Isolation and identification of Lactobacillus paracasei and Bacillus subtilis in goose feces / 中国生物制品学杂志

@#Objective To isolate and identify Lactobacillus paracasei and Bacillus subtilis from feces of northern white geese.Methods Lactobacillus paracasei and Bacillus subtilis,numbered S00834-8 and IS00439-2 respectively,were isolated from feces of 30 52-week-old northern white geese and cultured in specific medium,and identified by 16S DNA sequencing. The two strains were cultured for different time duration(0,2,4,6,8,10,12,14,16,18,20,22,24,36 and 48 h),at different temperature(34,36,37,38 and 40 ℃)and pH(5. 5,6. 0,6. 5,7. 0,7. 5 and 8. 0). The growth characteristics of the strains were evaluated according to the A600values measured by UV-Vis spectrophotometer. At the same time,the acid and bile salt tolerance(pH 2. 0,2. 5,3. 0,3. 5 and 7. 0,bile salt concentration 0,0. 3%,0. 5%,1. 0% and 1. 5%)and the antibiotic tolerance were tested. Results The strains S00834-8 and IS00439-2 were Lactobacillus paracasei subspecies tolerance H and Bacillus subtilis KA9,respectively. The strain S00834-8 cultured under the optimum condition of the culture medium with pH 6. 0,at 37 ℃,for 12 h,had good bile resistance and gastric acid resistance,which was moderately susceptible to piperacillin,norfloxacin,amikacin,gentamicin and tetracycline,and susceptible to levofloxacin,streptomycin and chloramphenicol. The strain IS00439-2 best cultured in the culture medium with pH 7. 5,at 36 ℃,for 8 h,showed good bile resistance and gastric acid resistance,which was moderately sensitive to levofloxacin and tetracycline,while sensitive to piperacillin,norfloxacin,streptomycin and chloramphenicol. Conclusion Goose-derived Lactobacillus paracasei subspecies tolerance H and Bacillus subtilis KA9 have good acid resistance and bile salt resistance,which are expected to be candidate strains for goose-derived microecological preparations.

Biofortified Vermicompost Mediated Induced Resistance Response in Tomato against Fusarium oxysporum f. sp. lycopersici

Tomato (Solanum lycopersicum) is a widely cultivated and versatile fruit. Tomato production faces various constraints, both biotic and abiotic stresses. Fusarium wilt, caused by the soil-borne fungus Fusarium oxysporum f. sp. lycopersici, is a devastating disease that affects tomato plants worldwide, causing significant economic losses in tomato cultivation. Inthis study the potential use of biofortified vermicompost for wilt management was investigated. The biological control agents viz. Trichoderma harzianum, Pseudomonas fluorescens and Bacillus subtilis were used to fortified the vermicpompost. The study monitored various antioxidants, plant growth indicators, and disease rates across designated treatments at different intervals. The findings revealed substantial differences: tomato plants treated with biofortified vermicompost showed notable reductions in disease occurrence, improved growth, increased yields, and heightened antioxidant activity compared to the control. Among these treatments, the most remarkable outcomes were observed in plants treated with T. harzianum fortified vermicompost, exhibiting the highest recorded values across the parameters studied. Data revealed that the Tomato plants treated with vermicompost fortified with Trichoderma showed maximum root length (14.95 cm) after 15 days of sowing followed by T-2 (11.25 cm) and T-3 (9.85 cm). Maximum dry weight was observed in plants treated with vermicompost fortified with Trichoderma. Maximum PAL, PO and PPO activity was recorded in leaves from plant grown in vermicompost fortified with T. harzianum (T1) at 48 h followed by T2, T3 and T4. The findings revealed tomato plants treated with biofortified vermicompost showed notable reductions in disease occurrence, improved growth, increased yields, and higher accumulation of defense enzyme activity compared to the control.

Influence of Organic Seaweed Pelleting on Seed Quality Parameters of Onion Seeds

Aim: Small and irregular seed shapes make precision sowing difficult. Seed pelleting, which helps to make seed handling easier during sowing, is converting such seeds into bold, spherical shapes with a smooth surface. Seaweeds are a rich source of nutrients that promote growth, such as IAA, kinetin, zeatin, and gibberellins; auxins and cytokinins; metabolic enhancers; macro and micro elements; amino acids; and vitamins. Their use in crop plants has been shown to have positive effects, including early seed germination and establishment, improved crop performance and yield, increased resistance to biotic and abiotic stress, and increased seed shelf life (Bluden).Study Design: The experiment was undertaken with three replications in a completely randomized block design. Eight treatmental combinations were used in this study. Statistical analysis was carried out by AGRES software. The Critical Differences (CD) was calculated at 5 per cent probability level.Place and Duration of Study: Department of Seed Science and Technology, Agriculture College and Research Institute, TNAU, Madurai, during the period of December 2020 and November 2021.Methodology: Organic seaweed pellet combinations can be successfully implemented to provide improved field emergence, crop establishment and production. Seeds were pelleted with different mixtures of Sargassum sp., Kappaphycus sp., Bacillus subtilis and Talc powder.Results: The results exposed that between the treatments seed pelleting with combination of T7- Sargassum sp + Kappaphycus sp + Bacillus subtilis+ Talc powder was found to be more efficient in comparison with other combinations.Conclusion: Pelleting with, Kappaphycus sp, Bacillus subtilis and Talc powder was found to be better in all aspects of seed quality and biochemical parameters.

Effect of Seed Treatment with Bioinoculants on Seed Yield and Quality Parameters in Chickpea (Cicer arietinum L.)

The field experiment, was conducted during Rabi 2022 at the field experimental center, Department of Genetics and Plant Breeding, SHUATS, Prayagraj, (U.P). The soil of the experimental plot was sandy loam in texture, nearly neutral in soil reaction (pH 7.1), low in organic carbon (0.36%), available N (171.48 kg/ha), available P (15.2 kg/ha) and available K (232.5 kg/ha). The experiment was laid out in Randomized Block Design with thirteen treatments including control which were replicated thrice. The treatments are as follows, T0- Control, T1 - Bacillus subtilis – 20 g/kg seeds, T2 – Rhizobium – 20 g/kg seeds, T3 – P.S.B – 20 g/kg seed, T4 - T. harzianum – 20 g/kg seed, T5 - T. viridae – 20 g/kg seed, T6 – B. subtilis + Rhizobium – (10 g + 10 g/kg seed), T7 - Rhizobium + P.S.B – (10 g + 10 g/kg seed), T8 - P.S.B + T. harzianum – (10 g + 10 g/kg seed), T9 – T. harzianum + T. viridae – (10 g + 10 g/kg seed), T10 - T. viridae + B. subtilis – (10 g + 10 g/kg seed), T11 - B. subtilis + Rhizobium + P.S.B – (10 g + 10 g+10 g/kg seed), T12 - P.S.B + T. harzianum + T, viridae – (10 g + 10 g+10 g/kg seed) respectively. The experiment results revealed that seeds treated with T7 – Rhizobium + P.S.B – 10 g + 10 g gave better than other treatments viz, days for 50% flowering (60.00), Days to 50% pod setting (75.00) and has matured earlier (99.00), Plant height (74.90 cm), number of pods per plant (52.33), number of seeds per pod (3.00), Seed yield per plot (522.80 g), Biological yield per plot (613.58), Seed index (32.24 gm). Were recorded significantly higher compared to other treatments.

Functional analysis of functional membrane microdomains in the biosynthesis of menaquinone-7 / 生物工程学报

Functional membrane microdomains (FMMs) that are mainly composed of scaffold proteins and polyisoprenoids play important roles in diverse cellular physiological processes in bacteria. The aim of this study was to identify the correlation between MK-7 and FMMs and then regulate the MK-7 biosynthesis through FMMs. Firstly, the relationship between FMMs and MK-7 on the cell membrane was determined by fluorescent labeling. Secondly, we demonstrated that MK-7 is a key polyisoprenoid component of FMMs by analyzing the changes in the content of MK-7 on cell membrane and the changes in the membrane order before and after destroying the integrity of FMMs. Subsequently, the subcellular localization of some key enzymes in MK-7 synthesis was explored by visual analysis, and the intracellular free pathway enzymes Fni, IspA, HepT and YuxO were localized to FMMs through FloA to achieve the compartmentalization of MK-7 synthesis pathway. Finally, a high MK-7 production strain BS3AT was successfully obtained. The production of MK-7 reached 300.3 mg/L in shake flask and 464.2 mg/L in 3 L fermenter.

Development of biosensors highly responsive to N-acetylneuraminic acid in Bacillus subtilis / 生物工程学报

Bacillus subtilis is recognized as a generally-regarded-as-safe strain, and has been widely used in the biosynthesis of high value-added products, including N-acetylneuraminic acid (NeuAc) which is widely used as a nutraceutical and a pharmaceutical intermediate. Biosensors responding to target products are widely used in dynamic regulation and high-throughput screening in metabolic engineering to improve the efficiency of biosynthesis. However, B. subtilis lacks biosensors that can efficiently respond to NeuAc. This study first tested and optimized the transport capacity of NeuAc transporters, and obtained a series of strains with different transport capacities for testing NeuAc-responsive biosensors. Subsequently, the binding site sequence of Bbr_NanR responding to NeuAc was inserted into different sites of the constitutive promoter of B. subtilis, and active hybrid promoters were obtained. Next, by introducing and optimizing the expression of Bbr_NanR in B. subtilis with NeuAc transport capacity, we obtained an NeuAc-responsive biosensor with wide dynamic range and higher activation fold. Among them, P535-N2 can sensitively respond to changes in intracellular NeuAc concentration, with the largest dynamic range (180-20 245) AU/OD. P566-N2 shows a 122-fold of activation, which is 2 times of the reported NeuAc-responsive biosensor in B. subtilis. The NeuAc-responsive biosensor developed in this study can be used to screen enzyme mutants and B. subtilis strains with high NeuAc production efficiency, providing an efficient and sensitive analysis and regulation tool for biosynthesis of NeuAc in B. subtilis.

Endophytic Bacillus subtilis P10 from Prunus cerasifera as a biocontrol agent against tomato Verticillium wilt / Bacillus subtilis P10 endofítico de Prunus cerasifera como agente de biocontrole contra murcha de Verticillium de tomate

Endophytic bacteria serve key roles in the maintenance of plant health and growth. Few studies to date, however, have explored the antagonistic and plant growth-promoting (PGP) properties of Prunus cerasifera endophytes. To that end, we isolated endophytic bacteria from P. cerasifera tissue samples and used a dual culture plate assay to screen these microbes for antagonistic activity against Verticillium dahliae, Botryosphaeria dothidea, Fusarium oxysporum, F. graminearum, and F. moniliforme. Of the 36 strains of isolated bacteria, four (strains P1, P10, P16, and P20) exhibited antagonistic effects against all five model pathogens, and the P10 strain exhibited the strongest antagonistic to five pathogens. This P10 strain was then characterized in-depth via phenotypic assessments, physiological analyses, and 16s rDNA sequencing, revealing it to be a strain of Bacillus subtilis. Application of a P10 cell suspension (1×108 CFU/mL) significantly enhanced the seed germination and seedling growth of tomato in a greenhouse setting. This P10 strain further significantly suppressed tomato Verticillium wilt with much lower disease incidence and disease index scores being observed following P10 treatment relative to untreated plants in pot-based experiments. Tomato plants that had been treated with strain P10 also enhanced defense-related enzymes, peroxidase, superoxide dismutase, and catalase activity upon V. dahliae challenge relative to plants that had not been treated with this endophytic bacterium. The results revealed that the P10 bacterial strain has potential value as a biocontrol agent for use in the prevention of tomato Verticillium wilt.

As bactérias endofíticas desempenham papel fundamental na manutenção da saúde e do crescimento das plantas. Poucos estudos até o momento, no entanto, exploraram as propriedades antagônicas e promotoras de crescimento de plantas (PGP) de endófitos de Prunus cerasifera. Para esse fim, isolamos bactérias endofíticas de amostras de tecido de P. cerasifera e usamos um ensaio de placa de cultura dupla para rastrear esses micróbios quanto à atividade antagonista contra Verticillium dahliae, Botryosphaeria dothidea, Fusarium oxysporum, F. graminearum e F. moniliforme. Das 36 cepas de bactérias isoladas, quatro (cepas P1, P10, P16 e P20) exibiram efeitos antagônicos contra todos os cinco patógenos modelo, e a cepa P10 exibiu o antagonista mais forte para cinco patógenos. Essa cepa P10 foi então caracterizada em profundidade por meio de avaliações fenotípicas, análises fisiológicas e sequenciamento de rDNA 16s, revelando ser uma cepa de Bacillus subtilis. A aplicação de uma suspensão de células P10 (1 × 108 UFC / mL) aumentou significativamente a germinação das sementes e o crescimento das mudas de tomate em casa de vegetação. Essa cepa P10 suprimiu ainda mais a murcha de Verticillium do tomate com incidência de doença muito menor e pontuações de índice de doença sendo observadas após o tratamento com P10 em relação a plantas não tratadas em experimentos baseados em vasos. As plantas de tomate que foram tratadas com a cepa P10 também aumentaram as enzimas relacionadas à defesa, peroxidase, superóxido dismutase e atividade da catalase após o desafio de V. dahliae em relação às plantas que não foram tratadas com essa bactéria endofítica. Os resultados revelaram que a cepa bacteriana P10 tem valor potencial como agente de biocontrole para uso na prevenção da murcha de Verticillium em tomate.

Identification by molecular techniques of halophilic bacteria producing important enzymes from pristine area in Campeche, Mexico / Identificação por técnicas moleculares de bactérias halofílicas que produzem enzimas importantes em área intocada de Campeche, México

Isla Arena is located in the coordinate 20° 70´ N - 90° 45´ W, from Campeche, Mexico. In these estuaries, the ocean mixes with fresh water, and ecosystems are concentrated where petenes and pink flamingos proliferate. Crustaceans and mollusks abound in the sea. Despite its enormous marine wealth, there are no studies carried out on which halophilic microorganisms are present in these waters. In this work, the diversity and structure of the microbial community was investigated through a metagenomics approach and corroborated for sequencing of 16S rRNA genes. It was found that the phylum Fimicutes predominates with more than 50%, in almost the same proportion of the class Bacilli and with almost 41% of relative abundance of the order Bacillales. The sequencing results showed that one of the samples presented a high percentage of similarity (99.75%) using the Nucleotide BLAST program with a peculiar microorganism: Bacillus subtilis. This microorganism is one of the best characterized bacteria among the gram-positive ones. Our results demonstrate that B. subtilis can be an efficient source of proteases, lipases and cellulases, from halophilic microbial communities located in poorly explored areas.

Isla Arena está localizada na coordenada 20°70’N - 90°45’W, de Campeche, México. Nesses estuários, o oceano se mistura com a água doce e os ecossistemas se concentram onde proliferam petenos e flamingos rosa. Crustáceos e moluscos abundam no mar. Apesar de sua enorme riqueza marinha, não há estudos realizados sobre a presença de microrganismos halofílicos nessas águas. Neste trabalho, a diversidade e estrutura da comunidade microbiana foram investigadas através de uma abordagem metagenômica e corroboradas para o sequenciamento de genes 16S rRNA. Verificou-se que o filo Fimicutes predomina com mais de 50%, quase na mesma proporção da classe Bacilli e com quase 41% de abundância relativa da ordem Bacillales. Os resultados do sequenciamento mostraram que uma das amostras apresentou alto percentual de similaridade (99,75%) pelo programa Nucleotide BLAST com um microrganismo peculiar: Bacillus subtilis. Nossos resultados demonstram que B. subtilis pode ser uma fonte eficiente de proteases, lipases e celulases, provenientes de comunidades microbianas halofílicas localizadas em áreas pouco exploradas.

Endophytic Bacillus subtilis P10 from Prunus cerasifera as a biocontrol agent against tomato Verticillium wilt

Abstract Endophytic bacteria serve key roles in the maintenance of plant health and growth. Few studies to date, however, have explored the antagonistic and plant growth-promoting (PGP) properties of Prunus cerasifera endophytes. To that end, we isolated endophytic bacteria from P. cerasifera tissue samples and used a dual culture plate assay to screen these microbes for antagonistic activity against Verticillium dahliae, Botryosphaeria dothidea, Fusarium oxysporum, F. graminearum, and F. moniliforme. Of the 36 strains of isolated bacteria, four (strains P1, P10, P16, and P20) exhibited antagonistic effects against all five model pathogens, and the P10 strain exhibited the strongest antagonistic to five pathogens. This P10 strain was then characterized in-depth via phenotypic assessments, physiological analyses, and 16s rDNA sequencing, revealing it to be a strain of Bacillus subtilis. Application of a P10 cell suspension (1×108 CFU/mL) significantly enhanced the seed germination and seedling growth of tomato in a greenhouse setting. This P10 strain further significantly suppressed tomato Verticillium wilt with much lower disease incidence and disease index scores being observed following P10 treatment relative to untreated plants in pot-based experiments. Tomato plants that had been treated with strain P10 also enhanced defense-related enzymes, peroxidase, superoxide dismutase, and catalase activity upon V. dahliae challenge relative to plants that had not been treated with this endophytic bacterium. The results revealed that the P10 bacterial strain has potential value as a biocontrol agent for use in the prevention of tomato Verticillium wilt.

Resumo As bactérias endofíticas desempenham papel fundamental na manutenção da saúde e do crescimento das plantas. Poucos estudos até o momento, no entanto, exploraram as propriedades antagônicas e promotoras de crescimento de plantas (PGP) de endófitos de Prunus cerasifera. Para esse fim, isolamos bactérias endofíticas de amostras de tecido de P. cerasifera e usamos um ensaio de placa de cultura dupla para rastrear esses micróbios quanto à atividade antagonista contra Verticillium dahliae, Botryosphaeria dothidea, Fusarium oxysporum, F. graminearum e F. moniliforme. Das 36 cepas de bactérias isoladas, quatro (cepas P1, P10, P16 e P20) exibiram efeitos antagônicos contra todos os cinco patógenos modelo, e a cepa P10 exibiu o antagonista mais forte para cinco patógenos. Essa cepa P10 foi então caracterizada em profundidade por meio de avaliações fenotípicas, análises fisiológicas e sequenciamento de rDNA 16s, revelando ser uma cepa de Bacillus subtilis. A aplicação de uma suspensão de células P10 (1 × 108 UFC / mL) aumentou significativamente a germinação das sementes e o crescimento das mudas de tomate em casa de vegetação. Essa cepa P10 suprimiu ainda mais a murcha de Verticillium do tomate com incidência de doença muito menor e pontuações de índice de doença sendo observadas após o tratamento com P10 em relação a plantas não tratadas em experimentos baseados em vasos. As plantas de tomate que foram tratadas com a cepa P10 também aumentaram as enzimas relacionadas à defesa, peroxidase, superóxido dismutase e atividade da catalase após o desafio de V. dahliae em relação às plantas que não foram tratadas com essa bactéria endofítica. Os resultados revelaram que a cepa bacteriana P10 tem valor potencial como agente de biocontrole para uso na prevenção da murcha de Verticillium em tomate.

Identification by molecular techniques of halophilic bacteria producing important enzymes from pristine area in Campeche, Mexico

Abstract Isla Arena is located in the coordinate 20° 70´ N - 90° 45´ W, from Campeche, Mexico. In these estuaries, the ocean mixes with fresh water, and ecosystems are concentrated where petenes and pink flamingos proliferate. Crustaceans and mollusks abound in the sea. Despite its enormous marine wealth, there are no studies carried out on which halophilic microorganisms are present in these waters. In this work, the diversity and structure of the microbial community was investigated through a metagenomics approach and corroborated for sequencing of 16S rRNA genes. It was found that the phylum Fimicutes predominates with more than 50%, in almost the same proportion of the class Bacilli and with almost 41% of relative abundance of the order Bacillales. The sequencing results showed that one of the samples presented a high percentage of similarity (99.75%) using the Nucleotide BLAST program with a peculiar microorganism: Bacillus subtilis. This microorganism is one of the best characterized bacteria among the gram-positive ones. Our results demonstrate that B. subtilis can be an efficient source of proteases, lipases and cellulases, from halophilic microbial communities located in poorly explored areas.

Resumo Isla Arena está localizada na coordenada 20°70N - 90°45W, de Campeche, México. Nesses estuários, o oceano se mistura com a água doce e os ecossistemas se concentram onde proliferam petenos e flamingos rosa. Crustáceos e moluscos abundam no mar. Apesar de sua enorme riqueza marinha, não há estudos realizados sobre a presença de microrganismos halofílicos nessas águas. Neste trabalho, a diversidade e estrutura da comunidade microbiana foram investigadas através de uma abordagem metagenômica e corroboradas para o sequenciamento de genes 16S rRNA. Verificou-se que o filo Fimicutes predomina com mais de 50%, quase na mesma proporção da classe Bacilli e com quase 41% de abundância relativa da ordem Bacillales. Os resultados do sequenciamento mostraram que uma das amostras apresentou alto percentual de similaridade (99,75%) pelo programa Nucleotide BLAST com um microrganismo peculiar: Bacillus subtilis. Nossos resultados demonstram que B. subtilis pode ser uma fonte eficiente de proteases, lipases e celulases, provenientes de comunidades microbianas halofílicas localizadas em áreas pouco exploradas.

Identification by molecular techniques of halophilic bacteria producing important enzymes from pristine area in Campeche, Mexico / Identificação por técnicas moleculares de bactérias halofílicas que produzem enzimas importantes em área intocada de Campeche, México

Abstract Isla Arena is located in the coordinate 20° 70´ N - 90° 45´ W, from Campeche, Mexico. In these estuaries, the ocean mixes with fresh water, and ecosystems are concentrated where petenes and pink flamingos proliferate. Crustaceans and mollusks abound in the sea. Despite its enormous marine wealth, there are no studies carried out on which halophilic microorganisms are present in these waters. In this work, the diversity and structure of the microbial community was investigated through a metagenomics approach and corroborated for sequencing of 16S rRNA genes. It was found that the phylum Fimicutes predominates with more than 50%, in almost the same proportion of the class Bacilli and with almost 41% of relative abundance of the order Bacillales. The sequencing results showed that one of the samples presented a high percentage of similarity (99.75%) using the Nucleotide BLAST program with a peculiar microorganism: Bacillus subtilis. This microorganism is one of the best characterized bacteria among the gram-positive ones. Our results demonstrate that B. subtilis can be an efficient source of proteases, lipases and cellulases, from halophilic microbial communities located in poorly explored areas.

Resumo Isla Arena está localizada na coordenada 20°70'N - 90°45'W, de Campeche, México. Nesses estuários, o oceano se mistura com a água doce e os ecossistemas se concentram onde proliferam petenos e flamingos rosa. Crustáceos e moluscos abundam no mar. Apesar de sua enorme riqueza marinha, não há estudos realizados sobre a presença de microrganismos halofílicos nessas águas. Neste trabalho, a diversidade e estrutura da comunidade microbiana foram investigadas através de uma abordagem metagenômica e corroboradas para o sequenciamento de genes 16S rRNA. Verificou-se que o filo Fimicutes predomina com mais de 50%, quase na mesma proporção da classe Bacilli e com quase 41% de abundância relativa da ordem Bacillales. Os resultados do sequenciamento mostraram que uma das amostras apresentou alto percentual de similaridade (99,75%) pelo programa Nucleotide BLAST com um microrganismo peculiar: Bacillus subtilis. Nossos resultados demonstram que B. subtilis pode ser uma fonte eficiente de proteases, lipases e celulases, provenientes de comunidades microbianas halofílicas localizadas em áreas pouco exploradas.

Endophytic Bacillus subtilis P10 from Prunus cerasifera as a biocontrol agent against tomato Verticillium wilt / Bacillus subtilis P10 endofítico de Prunus cerasifera como agente de biocontrole contra murcha de Verticillium de tomate

Abstract Endophytic bacteria serve key roles in the maintenance of plant health and growth. Few studies to date, however, have explored the antagonistic and plant growth-promoting (PGP) properties of Prunus cerasifera endophytes. To that end, we isolated endophytic bacteria from P. cerasifera tissue samples and used a dual culture plate assay to screen these microbes for antagonistic activity against Verticillium dahliae, Botryosphaeria dothidea, Fusarium oxysporum, F. graminearum, and F. moniliforme. Of the 36 strains of isolated bacteria, four (strains P1, P10, P16, and P20) exhibited antagonistic effects against all five model pathogens, and the P10 strain exhibited the strongest antagonistic to five pathogens. This P10 strain was then characterized in-depth via phenotypic assessments, physiological analyses, and 16s rDNA sequencing, revealing it to be a strain of Bacillus subtilis. Application of a P10 cell suspension (1×108 CFU/mL) significantly enhanced the seed germination and seedling growth of tomato in a greenhouse setting. This P10 strain further significantly suppressed tomato Verticillium wilt with much lower disease incidence and disease index scores being observed following P10 treatment relative to untreated plants in pot-based experiments. Tomato plants that had been treated with strain P10 also enhanced defense-related enzymes, peroxidase, superoxide dismutase, and catalase activity upon V. dahliae challenge relative to plants that had not been treated with this endophytic bacterium. The results revealed that the P10 bacterial strain has potential value as a biocontrol agent for use in the prevention of tomato Verticillium wilt.

Resumo As bactérias endofíticas desempenham papel fundamental na manutenção da saúde e do crescimento das plantas. Poucos estudos até o momento, no entanto, exploraram as propriedades antagônicas e promotoras de crescimento de plantas (PGP) de endófitos de Prunus cerasifera. Para esse fim, isolamos bactérias endofíticas de amostras de tecido de P. cerasifera e usamos um ensaio de placa de cultura dupla para rastrear esses micróbios quanto à atividade antagonista contra Verticillium dahliae, Botryosphaeria dothidea, Fusarium oxysporum, F. graminearum e F. moniliforme. Das 36 cepas de bactérias isoladas, quatro (cepas P1, P10, P16 e P20) exibiram efeitos antagônicos contra todos os cinco patógenos modelo, e a cepa P10 exibiu o antagonista mais forte para cinco patógenos. Essa cepa P10 foi então caracterizada em profundidade por meio de avaliações fenotípicas, análises fisiológicas e sequenciamento de rDNA 16s, revelando ser uma cepa de Bacillus subtilis. A aplicação de uma suspensão de células P10 (1 × 108 UFC / mL) aumentou significativamente a germinação das sementes e o crescimento das mudas de tomate em casa de vegetação. Essa cepa P10 suprimiu ainda mais a murcha de Verticillium do tomate com incidência de doença muito menor e pontuações de índice de doença sendo observadas após o tratamento com P10 em relação a plantas não tratadas em experimentos baseados em vasos. As plantas de tomate que foram tratadas com a cepa P10 também aumentaram as enzimas relacionadas à defesa, peroxidase, superóxido dismutase e atividade da catalase após o desafio de V. dahliae em relação às plantas que não foram tratadas com essa bactéria endofítica. Os resultados revelaram que a cepa bacteriana P10 tem valor potencial como agente de biocontrole para uso na prevenção da murcha de Verticillium em tomate.

Growth-promoting microorganisms in the development of orchid seedlings of Phalaenopsis, Cymbidium, and Dendrobium genera

The demand for sustainable agricultural production systems is increasing, and using growth-promoting microorganisms in plants has stood out because it decreases or even replaces chemical fertilizer use, reducing production costs. This study aimed to evaluate the response of some microorganisms applied to the seedlings of primary orchids cultivated in Brazil (Phalaenopsis sp. 'Taisuco Swan', Cymbidium atropurpureum, and Dendrobium secundum). The experimental design was completely randomized. There were seven treatments (absence of microorganisms ­ control, Trichoderma sp. in sodium alginate, Trichoderma sp. in clay, Trichoderma sp. in sodium alginate and clay, Trichoderma sp. in a liquid medium, Azospirillum brasilense + Bacillus subtilis in a liquid medium, and Bacillus pumilus in a liquid medium), four replications, and three plants per plot. The seedlings were grown in a greenhouse and evaluated 190 days after microorganism inoculation. The evaluation of morpho-physiological characteristics differed according to the particularities of each genus. The Bacillus pumilus and Azospirillum brasilense + Bacillus subtilis rhizobacteria in a liquid medium for Phalaenopsis sp. 'Taisuco Swan' and the Trichoderma sp. fungus in a liquid medium for Cymbidium atropurpureum increased seedling growth and development. Azospirillum brasilense + Bacillus subtilis in a liquid medium for the Dendrobium secundum orchid promoted more root biomass. Using beneficial microorganisms in orchid cultivation is promising, and seedling growth and development depend on their inoculation and the morpho-physiological characteristics of each plant.

Bioremediation Tofu Liquid Waste Based on Chemical Oxygen Demand (COD) Parameters

Background and Objective: The majority of liquid waste generated by the tofu industry in Semarang exceeds the recommended limit. This condition disturbs the ecology of the surrounding environment. so a solution is needed to manage it. The use of bacteria in bioremediation technology has been proven to reduce pollutants produced by the tofu industry. The purpose of this study was to identify specific bacteria and test their ability to bioremediate pollutants from tofu wastewater using the Chemical Oxygen Demand (COD) parameter. Methodology: Methodology: The study was exploratory and experimental in nature, with the variable ability of degradative bacteria degrading tofu industrial wastewater based on COD parameters. Bacterial identification was performed using the 16S-rRNA Gene Analysis method, genomic DNA extraction with the Quick-DNA Bacterial Miniprep Kit, PCR amplification, and bi-directional sequencing. Results: The results of the identification of the selected bacteria show that they are molecules of the type nucleic acid with a query length of 1394. with a lineage report and a 93% phylogenetic tree as Bacillus subtilis, Bioremediation test shows that tofu waste can be degraded. The degradation percentage is 17%. Conclusion: Bacillus subtillis, a bacterium that degrades tofu industrial wastewater, was found as an indigenous bacteria.

Antifungal potential of biosurfactants produced by strains of Bacillus spp. (Bacillales: Bacillaceae) selected by molecular screening / Potencial antifúngico de biosulfactantes producidos por cepas de Bacillus spp. (Bacillales: Bacillaceae) seleccionadas por tamizaje molecular

Introduction: Bacillus species are used as biological controllers for phytopathogenic fungi, and the mechanisms to produce controllers include biosynthesis of lipopeptide biosurfactants with antifungal activity. Objective: To evaluate the antifungal potential of the biosurfactants produced by Bacillus strains, selected by molecular screening, on Fusarium oxysporum. Methods: We selected four molecular markers, related to the biosynthesis of surfactin, fengicin, and lichenysin (srfA, spf, fenB, LichAA) in nine Bacillus strains. We used two mineral media with several culture conditions, for biosurfactant production, and a well diffusion test for antifungal potential. Results: Only the biosurfactant produced by UFAB25 inhibits the mycelial growth of F. oxysporum (44 % ± 13): this biosurfactant was positive for srfA, spf, and fenB genes involved in the synthesis of surfactin and fengicine. Antifungal activity depends on culture conditions and the strain. Conclusions: Genetic markers are useful to detect strains with antifungal potential, facilitating the selection of bio-controllers. The biosurfactant profile is influenced by the strain and by culture conditions.

Introducción: Especies de Bacillus han sido empleadas como controladores biológicos contra hongos fitopatógenos. Entre los mecanismos utilizados se destaca la biosíntesis de biosurfactantes lipopeptídicos con actividad antifúngica. Objetivo: Evaluar el potencial antifúngico de los biosurfactantes producidos por cepas Bacillus nativas, previamente seleccionadas mediante tamizaje molecular, sobre Fusarium oxysporum. Métodos: Se utilizaron cuatro marcadores moleculares, relacionados con la biosíntesis de surfactina, fengicina y liquenisina (srfA, spf, fenB, LichAA) sobre nueve cepas de Bacillus. Se utilizaron dos medios minerales con diferentes condiciones de cultivo para la producción del biosurfactante. Se evaluó el potencial antifúngico de los biosurfactantes mediante la prueba de difusión en pozos. Resultados: Se determinó que solo el biosurfactante producido por UFAB25 actúa como inhibidor del crecimiento micelial de Fusarium oxysporum (43.6 % ± 13), esta cepa es positiva para los genes srfA, spf y fenB, involucrados en la síntesis de surfactina y fengicina. La actividad antifúngica depende de las condiciones de cultivo y la cepa. Conclusiones: Los marcadores genéticos ayudan a detectar cepas con potencial antifúngico, facilitando la selección de biocontroladores. El perfil del biosurfactante está influenciado no solo por la cepa, sino también por las condiciones del cultivo.

Bacillus subtilis mejora el desarrollo de órganos digestivos, la morfología del intestino y el rendimiento productivo en pollos de engorde

La regulación en el uso de antibióticos promotores de crecimiento en la alimentación animal requiere de la búsqueda de alternativas nutricionales seguras, que mejoren la salud intestinal y el rendimiento productivo en los animales, mientras protegen la salud del consumidor. La investigación tuvo como objetivo evaluar el efecto de Bacillus subtilis sobre el desarrollo de órganos digestivos, la morfología del intestino y el rendimiento productivo, en pollos de engorde.192 pollos Cobb 500 de un día de edad fueron asignados aleatoriamente a una de tres dietas: basal libre de antibióticos (D1), basal adicionada con 10 ppm de avilamicina (D2) o basal adicionada con 50 ppm de esporas de B. subtilis (D3), durante 42 días. Los días 21 y 42 de edad, se determinó la conversión alimenticia (CA), la ganancia acumulada de peso (GAP) y se evaluó el desarrollo de los principales órganos digestivos y la histomorfología de cada segmento del intestino delgado, mediante el sacrificio de 48 aves (8 aves/dieta/día). El uso de B. subtilis incrementó significativamente la CA y la GAP. Por otra parte, B. subtilis aumentó la alométria del intestino en comparación con el uso de antibióticos, aunque no se evidenciaron diferencias significativas para el peso de órganos digestivos, entre ambos tratamientos; B. subtilis mejoró la altura de las vellosidades y disminuyó la profundidad de las criptas, significativamente. B. subtilis favorece el desempeño productivo en pollos de engorde, mejora el desarrollo de órganos digestivos y la histomorfología del intestino delgado.

Regulation in the use of growth promoters antibiotics in animal feed has led to the search for safe nutritional alternatives that improve intestinal health and growth performance in animal, while protecting consumer health. The aim was to assess the effect of Bacillus subtilis on the digestive organs development, intestinal morphology and growth performance in broilers. A total of 192 one-day old Cobb 500 chicks, were randomized and assigned to one of three diets: basal diet free of antibiotics (D1) or basal diet added with 10ppm of avilamicyn (D2) or basal diet added with 50ppm of B. subtilis's spores (D3) for 42 days. On days 21 and 42 the feed conversion ratio (FCR) and average body weight (ABW) were determined; also, eight birds per treatment were euthanized to assess the development of digestive organs and the histomorphology in the different sections of the small intestine. The use of B. subtilis significantly increased FCR and ABW. On the other hand, B. subtilis significantly small intestine length compared to antibiotic growth promoter, but there was no differences in the weight of the digestive organs between B. subtilis and antibiotic diet, in addition B. subtilis improve villus height and decreased crypt depth significantly. In conclusion, the use of B. subtilis improve growth performance, digestive organs development and small intestine histomorphology in broilers.

GC-MS Analysis and Antimicrobial Activity of the Extract and Fractions of Bacillus subtilis subsp. subtilis 168 Isolated from a River Bank in Nigeria

The menace of drug resistant pathogens is increasing and their level of evading conventional antimicrobials is rising. It is therefore important to discover new antimicrobials to counter the current challenges. Our preliminary investigation identified Bacillus subtilis subsp. subtilis 168 isolated from soil sample sourced from a river bank in Abuja, Nigeria, as the most potent antibiotic-producing bacteria among the other identified producers. The current study screened for the antimicrobial activity of the extract and fractions of the isolate by broth microdilution method. The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and the ratio of the MBC/MIC were determined. All the tested pathogens were susceptible to the ethyl-acetate extract (MIC between 28.70 mg/ml and 57.40 mg/ml). The extract displayed bactericidal activity against all tested pathogens (MBC/MIC between 1.00 and 2.00) while Proteus mirabilis was least susceptible. The extract was purified by vacuum liquid chromatography and the fractions challenged with pathogenic strains. The fraction E was the most potent (MIC between 0.09 mg/ml and 0.75 mg/ml) and also bactericidal against all the test microbes (MBC/MIC between 2.00 and 2.11). GC-MS analysis of the purified sub fraction obtained from fraction E identified 13 compounds with different Retention time and peak areas. Among these were three major compounds which include: (i) bis(2-ethylhexyl) phthalate (ii) 1,4-epoxynaphthalene-1(2H)-methanol, 4,5,7-tris(1,1-dimethylethyl)-3,4-dihydro- (iii) D:B-Friedo-B':A'-neogammacer-5-en-3-ol, (3.beta.)-. Our findings suggest that Bacillus subtilis subsp. subtilis 168 isolated locally could serve as a valuable source of lead compounds for pharmaceutical and biotechnological purposes.

Field evaluation of biosurfactants, surfactin and di-rhamnolipid produced by Bacillus subtilis subsp. subtilis (VCRC B471) and Pseudomonas fluorescens (VCRC B426) against immature stages of the urban malaria vector Anopheles stephensi

Background & objectives: Bacillus subtilis subsp. subtilis (VCRC B471) and Pseudomonas fluorescens (B426) produce mosquitocidal biosurfactant, surfactin and di-rhamnolipid. The objective of the study was to carry out a small-scale field evaluation of the two biosurfactants to determine the efficacy, application dosage, residual activity and frequency of application against Anopheles stephensi immatures in selected sites in Goa, India. Methods: Surfactin (VCRC B471) and di-rhamnolipid (VCRC B426) were formulated as aqueous suspensions (5% AS), and were applied at the dosages of 34, 51 and 68 mL/m2 and 27, 41 and 54 mL/m2 respectively. Two experiments were carried out with the two formulations. Results: Surfactin (VCRC B471) formulation was effective at all the dosages and there was sustained reduction (>80%) in immature density in the treated sites up to 18 days in experiment 1 and up to 15 days in experiment 2. No pupae were found in the treated sites throughout the study. Di-rhamnolipid (VCRC B426) formulation was also found to reduce the immature density in the treated sites up to 14 days in experiment 1 and up to 15 days in experiment 2. Interpretation & conclusion: For VCRC B471, the optimum application dosage determined was 51 mL/m2 and for VCRC B426, 27mL/m2 . The formulations are to be applied fortnightly for effective control of Anopheles. The application dosage determined in the present study can be used for large scale field evaluation to assess their suitability for use in public health programmes for the control of Anopheles mosquitoes vectoring malaria

Effect of homologous overexpression of nucleoside diphosphate kinase on the yield and molecular weight of hyaluronic acid / 中国药科大学学报

@#To investigate the effect of nucleoside diphosphate kinase (NDK) on the synthesis of hyaluronic acid, nucleoside diphosphate kinase gene (ndk) was overexpressed along with the hyaluronic acid-producing genes in recombinant B.subtilis. Two engineered strains named Hp8tg and Pn8tg were constructed.Uniform hyaluronic acid (HA) could be obtained from both engineered strains.HA produced by both recombinant strains was confirmed by monosaccharide composition analysis, Fourier transform infrared spectometry and nuclear magnetic resonance spectroscopy.Inducing conditions of HA fermentation were optimized by response surface methodology.Overexpression of ndk could increase the production and molecular weight of HA by 1.3-fold and 1.1-fold, respectively. This study revealed for the first time that overexpression of ndk could relieve the inhibition effect of uridine diphosphate (UDP) on Class II HA synthase and increase the production and molecular weight of HA, which proves to be an efficient strategy for the production of HA, and the preparation of other polysaccharides.

Advances in several important antimicrobial lipopeptids from Bacillus spp / 生物工程学报

Bacillus spp. are probiotics and can secrete a variety of natural antimicrobiol active substances, of which lipopeptides are an important class. Up to now, about 90 lipopeptides have been identified, and most of them are cyclic lipopeptides. surfactin, iturin, fengycin, bacillomycin and polymyxins are widely studied, and the first three have huge potential for application due to their properties of surfactants and anti-fungal, anti-bacterial, anti-viral, anti-tumor and anti-inflammatory functions. In this paper, the research progress in the structure, function, synthesis regulation, separation, purification and production of surfactin, iturin and fengycin was reviewed. Synthetic biology is a vital means to increase the yield of lipopeptides, and in the future, lipopeptides can be used in crop cultivation, animal farming, food, medicine and petroleum industries as well as environmental protection. Future research should be strengthened on the discovery of new lipopeptides, synthesis of high-activity lipopeptides, economical production of lipopeptides on a large scale and their safety evaluation.