan can interact with siRNA and create gene delivery system [22, 23].

Other applications of chitosan nanoparticles mentioned by Abere, and collaborators are wound healing, bone tissue engineering, biosensing, cancer diagnosis [23].

In Kazakhstan Zhatkanbayev et al. used chitosan, derived from the adult beetles Zophobas morio to make hydrogels with embedded mineral fertilizers. To obtain chitosan they diced beetles to 1mm and put it in 1M NaOH solution to deproteinize them. Then bleached chitin by immersing it in a 20 % hydrogen peroxide solution for 10 minutes, and dried at 70°C. Deacetylation of chitin was performed by treating it with NaOH solution. The reaction mixture was incubated at 80° C for 16 hours stirring constantly at a speed of 250 rpm. Then they proposed technique for hydrogel production by adding lactic or phosphoric acids, mineral fertilizers like ammonium nitrate, sodium hydrophosphate, or sodium humatesand and glutaric aldehyde. The reaction mixture then heated at temperatures ranging from 40°C to 90°C for 2–24 hours [24].

Authors introduced ammonium nitrate, phosphates, lactic acid and humates to stimulate the growth of Scots pine in in a quarter No.104 of the Semipalatinsk State Forestry Enterprise’s suburban forestry Semipalatinsk branch. Kazakhstani scientists found that the adaptability of seedlings increased from 21,6 % to 38,5 %. As a result, scientists proposed production of biodegradable hydrogels within fertilizers from insects’ chitosan, which is very useful for countries without access to the sea [24].

The vast diversity of arthropods gives opportunity for Kazakhstan in production of chitosan, and chitosan-based materials obtained from different insects. Unfortunately, there is particularly no data of Kazakhstani scientists on the chitosan and chitosan-based materials production.

6. Silk application in medicine

Silk is polymeric protein, mostly obtained from arthropods. Silk is a composite material made of 2 proteins: the central “rod” protein silk fibroin and sticky coating protein sericin [30].

Silk fibroin of Bombyx mori is made of 3 chains- heavy or H-chain, light chain and glycoprotein P25. The chemical structure of heavy chain (repetition of microcrystalline arrays and bulky residues) makes it hydrophobic. In contrast, light chains are more hydrophilic and elastic.

Basically, silk is obtained from larvae of Bombyx mori , but also species like Antherae mylitta, A. assama are used. Silk can be divided into 2 types: mulberry, obtained from Bombyx mori and non-mulberry. Non-mulberry silk has more Arg-Gly-Asp motifs, that are ligands for integrin proteins. As a result, this type of silk has better cohesion with cells. Also, non-mulberry silk is more mechanically stable and hydrophilic as it has poly (-Ala-) β sheets. In contrast, mulberry silk consists of poly (-Gly-Ala-) β sheets.

Sericin is a hydrophilic macromolecule that belongs to a family of glycoproteins. In contrast with silk fibroin, sericin may be in different 2 ^nd conformation like β-sheets or random coil. Also, its conformation may be changed easily and depends on the temperature, hydrolytic degradation and tensile forces [30].

Apart from sericin and silk fibroin there are several phytochemicals abandoned in cocoons of B.mori . These secondary metabolites are presented by flavonoids (quercetin, kaempferol), alkaloids, coumarin derivatives and some ethers like 3,4-dihydroxyphenyl-n-pentanyl ether and 2,3,4-trihydroxy-n-pentanyl ether.

Mahmood et al. found that cocoon silk extract can influence on cardiovascular system of Wistar rats by decreasing cardiac marker enzymes such as CK-MB or troponin, degree of myonecrosis, as well as improved cardiac antioxidant capacity and lipid peroxidation [31]. AliY et al. examined an antihyperlipidemic properties of sericin on rabbits with atherosclerosis and hyperglycemia. The result was that rabbits gained some weight and atherosclerosis plaques reduction was observed [32]. Lapphanichayakool et al. observed cholesterol-lowering feature of sericin-derived oligopeptides [33]. Okazaki et al. reported that sericin reduced serum triglycerides and cholesterol, free fatty acids, apolipoproteins rich in cholesterol, liver triglycerides, and the activities of known lipogenic enzymes glucose-6-phosphate dehydrogenase and malic enzyme [34].

You-Gui et al., studied a mice model of alcohol-induced liver damage and observed that sericin is able to decrease antioxidants such as GSH, GSH-PX, CAT, SOD to the normal level [35].

Yang et al. in vitro studied insulin resistant HepG2 cells and found that silk fibroin enhanced glucose and lipid metabolism [36]. Wang et al. found that flavonoid-rich ethanolic extract from the green cocoon shell of silkworm possess antihyperglycemic properties as it inhibits α-amylase and α-glycosidase [37]. Dong et al. and Park et al. reported that ethanolic extract from the sericin layer inhibited α-glucosidase. It resulted in the pancreatic cells proliferation and regeneration, expression of enzymes related to insulin metabolism and glycolysis such as glycogen synthase, GSK3β, GLK, PFK1, PKM2, AMPKα [38, 39].

Scientists also study impact of silk proteins on keratinocytes and fibroblasts. Zhang et al. designed semi-interpenetrating hybrid hydrogel containing sodium alginate functionalized with silk sericin and Ag nanoparticles (AgNPs) and applied it to treat wound injury [40].

Sutherland et al. have published a review on silks produced by insects and spiders, their glands and evolution of silk producing organs. They found that even though silks, made by insects possess different functions and evolution, some similarities such as same amino acid sequence and similar crystalline structure. Authors concluded that insect silks of different species have very good mechanical properties and may be used for production of silk fibers [41].

There is no available papers and articles on the investigations of above-said applications of silk fibroin and sericin in Kazakhstan, but as Sutherland et al. found silks of different species possess similar molecular structure, so we predict that it’s possible to use another species’ silk fibroins for treatment of several disease as was pointed above. Kazakhstan is rich and diverse for different insects and spiders, and it may be useful for scientists to investigate application of silks produced by Kazakhstani species.

7. Conclusion

On the basis of what was said above, Arthropods are a promising group for production of drugs and materials. There are too little works related to application of Kazakhstani species, so we listed some current achievements and gave some topics for future scientific works. We described application of arthropods for production of cysts of Artemia sp. , natural dyes, chitosan-based materials and application of silk. USAID labeled Kazakhstan as the most important country for biodiversity concentration among other Central Asian countries [4]. As a result of our work, we can conclude that Kazakhstan has great potential in production of natural red dyes based on carmine bugs and in growth of Artemia sp. cysts for world-wide application in aquaculture. We also predict that the use of different species of insects found in the vast territory of Kazakhstan can assist scientists in determining the role that chitosan-based nanoparticles play in medicine and in finding new ways of extracting chitosan and creating nanoparticles. Also, Kazakhstan has potential in silk production, but it still requires further investigations. We believe that in future Kazakhstani scientists will contribute in finding of new promising ways for utilization of Arthropoda.

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