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.

Phenotypic plasticity in adults of Anticarsia gemmatalis exposed to sub-doses of Bt-based bioinsecticide

Abstract Anticarsia gemmatalis Hünber, 1818 is one of the main defoliating species in the soybean crop. Bacillus thuringiensis Berliner, 1915, is a bacterium used in the biological control of this pest species. Resistant populations and their sublethal effects caused by the use of the bacteria have already been reported; however, there are no studies on phenotypic plasticity in adulthood exposed to Bt-based bioinsecticide sub-doses. This study aimed to evaluate the morphometry of A. gemmatalis adults under laboratory conditions submitted to the Bt-based bioinsecticide Dipel® over the three generations. The body segments mensuread were width, length, and area of the anterior and posterior wings, the weight of the integument, chest, abdomen, wings, and the whole adult of males and females. Among the treatments, LC5 in the first generation and LC10 in the second generation were those with lower thresholds in relation to the weight of the chest and abdomen, considering the proportions of the body smaller than the females. The females weight adulthood was reduced by 10% about males, and, only in the first generation. Males have larger body size and more pronounced phenotypic plasticity than females. Here, we demonstrate the first study assessing the phenotypic plasticity of A. gemmatalis adults.

Resumo Anticarsia gemmatalis Hünber, 1818 é uma das principais espécies desfolhadoras da cultura da soja. Bacillus thuringiensis Berliner, 1915, é uma bactéria utilizada no controle biológico dessa espécie de praga. Populações resistentes e seus efeitos subletais causados pelo uso da bactéria já foram relatados, no entanto, não há estudos sobre a plasticidade fenotípica na idade adulta exposta a subdoses de bioinseticida à base de Bt. Este trabalho teve como objetivo avaliar a morfometria de adultos de A. gemmatalis em condições de laboratório submetidos ao bioinseticida Dipel® ao longo de três gerações. Os segmentos corporais mensuráveis eram largura, comprimento e área das asas anterior e posterior, o peso do tegumento, tórax, abdômen, asas e todo o adulto de machos e fêmeas. Dentre os tratamentos, CL5 na primeira geração e CL10 na segunda geração foram aqueles com limiares mais baixos em relação ao peso do tórax e abdômen, considerando as proporções do corpo menores que as do sexo feminino. O peso da fêmea na idade adulta foi reduzido em 10% em relação aos machos e, apenas na primeira geração. Os machos têm tamanho corporal maior e plasticidade fenotípica mais pronunciada do que as fêmeas. Este estudo demonstra o primeiro estudo avaliando a plasticidade fenotípica de adultos de A. gemmatalis.

Phenotypic plasticity in adults of Anticarsia gemmatalis exposed to sub-doses of Bt-based bioinsecticide / Plasticidade fenotípica morfométrica em adultos de Anticarsia gemmatalis Hübner, 1818 expostos a subdoses de Bacillus thuringiensis Berliner, 1911

Anticarsia gemmatalis Hünber, 1818 is one of the main defoliating species in the soybean crop. Bacillus thuringiensis Berliner, 1915, is a bacterium used in the biological control of this pest species. Resistant populations and their sublethal effects caused by the use of the bacteria have already been reported; however, there are no studies on phenotypic plasticity in adulthood exposed to Bt-based bioinsecticide sub-doses. This study aimed to evaluate the morphometry of A. gemmatalis adults under laboratory conditions submitted to the Bt-based bioinsecticide Dipel® over the three generations. The body segments mensuread were width, length, and area of the anterior and posterior wings, the weight of the integument, chest, abdomen, wings, and the whole adult of males and females. Among the treatments, LC5 in the first generation and LC10 in the second generation were those with lower thresholds in relation to the weight of the chest and abdomen, considering the proportions of the body smaller than the females. The female's weight adulthood was reduced by 10% about males, and, only in the first generation. Males have larger body size and more pronounced phenotypic plasticity than females. Here, we demonstrate the first study assessing the phenotypic plasticity of A. gemmatalis adults.

Anticarsia gemmatalis Hünber, 1818 é uma das principais espécies desfolhadoras da cultura da soja. Bacillus thuringiensis Berliner, 1915, é uma bactéria utilizada no controle biológico dessa espécie de praga. Populações resistentes e seus efeitos subletais causados pelo uso da bactéria já foram relatados, no entanto, não há estudos sobre a plasticidade fenotípica na idade adulta exposta a subdoses de bioinseticida à base de Bt. Este trabalho teve como objetivo avaliar a morfometria de adultos de A. gemmatalis em condições de laboratório submetidos ao bioinseticida Dipel® ao longo de três gerações. Os segmentos corporais mensuráveis eram largura, comprimento e área das asas anterior e posterior, o peso do tegumento, tórax, abdômen, asas e todo o adulto de machos e fêmeas. Dentre os tratamentos, CL5 na primeira geração e CL10 na segunda geração foram aqueles com limiares mais baixos em relação ao peso do tórax e abdômen, considerando as proporções do corpo menores que as do sexo feminino. O peso da fêmea na idade adulta foi reduzido em 10% em relação aos machos e, apenas na primeira geração. Os machos têm tamanho corporal maior e plasticidade fenotípica mais pronunciada do que as fêmeas. Este estudo demonstra o primeiro estudo avaliando a plasticidade fenotípica de adultos de A. gemmatalis.

Use of probiotics in the catfish Sorubim cuspicaudus larviculture

Sorubim cuspicaudus, a migratory catfish distributed in the Magdalena, Sinú, and Catatumbo river basins, is categorized as vulnerable to extinction. Production of fingerlings in controlled environments stands as a strategic conservation approach, and larviculture is a critical phase in rearing this species. Probiotics are used for improvement in the critical stages of fingerling production. The study aimed to evaluate the use of probiotics (Bacillus subtilis and Bacillus licheniformis) during the larviculture phase of S. cuspicaudus. Larvae at 42 hours post-hatching (1.5±0.1mg, total length 5.7±0.4mm) were treated with four levels of probiotic inclusion in the water: 0, 5, 10, and 20ppm for 22 days. Water quality remained within suitable ranges for neotropical catfish species larviculture and the parameters assessed were weight gain (Gw), length gain (Gl), specific growth rate (G), survival rate (S), stress resistance (Sr), intestinal fold length (LF), and colony-forming units (CFU) count. Results showed higher Gl (22.23±3.5mm), Gw (40.0±12.6mg), G (14.9±1.5%/day), LP (205±72.7µm), and CFU (118.7±80.9) were found at 20 ppm (p0.05). The findings of this study suggest that probiotics (Bacillus subtilis and Bacillus licheniformis) could be used as an alternative to advance in the S. cuspicaudus larviculture.

Sorubim cuspicaudus, un bagre migratorio distribuido en las cuencas de los ríos Magdalena, Sinú y Catatumbo, se encuentra catalogado como una especie vulnerable a la extinción. La producción de alevinos en ambientes controlados es considerada como una estrategia crucial para su conservación, y la larvicultura es una fase crítica en la cría de esta especie. Para mejorar esta fase crítica de producción de alevinos se utilizan probióticos. El estudio tenía como objetivo evaluar el uso de probióticos (Bacillus subtilis y Bacillus licheniformis) durante la fase de larvicultura de S. cuspicaudus. Las larvas, a las 42 horas post eclosión (1,5±0,1mg, longitud total 5,7±0,4mm) fueron tratadas con cuatro niveles de inclusión de probióticos en el agua: 0, 5, 10 y 20ppm durante 22 días. La calidad del agua se mantuvo dentro de los rangos adecuados para la larvicultura de especies neotropicales de bagre y los parámetros evaluados fueron el aumento de peso (Gw), el aumento de longitud (GL), la tasa específica de crecimiento (G), la tasa de supervivencia (S), la resistencia al estrés (Sr), la longitud del pliegue intestinal (LF) y el recuento de unidades formadoras de colonias (UFC). Los resultados mostraron mayores GL (22,23±3,5 mm), Gw (40,0±12,6 mg), G (14,9±1,5%/día), LP (205±72,7 µm) y UFC (118,7±80,9) a 20 ppm (p0,05). Los resultados de este estudio sugieren que los probióticos (Bacillus subtilis y Bacillus licheniformis) podrían utilizarse como alternativa para avanzar en la larvicultura de S. cuspicaudus.

Enterotoxigenic profiles and submerged and interface biofilms in Bacillus cereus group isolates from foods / Perfiles enterotoxigénicos y biopelículas de superficie y sumergidas en aislados del grupo Bacillus cereus de alimentos

Abstract Biofilm formation by Bacillus cereus strains is now recognized as a systematic contaminaron mechanism in foods; the aim of this study was to evaluate the production of submerged and interface biofilms in strains of B. cereus group in different materials, the effect of dex-trose, motility, the presence of genes related to biofilms and the enterotoxigenic profile of the strains. We determine biofilm production by safranin assay, motility on semi-solid medium, toxin gene profiling and genes related to biofilm production by PCR in B. cereus group iso-lated from food. In this study, we observe strains used a higher production of biofilms in PVC; in the BHI broth, no submerged biofilms were found compared to phenol red broth and phenol red broth supplemented with dextrose; no strains with the ces gene were found, the enterotoxin profile was the most common the profile that includes genes for the three enterotoxins. We observed a different distribution of tasA and sipW with the origin of isolation of the strain, being more frequent in the strains isolated from eggshell. The production and type of biofilms are differential according to the type of material and culture medium used.

Resumen La formación de biopelículas por cepas de Bacillus cereus es reconocida como un mecanismo de contaminación sistemática en alimentos; el objetivo del estudio fue evaluar la producción de biopelículas sumergidas y de superficie en cepas del grupo de Bacillus cereus en diferentes materiales, el efecto de la dextrosa, la motilidad, la presencia de genes relacionados a biopelículas y el perfil enterotoxigénico de las cepas. Determinamos la producción de biopelículas por el ensayo de safranina, motilidad en medio sólido, perfil enterotoxigénico y genes relacionados a producción de biopelículas por PCR en aislados del grupo de Bacillus cereus de alimentos. En este estudio, observamos en las cepas utilizadas una alta producción de biopelículas en PVC; en caldo BHI, no se encontraron biopelículas sumergidas en comparación con el caldo rojo de fenol y caldo rojo de fenol suplementando con dextrosa; no se encontraron cepas con el gen ces, el perfil de enterotoxinas más común fue el perfil que incluía los genes de las tres enterotoxinas, también observamos una distribución diferente de tasA y sipW con relación al origen de la cepa, siendo más frecuente estos genes en las cepas aisladas de huevos. La producción y el tipo de biopelículas es diferente de acuerdo con el tipo de material y el medio de cultivo utilizado.

Caracterización bioinformática y producción de L-asparaginasa de Bacillus sp. M62 aislado de las salinas de Maras, Cusco, Perú

El objetivo del estudio fue realizar la caracterización bioinformática, así como optimizar la producción de L-asparaginasa extracelular de Bacillus sp. M62 aislada de las salinas de Maras (Cusco). Para ello, se verificó la producción de L-asparaginasa mediante el viraje del medio M9 modificado con azul de bromofenol 0.0075%, pH 7.4 a 37 °C por 72 h. A la vez, se extrajo el ADN genómico para amplificar los genes ribosómicos 16S y el gen ansA3. La secuencia aminoacídica codificada por el gen ansA3 se predijo mediante análisis bioinformático. La producción de L-asparaginasa intracelular y extracelular se evaluó a diferentes niveles de glucosa, L-asparagina, NaCl y pH en el medio M9 modificado. Adicionalmente, las actividades enzimáticas de L-asparaginasa y L-glutaminasa se determinaron mediante cuantificación del amonio liberado por el método de Nessler. Así, Bacillus sp. M62 produjo el viraje del medio M9 modificado, obtuvo alta similitud y cercanía evolutiva con Bacillus licheniformis, se encontró que el gen ansA3 amplificado codificaba para 319 aa, dentro de la cual se predijo una secuencia patrón del sitio activo (GFVITHGTDTM ) y 15 sitios inmunogénicos. La producción de L-asparaginasa extracelular fue superior a la intracelular, la que se optimizó de 0.37 U/mL (0.24 U/mg) a 2.15 ± 0.39 U/mL (0.63 U/mg). Finalmente, se encontró que Bacillus sp. M62 presenta L-asparaginasa extracelular con mínima actividad de L-glutaminasa.

The aim of this study was to perform bioinformatics characterization and optimize the production of extracellular L-asparaginase from Bacillus sp. M62, isolated from the Maras salt ponds (Cusco). To achieve this, the production of L-asparaginase was verified by the change in color of modified M9 medium, containing 0.0075% bromophenol blue, at pH 7.4 and 37°C for 72 hours. Genomic DNA was extracted to amplify the 16S ribosomal genes and the ansA3 gene. The amino acid sequence encoded by the ansA3 gene was predicted using bioinformatic analysis. The production of intracellular and extracellular L-asparaginase was evaluated at different levels of glucose, L-asparagine, NaCl, and pH in modified M9 medium. Additionally, the enzymatic activities of L-asparaginase and L-glutaminase were determined by quantifying the released ammonium using the Nessler method. Bacillus sp. M62 showed the change in color of the modified M9 medium, high similarity, and evolutionary closeness to Bacillus licheniformis. The amplified ansA3 gene was found to encode for 319 amino acids, with a predicted active site pattern (GFVITHGTDTM) and 15 immunogenic sites. The production of extracellular L-asparaginase was found to be higher than intracellular L-asparaginase and was optimized from 0.37 U/mL (0.24 U/mg) to 2.15 ± 0.39 U/mL (0.63 U/mg). Finally, it was found that Bacillus sp. M62 presents extracellular L-asparaginase with minimal L-glutaminase activity.

Infectious endocarditis due to Bacillus Cereus in a patient with a pacemaker: case report and literature review / Endocardite infecciosa por Bacillus Cereus em paciente portador de marcapasso: relato de caso e revisão de literatura

Introduction: Infective endocarditis (IE) is an infectious process of the cardiac endothelium, often related to the use of pacemakers and valve prostheses, which may facilitate microorganism" proliferation. Case Report: In this article, we describe the case of an 81-year-old man with infective endocarditis due to Bacillus Cereus related to the use of a pacemaker and perform a brief literature review. Discussion: Bacillus Cereus is a Gram-positive, aerobic, spore-forming, large, and generally motile bacterium that constitutes a rare cause of endocarditis, but few cases like this are described in the literature. Conclusion: Determining the etiology of IE through culture-guided methods plays a pivotal role in selecting appropriate antibiotic treatment. Maintain a high clinical suspicion for IE is paramount, especially when fever arises in patients with cardiac devices after surgical or dental procedures.

Introdução: A endocardite infecciosa é um processo infeccioso do endotélio cardíaco, muitas vezes relacionado ao uso de marca-passos e próteses valvares, que pode facilitar a proliferação de microrganismos. Relato de Caso: Neste artigo descrevemos o caso de um homem de 81 anos com endocardite infecciosa por Bacillus Cereus relacionada ao uso de marca-passo e realizamos uma breve revisão da literatura. Discussão: Bacillus Cereus é uma bactéria Gram-positiva, aeróbia, formadora de esporos, grande e geralmente móvel, que constitui uma causa rara de endocardite, com poucos casos descritos na literatura. Conclusão: A determinação da etiologia da EI através de métodos guiados por cultura desempenha um papel fundamental na seleção do tratamento antibiótico apropriado. Manter alta suspeita clínica de EI é fundamental, principalmente quando surge febre em pacientes portadores de dispositivos cardíacos após procedimentos cirúrgicos ou odontológicos.

Isolation,identification and virulence gene distribution and pathogenicity of three Bacillus cereus strains / 中国生物制品学杂志

@#Objective To isolate and identify the pathogenic bacteria in Tongliao area,Inner Mongolia Autonomous Region,China in 2019 and determine the virulence gene distribution and pathogenicity.Methods Three Gram positive isolates,FL1,FL2 and FL3,from the clinical specimens in Tongliao area were subjected to biochemical test,drug sensitivity test,16S rDNA sequencing,virulence gene test and pathogenicity test.Results Bio-chemical test proved the three isolates as Bacillus cereus. The strains FL1 and FL2 from bovine origin were resistant to penicillin and carbamazillin,while the strain FL3 from horse origin to Cefradine. Strains FL1 and FL2 showed the closest relationship to the isolate(KR063181. 1)from Hunan,China,while strain FL3 to the isolate(HM104658. 1)from Shandong,China. However,strain FL3 showed relatively far relationship to strains FL1 and FL2. All the distributions of eight virulence genes of B.cereus were observed in the three strains,while the distributions of four virulence genes of B.anthracoides were different. All the three isolates showed pathogenicity.Conclusion All the three isolates were pathogenic B.cereus,which showed a certain drug resistance. However,there were differences in virulence gene distri-bution between bovine and equine B.cereus.

Prokaryotic expression and biological activities of the hemolysin BL subunit of a pathogenic Bacillus cereus of cattle origin / 生物工程学报

Bacillus cereus belongs to Gram-positive bacteria, which is widely distributed in nature and shows certain pathogenicity. Different B. cereus strains carry different subsets of virulence factors, which directly determine the difference in their pathogenicity. It is therefore important to study the distribution of virulence factors and the biological activity of specific toxins for precise prevention and control of B. cereus infection. In this study, the hemolysin BL triayl was expressed, purified, and characterized. The results showed that the bovine pathogenic B. cereus hemolysin BL could be expressed and purified in the prokaryotic expression system, and the bovine pathogenic B. cereus hemolysin BL showed hemolysis, cytotoxicity, good immunogenicity and certain immune protection in mice. In this study, the recombinant expression of hemolysin BL triayl was achieved, and the biological activity of hemolysin BL of bovine pathogenic ceroid spore was investigated. This study may facilitate further investigating the pathogenic mechanism of B. cereus hemolysin BL and developing a detection method for bovine pathogenic B. cereus disease.

Using transporter to enhance the acid tolerance of Bacillus coagulans DSM1 / 生物工程学报

As the precursor of polylactic acid (PLA), optically pure l-lactic acid production is attracting increasing attention. The accumulation of lactic acid during fermentation inhibits strain growth. Therefore, it is necessary to improve the acid tolerance of lactic acid producers. In this study, comparative transcriptomic analysis was performed to investigate the effects of transporters on lactic acid tolerance of Bacillus coagulans DSM1, which is an l-lactic acid producer. The genes with more than two-fold up-regulation in transcriptional profile were further verified using real-time PCR. The transcriptional levels of RS06895, RS10595, RS10595, RS00500, RS00500, RS10635 and RS10635 were enhanced during lactic acid fermentation. Strain overexpressing RS10595 exhibited a retarded cell growth and low lactic acid production at pH 6.0, but an improved lactic acid production at pH 4.6. This study may facilitate the investigation of the acid tolerance mechanism in B. coagulans DSM1, as well as the construction of efficient lactic acid producers.

Advances in Bacillus Calmette-Guerin / 中国生物制品学杂志

@#Tuberculosis(TB) is a chronic and debilitating infectious disease caused by Mycobacterium tuberculosis(Mtb). It is a significant health concern that poses a serious threat to human well-being. Bacillus Calmette-Guerin(BCG)is the only approved vaccine for the prevention of TB. BCG is made from attenuated bovine Mtb and is highly effective in preventing tuberculous meningitis and granulomatous TB,which are particularly lethal in infants. However,BCG provides limited protection in both children and adults,and this protective effect decreases over time. Currently,there are ongoing improvements being made to BCG,which involve modifications in immunization routes and procedures. Additionally,researchers are working on the development of new TB vaccines. BCG is primarily used for the prevention of TB and the treatment of bladder cancer,although it also has other indications. This paper reviewed the recent research progress of BCG,focusing on its development history,improvements,and indications.

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.

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.

Isolation and identification of a polyester-polyurethane degrading bacterium Bacillus altitudinis YX8-1 / 生物工程学报

Although polyurethane (PUR) plastics play important roles in daily life, its wastes bring serious environmental pollutions. Biological (enzymatic) degradation is considered as an environmentally friendly and low-cost method for PUR waste recycling, in which the efficient PUR-degrading strains or enzymes are crucial. In this work, a polyester PUR-degrading strain YX8-1 was isolated from the surface of PUR waste collected from a landfill. Based on colony morphology and micromorphology observation, phylogenetic analysis of 16S rDNA and gyrA gene, as well as genome sequence comparison, strain YX8-1 was identified as Bacillus altitudinis. The results of high performance liquid chromatography (HPLC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) showed that strain YX8-1 was able to depolymerize self-synthesized polyester PUR oligomer (PBA-PU) to produce a monomeric compound 4, 4'-methylene diphenylamine. Furthermore, strain YX8-1 was able to degrade 32% of the commercialized polyester PUR sponges within 30 days. This study thus provides a strain capable of biodegradation of PUR waste, which may facilitate the mining of related degrading enzymes.

Isolation of a foodborne Bacillus cereus strain and its effect on intestinal mucosal immunity-associated factors and gut microbial community in mice / 生物工程学报

Bacillus cereus is a common foodborne pathogen. Accidently eating food contaminated by B. cereus will cause vomiting or diarrhea, and even death in severe cases. In the present study, a B. cereus strain was isolated from spoiled rice by streak culture. The pathogenicity and drug resistance of the isolated strain were analyzed by drug sensitivity test and PCR amplification of virulence-associated gene respectively. Cultures of the purified strain were injected intraperitoneally into mice to examine their effects on intestinal immunity-associated factors and gut microbial communities, to provide references for the pathogenic mechanism and medication guidance of these spoilage microorganisms. The results showed that the isolated B. cereus strain was sensitive to norfloxacin, nitrofurantoin, tetracycline, minocycline, ciprofloxacin, spectinomycin, clindamycin, erythrocin, clarithromycin, chloramphenicol, levofloxacin, and vancomycin, but resistant to bactrim, oxacillin and penicillin G. The strain carries seven virulence-associated genes including hblA, hblC, hblD, nheA, nheB, nheC and entFM, which are involved in diarrhea-causing toxins production. After infecting mice, the isolated B. cereus strain was found to cause diarrhea in mice, and the expression levels of immunoglobulins and inflammatory factors in the intestinal mucosae of the challenged mice were significantly up-regulated. Gut microbiome analysis showed that the composition of gut microbial community in mice changed after infection with B. cereus. The abundance of the uncultured_bacterium_f_Muribaculaceae in Bacteroidetes, which is a marker of body health, was significantly decreased. On the other hand, the abundance of uncultured_bacterium_f_Enterobacteriaceae, which is an opportunistic pathogen in Proteobacteria and a marker of dysbacteriosis, was significantly increased and was significantly positively correlated with the concentrations of IgM and IgG. These results showed that the pathogenic B. cereus carrying diarrhea type virulence-associated gene can activate the immune system by altering the composition of gut microbiota upon infection.

Research progress on the structure of crystal protein of Bacillus thuringiensis and its mechanism / 中国热带医学

@#Abstract: Mosquitoes are involved in the transmission of serious diseases such as malaria, dengue, chikungunya, yellow fever, Zika virus disease, and filariasis, and their prevention and control have always been a research hotspot. Currently, mosquito control methods mainly include physical control, chemical control and biological control. Physical control methods are environmentally friendly, but they are slow to take effect and have unsatisfactory control effects; although chemical control can quickly eliminate mosquitoes, it has been eliminated due to its high pollution, high residual, and easy drug resistance; biological control uses natural enemies or pathogens to kill mosquitoes and reduce their ability to transmit disease. Therefore, environmentally friendly biological control has become the main measure for controlling and preventing mosquitoes. In recent years, significant progress has been made in bacterial mosquito control agents, among which Bacillus thuringiensis has been the most extensively studied. Bacillus thuringiensis is a Gram-positive soil microorganism, which is the pathogenic bacterium of a variety of agricultural pests such as Lepidoptera, Coleoptera and Diptera. During the sporulation process, its strains produce a variety of insecticidal crystal proteins (ICPs) or δ-endotoxins with insecticidal activity against mosquito larvae. This review firstly introduces the crystal proteins of Bacillus thuringiensis, describes in detail the types and structures of crystal proteins in detail, and also reveals the mechanism of action of crystal proteins related to receptors.

Clinical treatment and observation of the endophthalmitis caused by Bacillus cereus / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To analyze the clinical characteristics and antibiotic sensitivity of traumatic endophthalmitis caused by Bacillus cereus and discuss the early diagnosis and treatment measures.METHODS: The data of 15 patients(15 eyes)with Bacillus cereus endophthalmitis admitted to the Eye Trauma Center of Shaanxi Provincial Eye Hospital from January 2019 to December 2021 were collected. The injury time, injury condition, preoperative visual acuity, corrected visual acuity in the last follow-up, operation method, bacterial culture and drug sensitivity test results were recorded.RESULTS: Among the 15 patients(15 eyes), 5 eyes with simple corneal perforating wound were treated with wound debridement and suture combined with intraocular antibiotic injection. And another 10 eyes with penetrating injuries combined with traumatic cataract and intraocular foreign bodies were treated with debridement, cataract extraction, removal of intraocular foreign bodies, vitrectomy combined with silicone oil tamponade and intraocular antibiotic injection. Among the 15 patients, 11 patients(73%)were treated effectively and the eyeball was preserved, and 4 patients(27%)were treated ineffectively and ocular evisceration was performed. Among the 11 patients with eyeball retention, 1(9%)had decreased vision, 1(9%)had unchanged vision, 4(36%)had improved vision by 1 level, and 5(45%)had improved vision by 2 levels, and the postoperative visual acuity was significantly improved. Correlation analysis showed that the course of disease was negatively correlated with corrected visual acuity(rs=-0.762, P=0.001). The cultures of vitreous humor and aqueous humor samples of patients were all grown in Bacillus cereus, and susceptibility tests were sensitive to vancomycin.CONCLUSIONS: Vancomycin is an effective drug for the treatment of Bacillus cereus endophthalmitis. Vitrectomy combined with silicone oil tamponade and intravitreal injection of vancomycin are effective method for the treatment of Bacillus cereus endophthalmitis.

Effect of hemX gene deletion on heme synthesis in Bacillus amyloliquefaciens / 生物工程学报

Heme, which exists widely in living organisms, is a porphyrin compound with a variety of physiological functions. Bacillus amyloliquefaciens is an important industrial strain with the characteristics of easy cultivation and strong ability for expression and secretion of proteins. In order to screen the optimal starting strain for heme synthesis, the laboratory preserved strains were screened with and without addition of 5-aminolevulinic acid (ALA). There was no significant difference in the heme production of strains BA, BAΔ6 and BAΔ6ΔsigF. However, upon addition of ALA, the heme titer and specific heme production of strain BAΔ6ΔsigF were the highest, reaching 200.77 μmol/L and 615.70 μmol/(L·g DCW), respectively. Subsequently, the hemX gene (encoding the cytochrome assembly protein HemX) of strain BAΔ6ΔsigF was knocked out to explore its role in heme synthesis. It was found that the fermentation broth of the knockout strain turned red, while the growth was not significantly affected. The highest ALA concentration in flask fermentation reached 82.13 mg/L at 12 h, which was slightly higher than that of the control 75.11 mg/L. When ALA was not added, the heme titer and specific heme production were 1.99 times and 1.45 times that of the control, respectively. After adding ALA, the heme titer and specific heme production were 2.08 times and 1.72 times higher than that of the control, respectively. Real-time quantitative fluorescent PCR showed that the expressions of hemA, hemL, hemB, hemC, hemD, and hemQ genes at transcription level were up-regulated. We demonstrated that deletion of hemX gene can improve the production of heme, which may facilitate future development of heme-producing strain.

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.