Identification of antarctic soil bacteria exhibiting antiproliferative activity against a colon cancer cell line / Identificación de bacterias de suelo antártico que presentan actividad antiproliferativa sobre una línea celular de cáncer de colon

SUMMARY: Cancer is the second leading cause of death in the world and colorectal cancer is the only cancer that has shown a sustained increase in mortality in the last decade. In the search for new chemotherapeutic agents against cancer, extremophilic microorganisms have shown to be a potential source to obtain molecules of natural origin and with selective cytotoxic action towards cancer cells. In this work we analyzed the ability of a collection of Antarctic soil bacteria, isolated on Collins Glacier from the rhizosphere of Deschampsia antarctica Desv plant, to secrete molecules capable of inhibiting cell proliferation of a colorectal cancer tumor line. Our results demonstrated that culture supernatants from the Antarctic bacteria K2I17 and MI12 decreased the viability of LoVo cells, a colorectal adenocarcinoma cell line. Phenotypic and genotypic characterization of the Antarctic bacteria showed that they were taxonomically related and nucleotide identity analysis based on the 16S rRNA gene sequence identified the bacterium K2I17 as a species belonging to the genus Bacillus.

El cáncer es la segunda causa de muerte en el mundo y el cáncer colorrectal es el único que presenta un aumento sostenido de la mortalidad en la última década. En la búsqueda de nuevos agentes quimioterapeúticos contra el cáncer, se ha propuesto a los microorganismos extremófilos como una fuente potencial para obtener moléculas de origen natural y con acción citotóxica selectiva hacia las células cancerígenas. En este trabajo analizamos la capacidad de una colección de bacterias de suelo antártico, aisladas en el glaciar Collins desde rizosfera de la planta de Deschampsia antarctica Desv, de secretar moléculas capaces de inhibir la proliferación celular de una línea tumoral de cáncer colorrectal. Nuestros resultados demostraron que los sobrenadantes de cultivo de las bacterias antárticas K2I17 y MI12 disminuyeron la viabilidad de la línea celular de adenocarcinoma colorrectal LoVo, en un ensayo de reducción metabólica de MTT. La caracterización fenotípica y genotípica de las bacterias antárticas, demostró que estaban relacionadas taxonómicamente y el análisis de la identidad nucleotídica en base a la secuencia del gen ARNr 16S identificó a la bacteria K2I17 como una especie perteneciente al género Bacillus.

Efficient production of L-asparaginase in Bacillus licheniformis by optimizing expression elements and host / 生物工程学报

L-asparaginase (L-ASN) is widely applied in the treatment of malignant tumor and low-acrylamide food production, however, the low expression level hampers its application. Heterologous expression is an effective strategy to increase the expression level of target enzymes, and Bacillus is generally used as the host for efficient production of enzymes. In this study, the expression level of L-asparaginase in Bacillus was enhanced through optimization of expression element and host. Firstly, five signal peptides (SPSacC, SPAmyL, SPAprE, SPYwbN and SPWapA) were screened, among which SPSacC showed the best performance, reaching an activity of 157.61 U/mL. Subsequently, four strong promoters (P43, PykzA-P43, PUbay and PbacA) from Bacillus were screened, and tandem promoter PykzA-P43 showed the highest yield of L-asparaginase, which was 52.94% higher than that of control strain. Finally, three Bacillus expression hosts (B. licheniformis Δ0F3 and BL10, B. subtilis WB800) were investigated, and the maximum L-asparaginase activity, 438.3 U/mL, was reached by B. licheniformis BL10, which was an 81.83% increase compared with that of the control. This is also the highest level of L-asparaginase in shake flask reported to date. Taken together, this study constructed a B. licheniformis strain BL10/PykzA-P43-SPSacC-ansZ capable of efficiently producing L-asparaginase, which laid the foundation for industrial production of L-asparaginase.

Antagonistic activity and application of Bacillus velezensis strain Bv-303 against rice bacterial-blight disease caused by Xanthomonas oryzae pv. oryzae / 生物工程学报

In this study, a new Bacillus velezensis strain Bv-303 was identified and its biocontrol effect against rice bacterial-blight (BB) disease caused by Xanthomonas oryzae pv. oryzae (Xoo) was investigated. Cell-free supernatant (CFS) of strain Bv-303 under different growth conditions were prepared to test the antagonistic activity and stability against Xoo by the Oxford-cup method in vitro. The antibacterial effect of strain Bv-303 to BB disease in rice were further analyzed in vivo by spraying the cell-culture broth (CCB), CFS and cell-suspension water (CSW), respectively, on the rice leaves inoculated with Xoo. Additionally, rice seeds germination rate and seedling growth under the strain Bv-303 CCB treatment were tested. The results showed that the strain Bv-303 CFS significantly inhibited Xoo growth by 85.7%‒88.0% in vitro, which was also stable under extreme environment conditions such as heat, acid, alkali and ultraviolet light. As tested in vivo, spraying the CCB, CFS or CSW of strain Bv-303 on the Xoo-infected leaves enhanced rice plant resistance to BB disease, with CCB showing the highest increase (62.7%) in disease-resistance. Notably, CCB does not have negative effects on rice seed germination and seedling growth. Therefore, strain Bv-303 has great potential for biocontrol of the rice BB disease.

Thermo-resistant enzyme-producing microorganisms isolated from composting / Microrganismos produtores de enzimas termo-resistentes isolados de compostagem

Organo-mineral fertilizers supplemented with biological additives are an alternative to chemical fertilizers. In this study, thermoresistant microorganisms from composting mass were isolated by two-step procedures. First, samples taken at different time points and temperatures (33 days at 52 ºC, 60 days at 63 ºC, and over 365 days at 26 ºC) were pre-incubated at 80 oC for 30 minutes. Second, the microbial selection by in vitro culture-based methods and heat shock at 60 oC and 100 oC for 2h and 4h. Forty-one isolates were able to grow at 60 °C for 4h; twenty-seven at 100 °C for 2h, and two at 100 °C for 4h. The molecular identification by partial sequencing of the 16S ribosomal gene using universal primers revealed that thirty-five isolates were from eight Bacillus species, one Brevibacillus borstelensis, three Streptomyces thermogriseus, and two fungi (Thermomyces lanuginosus and T. dupontii). Data from amylase, phytase, and cellulase activity assays and the enzymatic index (EI) showed that 38 of 41 thermo-resistant isolates produce at least one enzyme. For amylase activity, the highest EI value was observed in Bacillus licheniformis (isolate 21C2, EI= 4.11), followed by Brevibacillus borstelensis (isolate 6C2, EI= 3.66), Bacillus cereus (isolate 18C2, EI= 3.52), and Bacillus paralicheniformis (isolate 20C2, EI= 3.34). For phytase, the highest EI values were observed for Bacillus cereus (isolate 18C2, EI= 2.30) and Bacillus licheniformis (isolate 3C1, EI= 2.15). Concerning cellulose production, B. altitudinis (isolate 6C1) was the most efficient (EI= 6.40), followed by three Bacillus subtilis (isolates 9C1, 16C2, and 19C2) with EI values of 5.66, 5.84, and 5.88, respectively, and one B. pumilus (isolate 27C2, EI= 5.78). The selected microorganisms are potentially useful as a biological additive in organo-mineral fertilizers and other biotechnological processes.

Os fertilizantes organo-minerais suplementados com aditivos biológicos são uma alternativa aos adubos químicos. Neste estudo, microrganismos termoresistentes foram isolados de compostagem por procedimentos de duas etapas. Inicialmente, as amostras tomadas em diferentes períodos e temperaturas (33 dias a 52 ºC, 60 dias a 63 ºC e mais de 365 dias a 26 ºC) foram pré-incubadas a 80 oC por 30 minutos. Posteriormente, a seleção microbiana foi conduzida por métodos baseados em cultura in vitro e choque térmico a 60 oC e 100 oC por 2h e 4h. Quarenta e um isolados foram capazes de crescer a 60 °C por 4h; vinte e sete a 100 °C por 2h e dois a 100 °C por 4h. A identificação molecular por sequenciamento parcial do gene ribossomal 16S usando primers universais revelou que trinta e cinco isolados eram de oito espécies de Bacillus, um Brevibacillus borstelensis, três Streptomyces thermogriseus e dois fungos (Thermomyces lanuginosus e T. dupontii). Os dados dos ensaios de atividade de amilase, fitase e celulase e o índice enzimático (IE) mostraram que 38 dos 41 isolados termorresistentes produziram pelo menos uma enzima. Para a atividade da amilase, o maior valor de IE foi observado em Bacillus licheniformis (isolado 21C2, IE = 4,11), seguido por Brevibacillus borstelensis (isolado 6C2, IE = 3,66), Bacillus cereus (isolado 18C2, IE = 3,52) e Bacillus paralicheniformis (isolado 20C2, IE = 3,34). Para a fitase, os maiores valores de IE foram observados para B. cereus (isolado 18C2, IE = 2,30) e B. licheniformis (isolado 3C1, IE = 2,15). Em relação à produção de celulose, B. altitudinis (isolado 6C1) foi o mais eficiente (IE = 6,40), seguido por três Bacillus subtilis (isolados 9C1, 16C2 e 19C2) com valores de IE de 5,66, 5,84 e 5,88, respectivamente, e um B. pumilus (isolado 27C2, IE = 5,78). Pode-se inferir que os microrganismos selecionados são potencialmente úteis como aditivos biológicos em fertilizantes organo-minerais e outros processos biotecnológicos.

Isolation and screening of chromium resistant bacteria from industrial waste for bioremediation purposes / Isolamento e triagem de bactérias resistentes ao cromo de resíduos industriais para fins de biorremediação

Abstract Chromium (VI) a highly toxic metal, a major constituent of industrial waste. It is continuously release in soil and water, causes environmental and health related issues, which is increasing public concern in developing countries like Pakistan. The basic aim of this study was isolation and screening of chromium resistant bacteria from industrial waste collected from Korangi and Lyari, Karachi (24˚52ʹ46.0ʺN 66˚59ʹ25.7ʺE and 24˚48ʹ37.5ʺN 67˚06ʹ52.6ʺE). Among total of 53 isolated strains, seven bacterial strains were selected through selective enrichment and identified on the basis of morphological and biochemical characteristics. These strains were designated as S11, S13, S17, S18, S30, S35 and S48, resistance was determined against varying concentrations of chromium (100-1500 mg/l). Two bacterial strains S35 and S48 showed maximum resistance to chromium (1600 mg/l). Bacterial strains S35 and S48 were identified through 16S rRNA sequence and showed 99% similarity to Bacillus paranthracis and Bacillus paramycoides. Furthermore, growth condition including temperature and pH were optimized for both bacterial strains, showed maximum growth at temperature 30ºC and at optimum pH 7.5 and 6.5 respectively. It is concluded that indigenous bacterial strains isolated from metal contaminated industrial effluent use their innate ability to transform toxic heavy metals to less or nontoxic form and can offer an effective tool for monitoring heavy metal contamination in the environment.

Resumo O cromo (VI), metal altamente tóxico, é um dos principais constituintes dos resíduos industriais. É liberado no solo e na água, causa problemas ambientais e de saúde de crescente preocupação pública em países em desenvolvimento como o Paquistão. O objetivo básico deste estudo foi o isolamento e a triagem de bactérias resistentes ao cromo de resíduos industriais coletados em Korangi e Lyari, Karachi (24˚52'46,0"N 66˚59'25,7"E e 24˚48'37,5"N 67˚06'52,6"E). Do total de 53 cepas isoladas, sete cepas bacterianas foram selecionadas por enriquecimento seletivo e identificadas com base em características morfológicas e bioquímicas. Essas cepas foram designadas como S11, S13, S17, S18, S30, S35 e S48, apresentaram alta resistência aos metais contra concentrações variáveis (100-1500 mg / l) de cromo. Já as cepas S35 e S48 foram identificadas por meio da sequência 16S rRNA e apresentaram 99% de similaridade com Bacillus paranthracis e Bacillus paramycoides. Além disso, as condições de crescimento incluindo temperatura e pH foram otimizadas e ambas as cepas bacterianas apresentaram crescimento máximo na temperatura de 30 ºC, enquanto seu pH ótimo foi observado em 7,5 e 6,5, respectivamente. Conclui-se que o potencial de resistência dessas bactérias resistentes ao cromo pode ser efetivamente utilizado na remoção de cromo de efluentes industriais contaminados. Técnicas de base biológica usando bactérias ajudarão a fornecer métodos mais baratos e ecológicos de remoção, recuperação e desintoxicação de cromo.

Modulating the production of xylanase by Bacillus pumilus BS131 through optimization using waste fiber sludge / Modulando a produção de xilanase por Bacillus pumilus BS131 por meio da otimização usando lodo de fibra residual

Abstract In recent days, cheapest alternative carbon source for fermentation purpose is desirable to minimize production cost. Xylanases have become attractive enzymes as their potential in bio-bleaching of pulp and paper industry. The objective of the present study was to identify the potential ability on the xylanase production by locally isolated Bacillus pumilus BS131 by using waste fiber sludge and wheat bran media under submerged fermentation. Culture growth conditions were optimized to obtain significant amount of xylanase. Maximum xylanase production was recorded after 72 hours of incubation at 30 °C and 7 pH with 4.0% substrate concentration. In the nutshell, the production of xylanase using inexpensive waste fiber sludge and wheat-bran as an alternative in place of expensive xylan substrate was more cost effective and environment friendly.

Resumo Nos últimos dias, a fonte alternativa de carbono mais barata para fins de fermentação é desejável para minimizar o custo de produção. As xilanases têm se tornado enzimas atraentes como seu potencial no biobranqueamento da indústria de papel e celulose. O objetivo do presente estudo foi identificar a capacidade potencial na produção de xilanase por Bacillus pumilus BS131 isolado localmente usando lodo de fibra residual e farelo de trigo em meio de fermentação submersa. As condições de crescimento da cultura foram otimizadas para obter uma quantidade significativa de xilanase. A produção máxima de xilanase foi registrada após 72 horas de incubação a 30 °C e pH 7 com concentração de substrato de 4,0%. Resumindo, a produção de xilanase usando lodo de fibra residual de baixo custo e farelo de trigo como uma alternativa no lugar do substrato de xilano caro foi mais econômica e ecológica.

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.

The importance of environments and surfaces hygienization in pandemic times (Covid-19): a microbiological analysis in places for physical activity practice

The objective of this study was to analyse microbiological organisms in different locations and regions for physical activity in the city of João Pessoa, Brazil. Samples were collected on various objects used, such as: mattresses, drinking fountains, gloves, cell phones and others. The samples were collected in João Pessoa-PB, following the Standard Operating Procedure-SOP/ Microbiology of a specialized laboratory. The collection took place in the five macro-regions: North, South, East, West and Center. Foreach region samples were collected in one public place (square), a private one (gym) and one school (public or private), totaling fifteen collected sites and 450 samples. The following microorganisms were studied in all analyzed surfaces: Bacillus sp, Escherichia Coli, Klebsiella sppor Enterobacter sppand Coag. Neg. Staphylococcus.All regions had a high contamination level by some microorganism. The highest rates were found in the western, central and northern regions -96, 94 and 93% respectively. The Coag. Neg.Staphylococcus presented the highest and lowest incidence rates in the South and East regions, with 43.33 and 6.67%, respectively, as well as Klebsiella sppor Enterobacter spp, which presented high levels. It is concluded that there is a microorganisms' contamination in the most varied places and regions where physical activity practices are developed, with a predominance of Coag. Neg.Staphylococcusand Klebsiella sppor Enterobacter spp. These results lead to a warning about the hygiene importance in places for physical activity practice, especially in pandemic times (COVID-19), since almost all the evaluated surfaces were contaminated.

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.

Biodegradation of petroleum by bacteria isolated from fishes of Indian Ocean / Biodegradação do petróleo por bactérias isoladas de peixes do oceano Índico

In this study, oil degrading bacteria discovered from fish living near the oil ports at Karachi in Pakistan were characterized. The bacteria isolated from skin, gills, and gut in fish could consume crude oil as a source of carbon and energy. Total 36 isolates were tested using Nutrient Agar (NA) and MSA media with different crude oil concentrations (0.2%, 0.5%, 0.7%, 1%, 2%, and 5%) and 4 out of 36 isolates (two Gram positive and two Gram negative bacteria) were selected for further identification. 16S rRNA gene sequencing revealed that the isolates are related to Bacillus velezensis, Bacillus flexus, Pseudomonas brenneri and Pseudomonas azotoforman. Oil degrading potential of these bacteria was characterized by GC-MS analysis of degradation of oil components in crude oil as well as engine oil. We found that one (2, 6, 10, 14-Tetramethylpentadecane) out of 42 components in the crude oil was fully eliminated and the other oil components were reduced. In addition, 26 out of 42 oil components in the engine oil, were fully eliminated and the rest were amended. Taken together, these studies identify that B. velezensis, B. flexus, P. brenneri and P. azotoforman have high oil degrading potential, which may be useful for degradation of oil pollutants and other commercial applications.

Neste estudo, bactérias degradadoras de óleo descobertas em peixes que vivem perto dos portos de petróleo em Karachi, no Paquistão, foram caracterizadas. As bactérias isoladas da pele, guelras e intestinos dos peixes podem consumir petróleo bruto como fonte de carbono e energia. No total, 36 isolados foram testados usando Agar Nutriente (NA) e meio MSA com diferentes concentrações de óleo bruto (0,2%, 0,5%, 0,7%, 1%, 2% e 5%) e 4 de 36 isolados (dois Gram positivos e duas bactérias Gram negativas) foram selecionadas para posterior identificação. O sequenciamento do gene 16S rRNA revelou que os isolados estão relacionados a Bacillus velezensis, Bacillus flexus, Pseudomonas brenneri e Pseudomonas azotoforman. O potencial de degradação do óleo dessas bactérias foi caracterizado pela análise de GC-MS da degradação dos componentes do óleo no óleo cru, bem como no óleo do motor. Descobrimos que um (2, 6, 10, 14-tetrametilpentadecano) de 42 componentes do óleo cru foi totalmente eliminado e os outros componentes do óleo foram reduzidos. Além disso, 26 dos 42 componentes do óleo do motor foram totalmente eliminados e o restante corrigido. Juntos, esses estudos identificam que B. velezensis, B. flexus, P. brenneri e P. azotoforman têm alto potencial de degradação de óleo, o que pode ser útil para a degradação de poluentes de óleo e outras aplicações comerciais.

Endophytic bacteria Bacillus safensis and Pseudomonas hibiscicola and their ability to increase rice seedling growth

Endophytic bacteria Bacillus safensis RS95 and Pseudomonas hibiscicola RS121 were evaluated for their ability to promote the growth of rice seedlings and produce indole-acetic acid (IAA) and siderophores and to solubilize phosphates. 'Guri' rice seeds were immersed in bacterial endophyte cell suspensions (separated and two-strain mixed), as well as in Escherichia coli DH5α, phosphate-buffered saline (PBS) and water treatments (negative controls). Seeds were sown on agar-water in Petri plates placed vertically at an angle of 65°. The ability of plant growth-promoting endophytic bacteria (PGPEB) to produce IAA and siderophores was determined by Salkowski colorimetric and chrome azurol S (CAS) assays, respectively. Mineral phosphate solubilization activity was calculated by inoculating the endophytes onto medium containing insoluble phosphate. PGPEB showed a positive effect on the growth of rice seedlings, causing a mean growth of shoots and primary-roots of 60 and 67%, respectively. Bacterial strains also showed positive traits for IAA and siderophore production, as well as phosphate-solubilization activity

Biodegradation of petroleum by bacteria isolated from fishes of Indian Ocean / Biodegradação do petróleo por bactérias isoladas de peixes do oceano Índico

In this study, oil degrading bacteria discovered from fish living near the oil ports at Karachi in Pakistan were characterized. The bacteria isolated from skin, gills, and gut in fish could consume crude oil as a source of carbon and energy. Total 36 isolates were tested using Nutrient Agar (NA) and MSA media with different crude oil concentrations (0.2%, 0.5%, 0.7%, 1%, 2%, and 5%) and 4 out of 36 isolates (two Gram positive and two Gram negative bacteria) were selected for further identification. 16S rRNA gene sequencing revealed that the isolates are related to Bacillus velezensis, Bacillus flexus, Pseudomonas brenneri and Pseudomonas azotoforman. Oil degrading potential of these bacteria was characterized by GC-MS analysis of degradation of oil components in crude oil as well as engine oil. We found that one (2, 6, 10, 14-Tetramethylpentadecane) out of 42 components in the crude oil was fully eliminated and the other oil components were reduced. In addition, 26 out of 42 oil components in the engine oil, were fully eliminated and the rest were amended. Taken together, these studies identify that B. velezensis, B. flexus, P. brenneri and P. azotoforman have high oil degrading potential, which may be useful for degradation of oil pollutants and other commercial applications.

Neste estudo, bactérias degradadoras de óleo descobertas em peixes que vivem perto dos portos de petróleo em Karachi, no Paquistão, foram caracterizadas. As bactérias isoladas da pele, guelras e intestinos dos peixes podem consumir petróleo bruto como fonte de carbono e energia. No total, 36 isolados foram testados usando Agar Nutriente (NA) e meio MSA com diferentes concentrações de óleo bruto (0,2%, 0,5%, 0,7%, 1%, 2% e 5%) e 4 de 36 isolados (dois Gram positivos e duas bactérias Gram negativas) foram selecionadas para posterior identificação. O sequenciamento do gene 16S rRNA revelou que os isolados estão relacionados a Bacillus velezensis, Bacillus flexus, Pseudomonas brenneri e Pseudomonas azotoforman. O potencial de degradação do óleo dessas bactérias foi caracterizado pela análise de GC-MS da degradação dos componentes do óleo no óleo cru, bem como no óleo do motor. Descobrimos que um (2, 6, 10, 14-tetrametilpentadecano) de 42 componentes do óleo cru foi totalmente eliminado e os outros componentes do óleo foram reduzidos. Além disso, 26 dos 42 componentes do óleo do motor foram totalmente eliminados e o restante corrigido. Juntos, esses estudos identificam que B. velezensis, B. flexus, P. brenneri e P. azotoforman têm alto potencial de degradação de óleo, o que pode ser útil para a degradação de poluentes de óleo e outras aplicações comerciais.

Calidad microbiológica del aire en seis áreas de la microestación biológica- Zoocriadero de la Universidad Nacional Agraria de la Selva ­ Tingo María ­ Perú / Microbiological quality of air in six areas of the biological micro station - Zoocriadero of the Universidad Nacional Agraria de la Selva - Tingo Maria ­ Peru

El presente trabajo de investigación tiene como objetivo evaluar la calidad microbiológica del aire en seis áreas de la Microestación Biológica-Zoocriadero de la Universidad Nacional Agraria de la Selva-Tingo María-Perú; procediéndose de la siguiente manera: Se seleccionaron seis áreas de muestreo (A1: Boletería, A2: Caseta gallito de las rocas, A3: Zona de las tortugas, A4: Caseta del otorongo, A5: Caseta de los monos y A6: Cocina), de los cuales para la determinación de los parámetros físicos (PAS, temperatura, humedad relativa, precipitación y rosa de viento) se obtuvo información de la estación meteorológica de la UNAS y de lecturas directas en campo, para la determinación de las PAS se realizó por el método pasivo obteniéndose la mayor concentración de PAS en el área 1 "Boletería" (12.61 t/km2/mes, 11.08 t/km2/mes y 8.4 t/km2/mes) y la más baja en el área 5 "caseta de los monos" (2.80 t/km2/mes, 2.80 t/km2/mes y 3.2 t/km2/mes); para el análisis microbiológico se realizó por el método de borboteo en líquidos, identificándose 27 géneros entre bacterias y hongos, siendo las bacterias más frecuentes Bacillus sp., Staphylococcus sp., Enterobacter agglomerans, Klebsiella pneumoniae, Enterobacter hafniae y algunas bacterias patógenas para el hombre como Streptococcus sp., Clostridium perfringens, Bacillus anthracis. Los hongos de mayor frecuencia son Penicillium sp., Aspergillius sp., Geotrichum sp., algunos hongos patógenos identificados: Microsporum sp., Epirophyton sp. Por otro lado, para la percepción del público como trabajadores se aplicó encuestas, donde indicaron que la calidad microbiológica del aire en la Microestación Biológica-Zoocriadero es regular(AU)

The objective of this research paper is to assess the microbiological quality of the air in six areas of the Biologial Micro-station and animal breeding farm of the Peruvian Univeristy "Universidad Nacional Agraria de la Selva in Tingo María, Perú", proceeding as follows: Six sampling areas were selected (A1: Ticket Office, A2: Cock-of-the-Rock hut, A3: Turtle area, A4: Otorongo hut, A5: Monkey hut, A6: Kitchen), of wich for the determination of the phisical parameters (SBP, temperature, relative humidity, precipitation and wind rose) information was obtained from the UNAS meteorological station and direct readings in the field. For the determination of SBP was performed by the passive method obtaining the high concentration of SBP in area 1 "Ticket Office" (12.61 t/km2/month, 11.08 t/km2/month and 8.4 t/km2/month) and the lowest in area 5 "monkey house" (2.80 t/km2/month, 2.80 t/km2/month and 3.2 t/km2/month); microbiological analysis was performed by the liquid bubbling method, identifying 27 genera incluiding bacteria and fungi, with the most frequent bacteria being Bacillus sp., Staphylococcus sp., Enterobacter agglomerans, Klebsiella pneumoniae, Enterobacter hafniae and some bacteria pathogenic for humans such as Streptococcus sp., Clostridium perfringens and Bacillus anthracis. The most frequent fungi are Penicillium sp., Aspergillius sp. and Geotrichum sp.; some pathogenic fungi identified: Microsporum sp., Epirophyton sp. On the other hand,regarding the perception of the public and workers, surveys were applied, wich indicated that the microbiologial quality of the air in the Biological Micro-station and animal breeding farm is regular(AU)

Antimicrobial activity of bacteria isolated from tissue of the coral Palythoa caribaeorum (Zoantharia: Sphenopidae) from Paraíba, Brazil coastal reefs / Actividad antimicrobiana de bacterias aisladas del tejido del coral Palythoa caribaeorum (Zoantharia: Sphenopidae) de los arrecifes costeros de Paraíba, Brasil

Abstract Introduction: The coral-associated bacteria with antimicrobial activity may be important to promote the health of their host through various interactions, and may be explored as a source of new bioactive compounds. Objective: To analyze the antimicrobial activity of bacteria associated with the zoanthid Palythoa caribaeorum from the coral reefs of Carapibus, Paraiba state, Brazil. Methods: The phylogenetic analysis of the bacteria was conducted based on partial sequences of the 16S rRNA gene using molecular and bioinformatics tools. The antimicrobial activity of the 49 isolates was tested against four bacterial strains and one yeast strain: Bacillus cereus (CCT0198), Escherichia coli (ATCC 25922), Staphylococcus aureus (ATCC 25923), Pseudomonas aeruginosa and Candida albicans (ATCC 10231). The antibiosis and antibiogram assays were conducted and the Minimal Inhibitory Concentration (MIC) was determined by the microdilution method. Results: The bacterial isolates belonged to Firmicutes phylum (84 % of the isolates) and the Proteobacteria phylum (16 % of the isolates). Among the 49 isolates five genera were found, with the Bacillus genus being the most abundant (82 % of the isolates), followed by Vibrio (10 %), Pseudomonas (4 %), Staphylococcus (2 %) and Alteromonas (2 %). Antibiosis test revealed that 16 isolates (33 %) showed antimicrobial activity against one or more of five tested reference strains. The highest number of antagonistic bacteria were found in the Bacillus genus (12 isolates), followed by Vibrio (three isolates) and Pseudomonas (one isolate) genera. The B. subtilis NC8 was the only isolate that inhibited all tested strains in the antibiosis assay. However, antibiogram test with post-culture cell-free supernatant of NC8 isolate showed the inhibition of only B. cereus, S. aureus and C. albicans, and the lyophilized and dialyzed material of this isolate inhibited only B. cereus. The lyophilized material showed bacteriostatic activity against B. cereus, with a MIC value of 125 μg/μl, and in the cytotoxicity assay, the hemolysis value was of 4.8 %, indicating its low cytotoxicity. Conclusions: The results show the antimicrobial potential of some bacterial isolates associated with the P. caribaeourum tissue, especially those belonged to Bacillus genus.

Resumen Introducción: La actividad antimicrobiana realizada por las bacterias asociadas con los corales, además de promover la salud de su huésped, representa una fuente para obtener nuevos compuestos bioactivos. Objetivo: Analizar la actividad antimicrobiana de las bacterias asociadas con el zoantario Palythoa caribaeorum de los arrecifes de Carapibus, Paraíba, Brasil. Metodología: El análisis filogenético de la bacterias se realizó con base en secuencias parciales del gen RNAr 16S utilizando herramientas moleculares y de bioinformática. La actividad antimicrobiana de las cepas se probó contra cuatro cepas bacterianas y una cepa de levadura: Bacillus cereus (CCT0198), Escherichia coli (ATCC 25922), Staphylococcus aureus (ATCC 25923), Pseudomonas aeruginosa y Candida albicans (ATCC 10231), utilizando ensayos antibiosis y antibiograma, y la concentración inhibitoria mínima (CIM) que se determinó por el método de microdilución. Resultados: Las cepas bacterianas pertenecían a Firmicutes (84 %) y Gammaproteobacteria (16 %). Entre 49 cepas se encontraron cinco géneros de bacterias: Bacillus, Vibrio, Pseudomonas, Staphylococcus y Alteromonas. Un total de 19 cepas exhibieron actividad antimicrobiana, siendo el género Bacillus el responsable del mayor número de bacterias antagonistas, con 12 cepas positivas en el ensayo de antibiosis y cuatro en la prueba de antibiograma. El mayor número de bacterias antagonistas se encontró en Bacillus (12 aislamientos), seguido por Vibrio (tres aislamientos) y Pseudomonas (un aisladmiento). El NC8, clasificado como Bacillus subtilis, inhibió todas las cepas estándar en el ensayo de antibiosis y las cepas de B. cereus, S. aureus y C. albicans en la prueba de antibiograma. El material liofilizado del B. subtilis NC8 mostró acción bacteriostática contra B. cereus, con un valor de CIM de 125 μg/μl. En la prueba de citotoxicidad, el grado de hemólisis fue del 4.8 % para el material liofilizado a las concentraciones probadas, lo que indica su baja citotoxicidad. Conclusión: Los resultados muestran el potencial antimicrobiano de algunos aislamientos bacterianos asociados al P. caribaeourum, especialmente los pertenecientes al género Bacillus.

Antagonistic microorganisms and nitrogen fertilization in control of tomato southern blight

The present study assessed the efficacy of formulated biocontrol agents and nitrogen fertilization on southern blight control. Antagonism test in vitro was performed to assess the inhibitory activity of Bacillus methylotrophicus and Trichoderma asperellum against the growth of Sclerotium rolfsii. Tomato seedlings were transplanted into the substrate added with ammonium nitrate doses and inoculated with the formulated biocontrol agents Ônix (B. methylotrophicus) or Quality (T. asperellum). Subsequently, seedlings were inoculated with S. rolfsii. Plant mortality, shoot and root weight were assessed 11 days after the last inoculation. Agents had effective inhibitory activity against S. rolfsii; thus, they could reduce southern blight severity when combined with ammonium nitrate. However, plant mortality was not reduced by them.(AU)

Research progress in spore-based in-situ restoration technology of concrete with microorganisms / 生物工程学报

Concrete is the most widely used modern building material. It is easy to crack under the action of stress, which makes the concrete structure permeable, affecting the durability and integrity of the structure, and thus shortening its service life. Microbial in-situ remediation technology is a low cost, effective and green way for concrete crack repairing. Due to its excellent biocompatibility, service life elongation, economic losses and environmental pollution reduction, microbial in-situ remediation technology has been intensively investigated. Bacillus has attracted much attention because of its excellent biomineralization ability, extremely strong environmental tolerance and long-term survival ability of its spores. In order to promote the research, development and large-scale application of microbial in-situ healing of concrete, the paper reviews the mechanism of spore-based in-situ healing of concrete, the survival of spores exposed in concrete, the influence of spores and external additives on the mechanical properties of concrete, progress in research and development of healing agent as well as healing effects. Moreover, future research focuses such as improving the survival ability of spores in the harsh environment of concrete, reducing the influence of external additives on the mechanical properties of concrete, and strengthening the healing effect of actual field applications are also summarized.

Isolation and identification of phosphatolytic bacteria in Paris polyphylla var. yunnanensis / 中国中药杂志

The wild resources of Paris polyphylla var. yunnanensis, a secondary endangered medicinal plant, are severely scarce. Introduction and cultivation can alleviate market demand. To screen phosphatolytic bacteria in the rhizosphere soil of P. polyphylla var. yunnanensis and provide data support for the development of high-efficiency microbial fertilizer, in this study, the dilution plate coating method was used to isolate and screen the phosphorus solubilizing bacteria with the ability of mineralizing organic phosphorus from the rhizosphere soil of wild and transplanted varieties of P. polyphylla var. yunnanensis in 10 different locations in Yunnan, Sichuan and Guizhou. After separation and purification, the phosphatolytic capacity was analyzed by qualitative and quantitative analysis. Combined with physiological and biochemical experiments, the strains were identified using 16 S rDNA sequencing analysis. Forty one strains were selected from the rhizosphere soil of P. polyphylla var. yunnanensis from 10 different habitats. Among them, 21 strains were obtained from the rhizosphere soil of the wild variety P. polyphylla var. yunnanensis and 20 strains were obtained from the rhizosphere soil of the transplanted variety. And significance analysis found that 41 organophosphate solubilizing strains had significant differences in their ability to solubilize phosphorus. The amount of phosphate solubilizing was 0.08-67.61 mg·L~(-1), the pH value was between 4.27 and 6.82. The phosphatolytic amount of strain Y3-5 was 67.61 mg·L~(-1), and the phosphorus increase amount was 57.57 mg·L~(-1). All 41 strains were identified as Gram-positive Bacillus. Combining physiological characteristic and phylogenetic trees, Bacillus mobilis Y3-5 was finally selected as the candidate rhizosphere phosphatolytic bacteria of P. polyphylla var. yunnanensis. The distribution of phosphorus solubilizing bacteria in the rhizosphere soil of P. polyphylla var. yunnanensis was different, and there were significant diffe-rences in phosphorus solubility. Organophosphate-dissolving strain Y3-5 is expected to be a candidate strain of P. polyphylla var. yunnanensis microbial fertilizer.

Screening and Characterization of Bacillus Strains Producing Highly Thermostable Amylase from Various Hot Springs of Algeria

HIGHLIGHTS Screening extremophile Bacillus strains from various Hot Springs Characterization Of Bacillus Strains Producing Highly Thermostable Amylase Genetic identification of the best strains

Abstract Currently thermostable Amylase represents a broad biotechnological interest and desired by a various industries. In this study, forty-six bacterial strains belonging to the genus Bacillus were isolated from various hot springs in the North West of Algeria based on their ability to degrade starch and produce amylase in Starch Agar medium. The majority of isolates showed a positive amylolytic activity. In order to select the most thermostables amylase the effect of temperature on enzymes was estimated, therefore the study of amylase thermostability was culminated by the selection of Four Strains having an interesting optimum of activity and range of stability, reaching 75 °C for the strains HBH1-2, HBH1-3, HBH3-1and 85 °C for the strain HC-2, This indicates that the Enzyme produced by retained strains have optimum activity at high temperature. The identity of the selected strains was established on the basis of the morphological, biochemical characteristics and phylogenetic position as determined by 16S Ribosomal DNA gene sequencing. The whole strains belonged to the genus Bacillus and their phylogeny were also reported in this study.

Isolation and Characterization of Alkaline Pectinase Productive Bacillus tropicus from Fruit and Vegetable Waste Dump Soil

Abstract Alkaline pectinase is the utmost significant industrial enzyme of the bioscouring process. By considering bio scouring of cotton, 30 microbial isolates from fruit and vegetable waste rich dump soil of Solang Valley and Vasishta (Manali, Himachal Pradesh, India) were isolated and screened for the alkaline pectinase production in the current research work. Only four isolates P3, P16, P21, and P27 were capable to produce extracellular alkaline pectinase at pH 9. Further by applying submerged fermentation, the alkaline pectinase production was quantitatively screened. The most efficient isolate was P3 identified as Bacillus tropicus, based on morphological, biochemical, and molecular characterization. Molecular characteristics confirmed by 16S rDNA sequence analysis. The nucleotide sequence of the isolate was novel with a 97% similarity index and submitted to the GenBank with accession number MK332379. The Bacillus strain selected was active at broad pH range from 8-10.5 and a temperature range from 25-50 oC. Optimum pH and temperature observed were 9 and 37 oC respectively and can be suitably used for the bio scouring process for the pretreatment of the fabrics.

Screening of plant growth-promoting rhizobacteria and its effect on seed germination of Polygonum multiflorum / 中国中药杂志

In this study, the rhizobacteria and actinomycetes of Polygonum multiflorum were screened for the strains with indole acetic acid(IAA)-producing capacity by Salkowski method, the siderophore-producing strains by Chrome Azurol S(CAS) assay, and the strains with inorganic phosphorus-solubilizing capacity by PKO inorganic phosphorus medium. The strains were identified by morphological identification, physiological and biochemical characteristics, and 16 S rDNA sequences. Furthermore, the effect of growth-promoting strains on the seed germination and development of P. multiflorum was tested. The results showed that among 196 strains, two strains F17 and F42 were found to be capable of producing IAA and siderophore and solubilizing inorganic phosphorus simulta-neously. For F17 and F42, the results are listed below: 38.65 and 33.64 mg·L~(-1) for IAA production, 0.85 and 0.49 for siderophore-producing capacities(A_s/A_r), and 1.35 and 1.70 for inorganic phosphorus-solubilizing capacities(D/d), respectively. Comprehensive analysis revealed that strains F17 and F42 were identified as Pseudochrobactrum asacharolyticum and Bacillus aryabhattai, respectively, and both could significantly promote the seed germination of P. multiflorum.