OPPORTUNITIES OF MICRONUTRIENTAL AND PHYTOCORRECTION IN INCREASE OF RESISTANCE OF CATARRHAL DISEASES IN CHILDREN

A term «frequently ailing children» (FAC) characterizes increased morbidity with acute respiratory infections compared with healthy coevals, and this term is widely used by pediatrists in present time. unfavorable course of pregnancy, prematurity, defects in immune system are predisposing to this state. Rational nutrition, regular day regimen, sanitary measures are necessary to the normal living of such children. In some cases medicamental correction with use of poly vitamins, immunocorrectors and phytotherapy is needed. Deficiency of vitamins and minerals is frequently occurring in children from FAC group. «sanasol extravit», containing extracts from black currant, dog rose and strawberry, was developed for the deficiency of microelements and vitamins and for a correction of immune system.Key words: frequently ailing children, vitamins, immunity

DISTRIBUTION OF THIABENDAZOLE RESISTANT HELMINTHOSPORIUM SOLANI STRAIN

Helminthosporium solani strains were isolated from potato tubers that were grown in Russia, Germany and the Netherlands. Thiabendazole was effective against the sensitive strains of H. solani (EC50< 7.3 mg/l); however, 8 studied strains from Russia and the Netherlands were found to be extremely resistant to it (EC50 > 1000 mg/l). The sequence of their β-tubulin gene contained a SNP mutation in 198 codon or 200 codon, translating to Gln (CAG) instead of Glu (GAG) or Tyr (TAC) instead of Phe (TTC), respectively. Resistant to thiabendazole strains were found in distant regions of Russia (Moscow, Kostroma, Kaluga district) and in the Netherlands. Resistance to thiabendazole of the Russian, European, and American strains had the same genetic background and was conferred by the same mutations

Устойчивость Helminthosporium solani, Colletotrichum coccodes и Rhizoctonia solani к фунгицидам, используемым для обработки клубней картофеля

We evaluated the growth inhibition of colonies of phytopathogenic fungi Helminthosporium solani, Colletotrichum coccodes, and Rhizoctonia solani with fungicides difenoconazole (Skor), azoxystrobin (Kvadris), fludioxonil (Maksim), pencycuron (Prestizh), thiabendazole (Tekto), and colloidal silver (Zeroxxe). We used the isolates of C. coccodes, H. solani, R. solani from potato tubers, grown in 2013-2016 in the Russian Federation, as well as from seed potato from the Federal Republic of Germany (FRG) and the Netherlands. The type of isolates was determined by the culture-morphological features, the structure of the nuclear ribosomal gene section and internal transcribed spacers (ITS1-5.8S-ITS2). The isolates were cultured with wort agar and fungicides at concentrations of 0.1; 1; 10; 100 mg/L of an active substance, in the control - without fungicides. In the case of thiabendazole and pencycuron, a concentration of 1000 mg/L was also used. There were no strains with high resistance to difenoconazole (EC50 for H. solani was 0.05-0.13 mg/L, for R. solani - 4-70 mg/L, for C. coccodes - 0.05-0.12 mg/L) and colloidal silver (EC50 was 7.0-72.7, 0.75-7.8, 1.2-7.0 mg/L, respectively). Thiabendazole was highly effective (EC50 was 0.9-more than 100; 4.9-8.5; 0.9-50.3 mg/L, respectively), but it was revealed four highly resistant strains (EC50 was more than 1000 mg/L) of H. solani, which is due to the same mutations that of the European and American strains. Fludioxonil inhibited the growth of C. coccodes (EC50 was 0.60-0.90 mg/L) and R. solani(EC50 was 0.05-0.09 mg/L), but, when cultured for more than 20 days, strains formed stable sectors that grew faster than the original isolate. Pencycuron showed a good fungicidal effect only in relation to R. solani (EC50 was 0.05-more than 100 mg/L), however, three highly resistant strains of this fungus (EC50 was more than 1000 mg/L) were found. Resistance to azoxystrobin was very different, which is related to the work of alternative respiratory tracts when growing on a nutrient medium. In the field, its effectiveness may be higher.Оценивали ингибирование роста колоний фитопатогенных грибов Helminthosporium solani, Colletotrichum coccodes и Rhizoctonia solani фунгицидами дифеноконазол (Скор), азоксистробин (Квадрис), флудиоксонил (Максим), пенцикурон (Престиж), тиабендазол (Текто) и коллоидным серебром (Зерокс). Использовали изоляты C. coccodes, H. solani, R. solani из клубней картофеля, выращенных в2013-2016гг. в РФ, а также из семенного картофеля из ФРГ и Нидерландов. Вид изолятов определяли по культурально-морфологическим признакам, структуре участка ядерных рибосомных генов и внутренних транскрибируемых спейсеров (ITS1-5,8S-ITS2). Изоляты культивировали с сусло-агаром и фунгицидом в концентрациях 0,1; 1; 10; 100 мг/л действующего вещества, в контроле - без фунгицида. В случае тиабендазола и пенцикурона также использовали концентрацию 1000 мг/л. Не выявлено штаммов с высокой устойчивостью к дифеноконазолу (ЕС0для H. solani - 0,05-0,13 мг/л, для R. solani - 4-70 мг/л, для C. coccodes - 0,05-0,12 мг/л) и коллоидному серебру (ЕС50 7,0-72,7; 0,75-7,8; 1,2-7,0 мг/л соответственно). Тиабендазол отличался высокой эффективностью, но среди штаммов H. solani выявлены 4 высокоустойчивых (ЕС50 более 1000 мг/л), что обусловлено теми же мутациями, что у европейских и американских штаммов. Флудиоксонил ингибировал рост C. coccodes (ЕС50 0,60-0,90 мг/л) и R. solani(ЕС500,05-0,09 мг/л), но при культивировании более 20 дней штаммы образовывали устойчивые сектора, которые росли быстрее, чем исходный изолят. Пенцикурон показал хороший фунгицидный эффекттолько в отношении R. solani, однако в природных популяциях найдены 3 высокоустойчивых штамма этого гриба (ЕС50более 1000 мг/л). Устойчивость казоксистробину сильно различалась, что связано с работой альтернативных дыхательных путей при росте на питательной среде. В полевых условиях его эффективность может быть выше

Устойчивость Helminthosporium solani, Colletotrichum coccodes и Rhizoctonia solani к фунгицидам, используемым для обработки клубней картофеля

We evaluated the growth inhibition of colonies of phytopathogenic fungi Helminthosporium solani, Colletotrichum coccodes, and Rhizoctonia solani with fungicides difenoconazole (Skor), azoxystrobin (Kvadris), fludioxonil (Maksim), pencycuron (Prestizh), thiabendazole (Tekto), and colloidal silver (Zeroxxe). We used the isolates of C. coccodes, H. solani, R. solani from potato tubers, grown in 2013-2016 in the Russian Federation, as well as from seed potato from the Federal Republic of Germany (FRG) and the Netherlands. The type of isolates was determined by the culture-morphological features, the structure of the nuclear ribosomal gene section and internal transcribed spacers (ITS1-5.8S-ITS2). The isolates were cultured with wort agar and fungicides at concentrations of 0.1; 1; 10; 100 mg/L of an active substance, in the control - without fungicides. In the case of thiabendazole and pencycuron, a concentration of 1000 mg/L was also used. There were no strains with high resistance to difenoconazole (EC50 for H. solani was 0.05-0.13 mg/L, for R. solani - 4-70 mg/L, for C. coccodes - 0.05-0.12 mg/L) and colloidal silver (EC50 was 7.0-72.7, 0.75-7.8, 1.2-7.0 mg/L, respectively). Thiabendazole was highly effective (EC50 was 0.9-more than 100; 4.9-8.5; 0.9-50.3 mg/L, respectively), but it was revealed four highly resistant strains (EC50 was more than 1000 mg/L) of H. solani, which is due to the same mutations that of the European and American strains. Fludioxonil inhibited the growth of C. coccodes (EC50 was 0.60-0.90 mg/L) and R. solani(EC50 was 0.05-0.09 mg/L), but, when cultured for more than 20 days, strains formed stable sectors that grew faster than the original isolate. Pencycuron showed a good fungicidal effect only in relation to R. solani (EC50 was 0.05-more than 100 mg/L), however, three highly resistant strains of this fungus (EC50 was more than 1000 mg/L) were found. Resistance to azoxystrobin was very different, which is related to the work of alternative respiratory tracts when growing on a nutrient medium. In the field, its effectiveness may be higher.Оценивали ингибирование роста колоний фитопатогенных грибов Helminthosporium solani, Colletotrichum coccodes и Rhizoctonia solani фунгицидами дифеноконазол (Скор), азоксистробин (Квадрис), флудиоксонил (Максим), пенцикурон (Престиж), тиабендазол (Текто) и коллоидным серебром (Зерокс). Использовали изоляты C. coccodes, H. solani, R. solani из клубней картофеля, выращенных в2013-2016гг. в РФ, а также из семенного картофеля из ФРГ и Нидерландов. Вид изолятов определяли по культурально-морфологическим признакам, структуре участка ядерных рибосомных генов и внутренних транскрибируемых спейсеров (ITS1-5,8S-ITS2). Изоляты культивировали с сусло-агаром и фунгицидом в концентрациях 0,1; 1; 10; 100 мг/л действующего вещества, в контроле - без фунгицида. В случае тиабендазола и пенцикурона также использовали концентрацию 1000 мг/л. Не выявлено штаммов с высокой устойчивостью к дифеноконазолу (ЕС0для H. solani - 0,05-0,13 мг/л, для R. solani - 4-70 мг/л, для C. coccodes - 0,05-0,12 мг/л) и коллоидному серебру (ЕС50 7,0-72,7; 0,75-7,8; 1,2-7,0 мг/л соответственно). Тиабендазол отличался высокой эффективностью, но среди штаммов H. solani выявлены 4 высокоустойчивых (ЕС50 более 1000 мг/л), что обусловлено теми же мутациями, что у европейских и американских штаммов. Флудиоксонил ингибировал рост C. coccodes (ЕС50 0,60-0,90 мг/л) и R. solani(ЕС500,05-0,09 мг/л), но при культивировании более 20 дней штаммы образовывали устойчивые сектора, которые росли быстрее, чем исходный изолят. Пенцикурон показал хороший фунгицидный эффекттолько в отношении R. solani, однако в природных популяциях найдены 3 высокоустойчивых штамма этого гриба (ЕС50более 1000 мг/л). Устойчивость казоксистробину сильно различалась, что связано с работой альтернативных дыхательных путей при росте на питательной среде. В полевых условиях его эффективность может быть выше

The occurrence of thiabendazole-resistant isolates of Helminthosporium solani on potato seed tubers in Russia

Silver scurf is a potato tuber disease caused by Helminthosporium solani, which has become an increasing problem in many regions of Russia. The fungicide thiabendazole is widely used to control silver scab. Its target of action is the beta-tubulin gene. Single nucleotide mutations in this gene lead to a multiple increase in the resistance of the strain to the fungicide. The resistance of 37 Helminthosporium solani strains to the fungicide thiabendazole was tested in vitro. Tested strains were isolated from potato tubers grown in seven regions of Russia. In this study, the presence of thiabendazole-resistant strains was shown in populations of the European part of Russia, but not for the Far Eastern part. In the Far Eastern part, potatoes are grown from seeds of local production and there is practically no exchange with the European part of Russia. In the present study, we obtain sequences of the beta-tubulin gene and tested the resistance of strains affecting potatoes in the Far Eastern part of Russia to thiabendazole to show the absence of resistant strains. © 2020, Deutsche Phytomedizinische Gesellschaft

Resistance of Helminthosporium solani strains to selected fungicides applied for tuber treatment

Helminthosporium solani strains were isolated from potato tubers collected in Russia or taken from imported German and Dutch seed tubers. Sequences of the nuclear ribosomal genes and internal transcribed spacers (ITS) for all 24 tested strains were identical and had 100% similarity to the sequences from GenBank identified as Helminthosporium solani. The obtained molecular data confirmed the morphological identification based on the width and length of conidia, the shape of conidiophores and the colony morphology. Screening for resistance to the fungicides Score 250 SC (active ingredient difenoconazole 250 g/l), Quadris (azoxystrobin 250 g/l), Tecto 500 SC (thiabendazole 500g/l), Zeroxxe [colloidal silver particles (3 g/l) stabilized with amphoteric surfactant] was done. Agar blocks with pure cultures of the fungal strains were placed in the centre of Petri dishes containing malt agar amended with fungicide concentrations of 0.1, 1, 10, 100 and 1000 mg/l (accounted for the concentration of the active ingredient). Malt agar free of fungicide was used as the control. Growth inhibition of 50% (EC50) compared to the control was detected based on the dose-response curves. Difenoconazole (EC50 < 0.12mg/l) and colloidal silver (EC50 < 76 mg/l) were the most effective fungicides. No strains resistant to the aforementioned fungicides were found. In most cases, azoxystrobin was effective against H. solani (EC50 < 7 mg/l), but there were several strains with high resistance to this fungicide (EC50 > 100 mg/l). Thiabendazole appears to be effective against the sensitive strains of H. solani (EC50 < 7.3 mg/l); however, six studied strains from Russia and the Netherlands were found to be extremely resistant to it (EC50 > 1000 mg/l). The sequence of their β-tubulin gene contained a SNP mutation in the 198 codon or 200 codon, translating to Gln (CAG) instead of Glu (GAG) or Tyr (TAC) instead of Phe (TTC), respectively. Thus, the resistance to thiabendazole of the Russian, European and American strains had the same genetic background and was conferred by the same mutations. © 2017, Edizioni ETS. All rights reserved

Colletotrichum coccodes in potato and tomato leaves in Russia

Colletotrichum coccodes is a plant pathogenic fungus affecting different organs of potato, tomato, and some other plants. The leaf infection with C. coccodes may result in a rapid development of infection nidus and high losses caused by the infection of tomato fruits and potato tubers during harvesting. Our study showed the presence of C. coccodes on tomato and potato leaves with dry necrotic lesions. The presence of a DNA region specific for C. coccodes was detected in DNA samples isolated from infected tomato leaves collected in the Rostov region (7 and 1 samples from two distinct fields) and the Krasnodar Territory (5 samples from one field). In the case of potato leaf samples, the frequency of C. coccodes detection was lower than in tomato leaves. This fungus was revealed only in five potato leaf samples: one from the Northern Ossetia, one from the Kostroma region, and three from the Mariy El Republic. Pure cultures of two isolates were isolated from a naturally infected potato leaf. According to morphological criteria, size of spores and sclerotia, habitus of colonies, and species-specific DNA sequences, both strains completely corresponded to C. coccodes. © 2017, Deutsche Phytomedizinische Gesellschaft

Resistance of Helminthosporium solani strains to selected fungicides applied for tuber treatment

Helminthosporium solani strains were isolated from potato tubers collected in Russia or taken from imported German and Dutch seed tubers. Sequences of the nuclear ribosomal genes and internal transcribed spacers (ITS) for all 24 tested strains were identical and had 100% similarity to the sequences from GenBank identified as Helminthosporium solani. The obtained molecular data confirmed the morphological identification based on the width and length of conidia, the shape of conidiophores and the colony morphology. Screening for resistance to the fungicides Score 250 SC (active ingredient difenoconazole 250 g/l), Quadris (azoxystrobin 250 g/l), Tecto 500 SC (thiabendazole 500g/l), Zeroxxe [colloidal silver particles (3 g/l) stabilized with amphoteric surfactant] was done. Agar blocks with pure cultures of the fungal strains were placed in the centre of Petri dishes containing malt agar amended with fungicide concentrations of 0.1, 1, 10, 100 and 1000 mg/l (accounted for the concentration of the active ingredient). Malt agar free of fungicide was used as the control. Growth inhibition of 50% (EC50) compared to the control was detected based on the dose-response curves. Difenoconazole (EC50 < 0.12mg/l) and colloidal silver (EC50 < 76 mg/l) were the most effective fungicides. No strains resistant to the aforementioned fungicides were found. In most cases, azoxystrobin was effective against H. solani (EC50 < 7 mg/l), but there were several strains with high resistance to this fungicide (EC50 > 100 mg/l). Thiabendazole appears to be effective against the sensitive strains of H. solani (EC50 < 7.3 mg/l); however, six studied strains from Russia and the Netherlands were found to be extremely resistant to it (EC50 > 1000 mg/l). The sequence of their β-tubulin gene contained a SNP mutation in the 198 codon or 200 codon, translating to Gln (CAG) instead of Glu (GAG) or Tyr (TAC) instead of Phe (TTC), respectively. Thus, the resistance to thiabendazole of the Russian, European and American strains had the same genetic background and was conferred by the same mutations. © 2017, Edizioni ETS. All rights reserved

Colletotrichum coccodes in potato and tomato leaves in Russia

Colletotrichum coccodes is a plant pathogenic fungus affecting different organs of potato, tomato, and some other plants. The leaf infection with C. coccodes may result in a rapid development of infection nidus and high losses caused by the infection of tomato fruits and potato tubers during harvesting. Our study showed the presence of C. coccodes on tomato and potato leaves with dry necrotic lesions. The presence of a DNA region specific for C. coccodes was detected in DNA samples isolated from infected tomato leaves collected in the Rostov region (7 and 1 samples from two distinct fields) and the Krasnodar Territory (5 samples from one field). In the case of potato leaf samples, the frequency of C. coccodes detection was lower than in tomato leaves. This fungus was revealed only in five potato leaf samples: one from the Northern Ossetia, one from the Kostroma region, and three from the Mariy El Republic. Pure cultures of two isolates were isolated from a naturally infected potato leaf. According to morphological criteria, size of spores and sclerotia, habitus of colonies, and species-specific DNA sequences, both strains completely corresponded to C. coccodes. © 2017, Deutsche Phytomedizinische Gesellschaft