Aim. To conduct an express pest risk analysis of Grapevine Roditis leaf discoloration-associated virus (GRLDaV) for Ukraine, a virus that has been related to a grapevine disease and was included in the EPPO Alert List in 2018. Methods. The phytosanitary risk analysis was carried out on the basis of an analytical review of expert literature and in accordance with the EPPO Decision-support scheme for an Express Pest Risk Analysis (EPPO, 2012) and with methodological recommendations for Ukraine (Pylypenko et al, 2012). The possibility of further spread and the potential range of the virus were determined using modern software packages AgroAtlas (Afonin and Li, 2011; Shumilin and Li, 2009), MapInfo Pro15.0 (ESTIMap®) and IDRISI SELVA (Clarklabs®). Results. An express pest risk analysis of GRLDaV for Ukraine was carried out for the first time. Outbreaks of the virus were first detected in the 1980ies in Greece (Rumbos, Avgelis, 1989) and from 2014–2018 in: Italy (Chiumenti et al, 2015, 2016; Maliogka et al, 2015), Turkey (Adan, 2016; Serçe et al, 2018) and Croatia (Vončina et al, 2018). Climatic predictors were analyzed in the outbreaks of the countries where the causal agent of the disease was identified. The potential range of GRLDaV in Ukraine has been established in case of importing infected planting material with further spreading of the virus. Risk management measures are proposed, which envisage including GRLDaV in the list of the Regulated Non-Quarantine Harmful Organisms of Ukraine. Conclusions. There is a possibility of introduction, further spread, and harmfulness of Grapevine Roditis leaf discoloration-associated virus in Ukraine, which is due to the presence of the host plant (grapevine, Vitis vinifera ssp. vinifera L.) and the corresponding climatic conditions of the southern and (part of) western Ukraine, where grapevine is cultivated on an industrial scale. The introduction of GRLDaV into Ukraine is possible as a result of the import of GRLDaV-infected grapevine planting material from the countries where the virus has been reported and presumably is still present. The current Ukrainian phytosanitary measures cannot reliably prevent the risk of intoduction of GRLDaV into Ukraine. The inclusion of GRLDaV in the List of the Regulated Non-Quarantine Harmful Organisms of Ukraine with the recommendation of permission to import grapevine planting material certified for the absence of GRLDaV (from the countries where the virus is reported) or mandatory testing of imported grapevine planting material for the presence of GRLDaV (from areas where such certification is absent), can be an effective risk management measure. It requires the producers and importers of grapevine planting material to include GRLDaV virus in their certification schemes to prevent eventual further spreading of the virus. It is recommended to do a nation-wide survey to determine the likelihood of the presence of GRLDaV in the region. Further research to identify possible
{"title":"Grapevine roditis leaf Discoloration-associated virus: express pest risk analysis for Ukraine","authors":"Y. Klechkovskyi, L. Titova, O. Palagina, L. Janse","doi":"10.15407/agrisp9.01.039","DOIUrl":"https://doi.org/10.15407/agrisp9.01.039","url":null,"abstract":"Aim. To conduct an express pest risk analysis of Grapevine Roditis leaf discoloration-associated virus (GRLDaV) for\u0000Ukraine, a virus that has been related to a grapevine disease and was included in the EPPO Alert List in 2018. Methods. The\u0000phytosanitary risk analysis was carried out on the basis of an analytical review of expert literature and in accordance with the\u0000EPPO Decision-support scheme for an Express Pest Risk Analysis (EPPO, 2012) and with methodological recommendations\u0000for Ukraine (Pylypenko et al, 2012). The possibility of further spread and the potential range of the virus were determined\u0000using modern software packages AgroAtlas (Afonin and Li, 2011; Shumilin and Li, 2009), MapInfo Pro15.0 (ESTIMap®)\u0000and IDRISI SELVA (Clarklabs®). Results. An express pest risk analysis of GRLDaV for Ukraine was carried out for the\u0000first time. Outbreaks of the virus were first detected in the 1980ies in Greece (Rumbos, Avgelis, 1989) and from 2014–2018\u0000in: Italy (Chiumenti et al, 2015, 2016; Maliogka et al, 2015), Turkey (Adan, 2016; Serçe et al, 2018) and Croatia (Vončina\u0000et al, 2018). Climatic predictors were analyzed in the outbreaks of the countries where the causal agent of the disease was\u0000identified. The potential range of GRLDaV in Ukraine has been established in case of importing infected planting material\u0000with further spreading of the virus. Risk management measures are proposed, which envisage including GRLDaV in the list\u0000of the Regulated Non-Quarantine Harmful Organisms of Ukraine. Conclusions. There is a possibility of introduction, further\u0000spread, and harmfulness of Grapevine Roditis leaf discoloration-associated virus in Ukraine, which is due to the presence\u0000of the host plant (grapevine, Vitis vinifera ssp. vinifera L.) and the corresponding climatic conditions of the southern and\u0000(part of) western Ukraine, where grapevine is cultivated on an industrial scale. The introduction of GRLDaV into Ukraine\u0000is possible as a result of the import of GRLDaV-infected grapevine planting material from the countries where the virus has\u0000been reported and presumably is still present. The current Ukrainian phytosanitary measures cannot reliably prevent the risk\u0000of intoduction of GRLDaV into Ukraine. The inclusion of GRLDaV in the List of the Regulated Non-Quarantine Harmful\u0000Organisms of Ukraine with the recommendation of permission to import grapevine planting material certified for the absence\u0000of GRLDaV (from the countries where the virus is reported) or mandatory testing of imported grapevine planting material\u0000for the presence of GRLDaV (from areas where such certification is absent), can be an effective risk management measure. It\u0000requires the producers and importers of grapevine planting material to include GRLDaV virus in their certification schemes\u0000to prevent eventual further spreading of the virus. It is recommended to do a nation-wide survey to determine the likelihood\u0000of the presence of GRLDaV in the region. Further research to identify possible ","PeriodicalId":55933,"journal":{"name":"Agricultural Science and Practice","volume":" ","pages":""},"PeriodicalIF":0.4,"publicationDate":"2022-08-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43874517","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A. Lysytsya, P. Kryvoshyya, O. Kvartenko, O. Lebed
Aim. To investigate both toxic (hemolytic), and stimulating effects of two polymeric derivatives of guani- dine, in particular, polyhexamethylene guanidine (PHMG) and polyhexamethylene biguanidine (PHMB) both in the hydrochloride form, on eukaryotic cells depending on the concentration of the preparation; to study the possibility of using wound-healing and stimulating properties of these preparations in veterinary medicine. Methods. The hemolytic activity (toxicity) of PHMGH and PHMBH preparations in the concentration of 0.1% towards cattle and pig erythrocytes was determined by titration. Primary cell cultures of fetal kidney cells of calves and piglets were used to determine the influence of PHMGH and PHMBH both alone and in combination with the following biologically active substances: essential oils of Pinus sylvestris, Eucalyptus globulus, Citrus sinensis, Monarda didyma, ZnO nanoparticles (size c. 25 nm), and electrochemically acti- vated water – anolyte (Eh = –800 mV, pH 6.5–7.0). The concentration of the cells in the nutrient medium was determined via photocolorimetry. Results. It was found that depending on the concentration, PHMGH, and PHMBH preparations can cause the lysis of erythrocytes, and stimulate cell proliferative activity, including the formation of a monolayer of kidney cells of calves and piglets They cause hemolysis of cattle erythro- cytes in the concentrations commonly used for disinfection, i.e., about 0.1 %, in the average titers of 1 : 7 for PHMGH and 1 : 2.5 for PHMBH. Therefore, PHMBH shows greater hemolytic (biocidal) activity for cattle erythrocytes than PHMGH (in ≈2.8x). The high molecular weight fraction of PHMBH (M2 ≈ 2,000–7,000 Da) demonstrated a lower (in ≈2.4x) hemolytic activity than the low molecular weight basic fraction (M1 ≈ 500–2,000 Da). The experiments on the kidney cell cultures of pigs and cattle have shown that at non-toxic concentrations (10–5 %) PHMBH can effectively stimulate (from 27 to 65 % increase) the proliferative activ- ity of eukaryotic cells and accelerate the formation of a monolayer of cells. The combinations of PHMGH with some essential oils of medicinal plants also show a good effect (from 52 to 95 % increase), and PHMBH shows a good effect with oil of pine for pig kidney cells (20 % increase) and oil of horsemint for cattle kidney cells (67 % increase). Conclusions. PHMGH and PHMBH can possibly be used in agricultural production not only as disinfectants or antiseptics, but also in wound healing. Although their toxicity is also significant to eukaryotic cells, yet they can possibly be used in veterinary medicine in low concentrations (0.005–0.5 %) for the treatment of wounds of various origin, including burns, in the composition of ointments, gels, bandages, or plasters, which we have presently in investigation.
{"title":"Effect of hydrochloric polyhexamethylene guanidine (PHMGH) and polyhexamethylene biguanidine (PHMBH), also in combination with plant essential oils and ZnO nanoparticles on some eukaryotic cattle and pig cells","authors":"A. Lysytsya, P. Kryvoshyya, O. Kvartenko, O. Lebed","doi":"10.15407/agrisp9.01.015","DOIUrl":"https://doi.org/10.15407/agrisp9.01.015","url":null,"abstract":"Aim. To investigate both toxic (hemolytic), and stimulating effects of two polymeric derivatives of guani-\u0000dine, in particular, polyhexamethylene guanidine (PHMG) and polyhexamethylene biguanidine (PHMB) both\u0000in the hydrochloride form, on eukaryotic cells depending on the concentration of the preparation; to study the\u0000possibility of using wound-healing and stimulating properties of these preparations in veterinary medicine.\u0000Methods. The hemolytic activity (toxicity) of PHMGH and PHMBH preparations in the concentration of\u00000.1% towards cattle and pig erythrocytes was determined by titration. Primary cell cultures of fetal kidney\u0000cells of calves and piglets were used to determine the influence of PHMGH and PHMBH both alone and in\u0000combination with the following biologically active substances: essential oils of Pinus sylvestris, Eucalyptus\u0000globulus, Citrus sinensis, Monarda didyma, ZnO nanoparticles (size c. 25 nm), and electrochemically acti-\u0000vated water – anolyte (Eh = –800 mV, pH 6.5–7.0). The concentration of the cells in the nutrient medium was\u0000determined via photocolorimetry. Results. It was found that depending on the concentration, PHMGH, and\u0000PHMBH preparations can cause the lysis of erythrocytes, and stimulate cell proliferative activity, including\u0000the formation of a monolayer of kidney cells of calves and piglets They cause hemolysis of cattle erythro-\u0000cytes in the concentrations commonly used for disinfection, i.e., about 0.1 %, in the average titers of 1 : 7 for\u0000PHMGH and 1 : 2.5 for PHMBH. Therefore, PHMBH shows greater hemolytic (biocidal) activity for cattle\u0000erythrocytes than PHMGH (in ≈2.8x). The high molecular weight fraction of PHMBH (M2 ≈ 2,000–7,000\u0000Da) demonstrated a lower (in ≈2.4x) hemolytic activity than the low molecular weight basic fraction (M1 ≈\u0000500–2,000 Da). The experiments on the kidney cell cultures of pigs and cattle have shown that at non-toxic\u0000concentrations (10–5 %) PHMBH can effectively stimulate (from 27 to 65 % increase) the proliferative activ-\u0000ity of eukaryotic cells and accelerate the formation of a monolayer of cells. The combinations of PHMGH\u0000with some essential oils of medicinal plants also show a good effect (from 52 to 95 % increase), and PHMBH\u0000shows a good effect with oil of pine for pig kidney cells (20 % increase) and oil of horsemint for cattle kidney\u0000cells (67 % increase). Conclusions. PHMGH and PHMBH can possibly be used in agricultural production\u0000not only as disinfectants or antiseptics, but also in wound healing. Although their toxicity is also significant to\u0000eukaryotic cells, yet they can possibly be used in veterinary medicine in low concentrations (0.005–0.5 %) for\u0000the treatment of wounds of various origin, including burns, in the composition of ointments, gels, bandages,\u0000or plasters, which we have presently in investigation.","PeriodicalId":55933,"journal":{"name":"Agricultural Science and Practice","volume":" ","pages":""},"PeriodicalIF":0.4,"publicationDate":"2022-08-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41981967","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Aim. The aim of the study was to analyze chromosome sets of specific species of agricultural animals of Bovidae family using the techniques of differentiated G- and Ag-banding of chromosomes and to demonstrate the role of their variability in the evolution. Methods. The culture of lymphocytes and chromosome preparations were pre- pared by the method of Moorhead et al (1960). The preparations of metaphase chromosomes were analyzed by the G-banding methods of Barch M J et al (1997) and by Ag-banding method of Zhu JJ et al (2019). The chromosomal aberrations were classified according to the recommendations of the International System for Chromosome No- menclature of Domestic Bovids (2001). The dendrogram of phylogenetic relations between mammalian species was built using “STATISTICA 6.1”. Results. The comparative study of the chromosomal sets of cattle Bos taurus, domestic sheep Ovis aries and river buffalo Bubalus bubalis bubalis was conducted using G- and Ag-banding methods. The homology of the structural organization of chromosomes and the evolutionary changes in karyotypes of Bovidae family were analyzed, highlighting their sex chromosomes and the chromosomes with specific local- ized groups of gene linkage. A considerable homology of chromosome sites was found in the representatives of the investigated species by the G-banding profiles. To study the phylogenetic interrelations, the index of the number of active nucleolus organizer regions (NOR) in the chromosomes during the metaphase stage was used. Conclu- sions. The comparison of the morphology of chromosomes of agricultural animals of Bovidae family, Bos taurus, Ovis aries, and Bubalus bubalis bubalis confirms the association between the homology of some chromosomal sites and the formation of metacentric chromosomes due to the linkage of acrocentric ones. The species-specific morphological differences in sex chromosomes of the investigated animals were found in terms of the length and presence of pericentric inversions. The phylogenetic relations between the species of Bovidae family demonstrate that the value of distances, determined based on the variability of the number of active NOR, reflects the degree of their phylogenetic similarity.
目标本研究的目的是利用染色体的G和Ag带分化技术分析牛科特定农业动物物种的染色体组，并证明其变异性在进化中的作用。方法。淋巴细胞的培养和染色体制剂是通过Moorhead等人（1960）的方法制备的。采用Barch M J等人（1997）的G显带法和Zhu JJ等人（2019）的Ag显带法对中期染色体的制备进行了分析。根据国际家牛染色体无畸变系统（2001）的建议对染色体畸变进行了分类。利用“STATISTICA6.1”建立了哺乳动物亲缘关系的树状图。采用G带和Ag带方法对牛、绵羊和水牛的染色体组进行了比较研究。分析了牛科染色体结构组织的同源性和核型的进化变化，重点介绍了它们的性染色体和具有特定基因连锁局部化群的染色体。通过G-显带谱，在研究物种的代表中发现了相当大的染色体位点同源性。为了研究系统发育的相互关系，使用中期染色体中活性核仁组织区（NOR）的数量指数。结论。通过对牛科、牛头牛、绵羊和Bubalus bubalis bubalis的农业动物染色体形态的比较，证实了一些染色体位点的同源性与由于端部着丝粒染色体的连锁而形成的中心粒染色体之间的联系。研究动物性染色体的物种特异性形态差异主要表现在中心周围倒位的长度和存在性方面。牛科物种之间的系统发育关系表明，根据活跃NOR数量的可变性确定的距离值反映了它们系统发育相似性的程度。
{"title":"Comparative analysis of karyotypes of Вovidae family from the evolutionary aspect","authors":"V. Dzitsiuk, H. Bratytsia, T. Dyman","doi":"10.15407/agrisp9.01.027","DOIUrl":"https://doi.org/10.15407/agrisp9.01.027","url":null,"abstract":"Aim. The aim of the study was to analyze chromosome sets of specific species of agricultural animals of Bovidae\u0000family using the techniques of differentiated G- and Ag-banding of chromosomes and to demonstrate the role of\u0000their variability in the evolution. Methods. The culture of lymphocytes and chromosome preparations were pre-\u0000pared by the method of Moorhead et al (1960). The preparations of metaphase chromosomes were analyzed by the\u0000G-banding methods of Barch M J et al (1997) and by Ag-banding method of Zhu JJ et al (2019). The chromosomal\u0000aberrations were classified according to the recommendations of the International System for Chromosome No-\u0000menclature of Domestic Bovids (2001). The dendrogram of phylogenetic relations between mammalian species\u0000was built using “STATISTICA 6.1”. Results. The comparative study of the chromosomal sets of cattle Bos taurus,\u0000domestic sheep Ovis aries and river buffalo Bubalus bubalis bubalis was conducted using G- and Ag-banding\u0000methods. The homology of the structural organization of chromosomes and the evolutionary changes in karyotypes\u0000of Bovidae family were analyzed, highlighting their sex chromosomes and the chromosomes with specific local-\u0000ized groups of gene linkage. A considerable homology of chromosome sites was found in the representatives of the\u0000investigated species by the G-banding profiles. To study the phylogenetic interrelations, the index of the number\u0000of active nucleolus organizer regions (NOR) in the chromosomes during the metaphase stage was used. Conclu-\u0000sions. The comparison of the morphology of chromosomes of agricultural animals of Bovidae family, Bos taurus,\u0000Ovis aries, and Bubalus bubalis bubalis confirms the association between the homology of some chromosomal\u0000sites and the formation of metacentric chromosomes due to the linkage of acrocentric ones. The species-specific\u0000morphological differences in sex chromosomes of the investigated animals were found in terms of the length and\u0000presence of pericentric inversions. The phylogenetic relations between the species of Bovidae family demonstrate\u0000that the value of distances, determined based on the variability of the number of active NOR, reflects the degree\u0000of their phylogenetic similarity.","PeriodicalId":55933,"journal":{"name":"Agricultural Science and Practice","volume":" ","pages":""},"PeriodicalIF":0.4,"publicationDate":"2022-08-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42358586","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
H. Tsekhmister, E. Kopilov, O. Nadkernychna, A. Kyslynska
Aim. To investigate the phylogenetic relations of P. melonis strain 502 and to study the varietal sensitivity of cu- cumber plants to P. melonis strain 502. Methods. DNA was extracted using the enzymatic lysis buffer. The PCR was conducted following White et al. protocol (1990). The obtained PCR-products were determined by sequencing on the automatic capillary sequencer Applied Biosystems ABI Prism 3130. The sequence of the gene 5.8S rRNA of P. melonis strain 502 was compared to the sequences from the GenBank database using the BLAST analysis. The phy- logenetic analysis was conducted by the neighbor-joining method. The evolutionary distances were estimated by the method of Jukes & Cantor. The evolutionary analysis was conducted in MEGA7. The sensitivity of cucumber plants was determined during a vegetative experiment with artificial infection background (AIB), created by introducing the infectious material of fungus P. melonis strain 502 into the soil. The infectious material was introduced at a rate of 50 thousand CFU/per 1 g of soil. The damage to the root system was assessed after 14 days of cultivating plants on the AIB. The disease severity index (DSI) was estimated to determine the general sensitivity of the investigated varieties. The varieties, which received DSI
目标研究甜瓜502菌株的亲缘关系，研究黄瓜植株对甜瓜502菌株品种的敏感性。方法。使用酶解缓冲液提取DNA。PCR按照White等人的方案（1990）进行。通过在自动毛细管测序仪Applied Biosystems ABI Prism 3130上测序来测定获得的PCR产物。使用BLAST分析将甜瓜P.melonis菌株502的基因5.8S rRNA的序列与来自GenBank数据库的序列进行比较。采用邻域连接法进行物理成因分析。进化距离用Jukes&Cantor方法估计。在MEGA7中进行进化分析。在人工感染背景（AIB）的营养实验中，通过将真菌P.melonis菌株502的感染物质引入土壤中，测定了黄瓜植株的敏感性。感染性物质以每1g土壤5万CFU/的速率引入。在AIB上培养植物14天后评估对根系的损害。估计疾病严重程度指数（DSI）以确定所调查品种的总体敏感性。收到DSI的品种
{"title":"Phylogeny of Plectosphaerella melonis strain 502 and varietal sensitivity of cucumber plants","authors":"H. Tsekhmister, E. Kopilov, O. Nadkernychna, A. Kyslynska","doi":"10.15407/agrisp9.01.003","DOIUrl":"https://doi.org/10.15407/agrisp9.01.003","url":null,"abstract":"Aim. To investigate the phylogenetic relations of P. melonis strain 502 and to study the varietal sensitivity of cu- cumber plants to P. melonis strain 502. Methods. DNA was extracted using the enzymatic lysis buffer. The PCR was conducted following White et al. protocol (1990). The obtained PCR-products were determined by sequencing on the automatic capillary sequencer Applied Biosystems ABI Prism 3130. The sequence of the gene 5.8S rRNA of P. melonis strain 502 was compared to the sequences from the GenBank database using the BLAST analysis. The phy- logenetic analysis was conducted by the neighbor-joining method. The evolutionary distances were estimated by the method of Jukes & Cantor. The evolutionary analysis was conducted in MEGA7. The sensitivity of cucumber plants was determined during a vegetative experiment with artificial infection background (AIB), created by introducing the infectious material of fungus P. melonis strain 502 into the soil. The infectious material was introduced at a rate of 50 thousand CFU/per 1 g of soil. The damage to the root system was assessed after 14 days of cultivating plants on the AIB. The disease severity index (DSI) was estimated to determine the general sensitivity of the investigated varieties. The varieties, which received DSI","PeriodicalId":55933,"journal":{"name":"Agricultural Science and Practice","volume":"1 1","pages":""},"PeriodicalIF":0.4,"publicationDate":"2022-08-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41380126","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
This review presents the scientific studies data on the special characteristics of developmental biology and epizo- otiology of Cryptocotyle trematodes, which belong to Heterophyidae family and pose a threat to the health of the endotherms, including humans, i.e., it is a typical zoonosis. These trematodes are predominantly widespread in the Mediterranean region, namely, in the western Mediterranean region and the Aegean province. The trematodes of Cryptocotyle genus are found in Europe, Asia, North and South America, and Antarctica. They are typical bio- helminths, i.e., they have a complicated life cycle, where the initial intermediate hosts are mollusks, the second ones – fish of different species, the definitive and final hosts are piscivorous birds, carnivorous animals (foxes, wolves, dogs, cats, etc.), and humans. Cryptocotylosis is remarkable for its seasonal prevalence, which depends on the climatic zoning of territories. For instance, in the territorial waters of Ukraine, the highest indices of crypto- cotylosis invasion among Agonidae fish are mainly observed in summer and autumn, but the peak of the invasion comes in autumn. The parasitizing of Cryptocotyle trematodes in the organism of mollusks impacts the reproduc- tion ability and behavioral specificities (motility) of the latter. The invaded fish have black pigment spots on the surface of their bodies – these are metacercariae. In the organism of definitive hosts, the agent is localized in the gastrointestinal tract and may cause inflammatory processes in the mucous membrane of the intestines and changes in parenchymatous organs, which demonstrates the toxic effect of the parasite on the host organism. The diagnos- tics of cryptocotylosis is based on detecting the agent in the host organism and its further taxonomic identification by its anatomic and morphological specificities. The pollution of the aqueous medium with organic and inorganic residues impacts the organisms of both hosts and parasites.
{"title":"Cryptokotyle lühe, 1899 (trematoda: heterophyidae): special characteristics of developmental biology and epizootiology","authors":"S. Honcharov, N. Soroka, A. Dubovyi, M. Galat","doi":"10.15407/agrisp9.01.050","DOIUrl":"https://doi.org/10.15407/agrisp9.01.050","url":null,"abstract":"This review presents the scientific studies data on the special characteristics of developmental biology and epizo-\u0000otiology of Cryptocotyle trematodes, which belong to Heterophyidae family and pose a threat to the health of the\u0000endotherms, including humans, i.e., it is a typical zoonosis. These trematodes are predominantly widespread in\u0000the Mediterranean region, namely, in the western Mediterranean region and the Aegean province. The trematodes\u0000of Cryptocotyle genus are found in Europe, Asia, North and South America, and Antarctica. They are typical bio-\u0000helminths, i.e., they have a complicated life cycle, where the initial intermediate hosts are mollusks, the second\u0000ones – fish of different species, the definitive and final hosts are piscivorous birds, carnivorous animals (foxes,\u0000wolves, dogs, cats, etc.), and humans. Cryptocotylosis is remarkable for its seasonal prevalence, which depends on\u0000the climatic zoning of territories. For instance, in the territorial waters of Ukraine, the highest indices of crypto-\u0000cotylosis invasion among Agonidae fish are mainly observed in summer and autumn, but the peak of the invasion\u0000comes in autumn. The parasitizing of Cryptocotyle trematodes in the organism of mollusks impacts the reproduc-\u0000tion ability and behavioral specificities (motility) of the latter. The invaded fish have black pigment spots on the\u0000surface of their bodies – these are metacercariae. In the organism of definitive hosts, the agent is localized in the\u0000gastrointestinal tract and may cause inflammatory processes in the mucous membrane of the intestines and changes\u0000in parenchymatous organs, which demonstrates the toxic effect of the parasite on the host organism. The diagnos-\u0000tics of cryptocotylosis is based on detecting the agent in the host organism and its further taxonomic identification\u0000by its anatomic and morphological specificities. The pollution of the aqueous medium with organic and inorganic\u0000residues impacts the organisms of both hosts and parasites.","PeriodicalId":55933,"journal":{"name":"Agricultural Science and Practice","volume":" ","pages":""},"PeriodicalIF":0.4,"publicationDate":"2022-08-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48334232","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Aim. To study the specificities of population genetic structure of Ukrainian Black-and-White dairy breed, Ukrainian Red-and-White dairy breed, and Ukrainian Grey cattle breed by polymorphism of TLR1, TLR4, and SLC11A1 genes. Methods. The study was conducted using the method of polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). Results. The results of the study demonstrated that in all the experimental populations, the locus TLR4 by mutations 8732G>A, 8834G>C, and 2021C>T was monomorphic, while loci TLR1 (1596G>A) and SLC11A1 (7400C>G and 7808A>T) were polymorphic. For the TLR4 gene by 8732G>A mutation, only individu- als with genotype BB were found; by 8834G>C – with genotype GG; by 2021C>T – with genotype CC. By BclI- polymorphism in the first exon of TLR1 (1596G>A) the A and G allelic variants were found in each experimental population. The deviation from the Hardy-Weinberg equilibrium state was revealed for the populations of Ukrainian Black-and-White and Ukrainian Grey cattle breeds. A considerable excess of heterozygous individuals was fixed in both cattle populations (31 and 39 % respectively). Considering PstI-polymorphism in exon 11 of SLC11A1 gene, allelic variants C and G were found for SNP5 (7400C>G), as well as A and T for SNP6 (7808A>T) in all experimen- tal populations. As for SNP5 (7400C>G), a considerable excess (from 15 to 30 %) of heterozygous individuals was determined for all experimental groups. Unlike other breeds, there was no excess (Fis = 0,04) of heterozygotes for SNP6 (7808A>T) in the population of Ukrainian Grey cattle. Conclusions. The parameters of genetic variability in the different cattle populations of Ukrainian selection by TLR1, TLR4, and SLC11A1 loci were studied. The results of the study showed the impossibility of using TLR4 locus by 8732G>A, 8834G>C and 2021C>T mutations in the programs of marker-associated selection of the investigated cattle breeds due to its monomorphic nature. The analysis of the allele and genotype distribution by TLR1 and SLC11A1 loci (presence of individuals with different genotypes in all experimental cattle breeds) indicates the possibility of using different allelic variants of these genes in the breeding programs for the studied cattle populations.
{"title":"Polymorphism of TLR1, TLR4, and SLC11A1 genes in populations of different cattle breeds of Ukrainian selection","authors":"R. Kulibaba, Y. Liashenko, O. Ivashchenko","doi":"10.15407/agrisp8.03.025","DOIUrl":"https://doi.org/10.15407/agrisp8.03.025","url":null,"abstract":"Aim. To study the specificities of population genetic structure of Ukrainian Black-and-White dairy breed, Ukrainian\u0000Red-and-White dairy breed, and Ukrainian Grey cattle breed by polymorphism of TLR1, TLR4, and SLC11A1 genes.\u0000Methods. The study was conducted using the method of polymerase chain reaction and restriction fragment length\u0000polymorphism (PCR-RFLP). Results. The results of the study demonstrated that in all the experimental populations,\u0000the locus TLR4 by mutations 8732G>A, 8834G>C, and 2021C>T was monomorphic, while loci TLR1 (1596G>A)\u0000and SLC11A1 (7400C>G and 7808A>T) were polymorphic. For the TLR4 gene by 8732G>A mutation, only individu-\u0000als with genotype BB were found; by 8834G>C – with genotype GG; by 2021C>T – with genotype CC. By BclI-\u0000polymorphism in the first exon of TLR1 (1596G>A) the A and G allelic variants were found in each experimental\u0000population. The deviation from the Hardy-Weinberg equilibrium state was revealed for the populations of Ukrainian\u0000Black-and-White and Ukrainian Grey cattle breeds. A considerable excess of heterozygous individuals was fixed in\u0000both cattle populations (31 and 39 % respectively). Considering PstI-polymorphism in exon 11 of SLC11A1 gene,\u0000allelic variants C and G were found for SNP5 (7400C>G), as well as A and T for SNP6 (7808A>T) in all experimen-\u0000tal populations. As for SNP5 (7400C>G), a considerable excess (from 15 to 30 %) of heterozygous individuals was\u0000determined for all experimental groups. Unlike other breeds, there was no excess (Fis = 0,04) of heterozygotes for\u0000SNP6 (7808A>T) in the population of Ukrainian Grey cattle. Conclusions. The parameters of genetic variability in\u0000the different cattle populations of Ukrainian selection by TLR1, TLR4, and SLC11A1 loci were studied. The results\u0000of the study showed the impossibility of using TLR4 locus by 8732G>A, 8834G>C and 2021C>T mutations in the\u0000programs of marker-associated selection of the investigated cattle breeds due to its monomorphic nature. The analysis\u0000of the allele and genotype distribution by TLR1 and SLC11A1 loci (presence of individuals with different genotypes in\u0000all experimental cattle breeds) indicates the possibility of using different allelic variants of these genes in the breeding\u0000programs for the studied cattle populations.","PeriodicalId":55933,"journal":{"name":"Agricultural Science and Practice","volume":" ","pages":""},"PeriodicalIF":0.4,"publicationDate":"2021-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46499590","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
H. Tsekhmister, A. Kyslynska, E. Kopilov, O. Nadkernychna
Aim. To investigate the ability of our phytopathogenic fungal strain 502, earlier preliminarily identified as the phytopathogen Plectosphaerella melonis (syn. Acremonium cucurbitacearum), to have phytotoxic and/or plant growth regulatory activity. Methods. The phytotoxicity of strain 502, was studied by bioassays using the test cultures of corn (Zea mays L.), garden cress (Lepidium sativum L.), cucumber (Cucumis sativus L.), and onion (Allium cepa L.). The cytotoxicity and genotoxicity of the fungus were estimated using the Allium cepa-test. The mitotic index of the, the duration of mitosis phases, and the frequency of aberrant ana-telophases of Allium cepa L. roots meristem was also investigated. For this purpose, strain 502, was grown in the following culture media: synthetic Raulin-Thom medium for 10 days at 26 ± 2 °С. Cell-free filtrate (culture fluid) was used for the study. Ethylene production was quantified in culture filtrate using gas-chromatography meth- od. Ethylene measurement was performed every 7 days during 8 weeks. The determination was carried out using a gas chromatograph «Agilent Technologies 6850» (USA) fitted with a flame ionization detector, using commercial ethylene as a standard for identification and quantification Every experiment had three repeats. The reliability of experimental data was assessed by statistical methods using Statistica 12 (Stat-Soft Inc., USA). Results. Undiluted culture fluid (obtained by growing the fungus on liquid wort) of our strain 502 inhibited the growth of Z. mays seedlings by 14 %, L. sativum seedlings by 18 % (1 : 100 dilution) and stimulated the growth of L. sativum roots by 54 and 41 % (1 : 10 and 1 : 100 dilutions, respectively). The culture fluid, obtained by growing the fungus on Raulin-Thom’s synthetic agar, demonstrated a slight inhibitory effect on the seedlings and roots of L. sativum, and at the dilution of 1 : 1000 stimulated growth by 30 %. Insignificant changes in the mitotic index of the meristem of A. cepa roots were revealed at the effect of the culture fluid of P. melonis, strain 502, diluted at the ratio of 1 : 100 and 1 : 1000. At the same time, the number of cells at the prophase stage decreased 1.7 times (1 : 100 dilution). There is a significant increase in the number of cells at the metaphase stage – 1.3 and 1.4 times (dilution 1 : 100 and 1 : 1000, respectively), the anaphase stage – 2.1 and 1.8 times (dilution 1 : 100 and 1 : 1000, respectively) and the telophase stage – 1.8 times (1 : 100 dilution), as compared with the positive control (culture medium). The frequencies of aberrant ana-telophases in the apical meristems of the initial roots were 5.0 and 2.2 % (at the culture fluid dilution of 1 : 100 and 1 : 1000, respectively). We researched the abil- ity of P. melonis 502 to synthesize ethylene and the highest level of it was registered after 5 weeks of cultivation (111.78 nmol/h g). Conclusions: It was demonstrated by us that the culture fluid of strain 502 showe
{"title":"Plant growth regulatory activity in the phytopathogenic fungus Plectosphaerella melonis strain 502","authors":"H. Tsekhmister, A. Kyslynska, E. Kopilov, O. Nadkernychna","doi":"10.15407/agrisp8.03.013","DOIUrl":"https://doi.org/10.15407/agrisp8.03.013","url":null,"abstract":"Aim. To investigate the ability of our phytopathogenic fungal strain 502, earlier preliminarily identified as the phytopathogen\u0000Plectosphaerella melonis (syn. Acremonium cucurbitacearum), to have phytotoxic and/or plant growth regulatory activity.\u0000Methods. The phytotoxicity of strain 502, was studied by bioassays using the test cultures of corn (Zea mays L.), garden\u0000cress (Lepidium sativum L.), cucumber (Cucumis sativus L.), and onion (Allium cepa L.). The cytotoxicity and genotoxicity\u0000of the fungus were estimated using the Allium cepa-test. The mitotic index of the, the duration of mitosis phases, and the\u0000frequency of aberrant ana-telophases of Allium cepa L. roots meristem was also investigated. For this purpose, strain 502,\u0000was grown in the following culture media: synthetic Raulin-Thom medium for 10 days at 26 ± 2 °С. Cell-free filtrate\u0000(culture fluid) was used for the study. Ethylene production was quantified in culture filtrate using gas-chromatography meth-\u0000od. Ethylene measurement was performed every 7 days during 8 weeks. The determination was carried out using a gas\u0000chromatograph «Agilent Technologies 6850» (USA) fitted with a flame ionization detector, using commercial ethylene\u0000as a standard for identification and quantification Every experiment had three repeats. The reliability of experimental data\u0000was assessed by statistical methods using Statistica 12 (Stat-Soft Inc., USA). Results. Undiluted culture fluid (obtained\u0000by growing the fungus on liquid wort) of our strain 502 inhibited the growth of Z. mays seedlings by 14 %, L. sativum\u0000seedlings by 18 % (1 : 100 dilution) and stimulated the growth of L. sativum roots by 54 and 41 % (1 : 10 and 1 : 100\u0000dilutions, respectively). The culture fluid, obtained by growing the fungus on Raulin-Thom’s synthetic agar, demonstrated\u0000a slight inhibitory effect on the seedlings and roots of L. sativum, and at the dilution of 1 : 1000 stimulated growth by\u000030 %. Insignificant changes in the mitotic index of the meristem of A. cepa roots were revealed at the effect of the culture\u0000fluid of P. melonis, strain 502, diluted at the ratio of 1 : 100 and 1 : 1000. At the same time, the number of cells at the prophase\u0000stage decreased 1.7 times (1 : 100 dilution). There is a significant increase in the number of cells at the metaphase stage –\u00001.3 and 1.4 times (dilution 1 : 100 and 1 : 1000, respectively), the anaphase stage – 2.1 and 1.8 times (dilution 1 : 100 and\u00001 : 1000, respectively) and the telophase stage – 1.8 times (1 : 100 dilution), as compared with the positive control\u0000(culture medium). The frequencies of aberrant ana-telophases in the apical meristems of the initial roots were\u00005.0 and 2.2 % (at the culture fluid dilution of 1 : 100 and 1 : 1000, respectively). We researched the abil-\u0000ity of P. melonis 502 to synthesize ethylene and the highest level of it was registered after 5 weeks of cultivation\u0000(111.78 nmol/h g). Conclusions: It was demonstrated by us that the culture fluid of strain 502 showe","PeriodicalId":55933,"journal":{"name":"Agricultural Science and Practice","volume":" ","pages":""},"PeriodicalIF":0.4,"publicationDate":"2021-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41366608","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Aim. To determine the tendencies in the changes in air temperature and their influence on the productivity of crops during the vegetative cycle periods, especially in soil-climatic zones of Ukraine for the 1981–2010 climate normals period. Methods. The analytical and synthetic, statistical, climatic methods, simulation (model of V.P. Dmitrenko “Weather-yield” (Dmitrenko VP et al, 2017, 2010), used to forecast the productivity of grains in the Ukrainian Hy- drometeorological Center since 1970), abstract-logical method. Results. The rising air temperatures were determined throughout the whole vegetative period of growing corn and spring barley over the period of 1981–2010. It was found that this rise in different phases of crop development was of different magnitude and relevance in all regions and soil-climatic zones of Ukraine. The reliable changes in the surface air temperature were noted in the phases of the third leaf, panicle emergence, and blossoming of corn in Polissia, Forest-Steppe, and especially Steppe (0.7– 0.8 °С/10 years, 0.8–0.9 °С/10 years and 0.9–1.1 °С/10 years, respectively). During the pre-sowing period, the periods of corn sowing and seedlings, the velocity of changes was twice lower in the whole territory of the country, and during the pe- riods of milky ripeness and middle dough – in the eastern Forest-Steppe and dry Steppe, amounting to 0.4–0.5 °С/10 years. A considerable rise in the temperature during the period of the third leaf, panicle emergence, and blossoming promoted the decrease in the influence of temperature during these phases of crop development, especially in the Steppe (up to 10–15 % in 10 years). Only the rise in the temperature during the pre-sowing period promoted the 3–6 % increase in the whole terri- tory of the country, and during the periods of milky ripeness and middle dough of corn – up to 8 % in 10 years in the Forest- Steppe and Steppe. Generally, the thermal conditions for corn cultivation deteriorated considerably but remained favorable in Polissia, satisfactory – in the Forest-Steppe and northern Steppe, and unsatisfactory – in the south, in the dry Steppe. The most intense changes in the air temperature during the vegetation period of spring barley were noted in the phase of milky ripeness and middle dough in all soil-climatic zones, amounting to 0.8–1.1 °С/10 years. During the sowing period, the phases of the third leaf, stem elongation, and ear formation, they were 0.6–0.7 °С/10 years, and during the pre-sowing period – 0.3– 0.4 °С/10 years. During the spring barley tillering phase, the change in the air temperature was insignificant in the whole territory of the country. A considerable increase in the air temperature was unfavorable for crop cultivation in all the soil- climatic zones of Ukraine during the vegetative cycle of spring barley, especially during the phases of milky ripeness and middle dough, and promoted the decrease in its productivity in Polissia, Forest-Steppe, and Steppe by 5
目标确定1981–2010年气候正常期内，特别是乌克兰土壤气候区的气温变化趋势及其对作物生产力的影响。方法。分析和综合、统计、气候方法、模拟（V.P.Dmitrenko“天气产量”模型（Dmitrenko-VP et al，20172010），自1970年以来用于预测乌克兰水文中心的粮食生产力），抽象逻辑方法。后果1981年至2010年期间，在玉米和春大麦的整个营养期内，气温都在上升。研究发现，在乌克兰的所有地区和土壤气候区，这种在作物发展的不同阶段的增长具有不同的幅度和相关性。在Polisia、森林草原，尤其是草原（分别为0.7–0.8°С/10、0.8–0.9°С/10和0.9–1.1°С/10年）的玉米第三叶、穗部出苗和开花阶段，地表气温发生了可靠的变化。在播种前、播种期和幼苗期，全国范围内的变化速度低两倍，在乳熟期和中熟期，东部森林草原和干燥草原的变化速度为0.4–0.5°C/10年。在第三片叶子、圆锥花序出现和开花期间，温度的显著升高促进了作物发育的这些阶段温度的影响的降低，尤其是在草原（10年内高达10-15%）。只有播种前温度的升高才促进了该国整个地区3-6%的增长，在玉米乳熟期和中间面团期，在森林草原和大草原，10年内最高可达8%。总体而言，玉米种植的热条件显著恶化，但在波利斯仍然有利，在森林草原和北部草原令人满意，在南部干旱草原令人不满意。在所有土壤气候区，春大麦植被期的气温变化最为剧烈，发生在乳绿质和中间面团阶段，达0.8–1.1°С/10年。在播种期，第三叶、茎伸长和穗形成期为0.6–0.7°С/10年，在播种前为0.3–0.4°С/10年间。在春大麦分蘖期，全国气温变化不大。在春大麦的营养周期期间，特别是在乳白色成熟期和中等面团期，气温的显著升高不利于乌克兰所有土壤气候区的作物种植，并在10年内促使其在波利斯西亚、森林草原和草原的生产力分别下降5%、7.5%和10%。总的来说，气温的升高对春大麦栽培热条件的恶化起到了调节作用，但在Polisia和Forest Steppe，在播种前和营养周期，它们仍然是有利的或令人满意的。
{"title":"Influence of changes in air temperature on crop productivity formation in Ukraine at the turn of XX–XXI centuries (1981–2010)","authors":"V. Balabukh, O. Tarariko, T. Ilienko, V. Velychko","doi":"10.15407/agrisp8.03.071","DOIUrl":"https://doi.org/10.15407/agrisp8.03.071","url":null,"abstract":"Aim. To determine the tendencies in the changes in air temperature and their influence on the productivity of crops\u0000during the vegetative cycle periods, especially in soil-climatic zones of Ukraine for the 1981–2010 climate normals\u0000period. Methods. The analytical and synthetic, statistical, climatic methods, simulation (model of V.P. Dmitrenko\u0000“Weather-yield” (Dmitrenko VP et al, 2017, 2010), used to forecast the productivity of grains in the Ukrainian Hy-\u0000drometeorological Center since 1970), abstract-logical method. Results. The rising air temperatures were determined\u0000throughout the whole vegetative period of growing corn and spring barley over the period of 1981–2010. It was\u0000found that this rise in different phases of crop development was of different magnitude and relevance in all regions\u0000and soil-climatic zones of Ukraine. The reliable changes in the surface air temperature were noted in the phases of\u0000the third leaf, panicle emergence, and blossoming of corn in Polissia, Forest-Steppe, and especially Steppe (0.7–\u00000.8 °С/10 years, 0.8–0.9 °С/10 years and 0.9–1.1 °С/10 years, respectively). During the pre-sowing period, the periods of\u0000corn sowing and seedlings, the velocity of changes was twice lower in the whole territory of the country, and during the pe-\u0000riods of milky ripeness and middle dough – in the eastern Forest-Steppe and dry Steppe, amounting to 0.4–0.5 °С/10 years.\u0000A considerable rise in the temperature during the period of the third leaf, panicle emergence, and blossoming promoted the\u0000decrease in the influence of temperature during these phases of crop development, especially in the Steppe (up to 10–15 %\u0000in 10 years). Only the rise in the temperature during the pre-sowing period promoted the 3–6 % increase in the whole terri-\u0000tory of the country, and during the periods of milky ripeness and middle dough of corn – up to 8 % in 10 years in the Forest-\u0000Steppe and Steppe. Generally, the thermal conditions for corn cultivation deteriorated considerably but remained favorable\u0000in Polissia, satisfactory – in the Forest-Steppe and northern Steppe, and unsatisfactory – in the south, in the dry Steppe. The\u0000most intense changes in the air temperature during the vegetation period of spring barley were noted in the phase of milky\u0000ripeness and middle dough in all soil-climatic zones, amounting to 0.8–1.1 °С/10 years. During the sowing period, the phases\u0000of the third leaf, stem elongation, and ear formation, they were 0.6–0.7 °С/10 years, and during the pre-sowing period – 0.3–\u00000.4 °С/10 years. During the spring barley tillering phase, the change in the air temperature was insignificant in the whole\u0000territory of the country. A considerable increase in the air temperature was unfavorable for crop cultivation in all the soil-\u0000climatic zones of Ukraine during the vegetative cycle of spring barley, especially during the phases of milky ripeness and\u0000middle dough, and promoted the decrease in its productivity in Polissia, Forest-Steppe, and Steppe by 5","PeriodicalId":55933,"journal":{"name":"Agricultural Science and Practice","volume":" ","pages":""},"PeriodicalIF":0.4,"publicationDate":"2021-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42860343","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Aim. The aim of this study was to develop a loop-mediated isothermal amplification (LAMP) assay for African swine fever virus (ASFV) detection. Methods. Primer design was performed using publicly available full genome sequences of ASFV. A panel of heterologous DNA samples and reference ASFV DNA samples were used for the assay specificity testing. The limit of detection (LOD) was assessed using purified and quantified serial dilution of the amplified target sequence. LAMP product detection was performed via gel-electrophoresis and via ethidium bromide fluorescence under UV after adding the ethidium bromide directly to the tube with the LAMP product. Results. Three primer sets amplifying different regions of ASFV gene C962R were developed, of which the set № 2 providing the most intense product synthesis with the most vivid and clear pattern was selected for further studies. The optimal concentration of reaction mix components for the most effective primer set was established. In the final protocol the LAMP reaction was carried out at 60 °C for 40 min. The limit of detection (LOD) of the assay was 50 copies of the target sequence per reaction. In a preliminary testing the assay proved specific, using 10 reference and 4 heterologous viral and two bacterial DNA samples. Our LAMP assay detected ASFV genotypes I and II that are currently spread in Europe, Asia, and the Pacific and IX, occurring in Africa. Conclusion. A LAMP assay was developed based on the C962R gene that proved in preliminary validation to be specific and sensitive and was able to detect down to 50 copies per reaction of purified target gene within 40 minutes. Classical gel electrophoresis and direct staining using ethidium bromide were used for product visualisation in this study. Colorimetric approaches or the use of lateral flow devices in the visuali- sation step could make the assay less equipment dependent. Further validation of the assay, determining analytical specificity, selectivity and reproducibility performance characteristics also using clinical samples under field condi- tions and inclusion of an internal control would possibly enable its use as a test of choice at point-of-care and at low resource laboratories.
{"title":"Development of a loop-mediated isothermal amplification (LAMP) assay based on the C962R gene for african swine fever virus detection","authors":"M. Kit, J. Schwarz, A. Gerilovych","doi":"10.15407/agrisp8.03.003","DOIUrl":"https://doi.org/10.15407/agrisp8.03.003","url":null,"abstract":"Aim. The aim of this study was to develop a loop-mediated isothermal amplification (LAMP) assay for African swine\u0000fever virus (ASFV) detection. Methods. Primer design was performed using publicly available full genome sequences\u0000of ASFV. A panel of heterologous DNA samples and reference ASFV DNA samples were used for the assay specificity\u0000testing. The limit of detection (LOD) was assessed using purified and quantified serial dilution of the amplified target\u0000sequence. LAMP product detection was performed via gel-electrophoresis and via ethidium bromide fluorescence\u0000under UV after adding the ethidium bromide directly to the tube with the LAMP product. Results. Three primer sets\u0000amplifying different regions of ASFV gene C962R were developed, of which the set № 2 providing the most intense\u0000product synthesis with the most vivid and clear pattern was selected for further studies. The optimal concentration of\u0000reaction mix components for the most effective primer set was established. In the final protocol the LAMP reaction\u0000was carried out at 60 °C for 40 min. The limit of detection (LOD) of the assay was 50 copies of the target sequence\u0000per reaction. In a preliminary testing the assay proved specific, using 10 reference and 4 heterologous viral and two\u0000bacterial DNA samples. Our LAMP assay detected ASFV genotypes I and II that are currently spread in Europe, Asia,\u0000and the Pacific and IX, occurring in Africa. Conclusion. A LAMP assay was developed based on the C962R gene that\u0000proved in preliminary validation to be specific and sensitive and was able to detect down to 50 copies per reaction of\u0000purified target gene within 40 minutes. Classical gel electrophoresis and direct staining using ethidium bromide were\u0000used for product visualisation in this study. Colorimetric approaches or the use of lateral flow devices in the visuali-\u0000sation step could make the assay less equipment dependent. Further validation of the assay, determining analytical\u0000specificity, selectivity and reproducibility performance characteristics also using clinical samples under field condi-\u0000tions and inclusion of an internal control would possibly enable its use as a test of choice at point-of-care and at low\u0000resource laboratories.","PeriodicalId":55933,"journal":{"name":"Agricultural Science and Practice","volume":" ","pages":""},"PeriodicalIF":0.4,"publicationDate":"2021-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48944507","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A. Shelyov, K. Kopylov, Y. Vdovychenko, S. Kramarenko, O. Kramarenko
Aim. Our work was aimed at investigating the specificities in the formation of the genetic structure of populations depending on the productivity direction of cattle, bred in Ukraine, using single locus DNA fragments, and studying the impact of the parental form on genetic polymorphism of modern intensive specialized breeds as a factor. Methods. The following methods were used in the work: veterinary methods (peripheral blood sampling); molecular-genetic meth- ods (the isolation and genotyping of DNA samples of dairy (83 animals), meat (192 animals) and aboriginal (43 ani- mals) cattle, bred in Ukraine, were performed by 10 microsatellite loci from the list, recommended by the International Society for Animal Genetics (ISAG); the complex of statistics methods was used for mathematic-statistical analysis, using modern software. Results. The analysis by 10 microsatellite loci demonstrated the specificities of genetic dif- ferentiation and the similarities between the investigated cattle populations, bred in Ukraine. Our results provide new information about the impact of artificial selection factors on single locus DNA fragments under the specialization of cattle breeds. The impact of the factor of ancestral form on the genetic structure was determined and confirmed by the same polymorphism spectra of the investigated DNA fragments in the maternal and derivative breeds. Another con- firmation was found in the differences, observed in animals of different productivity directions, which are a probable result of the breed-forming process, demonstrated by the results of the mathematic calculations of the data obtained. It was shown that microsatellite DNA loci are highly informative markers of genetic processes, occurring in domestic cattle populations. Conclusions. The specificities in the formation of the genetic structure of populations depending on the productivity direction of animals were determined. The impact of the parental form on genetic polymorphism of modern intensive specialized breeds was noted. It was found that among 10 microsatellite loci, used by us, there were loci in each group of animals, regarding which the hypothesis about their neutrality was reliably rejected accord- ing to the results of Ewens-Watterson test: for dairy cattle (INRA023, ETH3, ETH225, BM1824, BM2113, ETH10 and SPS115), for meat cattle (TGLA122 and ETH225), and for aboriginal cattle (TGLA126, INRA023 and TGLA227). We determined a high level of genetic diversity, remarkable for each investigated cattle population, bred in Ukraine, and general tendencies of differentiation in the selected populations depending on the targeted breeding work, on the level of polymorphism of microsatellite DNA loci (Friedman’s test: P < 0.01), and a similar genetic picture for a number of loci of investigated DNA fragments, which may be related to the history of creating these breeds.
{"title":"Formation of the genetic structure of cattle populations by single locus DNA fragments depending on their productivity direction and origin","authors":"A. Shelyov, K. Kopylov, Y. Vdovychenko, S. Kramarenko, O. Kramarenko","doi":"10.15407/agrisp8.03.035","DOIUrl":"https://doi.org/10.15407/agrisp8.03.035","url":null,"abstract":"Aim. Our work was aimed at investigating the specificities in the formation of the genetic structure of populations\u0000depending on the productivity direction of cattle, bred in Ukraine, using single locus DNA fragments, and studying the\u0000impact of the parental form on genetic polymorphism of modern intensive specialized breeds as a factor. Methods. The\u0000following methods were used in the work: veterinary methods (peripheral blood sampling); molecular-genetic meth-\u0000ods (the isolation and genotyping of DNA samples of dairy (83 animals), meat (192 animals) and aboriginal (43 ani-\u0000mals) cattle, bred in Ukraine, were performed by 10 microsatellite loci from the list, recommended by the International\u0000Society for Animal Genetics (ISAG); the complex of statistics methods was used for mathematic-statistical analysis,\u0000using modern software. Results. The analysis by 10 microsatellite loci demonstrated the specificities of genetic dif-\u0000ferentiation and the similarities between the investigated cattle populations, bred in Ukraine. Our results provide new\u0000information about the impact of artificial selection factors on single locus DNA fragments under the specialization of\u0000cattle breeds. The impact of the factor of ancestral form on the genetic structure was determined and confirmed by the\u0000same polymorphism spectra of the investigated DNA fragments in the maternal and derivative breeds. Another con-\u0000firmation was found in the differences, observed in animals of different productivity directions, which are a probable\u0000result of the breed-forming process, demonstrated by the results of the mathematic calculations of the data obtained.\u0000It was shown that microsatellite DNA loci are highly informative markers of genetic processes, occurring in domestic\u0000cattle populations. Conclusions. The specificities in the formation of the genetic structure of populations depending\u0000on the productivity direction of animals were determined. The impact of the parental form on genetic polymorphism\u0000of modern intensive specialized breeds was noted. It was found that among 10 microsatellite loci, used by us, there\u0000were loci in each group of animals, regarding which the hypothesis about their neutrality was reliably rejected accord-\u0000ing to the results of Ewens-Watterson test: for dairy cattle (INRA023, ETH3, ETH225, BM1824, BM2113, ETH10 and\u0000SPS115), for meat cattle (TGLA122 and ETH225), and for aboriginal cattle (TGLA126, INRA023 and TGLA227). We\u0000determined a high level of genetic diversity, remarkable for each investigated cattle population, bred in Ukraine, and\u0000general tendencies of differentiation in the selected populations depending on the targeted breeding work, on the level\u0000of polymorphism of microsatellite DNA loci (Friedman’s test: P < 0.01), and a similar genetic picture for a number of\u0000loci of investigated DNA fragments, which may be related to the history of creating these breeds.","PeriodicalId":55933,"journal":{"name":"Agricultural Science and Practice","volume":" ","pages":""},"PeriodicalIF":0.4,"publicationDate":"2021-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48831893","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
京公网安备 11010802042870号
京ICP备2023020795号-1
扫码关注我们
求助内容：
应助结果提醒方式：