Anacyclus pyrethrum L., a threatened medicinal species endemic to Morocco, we are the first to study the reproductive biology of two varieties of Anacyclus pyrethrum L.: Anacyclus pyrethrum var. pyrethrum (L.) Link and Anacyclus pyrethrum var. depressus  Maire. The reproductive biology of the two varieties was examined in detail by studying the development of the inflorescence, phenology of flowering, breeding system, pollinators, production, and seed dispersal. The experimental results described in this work suggest that Anacyclus pyrethrum L. is a gynomonic species, with a mixed autogamy–allogamy reproductive regime with a high predominance of allogamy. It appears to be partially self-incompatible, with allogamy rates for Anacyclus pyrethrum var. depressus  Maire and Anacyclus pyrethrum var. pyrethrum (L.) Link of 78.70% and 79.01%, respectively. It depends on pollination vectors to produce a large number of seeds. This study on the breeding system of Anacyclus pyrethrum L. provides a tool for developing management strategies and adequate conservation measures.The reproductive system is essential for the structuring and transmission of genetic diversity. Understanding the reproductive biology of threatened endemic species is considered to be a crucial element for the implementation of effective conservation strategies. Given the lack of information and the insufficient state of knowledge on the reproductive system of  Reproductive biology is an important and critical trait as it characterises growth and development, and plays a role in the implementation of conservation strategies ,2. PlantFor a long time, the diversity of reproductive systems observed in plants has fascinated biologists ,10,11. THistorically, the identification of Asteraceae has been based on their capitula, with flowers closely arranged on a receptacle surrounded by bracts ,15,16,17Anacyclus pyrethrum L. is a medicinal species endemic to Morocco, Algeria, and Spain belonging to the Asteraceae family genus Anacyclus. In 1979, the species was identified with two varieties: Anacyclus pyrethrum var. pyrethrum (L.) Link (A.P var. pyrethrum) and Anacyclus pyrethrum var. depressus  Maire (A.P var. depressus) [pressus) ,21,22. Tpressus) ,24,25, apressus) ,29,30,31pressus) ,33, an apressus) , and a mpressus)  as well pressus) ,37,38,39pressus) ,41, antipressus) , insectipressus) ,44,45, lpressus) , anti-inpressus) ,48,49, apressus) ,50,51,52pressus) ,54, antipressus) ,55,56,57pressus) , antiamnpressus)  propertipressus) .The excessive overexploitation of the roots of the plant for trade makes it very local, uncommon, and quite rare in most of its known sites. Recently, it has been placed in the ‘vulnerable’ category according to the IUCN red list and the red list of vascular plants in Morocco. Anacyclus pyrethrum L., the objective of this study was to analyse its reproductive biology; specifically, to examine the development of the inflorescence, the phenology of flowering, the reproductive system, and the production and dispersal of seeds as well as the insect pollinators. The ultimate aim was to understand the implications of these data in evaluating future breeding and conservation strategies for the studied species. The questions we attempted to answer were:Anacyclus pyrethrum L.?− What is the reproductive system of − Are pollinating insects necessary for the two varieties to be successful in producing seeds?Given the insufficient state of knowledge and the lack of information on the reproductive biology of the two varieties of After setting up quadrats, the number of individuals within the limits of each quadrat was counted. The size of each population is represented in The results showed that the individuals constituting each population were randomly dispersed according to an aggregate dispersal pattern, and that the density of plants in 2019 was significantly lower than observed in 2018, with the exception of the Ifrane station where we observed an increase in the number of individuals. This increase was explained by the protection of this population due to its proximity to the royal palace. This confirmed that the number of individuals increased in legally protected populations.A.P var. depressus being more frequent than A.P var. pyrethrum. The plant density in 2019 was low compared with 2018; these differences were mainly due to the collection process. In recent years, local collectors have reported a significant decline in both varieties of Anacyclus pyrethrum L., aggravated by premature harvesting ; A.P var. pyrethrum is the variety most affected by this overexploitation. Our results on the decline of A.P var. pyrethrum in the Middle Atlas were consistent with those of Ouarghidi, who showed a significant decline in A.P var. pyrethrum populations in southern Morocco  [A comparison of the different quadrats revealed that the plant density was significantly higher in the quadrats closest to habitations, indicating that individuals closer to habitations were more protected. The plant density per quadrat was also significantly different between the two varieties, with  valley) ,22.The natural populations remain dormant as underground rhizomes throughout the winter; they start to develop from the fourth week of March and continue until the end of the vegetative phase and the beginning of the sexual phase (inflorescence) in the third week of April. The formation of floral buds continues until the fourth week of June; the flowers of the same plant do not flower at the same time. At the capitulum level, the flowering is centripetal; the florets open from the periphery of the floral ray toward the centre and they remain fully open for about 4 weeks after the emergence of the floral bud. Senescence takes place during July and August; during these months, all aerial parts dry out and the dry flower heads are dispersed. Seeds do not immediately disintegrate from the capitula. Under greenhouse conditions, the activation of the vegetative phase starts in the first week of March and enters the sexual phase (appearance of the first flowers) at the latest from the fourth week of May until the first or second week of July. The beginning of flowering is signalled by the centripetal blooming of the peripheral ray florets and the continuation of the tubular florets. The plants begin to senesce by the latest at the end of July and continue until the second week of August.A.P var. depressus and from 13 mm to 23 mm for A.P. var. pyrethrum  (The results showed that the variety ge (90%) .Under the experimental conditions in a greenhouse, the bagged capitula failed to form seeds. The capitula of bagged individuals only produced seeds from the female peripheral flowers whereas the non-bagged individuals produced seeds not only from the central (hermaphroditic), but also from the peripheral  flowers .A.P. var. depressus and A.P. var. pyrethrum, respectively, for the lot of bagged individuals (lot 2). The fructification index was 0.75 for A.P. var. depressus and 0.74 for A.P. var. pyrethrum for the free pollination lot (lot 3). The allogamy rate was 100% for lot 1; for lot 2, the allogamy rates for A.P. var. depressus and A.P. var. pyrethrum were 78.70% and 79.01%, respectively. Therefore, Anacyclus pyrethrum L. had a mixed self-pollinated–allogamous reproductive regime with a strong predominance of allogamy. We could deduce from these results that the absence of seeds in the bagged capitula lot showed that there was a self-incompatibility between the female peripheral flowers and the hermaphrodite central flowers in the capitula. In the case of the bagged individuals, only the peripheral female flowers produced seeds, showing intra-individual self-compatibility between the peripheral female flowers and the central hermaphrodite flowers, and intra-individual self-incompatibility between the central hermaphrodite flowers of the same individual.The fructification index was nil for both varieties for the bagged capitula lot (lot 1), and 0.16 and 0.15 for Anacyclus pyrethrum L. and show that the two varieties, A.P. var. depressus and A.P. var. pyrethrum, are able to reproduce in xenogamy with a fructification index of 0.75 and 0.74, respectively. In geitonogamy, only the peripheral flowers produce fruit; we observed a fructification index of 0.16 in A.P. var. depressus and 0.15 in A.P. var. pyrethrum. However, fruit production is significantly higher in allogamy than in autogamy. In light of these results, we can state that Anacyclus pyrethrum L. is a gynomonoecious species and partially self-incompatible, with geitonogamous peripheral flowers that have adapted to the absence of pollinators and xenogamous central flowers. These results were in alignment with those of Humphries, who found that four populations of A.P. var. pyrethrum and A.P. var. depressus were self-incompatible; all attempts to cross A.P. var. pyrethrum or A.P. var. depressus with annual taxa of the same genus ended in failure [Anacyclus pyrethrum L. can reproduce asexually from root cuttings [In Asteraceae, mutations probably related to organ identity genes may have occurred independently during angiosperm evolution, leading to gynomonoecy as the main sexual system in all species with radiate capitula ,62,63. I failure . A few s failure . Self-po failure ,70. Howe failure . Differecuttings ,21,22,36At anthesis, the stamens release a large quantity of pollen grains. Microscopic observations showed spheroidal pollen grains that were radially symmetrical, circular-polar, and equatorial in outline; they were echinulate with triangularly outlined spines tapering toward the tip and broad at the base, and densely arranged . This poA.P. var. pyrethrum and A.P. var. depressus , the seedlings were placed in the field and monitored until flowering. Flower heads emerged between 12 and 14 months after sowing, from which we determined, under experimental conditions, the modalities of reproduction. In particular, we quantified the production of seeds under self-pollination and open pollination.A total of 20 capitula bagged at the bud stage to test self-pollination (Lot 1).A total of 10 individuals bagged at the bud stage to test intra-individual self-compatibility (Lot 2).A total of 20 individuals not bagged to test seed formation in free pollination (Lot 3).The sample size was 50 plants. The observations on the reproductive biology were made in 12–14-month-old plantations. At the beginning of the flowering period, the capitula/individuals, in bud, were bagged in a fine mesh netting in order to allow abundant entry of light, humidity, and gas exchange with the outside environment and to prevent visits from pollinators. The bags were regularly shaken during the day to encourage the deposition of pollen on the stigmas. The bags were not removed until the fruit had formed or the flowers were falling. The number of mature and non-mature seeds per capitula was recorded for each treatment:The fructification index (FI) was calculated for each treatment according to the following formula :(1)IF=The difference between the fruiting indices of the different treatments was used to determine the rates of allogamy (TC) and autogamy (TA), which were calculated according to the following formulae ,85:TC=IThe pollen was mounted in glycerine water and was observed by light microscopy to determine the structure and diameter. Photographs were taken to document the morphology of the pollen.The capturing of pollinators was favoured during days without strong winds and significant cloud cover. The insect specimens encountered at each site were captured and preserved in ethyl acetate. The taxonomic identification of the insect specimens was carried out by an entomology specialist.p < 0.05.The results were analysed using Graph Prism version 7 and Microsoft Excel 2010 in order to highlight the effect of the various factors and their possible interactions. Statistical processing was performed through an analysis of variance, followed by a Tukey multiple comparison test. A significant difference was considered at Anacyclus pyrethrum L. is a gynomonoecious, partially self-incompatible species with geitonogamous peripheral flowers and xenogamous central flowers. The pollinators of the two varieties, A.P var. depressus and A.P var. pyrethrum, mainly belonged to Coleoptera and Hymenoptera.The results of the experiments described in this work revealed that


Capparis spinosa L. distributed on Calcisols formed on eroded alluvial-proluvial gravel textured rocks in the south of the Fergana Valley . The predominance of immature plants in the cenopopulation was detected in the Arsif hills massive, and quantitative indicators of micronutrients in the vegetative and generative organs of C. spinosa L. were determined. The study of biomorphological characteristics of the plant during the growing season (April-October) was carried out in the identified 10 observational experimental field populations. The cenopopulation dynamics and plant development patterns of Capparis spinosa L. were characterized for environmental conditions of south Uzbekistan for the first time. Soil, plant element analysis was performed by neutron-activation method. In this case, the samples were irradiated in a nuclear reactor with a neutron flux of 5 × 1013 neutrons/cm2 s, and their quantities were determined in accordance with the half-life of chemical elements. It has also been compared with research materials conducted by world scientists on the importance and pharmacological properties of botanicals in medicine and the food industry, as well as their botanical characteristics. The plant can serve to conserve soil resources, as it prevents water and wind erosion of dense clay soils in the dry subtropical climate of Central Fergana and could be considered an effective agent of destroyed soils remediation. The development of this plant will contribute to the diversification of agriculture in Uzbekistan  and the development of the food industry and pharmacology.The article provides an analysis of the cenopopulation and tissues element composition of the medicinal caper plant  Mankind has been using various herbs for thousands of years to find a cure for food support, and people have been particularly well aware of the properties of medicinal plants and have been able to use them effectively. Unfortunately, nowadays, natural medicinal plants have been virtually non-existent. If one looks at the countries of the world, the propagation and processing of medicinal plants are widely developed in countries such as China, India, Canada, and the United States. Trade-in medicinal plants is growing in volume and in exports. It is estimated that the global trade in medicinal plants is US$800 million per year . The annC. spinosa L. plant.One of the most pressing issues of today is the development of technology for the cultivation and cultivation of natural flora and medicinal plant species belonging to foreign flora, targeted research to obtain environmentally friendly products, introduced into medical practice. One such naturally occurring medicinal plant species is the C. spinosa L.), a drought-tolerant plant belonging to the genus Capparis of the family Capparidaceae is mainly distributed in arid and semi-arid regions of the tropical and subtropical environments. The plant, as a potential source of valuable nutrients such as vitamins  to investigate biogeochemical features of soil and various parts of the C.spinosa L. Plants.In the following period, population growth dramatically increased the demand for medicinal plants. As a result, due to the unplanned use of medicinal plants, their natural resources are reduced, and even some species have to be considered as species with protected status. Therefore, the cenopopulation analysis, which reveals the soil-climatic conditions of each species and the laws of natural regeneration, is of great scientific and practical importance. Thus, the aim of this study was to investigate the dynamics of the cenopopulation of caper and geochemical peculiarities of the environments of caper in Fergana valley. Following objectives were formulated: (1) to evaluate the dynamics of cenopopulation within the onthogenesis of plants in one season, (2) to identify soil types, key soil properties and to characterize environments, suitable for plantation of Cenopopulation analysis of biomorphological properties of plants during ontogenesis was studied using the methods of O.V.Smirnova and others , root syC. spinosa L., which is widespread in this area. Soil profile consists of the following horizons: A—humus accumulative with high roots content, BCA—horizon of secondary accumulation of carbonates, C—pebble gypsum parent materials. Detailed soil characteristics of this region are provided in [The object of study is the light Calcisols and irrigated soils (old irrigated hydromorphic soil (7 A) and recently irrigated hydromorphic soils (6 A) formed on the eroded weakly skeletal alluvial-proluvial rocks of southern Fergana, and the plant vided in . In geneThe location of plots is given in Phenological and morphological methods were used  methods C. spinosa L. [The morphogenetic methods of V.V.Dokuchaev, pedogeochemical approaches of M.A. Glazovskaya, and A.I.Perelman were used as the main methods in the study of soil properties of inosa L. . Soil prinosa L. .The number of soil transects was 10, of which 8 represented automorphous Calcisols and 2 represented semi-hydromorphic soils. In addition, according to the methodology of the field survey around each reference soil section, samples were taken from four small sections for soil description and sampling. For chemical analysis 20 plants were selected and each plant was analyzed in 5 replicates. Roots, stems, leaves, buds, flowers, and fruits were sampled.Mathematical processing was performed according to the method of Kuziev R.K., Yuldashev G. et al. ,33. Stat13 neutron/cm2 s. When irradiated with a neutron flux, natural chemical elements present in the samples under study are converted into radioactive isotopes with different half-lives. It is convenient to measure the content of some elements by short-lived radioisotopes, some by medium-lived radioisotopes, and some by long-lived isotopes. Therefore, samples under study are usually irradiated in three modes: for analysis of short-lived, medium-lived, and long-lived radioisotopes. Short-lived ones have a half-life from a few seconds to several hours, medium-lived from several hours to several days, and long-lived have a half-life of months, years after irradiation in a nuclear reactor.The neutron activation method was used for elemental analysis at the Institute of Nuclear Physics, ANRUz. The Microelement software was applied for statistical processing with the methodology by Kuziev et al. , Kuziev 2SO4 and HClO4 mixture. After 30 min of reaction, the mixture was boiled on the electric stove. The mixture was poured into the measuring flask and passed through the filter. A 2 mm aliquot was taken and poured into a 50 mL flask and further amended by Rochelle’s salt. Then the mixture was neutralized by sodium hydroxide, the final solution was used for colorimetrical determination of the oxides with a coloring reagent (Nessler solution) The fine earth samples were prepared by the grounding of the bulk soil mass and passing the soil through a 2 mm sieve. Soil chemical analyses were performed according to the routine methods of soil chemistry : total cC. spinosa L cenopopulation and its conditions of existence in the dry subtropics of southern Uzbekistan was carried out. Pictures of vegetative C. spinosa L. are giver on A study of the C. spinosa L. roots, stems, leaves, buds, flowers, fruits also expands the use of this plant species in phytobars, food and pharmaceutical industries. In biogeochemical research, the identification of correlations between the chemical, more precisely the elemental composition of plants and the elemental composition of the soils in which they grow is of great scientific and practical importance in establishing programmed yields in agriculture. In particular, knowledge of the exact amount of macro and micronutrients in the generative and vegetative organs of the plant C. spinosa L. belongs to Capparaceae. Capparidaceae is a family of two genus plants with 40 genera and 850 species. Most of the plants belonging to the Capparis family are wild species, which are mainly distributed in arid regions of tropical and subtropical regions [C. spinosa L. in Uzbekistan depends on different geographical conditions [The study of ontogenetic and phenological properties of plants is one of the most convenient and effective methods to determine changes in different phases of the observed plant species, their resistance to environmental conditions, productivity, as well as the rhythm of life processes in them.  regions . The Fer regions ,38. The nditions . The patnditions .C. spinosa L. to soil and air drought allows it to grow in arid areas where water is scarce and in soils with high concentrations of water-soluble salts. Thus, environments of C. spinosa L. habitat were characterized in detail for Souht Uzbekistan for the first time.During our studies, it was found that this species has entered the desert and semi-desert zone, in the foothills and lower mountain regions, sometimes up to the middle zone of the mountains. The study of the biological and ecological properties of any plant requires, first of all, the study of its condition under natural conditions. The natural adaptation of C. spinosa L., has been observed by Saksali et al. as a promising plant that can grow in arid and strongly saline soils with nutrient deficiencies as well as in high-temperature regions [ regions . While tThe length of the stem of plants distributed in the study areas reached 70–170 cm depending on the growing conditions. The inside of the newly formed young stems is covered with fine short hairs, but the hairs fall off as the branch grows during the growing season. The color of the stem is green, there are twisted spines on the underside of the leaf bundle. The number of side branches was 2–6, depending on the stage of development, and was 10–15 cm long. Poya diameter 7–12 mm. The leaves on the stem of the plant differ in shape, width, and length. Usually the leaf shape is round, inverted ovate or elliptical, 3–6 cm long, green, hairless or the lower side has scattered hairs, arranged in series on the main stem and lateral branches through a short leaf band.The flowers are solitary, slightly zygomorphic, 5–8 cm in size, fragrant, located in the axils of one leaf, the petals are 4, curved, ovoid, green, covered with small short hairs on the outside. The petals are 4, but 2 are up to half, white or light pink, many paternal pollen, varying in length, pollinated, brown (flowers turn red after pollination). The flowers are 4–6 cm long. Normally, it blooms in April–May, depending on the amount of precipitation in the study area. The fruit is a multi-seeded berry. The color is green, with long white stripes. The shape is inverted ovate, oblong, walnut or round, many-seeded, or elongated. The outside is smooth, the inside is dark red. The fruit resembles the appearance of a watermelon. When the fruit was ripe, the fruit peel turned outwards and opened. Fruits are 3–5 cm long and 1.3–2.7 cm wide.C. spinosa L. to the diet helps to relieve rheumatic pains. Currently, all parts of the plant are used in modern medicine and folk medicine in the treatment of meteorism, goiter, dentistry , cardiovascular diseases, as well as hypertension, pruritus, jaundice, neurosis, brucellosis [Nowadays, the regular addition of cellosis . Biochemcellosis ,41. C. spinosa L., the need for in-depth study of its biogeochemistry and agroecology was put on the agenda, given the special attention paid to its export potential. It is important to determine the position of the C. spinosa L. in the vegetation cover, the status, ontogeny, and viability of the populations that determine its natural recovery, and thus its current and future raw material reserves.Given the growing demand for raw materials of C. spinosa L. cenopopulation in 10 experimental observation sites (100 m2 each) in Arsif, Satkak, Chimgan, and Altiariq hills were as follows (p) plants averaged 6.0, 5.0 plants belonging to the juvenile (j) state, 10.8 plants belonging to the immature (im) stage, 5.5 plants belonging to the virginil (v) state, 3.5 plants belonging to the generative (g) period, plants typical of the senile (s) period were 2.3.Results of the analysis of  follows : grass  order. It was observed that 60–70% of plants belonging to the immature stage go to the virginil stage in late June and early July, and 10–15% go to the virginil stage in early May after the winter dormancy period. The duration of the immature phase lasts from 20–25 days to 10 months.C. spinosa L., plants belonging to the juvenile and immature stages, are resistant to drought, but most of them die due to the crushing of livestock.Seedlings of Plants belonging to the virginil stage are observed in late June to early July, the length of their main stem reaches 40–80 cm, and it branches up to 2 orders. The root reaches 90–110 cm and branches in 3–4 orders. At this stage is characterized by the formation of thorns on the stems and the thickening of the main root (diameter 4–5 mm). The duration of the virginil state depends in many respects on external environmental factors. It was observed that 15–20 per cent of virginil plants enter the full generative period in the first year and the rest in the second year.Vegetation of plants belonging to the middle-aged generative stage in the Arsif hills lasted from April to December. The length of the main generative stem is 70–170 cm, branched to 2–3 rows, with leaves 4 × 3 cm. In one bush formed an average of 9–14 generative stems. The growing of generative plants was observed in May, flowering in late May, and the formation of fruits began in the second half of June.C. spinosa L. lasted from June to October. One bush, C. spinosa L., produced an average of more than 80 fruits (180–210 in the Arsif and Satkak hills) on the Chimgan hills, and 150 on some bushes. An average of 220–235 seeds were observed in each fruit, the absolute weight of 1000 seeds was 7.25 g. Seed length was about 1–3 mm [The fruiting process of C. spinosa L. in Uzbekistan [C. spinosa L. plant, the migration of chemical elements in the plant-soil chain, and the biogeochemical properties were studied. The population studies of C. spinosa L. have been carried out in a neighbouring country, Iran. It was found that the present population of the plant is very heterogeneous at the genetic level, which is due, among other things, to environmental factors [A few works were published previously about ecology of bekistan ,17. Howe factors .C. spinosa L. contains: phosphorus (679 mg/g), sodium (652 mg/g), calcium (419 mg/g), magnesium (213 mg/g), potassium (157 mg/g), macro and micronutrients such as iron (6.8 mg/g), zinc (5.5 mg/g), manganese (3.30 mg/g) have been reported [This plant is valued by many peoples of the world as a potential source of nutrients, vitamins, phenolic compounds, flavonoids, nutrients in its organs, as well as its strong antioxidant properties and ability to grow in arid conditions. According to the data , 100 g ootassium 7 mg/g, mC. spinosa L. buds contain elements of vitamin K, potassium, calcium and magnesium, which strengthen the bones and prevent the development of osteoporosis. C. spinosa L. prevents hair loss due to the presence of iron and B vitamins, and makes hair grow beautiful and shiny [C. spinosa L. has aroused great interest among scientists. This substance has strong antimetastatic properties and is used in the treatment of prostate cancer. This substance stops the growth and development of cancer cells. This scientific breakthrough is important in the development of anti-cancer drugs. The use of this plant in bowel cleansing and prevention of colon cancer plays an important role [nd shiny . The recant role .C. spinosa L. may be problematic in humans when consumed in the existence of the following diseases: hypotension, constipation, and is not recommended for pregnant women because high sodium levels have been shown to affect the fetus, in individual cases, and in some cases to cause allergies [The high content of sodium in the vegetative and generative organs of plant species, especially in salinized soils of desert areas, requires caution in some diseases. llergies . Althougllergies ,46 and bllergies .The sodium content in the genetic layers of Calcisols ranges from 0.68–0.94 μg/g, which indicates its accumulation in 1.08–1.49 times higher than in Phaeozems of subboreal steppe ecosystem. The medicinal plant C. spinosa L. absorbs in its organs an average of 439 μg/g sodium .The granulometric composition of the soil investigated is characterized by the amount of sand, silt and clay particles in the fine earth. The amount of each particle, together with the amount of other particles, affects the number of properties of the soil. In particular, small particles allow the particles of soil to stick together and grow, and participate in the formation of structural aggregates in the soil. Larger particles are involved in the formation of soil skeletons, improving water and air permeability. Data obtained on soil texture vertical distribution  are in gIt is known that humus is a complex structural substance with a variable composition, the composition of which is constantly changing and renewing. This dynamic condition applies primarily to humic substances, such as humic and ulminic acids, fulvic acids, hyimatomelane acid, etc., along with humic substances, carbohydrates, organic acids, alcohols, hydrocarbons, ethers, aldehydes, nitrogenous substances, and others. Calcisols investigated do not show high content of organic carbon, which well corresponds with data, published recently . The dynIn the soils of Central Fergana, the root system of natural and cultivated plants is located in the very superficial layers of the soil, so the humus layer is weak in these soils. This is true even for soils with arzyk-shokh and shokh-arzyk horizons at different depths .Based on the data, given in the In the upper driving layers of newly irrigated soils, humus content is about 0.8–0.9%.The total amount of nitrogen in the soil humus is correlated, so the law of cross-sectional changes in the amount of nitrogen in the studied soils is reminiscent of humus. The C:N ratio in the soil is a relative measure of the nitrogen content of humus, a characteristic of most irrigated soils. This ratio Kuziyev R.K. data has been found to vary from 7.2 to 13.5, especially in gray-oasis soils. For gray soils, this value is 8 and indicates that nitrogen is rich in nitrogen . The C:NThe total amount of phosphorus and potassium in these soils is not high. For example, gross phosphorus fluctuates around 0.03–0.125% in soil sections. The amount of total potassium fluctuates around 0.80–2.45%, of course 0.80–0.90% corresponds to the man-made strata, while 2.45% corresponds to the driving layer of old irrigated soils. According to the amount of mobile nutrients, these soils belong to the low-income group.In this regard, we can conclude that the total phosphorus and potassium content in the soils of the old irrigated deep 7A section (93–111 cm) and the newly irrigated hydromorphic 6A section (32–55 cm) also decreased sharply as humus.+, Mg2+, Fe2+ and Zn2+ which could be problematic in desert and dry conditions. The regulation of uptake is possible by adding native arbuscular mycorrhizal fungi inoculum into soil under plantation of capper. In any case numerous eco-engeniring strategies could be elaborated with the aim to increase the productiveness and yield of C. spinosa [It is known that the amount of chemical elements and substances varies in different soil types  Table 4Table 4.  spinosa . According to the data in Strengthening the process of hydromorphism leads to an increase in the amount of Mn, Zn, Mo in the soil and its layers from Calcisols to meadow soils. This can also be explained by the fact that the geochemical migration of these elements towards a naturally dependent landscape is accumulated with taking into account that fact that investigated soil were not amended be any fertilizers.C. spinosa L. which is widespread in the Calcisols of South Fergana, changes under the influence of soil properties, plant type, natural climatic conditions and other factors. It was observed that the amount of elements in the composition of C. spinosa L. varies several thousand times depending on the physiological properties of plant organs.The elemental composition of the Our data shows that Mo belongs to the group of strong and very strong aggregates by root bark and fruit. According to the range of biological absorption coefficients, the elements Co, Zn, and Mn belong to the group of very weak, weak, and moderately biodegradable, respectively.There is a positive correlation between the content of trace elements in the soil and their amount in the organs of medicinal plants. The range of correlation is from 0.80 to 0.95 . The higThis, in turn, satisfies the need for certain macro and micronutrients by consuming a biologically active supplement made from this medicinal plant as well as the daily norm of food. The study of the correlation between the elemental composition of the plant and the elemental composition of the soil in which it grows expands the scope of its use in folk medicine, phytobars, modern medicine, and the pharmaceutical industry. This makes a huge contribution to socio-economic development.C. spinosa L. is related to the types of products made from it. In particular, pickled flower buds, known as “capers” or “caper berry”, are the main subject of trade in international markets. In recent years, the annual growth rate of production from C. spinosa L. has increased by 6%. Currently, pumpkin is valued as an important consumer product in the United States and about 60 countries around the world, where the cost of 1 kg of ready-to-eat product is $25. From this round, the Chinese earn three million a year. They are making a profit in the amount of USD. Today, the Kingdom of Saudi Arabia, Lebanon, Syria, and the Mediterranean countries have proposed C. spinosa L. as the main crop type to raise the socio-economic level [C. spinosa L.  are used as anti-aging, skin protection, or anti-inflammatory agents. was commercialized and put up for sale [The main economic importance of ic level . Howeverfor sale .C. spinosa L. It should be noted that until recent years, this plant has been neglected, whereas C. spinosa L. is a very valuable raw material in the pharmaceutical, and food industry. In 2019, in the Jizzakh region, this plant was harvested from existing natural resources and cultivated. 1909 tons of ready-to-eat products were made from its flower buds and fruits. It exported $3.2 million worth of goods to Turkey and Spain. In addition to productivity and yield parameters C. spinosa L. this plant species is characterized by good adaptation to conditions of marginal environments [In the Jizzakh region of the country, 12 enterprises have launched the export of ronments  and, thuC. spinosa L. plantation located on dry subtropical Calcisols of Central Fergana provided for the first time for Central Asia on the territory of the Former Soviet Union (NIS). It was shown, that C. spinosa L. is well adapted to dry soils with alkaline reaction, low humus percentages, and middle key nutrients content. The increasing of soils humidity is not a critical factor for growing C. spinosa L. dynamics. Soil-plant interaction in a case study of C. spinosa L. in a field experiment in case of the various soils was conducted. According to the observations, the viability, and drought tolerance of C. spinosa L. populations is relatively high, and studied plants of the population belonging to the immature and virginil stage are 5–10 times more than senile. This shows that it is possible to collect raw materials of the caper from the southern Fergana hills with severe climatic conditions on a regular basis. Determining the quantitative supply of nutrients and medicinal substances in C. spinosa L. and other medicinal plants will further increase the productivity and increase the medical features of vegetated medical plants in the Central Fergana oasis.Cenopopulational observation of 0, 0n → 0,n → n → 10n, depending on the amount of elements studied. Molybdenum showed features of very strong biological accumulation in plants, while manganese, Zinc, and Cobalt occupy a medium, weak, and very weak retention line.The series of biological absorption is assimilated in the form of C. spinosa L. depending on the composition of the soil, it is possible to assess its sanitary and hygienic characteristics, as well as the level of safety in pharmaceutical use and food quality. Because this plant has anti-cancer, anti-microbial, and anti-viral effects, further surveys of its chemical composition and biochemical specificity should be provided. C. spinosa L. can be used as a raw material in the creation of new medicines.By studying the amount of chemical elements in the organs of the plant C. spinosa L. the global increase in its use in the food industry, and modern medicine, the increase in natural products made from it in the pharmaceutical and cosmetic industries, in turn, increase the pressure on natural resources of this plant species. This would jeopardize the natural reserves of C. spinosa L. in the nearest future. Thus, further research of C. spinosa L. population dynamics in various environmental conditions should be conducted together with a simultaneous determination of the productivity of the plant species.The intensively increasing demand for natural and environmentally friendly products made from


Probiotic microorganisms that are potentially beneficial to the health of the host are commercially available in a great variety of products. Not all microorganism strains present in products have proven beneficial to the health properties. These products include not only foodstuffs but also dietary supplements, food for special medical purposes, medicinal products, as well as cosmetics and medical devices. These products contain from one to a dozen bacterial strains of the same or different species and sometimes also fungal strains. Since the pro-health effects of probiotics depend on a specific strain, the number of its cells in a dose, and the lack of pathogenic microorganisms, it is extremely important to control the quality of probiotics. Depending on the classification of a given product, its form, and its content of microorganisms, the correct determination of the number of microorganisms and their identification is crucial. This article describes the culture-dependent and culture-independent methods for testing the contents of probiotic microorganisms, in addition to biochemical and genetic methods of identification. The microbiological purity requirements for various product categories are also presented. Due to numerous reports on the low quality of probiotic products available on the market, it is important to standardise research methods for this group of products and to increase the frequency of inspections of these products. Recently, interest in biologically active products with potentially beneficial effects on the patient or consumer has increased significantly. Some of the products containing probiotic microorganisms can be used for both therapeutic and prophylactic purposes. Depending on the indications, probiotic microorganisms are applied to humans in the form of foodstuffs, mainly fermented food, dietary supplements, foods for special medical purposes, medicinal products, or medical devices. They can also be found in cosmetics, most commonly in the forms of creams or serums . In addiLarge and important sources of probiotics also include yoghurts, kefirs, and fermented food, most often in the form of cheeses, vegetables , and meaProbiotics are used not only by humans but also in animal production. In recent years, due to the limitation of the use of antibiotics, especially growth promotor factors, the use of LAB on farms to intensify meat production and prevent the development of certain pathogenic microorganisms has started . ProbiotTaking into account the large variety of probiotic products available on the market, some of which are of low microbiological quality, it is unclear how the quality of these products is controlled and whether it is possible to standardise these methods to ensure a safe product with health-promoting properties.An extensive analysis of the available literature describing numerous different methods of determining the contents of probiotic microorganisms and their identification was performed. The aim of this review is also to present the variety of available normative methods for determining the contents, purity and identities of microorganisms in probiotic products. In addition, this review presents the advantages and disadvantages of the presented methods.Lactobacillaceae family, in particular L. acidophilus and L. rhamnosus, as well as L. plantarum, L. casei, L. paracasei, and L. salivarius [Bifidobacterium  [Lactococcus, Bacillus or strains of Streptococcus thermophiles. Strains of yeast species, such as Saccharomyces boulardii, may also be present in these preparations , Staphylococcus aureus, Campylobacter jejuni, or Clostridioides difficile. They prevent the adhesion of these pathogens to the intestinal mucosa through competition for receptors, but they also inhibit their multiplication by competing for nutrients or producing antibacterial substances such as organic acids, hydrogen peroxide, and/or bacteriocins [The group of probiotic organisms includes bacteria and fungi which, when administered in appropriate amounts, may exert a beneficial effect on the host’s health . The moslivarius . Frequen. breve) , Lactocoarations . Lactobaeriocins ,17,18,198–1011 CFU (colony-forming units) of bacterial or yeast cells in the daily dose [Both the FAO and the WHO ,20 definily dose .To assess the safety of probiotics application, the following factors should also be taken into account: a large variety of probiotic strains, the risks associated with the use of strains which do not have GRAS or QPS  status, as well as the possibility of interaction between the probiotic strains and the host microbiota. Probiotics may be responsible for systemic infections; excessive immune stimulation, especially in immunocompromised individuals; deleterious metabolic effects; and gene transfer .Enterococcus, namely E. durans, E. faecium and E. faecalis, classified as probiotic bacteria , although they are opportunistic microorganisms capable of causing infections in humans. Numerous studies indicate the increasing importance of multidrug-resistant Enterococcus sp., especially those resistant to vancomycin, and the possibility of transferring resistance genes through horizontal gene transfer to other bacterial genera [E. faecalis DSM 16431 is a compound of a drug called Symbioflor 1 and is used in acute and recurrent sinusitis and bronchitis [E. faecium M74 and E. faecium SF-68 are included in dietary supplements such as FortiFlora and Cernivet, which are considered effective and safe [Some concerns have been raised regarding strains of the genus l genera . Howeverl genera ,25,26,27onchitis . On the The probiotic preparations available on the market are present in a variety of forms. Without consideration of fermented foods such as yoghurts and kefirs as probiotics, present in almost every supermarket, the most common pharmaceutical forms of probiotics are lyophilised capsules  and oral drops. Recently, however, it has become possible to frequently encounter microencapsulated lyophilisates, which are designed to preserve the stability of probiotics during storage, protect them from harsh conditions in the upper gastrointestinal tract, release them in the colon, and facilitate probiotic microorganisms to colonise the mucosal surface ,38,39. TMoreover, lyophilised probiotics are also available in the form of ampoules, vials, or sachets. Probiotics in the form of tablets, as well as chocolate tablets in various forms , or even lollipops, are also on sale.S. salivarius K-12, S. salivarius M-18 and L. plantarum 8P-A3), which, by producing bacteriocins, could inhibit the growth of bacteria present on the surface of the skin as well as pathogens that cause wound infections  [Probiotic microorganisms are also included in cosmetic products. The most common products of this group found on the market are creams, serums, masks, and gels, but also exfoliants, cleansers, foundations, soaps, lotions, shampoos, toothpaste, or deodorants ,42. Mostuginosa) .Determination of the microbial content in probiotic products can be performed using various methods presented in the literature. The most common ones are cultivation methods with the use of appropriate media, as well as the increasingly popular method of flow cytometry. Other methods are also described, such as fluorescence in situ hybridisation (FISH)  or nucleBifidobacterium and E. coli present in the same product, using Blaurock medium with sodium azide and Endo Agar [Cultivation methods, such as plate count methods, are the gold standard . The plando Agar .In analytical tests carried out in various laboratories on the contents of probiotic microorganisms in products, described in the literature, the following procedure can be recommended. The weighed sample is dissolved in a peptone buffer ,3,7 or pBifidobacterium. However, it is necessary to apply appropriate sterilisation conditions during the preparation of the medium. According to the information provided by the manufacturer, MRS agar should be sterilised at 121 °C for 15 min. Moreover, if the growth of the Bifidobacterium spp. is desired, a temperature of 118 °C for 15 min should be applied [Bifidobacterium strains, it is more practical to use two different media, e.g., MRS agar for quantifying lactobacilli and TOS-MUP for quantifying Bifidobacterium [Bifidobacterium bacteria on the MRS agar medium is excluded. The problem grows when products containing even more types of probiotic microorganisms are tested. The media should be selected in such a way that only one type of microorganism will grow on each of them. For instance, the most common medium used for the cultivation of S. thermophilus and Lc. lactis is the M17 medium. However, to obtain the growth of only the desired group of microorganisms or to determine the number of cells of each type in the product separately, it is necessary to apply different incubation conditions  and ciprofloxacin (10.0 mg/L) to the medium is recommended. These antibiotics inhibit the growth of the mentioned species, except for L. acidophilus (ISO 20128:2012) [To mark the microorganisms of the species 28:2012) .Bifidobacterium consists of strictly anaerobic bacteria which grow on the agar surface in the form of round, whitish colonies, some of them star-shaped or triple-lobed [L. delbrueckii subsp. bulgaricus forms lenticular colonies with sharply defined contours on the acidified MRS agar [L. acidophilus grows in the form of flat, opaque grey or whitish colonies with uneven edges [S. thermophilus, however, grows on this agar medium in the form of lenticular colonies [In some cases, it is possible to distinguish bacterial colonies of different species remaining on the same Petri dish. The genus le-lobed . In contMRS agar , and L. en edges . The specolonies .Although the direct plating on Petri dishes with agar medium is the most popular method, it also has its limitations. For example, sample preparation—the rehydration of lyophilised probiotics —is extreThe analytical, flow cytometry method enables the qualitative and quantitative determination of microorganisms in the tested sample within a very short time, which is an advantage compared to culturing methods. The study uses fluorescent dyes, which enable the assessment of parameters related to the surface, structure, and size of cells . By usinIn a study using flow cytometry, the number of bacterial cells was determined directly in the test sample after the addition of an appropriate dye. Bacteria capture the dye, which, under the influence of intracellular enzymes, splits and releases molecules capable of fluorescence . DependiThe results obtained with the cytometry method are expressed in units of fluorescence activity (Active Fluorescent Unit AFU/g). Additionally, it is possible to quantify the value of the non-Active Fluorescent Unit (n-AFU/g), which represents damaged and dead bacterial cells, stained with PI, as it enters cells with an intact membrane and binds to DNA. The Total Fluorescent Unit (TFU/g) represents the total number of cells as the sum of AFU and n-AFU .Bacillus subtilis, B. licheniformis, B. pumilus and B. velezensis strains with the use of SYTO 24 and LDS 751 (Laser Dyes Styryl)—cell permeant nucleic acid stain. The obtained results indicate no statistically significant differences in the determination of the number of spores by flow cytometry and the use of the plating methods [Genovese et al.  used flo methods .B. longum strain was an exception, for which a greater number of colony-forming units was demonstrated with classical microbiological methods. On the other hand, a study conducted by Michelutti et al. [B. animalis and L. acidophilus in probiotic products. Moreover, the flow cytometry method was characterised by greater repeatability and better precision. However, the flow cytometry method is used to quantify all microbial cells in the tested sample, not only probiotic cells but also microbes contaminating the preparation, which may cause false-positive results.Comparative studies of the two methods used for the quantification of probiotic bacteria, the culture method and flow cytometry, conducted by Chiron et al. , demonsti et al.  showed aP. acidilactici, P. pentosaceus and L. plantarum as well as spores of B. subtilis. The number of cells estimated in the presented method was higher than the manufacturer’s declaration for all the species mentioned. The second product contained a mixture of spores of four Bacillus species: B. subtilis, B. amyloliquefaciens, B. licheniformis and B. pumilus. The number of estimated Bacillus spores was consistent with the declaration on the label [Other methods of microorganism counting are much less frequently used . Fluoreshe label .P. acidilactici, P. pentosaceus, L. plantarum, and B. subtilis in 20 samples of a complex probiotic product. In their study, flow cytometry and the qPCR method gave similar results, which were, however, significantly higher compared those provided by the plate method, especially in determinations performed in the later storage periods. These results suggest that some bacteria in the population entered the VBNC state and could only be counted by flow cytometry and qPCR methods [Other methods, namely molecular techniques based on the detection of nucleic acid sequences, can also be used to count bacterial cells. These methods include, e.g., polymerase chain reaction (PCR), reverse transcriptase PCR (RT-PCR), or real time-quantitative polymerase chain reaction (RT-qPCR or qPCR). Quantitative polymerase chain reaction (qPCR) is a technique that enables the quantitative assessment of the microbial population using appropriate dyes and probes. Appropriate equipment is necessary, and the method facilitates the monitoring of the increase in DNA in each subsequent reaction cycle . Gorsuch methods .P. acidilactici, P. pentosaceus, L. plantarum, and B. subtilis in 20 samples of a complex probiotic product. In their study, flow cytometry and the qPCR method gave similar results, which were, however, significantly higher compared those provided by the plate method, especially in determinations performed in the later storage periods. These results suggest that some bacteria in the population entered the VBNC state and could only be counted by flow cytometry and qPCR methods [Other methods, namely molecular techniques based on the detection of nucleic acid sequences, can also be used to count bacterial cells. These methods include, e.g., polymerase chain reaction (PCR), reverse transcriptase PCR (RT-PCR), or real time-quantitative polymerase chain reaction (RT-qPCR or qPCR). Quantitative polymerase chain reaction (qPCR) is a technique that enables the quantitative assessment of the microbial population using appropriate dyes and probes. Appropriate equipment is necessary, and the method facilitates the monitoring of the increase in DNA in each subsequent reaction cycle . Gorsuch methods .Identification of probiotics is usually carried out by known standard microbiological methods. According to the FAO/WHO  recommenLactococcus, the API 50 CHL test. Boyd et al. [Bifidobacterium should be performed using the API 20A kit, dedicated to anaerobic bacteria. On the other hand, the identification of S. boulardii yeasts can be performed with the API 20C AUX test, dedicated to yeasts. In addition, the API ZYM Kit can be used to help identify bacteria and determine the potential of probiotic microorganisms [Initial characterisation of probiotic bacteria consists of the determination of the cells shape after staining with the Gram staining method as well as the assessment of mobility and the ability to produce catalase . The easd et al.  correctlrganisms .Bifidobacterium only to the genus level. However, the test results obtained are often inconclusive [The VITEK system (bioMérieux) is characterised by a similar principle of operation, where microorganisms can be identified on the basis of biochemical reactions. The VITEK 2 ANC system card allows for identifying lactobacilli to the species level and nclusive ., Lactococcus, and Bifidobacterium [E. faecalis, four E. durans, two Streptococcus spp., and one Lc. lactis; twelve isolates were classified as other species, whereas five were not identified at all. Analysing the same samples using API 50 CHL tests resulted in fourteen L. plantarum, six L. paracasei, six Lc. lactis, and one Lactobacillus spp.; one sample was classified as a different species, and one was not identified at all. The authors of this publication confirmed their results using molecular identification—16S rDNA sequencing, in which 20 results were obtained, identifying the tested microorganisms as Enterococcus spp., five L. plantarum, three Lc. lactis, and one Streptococcus spp. [Another method of microbial identification is the BIOLOG system , which is used to identify species of aerobic as well as anaerobic bacteria, yeasts, and fungi , includiacterium . This syacterium . Moraes acterium  identificus spp. .Databases for the analysis of phenotypic results often do not take into account the latest taxonomic changes or newly described species, and therefore, the interpretation of the obtained results is not accurate.B. animalis, B. breve, B. longum, B. bifidum, B. infantis, E. faecium, L. acidophilus, L. casei, L. gasseri, L. paracasei, L. plantarum, L. reuteri, L. rhamnosus, L. salivarius, Lc. lactis, S. thermophilus, and S. cerevisiae. The accuracy of this identification was also confirmed with the PCR method. Nevertheless, when the MALDI-TOF method is used, errors also occur in the identification of lactic acid bacteria, especially in the case of closely related species such as L. casei and L. paracasei [Another method enabling accurate and prompt identification of the tested microorganism is the MALDI-TOF MS—Matrix—Assisted Laser Desorption/Ionisation—Time of Flight—Mass Spectrometry technique. This method allows a comprehensive analysis of the protein panel of a given microorganism. The test consists of analysing the spectral distribution of proteins directly in bacterial cells ,80. Protaracasei ,78,82. CLactobacillaceae: L. acidophilus, L. delbrueckii, L. casei, L. gasseri, L. plantarum, L. reuteri, and L. rhamnosus [L. acidophilus group . In this case, it is impossible to distinguish particular species [L. acidophilus, L. plantarum and L. casei. The obtained results of the analysis of 17 probiotic products showed that not all products contain bacterial species corresponding with the information provided on the package.The problem with unequivocal identification of a species using phenotypic methods is related to the common phenomenon of phenotypic variability, resulting, among other things, from changes in gene expression under the influence of environmental conditions. Molecular biology methods, based on the analysis of the genetic material of bacteria, are much more accurate, sensitive, and reproducible. These methods are less reliant on the growth conditions of the bacteria, allowing the microorganism to be identified not only down to the species level but even to the strain. Many different molecular biology methods are used to identify and differentiate probiotic microorganisms. The most commonly used molecular assays for the identification of lactic acid bacteria are nucleic acid amplification tests. The PCR-based research is characterised by high sensitivity and specificity. The process of identifying lactic acid bacteria is performed based on gene sequences which encode ribosomal RNA , amplification of ITSs (intergenic spacer regions) located between the genes encoding the 16S and 23S rRNA (ITS-PCR) and amplification of regions between genes encoding tRNA (tDNA PCR), sequence analysis of the genes encoding 16S rRNA, 23S rRNA, or ITS, restriction analysis of the rDNA gene amplification product ARDRA , ribotyping and DGGE/TGGE (Denaturing Gradient Gel Electrophoresis/Temperature Gradient Gel Electrophoresis) ,80. In mhamnosus . However species . Kim et  species  determinL. helveticus, L. sake, L. plantarum and L. delbrueckii subsp. bulgaricus [L. casei group: L. rhamnosus, L. paracasei subsp. tolerans, and L. zeae.Another technique used for probiotics identification based on DNA amplification is RAPD (Random Amplified Polymorphic DNA). This method is based on amplification with the use of a short primer , where the ratio of G-C to A-T pairs is taken into account. The primer bonds with numerous homologous sequences in the analysed chromosomal DNA of a given species ,86. The lgaricus  can be slgaricus , with thL. acidophilus and L. crispatus; L. casei and L. rhamnosus, as well as L. acidophilus, L. helveticus, and L. amylovorus. Moreover, with the additional use of Sau3AI or Mse I restrictase, they were able to distinguish L. plantarum and L. pentosus species.Among the methods that use restrictive analysis to identify probiotic bacteria, the RFLP (Restriction Fragment Length Polymorphism) method may be employed. In this method, the differences in the band patterns reflect changes in the DNA sequence, which result in the absence or an additional restriction locus recognised by the restriction enzyme used . BlaiottThe T-RFLP technique  is a modification of the PCR-RFLP technique, in which the 5′-end primer is labelled with a fluorescent dye  so that only the labelled terminal restriction fragments are detected. This method does not require conducting a culture to identify a species from a mixed bacterial population; moreover, its accuracy can be increased by using more restriction enzymes . This meL. plantarum, L. pentosus and L. pseudoplantarum. Jarocki et al. [L. casei group.Another molecular biology method used to identify probiotic strains is the AFLP (Amplified Fragment Length Polymorphism) method, based on the analysis of the entire bacterial genome. This technique employs the phenomenon of the ligation of nucleotide adapters and the selective amplification of restriction fragments. In the AFLP technique, the following restriction enzymes are used: frequently cutting  and rarely cutting , leaving sticky ends ,86,88. TL. casei group: L. casei, L. paracasei, L. rhamnosus, and L. zeae. Xu et al. [L. rhamnosus, L. paracasei, L. plantarum, L. acidophilus, L. fermentum, L. curvatus, and L. delbrueckii subsp. lactis.The method in the differentiation and relationship searching of strains, recommended by the FAO/WHO, is Pulsed Field Gel Electrophoresis (PFGE) . This meu et al.  digestedThe gold standard method that serves both to identify and to determine the LAB drug resistance profile is the whole genome sequencing method (WGS). The identification of strains using WGS can be performed using one of the available methods—single nucleotide polymorphism (SNP) analysis or the gene-by gene analysis method. The SNP method consists in comparing the genome of a given bacterium with a reference genome, as a result of which information about nucleotide differences is obtained. In turn, the gene-based method can be used to analyse the genetic relationship between LAB strains . SpecialE. coli or Salmonella sp. Depending on the consumer groups for these products, consideration should also be given to excluding the presence of other pathogens such as S. aureus, P. aeruginosa, Listeria monocytogenes, Clostridioides or Cronobacter sakazakii in infant products. The requirements to be met by individual product groups are regulated by the European Pharmacopoeia (Ph. Eur.) [Probiotic products, both those classified as medicinal products and food, should meet several quality requirements which are regulated depending on the status of the product. Regardless of the classification of the product as food or medicine, the product should not be contaminated with pathogenic bacteria, such as h. Eur.) ,98,99, th. Eur.) ,101,102,h. Eur.)  and the h. Eur.) .Bacillus clausii spores, yeast S. cerevisiae var. boulardii), various media and incubation conditions should be used, tailored to the specifics of the test product and the presence of the microorganisms in it  should be excluded , there are maximum permissible counts of contaminating microorganisms [E. coli or Salmonella sp. are specified. If there is a risk of contaminating raw materials or the finished product, the presence of L. monocytogenes, S. aureus and P. aeruginosa should also be excluded, whereas in the products intended for infants, bacteria such as Clostridium perfringens and Cronobacter sakazakii must also be excluded [The US Pharmacopeia specifies the microbial purity requirements for products classified as both medicinal products and dietary supplements . Dependirganisms ,100. In excluded ,101.Salmonella [Cronobacter spp. [E. coli [Enterobacteriaceae [Enterobacter sakazakii [L. monocytogenes [Documents published by the FDA and the European Commission  regardinlmonella ,104, Croter spp. , E. coli[E. coli , Enteroberiaceae , Enterob3 CFU in 1 mL or 1 g of the product; preparations used in the vicinity of the eyes are an exception [2 CFU per 1 g [2 CFU per 1 g or 1 mL [S. aureus [P. aeruginosa [Streptococcus pyogenes [Klebsiella pneumoniae [E. coli [C. albicans [The FDA , like thxception ,106. Whe per 1 g , and in . aureus ,106, P. ruginosa ,106, Strpyogenes , Klebsieeumoniae , E. coliE. faecium, even at the level of 109 CFU/dose. In addition, contamination with microorganisms such as Acinetobacter baumannii (1011 CFU/dose), Lysinibacillus fusiformis (1011 CFU/dose), B. cereus (1010 CFU/dose), Bacillus leantus (109 CFU/dose), and Staphylococcus spp. (102 CFU/dose) was detected [Mazzantini et al.  collectedetected .An increasing number of probiotic products appear on the market. In the literature, there is a large amount of information about the incorrect number of probiotic microorganisms, contamination of the tested products at a very high level, and the lack of proper labelling of the strains included in the composition of the preparations.There is no doubt that methods of testing the contents of probiotic products, especially the proper preparation of the sample and the selection of the appropriate method for counting and identification of microorganisms, are necessary. However, there are no detailed, universal guidelines for testing these products, especially when they contain many different types of microorganisms (strains of the same or different species and genera), which may cause differences in the results obtained by laboratories. The different survival times of microorganisms in the product also affect the identification of the strains declared by the manufacturers, which often, during the shelf life of the product, are found in very low numbers that are not sufficient to provide the health benefits to the host in any way, which is the basic task of probiotics.Besides, the probiotics contaminants may include pathogenic microorganisms, which suggests that products containing live microorganisms, regardless of whether they belong to the category of medicinal products, dietary supplements, food or cosmetics, may not be safe and should be subject to strict quality control.Taking into account reports on the poor survival of microorganisms in products, they should be also subjected to stability tests, similarly to medicinal products, to eliminate poor-quality preparations or to shorten the validity period. Detailed information about the strains contained in particular product should be provided by the manufacturers on the product package or in informational materials. Not only the generic or species name but also strain designation should be stated, because the properties of probiotics are strain-dependent.Based on this review, a substantial amount of work needs to be done to ensure that the probiotic products available on the market are of good quality, safe and fulfil a pro-health function.


Oedaleus decorus asiaticus (Bey-Bienko) populations are highly concentrated in Xilingol League of the Inner Mongolia Autonomous Region where they regularly harm the ecosystems protecting northern China. In order to prevent the occurrence of O. d. asiaticus in the Xilingole grassland, the establishment of a high-precision method and extracting the area of occurrence for O. d. asiaticus is required. The primary objectives of this study were: (1) to project the potential distribution of O. d. asiaticus by MaxEnt model and remote sensing data, and (2) to explore the responses and relative contributions of habitat factors to the distribution of O. d. asiaticus. The study will help guide managers and decision-makers to prevent and control the occurrence of O. d. asiaticus early on and this work may facilitate meaningful reductions in pesticide application.Oedaleus decorus asiaticus (Bey-Bienko) (O. d. asiaticus) is the most serious species in Xilingol League of the Inner Mongolia Autonomous Region. The region is not only an important grassland but also a site of agricultural heritage systems in China. Therefore, projecting the potential geographic distribution of O. d. asiaticus to provide an early warning is vital. Here, we combined temperature, precipitation, soil, vegetation, and topography with remote sensing data to screen the predictors that best characterize the current geographical distribution of O. d. asiaticus. A MaxEnt model approach was applied to project the potential suitable distribution of O. d. asiaticus in Xilingol League (the Inner Mongolia Autonomous Region of China) combined with a set of optimized parameters. The modeling results indicated that there were six main habitat factors that determined the suitable distribution of O. d. asiaticus such as the soil type (ST), grassland type (GT), elevation, precipitation during the growing period (GP), precipitation during the spawning period (SP), and normalized difference vegetation index during the overwintering period (ONDVI). The simulated result was good, with average AUC and TSS values of 0.875 and 0.812, respectively. The potential inhabitable areas of grasshoppers were 198,527 km2, distributed mainly in West Urumqi, Xilinhot City, East Urumqi, Abaga Banner, and Xianghuang Banner of Xilingol League. This study is valuable to guide managers and decision-makers to prevent and control the occurrence of O. d. asiaticus early on and this study may facilitate meaningful reductions in pesticide application.Grasshopper populations can quickly grow to catastrophic levels, causing a huge amount of damage in a short time.  Oedaleus decorus asiaticus (O. d. asiaticus) populations are highly concentrated in Xilingol League of the Inner Mongolia Autonomous Region where they regularly harm the ecosystems protecting northern China. Moreover, previous research has explored the relationships between O. d. asiaticus and climate and showed that the O. d. asiaticus would occur with a high density since the El Niño phenomenon happens [O. d. asiaticus is not only valuable for the construction of ecological grasslands but it is also vital to maintaining healthy grass ecosystems and supporting the livestock depending on grass.The  happens ,2. ThereDisasters caused by locusts and grasshoppers have occurred in China over thousands of years. Analyzing the occurrence patterns of locusts and grasshoppers and their drivers is the basis of related prediction researches ,4. PreviThe methods for extracting the habitat factors mainly consist of species distribution models and expert empirical scoring. However, the expert empirical method is inadequate in the quantification of indicators due to the subjective experiences of researchers . The speO. d. asiaticus, Dasyhippus barbipes , and Bryodema luctuosum. Here, the O. d. asiaticus was selected due to the most serious species of disaster. O. d. asiaticus eggs are laid from July to September, remain in the soil during the cold winter period (October–March), before hatching during the period of April–May in the following year, and finally grow to maturity in June. Therefore, we divided the reproductive periods into four stages, such as the spawning period (July–September 2019), overwintering period (January–February 2020), incubation period (April–May 2020), and growing period (June 2020). Note that these stages are correspond to the habits of O. d. asiaticus  to select a MaxEnt model with the optimal combination of parameters to best predict the potential distribution of O. d. asiaticus; (2) to explore the responses and relative contributions of habitat factors to the distribution of O. d. asiaticus; and (3) to characterize the areas in Xilingole, which are suitable for O. d. asiaticus. The study can guide managers and decision-makers in providing an early warning for the disaster occurrence of O. d. asiaticus and may further help the precise control by using pesticides and reducing the amount of pesticide application.In summary, in order to prevent the occurrence of grasshoppers in the Xilingole grassland, a high-precision method needs to be established and a suitable area of the occurrence of O. d. asiaticus populations are concentrated in Xilingol League of China, containing existing areas of grasshoppers’ occurrence of nearly 300 km2 across 13 banners . Hundreds of tons of chemical pesticides have been applied to prevent and control O. d. asiaticus disasters in this region [O. d. asiaticus damaged grasslands in an area of 235.10 km2 in the Xilingol League, mainly distributed in the West Ujimqin Banner, Xilinhot city, East Ujimqin Banner, and Abaga Banner . O. d. asiaticus surveys were conducted from June to August 2020. If the grasshopper densities were above 15 specimens per square meter in the grassland, the location was marked as a grasshopper distribution site, and a total of 310 occurrence points were collected . The topography indicator (elevation data) was derived from the ASTER GDEM V3 data, obtained from China’s Geospatial Data Cloud .The soil indicator (soil type data) was obtained from the 1:1 million China soil map . To describe the vegetation condition, MOD13A2 V6 was used to calculate NDVI .The vegetation indicators, including the vegetation type and normalized difference vegetation index (NDVI), were obtained from China’s Resource and Environment Science and Data Center . To describe the precipitation conditions, this study used the Climate Hazards Group InfraRed Precipitation with Station data (CHIRPS), which is a quasi-global precipitation dataset . The meteorological indicators, including the land surface temperature and precipitation, were inverted by remote sensing. To describe the temperature conditions, MOD11A2 V6 was applied to determine the average 8-day land surface temperatures (LST) . Data during the period of July 2019–July 2020 were extracted, covering the growth and reproduction periods of the O. d. asiaticus. The whole data were further processed following these steps: (1) the resolution of all environmental variables were either at 1 km or were resampled to 1 km by using the nearest-neighbor method; (2) the World Geodetic System 1984 benchmark was used to project all the geographic data into a geographic coordinate system. A batch processing code based on Python in ArcGIS was used to pre-process to a unified data coordinate system, spatial resolution, and data analysis range, so as to facilitate the subsequent modeling analysis.We downloaded these data for each reproductive period from the Google Earth Engine  [Moreover, we used the subsampling replicated run type, and 70% of the sample points were randomly selected for the training dataset, while the remaining 30% were set aside as the test dataset. The convergence threshold was set to 100 and 1) .We used the area under the receiver operating characteristic curve (ROC) of the test data, i.e., the AUC, to evaluate the quality of MaxEnt models. The AUC value ranges within 0–1.0. The overall accuracy of the developed model was divided into excellent (AUC > 0.9), good (0.8 < AUC ≤ 0.9), fair (0.7 < AUC ≤ 0.8), poor (0.6 < AUC ≤ 0.7), and fail (AUC ≤ 0.6) . MeanwhiThe environmental variables of the same category may be correlated with each other, reducing the predicted accuracy of the model . The “CoAdditionally, the percent contribution and permutation importance are the contribution and dependence  of each environmental variable. The high value of the percent contribution and permutation importance represent the large contribution and dependence .p < 0.05, 0.05 ≤ p < 0.33, 0.33 ≤ p < 0.66, and p ≥ 0.66 represent a low, moderately low, moderately high, and high suitability area, respectively [Combining the sample dataset with the optimal combination of environmental variables affecting the distributions of grasshoppers, the MaxEnt model was used to reconstruct their relationship. The existence probability (P) of grasshoppers was graded based on the expert empirical method at each grid point, where ectively ,33.The correlation of all the habitat factors was calculated using the Spearman’s rank correlation coefficients to eliminate the influence of covariance on the modeling process and the interpretation of the results. The results indicated that the correlation coefficients of all the habitat factors were below 0.8, which implied that the combination of all the habitat factors can be used to develop the MaxEnt model a. The RO2, 73,575 km2, 53,005 km2, and 7442 km2, respectively, accounting for 32.49%, 37.06%, 26.70%, and 3.75% of the grassland areas  Keng, Stipa krylovii Roshev, and Leymus chinensis (Trin.) Tzvel were all highly suitable with values of 0.87, 0.77, 0.62, and 0.58, respectively. The elevation ranged from 800 m to 1400 m with a suitability value of above 0.5. The relationship between the suitability of O. d. asiaticus and precipitation was positive when the precipitation was below 1.3 mm/day during the growing period, and negative if the precipitation was over 2.0 mm/day. However, the precipitation during 1.3–2.0 mm/day was the most suitable. In addition, the suitability of O. d. asiaticus increased when the mean precipitation during the incubation period was less than 0.5 mm/day, while the suitability decreased when the value of the precipitation was over 0.7 mm/day. The suitability is high when the ONDVI was between −0.1 and 0.1. The relationship between the suitability of O. d. asiaticus and ONDVI during the overwintering period was positive when the value of ONDVI was below 0, while the suitability decreased when the value of ONDVI was over 0.The response curves of dominant habitat factors are shown in O. d. asiaticus distribution. This study used MODIS remote sensing products from assimilation systems as the main data sources of the environmental factors from September 2019 to June 2020 in Xilingol League. The addition, the remote sensing data improved the spatial and temporal resolutions of the potential distributions and increased the accuracy of the simulated results. To date, remote sensing has been widely applied in monitoring plant diseases and pests [O. d. asiaticus in the coming decades. For example, it will be used to monitor the severity of grassland disasters caused by pests based on the changes in forage biomass. Meanwhile, it will be applied to narrow the scope of the control through a remote sensing inversion of the host plants, further improving the precision of the preventative measures. Currently, the grassland grazing activities by pastoralists resulted in a large variability in the growing periods of the pastures. Likewise, cloud and foggy weather frequently interfered with the coefficient of crop remote sensing inversion in Xilingol League. Under these circumstances, the vegetation index, as a relatively convenient and efficient method, can reflect the vegetation conditions and improve the accuracy.Remote sensing data enable the possibility of real-time monitoring, allowing short-term predictions of nd pests . FurtherO. d. asiaticus [O. d. asiaticus, the influences of vegetation and soil cannot be ignored.This study showed that the majority of potentially habitable areas for grasshoppers were distributed in West Urumqi, Xilinhot City, East Urumqi, Abaga Banner, and Xianghuang Banner of Xilingol League. This analysis also identified the key habitat factors determining the suitability for grasshoppers. To comprehensively consider the impact of the environmental factors, this work used a total of six environmental factors from three categories, including meteorologic, vegetation, and soil. However, most previous studies only considered the impact of meteorological factors on the suitability for siaticus ,42,43. AO. d. asiaticus distribution for the soil and vegetation were 31.3% and 19.7%, respectively. Both the type and composition of vegetation affected the O. d. asiaticus feeding. In addition, the growth of vegetation affects the near-surface hydrothermal conditions, which ultimately affects the survival and development of O. d. asiaticus. A previous study indicated that needlegrass and sheepgrass survived well in warm and dry climatic conditions, which agreed with the results of our study [O. d. asiaticus because it affects its growth and development [O. d. asiaticus distribution.The temperature, texture, moisture content, and salt content of soil influence the growth and reproduction of grasshoppers in grasslands . The phyur study . AdditioO. d. asiaticus was when the ONDVI value was less than 0. This is because an ONDVI value of less than 0 represents that there is a snow cover [O. d. asiaticus, directly affecting the O. d. asiaticus growth, changing the grassland vegetation to indirectly influence its growth. We manifest that GP accounted for 11.0% among the whole environmental factors. The relationship between the suitability of O. d. asiaticus and the precipitation was positive when the precipitation was below 1.3 mm/day during the growing period, and negative if the precipitation was over 2.0 mm/day. Meanwhile, GP with the value of 1.3–2.0 mm/day was most suitable for O. d. asiaticus. The reason is that the hatching rate of O. d. asiaticus was reduced when they were immersed in warm and wet soil for a long period, which was consistent with other studies [O. d. asiaticus distribution is higher than the other climatic factors.Moreover, the land surface temperature, precipitation, and NDVI factors were divided into four periods according to the different growth stages of grasshoppers in this region. We concluded that the highest suitability for ow cover . The mea studies ,45. TherO. d. asiaticus. Certainly, a further data collection should follow to improve the accuracy of the future modeling predictions. Furthermore, the selection of influence factors was limited, and other factors, such as the grasshopper control situation, grazing situation, and the migration of grasshoppers from external areas, are not considered. It is well known that the modeling precision with species distribution model ensembles is better than a single species distribution model due to the uncertainties of each model. Therefore, the next step is to predict suitable grasshopper areas with multispecies distribution models.In general, this study only extracted a suitable distribution using grasshopper occurrence data from a short time series; thus, this research could provide methodological support for an early warning and the efficient prevention and control of


Proc. R. Soc. B. 275, 2831–2899. (Published online 26 August 2008). (https://doi.org/10.1098/rspb.2008.0876)Due to an error in their calculation, the standard errors reported in if chemical cues are used in habitat selection, rather than determining a detection threshold for this species of a concentration gradient.Additionally, we would like to clarify that the strong treatment effects found in this experiment are likely the result of a combination of factors including, but not limited to, the ecology of the focal species, the flume apparatus, the chemical comparisons being tested as well as the concentration of the chemical cues tested. The chemical cue concentration is likely higher than the organisms would experience in nature and therefore likely a form of supernormal stimuli. Simply put, supernormal stimuli are bigger and more intense than normal, and elicit a larger than normal response from the animal . Here, t


Gossypium has important ethnobotanical and economic value for Amazonian Native Communities (A.N.C.). However, little research has been undertaken on the distribution and genetic diversity of cotton populations maintained in the Peruvian rainforest. This work aims to present the first report on the genetic diversity of Gossypium spp. populations in the A.N.C. of the province of La Convención, Cusco-Peru. The methodology was based on exploring, collecting, identifying, and characterizing the Gossypium populations present in the A.N.C. Twenty-six descriptors were evaluated , and with this information, distribution, correlation, and principal component (PC) analyses were carried out. As a result, plants of two species [G. barbadense L. (44 samples) and G. hirsutum L. (19 samples)], one variety [G. barbadense var. brasiliensis (75 samples)], and three previously unidentified variations (9 samples) were identified. Altogether, 147 samples were collected. G. barbadense var. brasiliensis, which was always found in association with other economic crops within an altitude range of 338 to 1086 m, was the most predominant (51%), distributed in eleven A.N.C. and always in small plots (up to 2 ha). G. barbadense L. was cultivated between 397 and 1137 m of altitude in eight A.N.C. in plots of up to 3 ha in marginal lands. G. hirsutum L., with a smaller distribution (13%), was found between 334 and 497 m of altitude in only three communities; this species is cultivated in marginal areas throughout the year. The variability found for the first two PCs when considering the quantitative and qualitative descriptors was high (74.7%) and moderate (48.2%), respectively. When combining all the descriptors, the analysis showed that the first two PCs accounted for 51.8% of the total variability of the data. The PCs of the two types of data and their combination confirmed that the three populations found were grouped. The nine undefined samples were close to or intermediate between the described ones, showing that these samples may be the result of spontaneous crosses; as such, these samples need to be better evaluated with other tools for further definition. The information obtained shows that in the A.N.C. of Cusco-Peru, there is variability conserved by the inhabitants, who have been able to maintain and use these genotypes, even from their Amazonian indigenous ancestry, and the environment has been able to generate variability among the species, as will be highlighted in future works.The genus  Gossypium spp. in some Peruvian Amazonian regions. To date, some species cultivated on the Peruvian coast have been described and found in more than 68 countries  and one botanical variety [G. barbadense L. var. brasiliensis (51%)] were identified and characterized from a total of 147 samples collected in the two districts of La Convención Province, distributed in 12 A.N.C.Two species  (G. barbadense L. var. brasiliensis (red) and G. hirsutum L. (Green), G2 was located close to G. hirsutum L. (Green), and G3 was located close to the group formed by the samples of G. barbadense L. var. brasiliensis (red)  in the PCs, we found that 51.8% of the variability of the data was retained in the first two components . As obse(green)] . For theis (red) .Gossypium species, which prevents clearer information about this genus from being made available [G. barbadense L. (30%) and populations of the introduced species G. hirsutum L. (13%)], one botanical variety G. barbadense L. var. brasiliensis (51%), and in 6% of the total samples collected , the evaluated characteristics did not allow us to identify the samples at the species level. However, this is the first report of the presence of G. hirsutum L. in the Peruvian Amazon. The conservation and distribution levels found for G. barbadense L. and G. hirsutum L. showed the economic importance they represent for Amazonian communities, owing to the quality of their fibers, as already described by Ozyigit [Currently, there are difficulties in identifying and determining the exact distribution of the cotton populations of the vailable . There available ,3,24, bu Ozyigit  and Ahme Ozyigit .Gossypium comprises more than 50 recognized species distributed in arid and semiarid zones in the tropics and subtropics [G. arboreum, G. herbaceum, G. hirsutum, and G. barbadense [G. hirsutum L. in the region of Cusco; however, in the regions of Cajamarca, Lambayeque, and Pasco, these species can be found. On the other hand, it is known that G. hirsutum L. is an introduced and cultivated species [The genus btropics . Four sprbadense ,27. The  species ; its ori species , and its species ,21. The G. barbadense L. has been reported in 24 regions in Peru [ in Peru ,29. The  in Peru . However in Peru  and MINAG. barbadense L. var. brasiliensis is of the Amazonian type, with an undetermined origin. It has been noted that it may have spread through the Apure River to the northwest of Colombia, being of interest for the study of the west and east of the Andes. This variety within G. barbadense L. was more prevalent (51% of the collected samples) in the districts of Echarate and Megantoni. This result agrees with the studies of Lazo [G. barbadense L. var. brasiliensis has peculiar characteristics, i.e., its seeds are welded or kidney-shaped, as also observed in our research  and in the Miaría, Timpía, Shivankoreni, Kirigueti, Nuevo Mundo, Sensa, Ticumpinía and Camisea communities of the Megantoni district, located between the coordinates UTM WGS-84 Zone: 18 South (724020 E and 8703864 N) Gossypium spp.) were collected in 12 native Amazonian communities in the districts of Echarati and Megantoni. The type of sampling was nonprobabilistic purposive, by convenience [Gossypium spp. was cultivated or found naturally, according to the collection and pressing procedures recommended by MINAM [The collections and evaluations were conducted during the period from April to December 2021. Permission was requested from the native Amazonian communities for the collection. Samples of cotton ; for FLOWERs: the presence of spots, color of the flower spots, flower color, and position of the stigma in relation to the anthers; for SEEDS: arrangement, number, main characteristics, color, and shape; for BRACTS: types of serration and color; for CAPSULES: shape in longitudinal section, characteristics and prominence of the tip; and finally, for LEAF: shape and presence of pubescence. All these descriptors were evaluated following the recommendations of Manco Céspedes et al.  and MINAAll the descriptors were tabulated, and different classes were established for the different species, which allowed us to undertake a descriptive study of the collection sites and the distribution of the main cotton species collected in each locality.The length and width of the flowers, bracts, capsules, and leaves were determined for all samples. All these measurements were made in cm using a ruler. The height of the plants was also determined in meters with the aid of a ruler. The evaluations followed the recommendations of Manco Céspedes et al.  and MINA® (Systat Software Inc.) were used for the elaboration of the graphs, and R [To verify the relationship between the quantitative descriptors and the groups of species, ANOVA was initially performed considering the unbalanced treatments (species), and when significant, the means were compared by Tukey’s test at 1% probability. Using the quantitative data, a scatter plot was made showing the Pearson correlations and the dispersion of each characteristic. The quantitative and qualitative data were used to perform a multivariate analysis using the PCA method to see the overall variability of the two individual datasets and combined for the various groups formed, as well as the trends of the groupings and descriptors. Rbio software  and Sigms, and R  with theGossypium spp. (G. barbadense L. and G. hirsutum L.) and a botanical variety G. barbadense L. var. brasiliensis, located in the districts Echarati and Megantoni, along with three groups of samples that could not be identified using the qualitative and quantitative descriptors evaluated. This report contributes to the information that has been presented for this region of the Peruvian Amazon, verifying that there is phenotypic variability for the species Gossypium spp. and that conservation efforts are being made by the indigenous people who populate these places. The multivariate analysis tools and the evaluation of different descriptors allowed us to affirm that variability exists within the described species and that the new groups identified may be the result of spontaneous crosses between the described species of Gossypium spp.; further studies will be necessary to confirm this hypothesis. The results also show the existence of G. hirsutum L. in this zone, and the greatest distribution was observed for the species G. barbadense L.The present study allowed us to identify two species of


Corydoras pygmaeus Knaack, 1966, is the smallest member of the genus Corydoras, belonging to the family Callichthyidae and order Siluriformes. The complete mitochondrial genome of C. pygmaeus was sequenced and assembled using next-generation sequencing technology, and phylogenetically compared with those of other species of this genus. The mitochondrial genome of C. pygmaeus is a circular DNA molecule with a size of 16,840 bp (GenBank no. ON729306). A phylogenetic tree was constructed based on 13 protein-coding genes of C. pygmaeus and 13 species of the family Callichthyidae, which showed that C. pygmaeus clustered with other species of this genus, but was the first branch to differentiate. These results could provide basic data for phylogenetic analysis and population genetic diversity protection of Corydoras and Callichthyidae fish in the future.The Pygmy corydoras  Aspidoras, Brochis, and Corydoras of the subfamily Corydoradinae. They have two cute ‘moustaches’ next to their mouths and look like small mice swimming in water, and thus the name ‘mouse fish.’ Mouse fish are typical benthic and migratory fish with omnivorous feeding habits. They are found in South America. Almost all the main and tributary water systems of the Amazon River contain pikes, but the composition of species in the watersheds where various species are distributed is different  is maternally inherited, undergoes rapid evolution, and small in size, making it an important molecular marker for research on fish evolutionary genetics, molecular ecology, species identification, and conservation biology .The sample was collected from Duyun City, Guizhou, China  on June 10, 2021. This study was reviewed and approved by the Animal Ethics Committee of the Qiannan Normal University for Nationalities. Voucher specimens were stored at the School of Tourism and Resource Environment, Qiannan Normal University for Nationalities, Guizhou, China, under the voucher number C_pygmaeus_A (http://mitofish.aori.u-tokyo.ac.jp/annotation/input.html) and MITOS (http://mitos.bioinf.uni-leipzig.de/index.py). The tRNAscan-SE 1.21 (Lowe and Eddy http://lowelab.ucsc.edu/tRNAscan-SE/index.html) online tool was used to predict the secondary structure of tRNA, which was corrected manually and finally submitted to GenBank (Accession no. ON729306). To determine the phylogenetic significance of C. pygmaeus in the genus Corydoras, 11 other species of this genus were used to reconstruct the phylogenetic tree based on 13 protein-coding genes, using Hemigrammus erythrozonus as the outgroup  were observed for most protein-coding genes, except for incomplete terminal codons T for ND2, COII, COIII, ND3, and ND4; AGA for ND5; and AGG for COI.The complete mitochondrial genome of n length , with a Brochis multiradiatus clustered with other species of the genus Corydoras before C. pygmaeus (C. pygmaeus clustered with other species of this genus, but was the first branch to differentiate.Phylogenetic analysis showed that Callichthyidae species formed a branch and were well separated from the outgroup, and that pygmaeus . The tarC. pygmaeus mtDNA were predicted and analyzed to provide basic data for phylogenetic analysis and population genetic diversity protection of Corydoras and Callichthyidae fish in the future.In this study, the composition and structure of