In order to improve the spore yield of compound Bacillus spp. (B. amyloliquefaciens, B. laterosporus and B. megaterium), the effects of nutrient conditions including carbon source, nitrogen source, mineral salt and fe...In order to improve the spore yield of compound Bacillus spp. (B. amyloliquefaciens, B. laterosporus and B. megaterium), the effects of nutrient conditions including carbon source, nitrogen source, mineral salt and fermentation conditions including the inoculum age, inoculation amount, loading volume of liquid and initial pH on the spore yield were studied. The results indicated that the optimized medium was glucoses 20 g/L, soybean meal 30.0 g/L, K2HPO4 1.0 g/L;fermentation temperature is 37℃, the inoculum age 12 h, initial pH 7.0, 2% inoculation amount, loading volume of liquid 20 mL/250 mL. Under the optimized conditions of culture medium and fermentation for compound Bacillus spp., spore yield was 10.24 times more than the initial medium, and the spore formation rate reached more than 90%.展开更多
The four effective antagonistic Bacillus strains, isolated from the rhizosphere soil of cucumber in a greenhouse, produced antifungal volatiles. These volatiles strongly inhibited the growth of the most tested pathoge...The four effective antagonistic Bacillus strains, isolated from the rhizosphere soil of cucumber in a greenhouse, produced antifungal volatiles. These volatiles strongly inhibited the growth of the most tested pathogenic fungi with wide host plants, induced the mycelial morphological abnormalities, and decreased the sclerotoid production of Sclerotinia sclerotiorum in sealed plates. Spores of Botrytis cinerea exposed to these volatiles for 24-48 h in cavity slides cracked and the sporaceous inclusion became brown and effused to the suspension. An interesting phenomenon observed was that all the bacterial volatiles exhibited intense inhibitory activities against the pigment formation of tested pathogenic fungi, including Ascochyta citrullina, Alternaria solani, Alternaria brassicae, and so on. Interactions mediated by microbial volatiles could be widespread in soils, and volatiles may play an important role in reducing disease levels. A phylogenetic analysis based on 16S rDNA sequence placed the four bacteria in three species Paenibacillus polymyxa （BMP-11）, Bacillus subtilis （BL02）, and Bacillus pumilus （BSH-4 and ZB 13）. Through headspace sampling and GC-MS analysis, a rich profile was found from B. subtilis and overlapping volatile patterns could be found among the different species. Studies are under the way to find the possible action mechanisms and to seek the effective application of bacterial volatiles in greenhouse.展开更多
Background:Previous research has shown that dietary supplementation of Bacillus spp.probiotics exerts beneficial effects on animals’growth.However,limited studies have evaluated the efficacy of Bacillus spp.on weaned...Background:Previous research has shown that dietary supplementation of Bacillus spp.probiotics exerts beneficial effects on animals’growth.However,limited studies have evaluated the efficacy of Bacillus spp.on weaned pigs and their effects on host gut health and microbiome,and systemic immunity using a disease challenge model.The objective of this experiment was to investigate the effects of two Bacillus spp.strains(Bacillus subtilis DSM 32540 and Bacillus pumilus DSM 32539)on growth performance,diarrhea,intestinal health,microbiome,and systemic immunity of weaned pigs experimentally infected with an enterotoxigenic Escherichia coli(ETEC).Results:Pigs in PRO1(Bacillus subtilis DSM 32540)had greater(P<0.05)body weight on d 7 and 14 PI,greater(P<0.05)ADG from d 0 to 7 and d 7 to 14 PI,compared with pigs in CON(Control).Pigs in PRO1 had milder(P<0.05)diarrhea on d 2 and 3 PI compared with pigs in CON.However,no differences were observed in growth performance and diarrhea score between PRO2(Bacillus pumilus DSM 32539)and CON groups.Supplementation of PRO1 decreased(P<0.05)lymphocyte counts on d 7 and 14 PI,compared with CON.Supplementation of PRO1 and PRO2 both reduced(P<0.05)total coliforms in mesenteric lymph nodes on d 21 PI.Pigs in PRO2 had greater(P<0.05)goblet cell number and sulfomucin percentage in duodenal villi and greater(P<0.05)sialomucin percentage in jejunal villi than pigs in CON.Supplementation of PRO1 up-regulated(P<0.05)MUC2 gene expression in jejunal mucosa and reduced(P<0.05)PTGS-2 and IL1B gene expression in ileal mucosa on d 21 PI,compared with CON.Pigs in PRO1 had reduced(P<0.05)relative abundance of families Lachnospiraceae,Peptostreptococcaceae and Pasteurellaceae in the ileum.Conclusions:Supplementation of Bacillus subtilis DSM 32540 improved growth performance,alleviated diarrhea severity,enhanced gut health,and reduced systemic inflammation of weaned pigs infected with ETEC F18.Although Bacillus pumilus DSM 32539 was able to alleviate systemic inflammation,it had limited impacts on growth performance and severity of diarrhea of ETEC F18 challenged weaned pigs.展开更多
A compound bioflocculant CBF,produced by mixed culture of Rhizobium radiobacter F2 and Bacillus sphaeicus F6, was investigated with regard to its production and flocculating properties. The optimization of the culture...A compound bioflocculant CBF,produced by mixed culture of Rhizobium radiobacter F2 and Bacillus sphaeicus F6, was investigated with regard to its production and flocculating properties. The optimization of the culture medium constituents including carbon source,nitrogen source and C / N ratio,metal ions and ionic strength on CBF production were studied. Flocculating properties of CBF were examined by a series of experiments and CBF had good flocculating activities in kaolin suspension with divalent cations and stable over wide range of p H. Studies of the flocculating properties revealed that the flocculation could be stimulated by cations Ca2+,Mg2+,Fe2+,Al3+and Fe3+. In addition,it was stable at 4-30 ℃ in the presence of Ca Cl2. It was found to be effective for flocculation of a kaolin suspension under neutral and weak alkaline conditions( p H 7. 0- 9. 0),and flocculating activities of higher than 95% were obtained when the CBF concentrations among 6- 14 mg / L at p H 8. 0. The results of this study indicate that CBF is a potential replacement of conventional synthetic flocculants and is widely applied in water treatment and downstream processing of food and fermentation industries.展开更多
[ Objective] The paper was to screen the antagonistic Bacillus spp. strains against Phytophthora infestans （Mont.） de Bary and analyze its lipepeptide substances. [ Methods] Using flat isolation and culture method, ...[ Objective] The paper was to screen the antagonistic Bacillus spp. strains against Phytophthora infestans （Mont.） de Bary and analyze its lipepeptide substances. [ Methods] Using flat isolation and culture method, the bacteria that could produce Bacillus spp. were isolated from rhizosphere soil, and carried out primary and secondary screening by confrontation culture. The lipopeptide substances of Y-3 strain with better antifungal effect were carried out I-IPLC analysis. [Result] Nine strains of bacteria had certain antagonistic effects against P. infestans in primary screening, Y-1 and Y-3 strains had significant inhibition effect against the growth of P. infestans in secondary screening, with inhibition rate higher than 80% ; the crude extract of lipepeptide produced by Y-3 strain had better antagonistic effect against P. infestans, with inhibition rate higher than 90%. HPLC analysis showed that the crude extract of lipepeptide mainly contained 8 components. [ Conclusion] The lipopeptide substances produced by Y-3 strain had significant inhibition effect, which had potential application value for biological control of potato late blight.展开更多
The present study was taken up with a view to ascertain the possibility of introduction of alcohol resistant bacteria in vitro through the aseptically raised watermelon (Citrullus lanatus) seedlings in the backdrop of...The present study was taken up with a view to ascertain the possibility of introduction of alcohol resistant bacteria in vitro through the aseptically raised watermelon (Citrullus lanatus) seedlings in the backdrop of isolating such organisms from micropropagated watermelon stocks. Watermelon cv. Arka Manik seedlings grown in vitro from surface-sterilized seeds with the intact seed coat on MS medium appeared visibly clean largely, but upon subjecting them to tissue-indexing, the segments from the collar or root tissue showed bacterial colony growth on Nutrient Agar (NA) from 72% of such healthy seedlings and the cotyledon and hypocotyl tissue of 44% seedlings. The pooled colony growth from NA upon challenge with 90% alcohol yielded 10 distinct colony types, identified as B. pumilus (4×), B. subtilis (4×), B. cereus (1×) or B. safensis (1×) based on partial 16S rRNA sequence analysis. The shoot-tip tissue from the healthy index-negative seedlings cultured on watermelon proliferation medium partly turned index-positive within 2 - 4 sub-culture cycles while being apparently clean. On the other hand, those from the previously index-positive cultures tended to show obvious bacterial growth during subsequent in vitro culturing. The observations suggested the possibility of introduction of spore-forming Bacillus spp. through surface-sterilized seeds, their gradual emergence in vitro in visibly clean seedlings, possible transmittal of spores to the alcohol through tissue-culturing tools and the survival therein with the chances of unsuspected lateral spread. Seed coat removal followed by surface sterilization with sodium hypochlorite facilitated the raising of clean seedlings with no detectable bacterial association.展开更多
Volatile organic compounds (VOCs) are harmful to human health and the environment. Recently, loess (Hwangtoh) was used as an eco-friendly interior paint formulation in Korea. It is used even more commonly as a filter ...Volatile organic compounds (VOCs) are harmful to human health and the environment. Recently, loess (Hwangtoh) was used as an eco-friendly interior paint formulation in Korea. It is used even more commonly as a filter carrier to remove VOCs. In this study, we isolated Bacillus strains from a loess filter. The strains that were tolerant to VOCs were labeled according to the series VOC01 to VOC35. Four strains—VOC03, VOC11, VOC18, and VOC30—were investigated for their ability to degrade cyclohexane and toluene. Strain VOC18 best degraded both VOCs, whereas VOC03 demonstrated no ability to degrade VOCs. In keeping with this, VOC18 grew best on cyclohexane or toluene as the sole carbon source. The strains were identified by their physiochemical and phylogenetic characteristics. Strain VOC18 was determined as a strain of Bacillus cereus;VOC11 and VOC30 were determined as differentiated strains of B. thuringiensis. Strain VOC03, which demonstrated high tolerance but no ability to degrade VOCs, was identified as a strain of B. megaterium.展开更多
Phytophthora species are particularly aggressive plant pathogens and are often associated with the decline of many tree species, including oak and beech. Several fungi and bacteria species are known as potential antag...Phytophthora species are particularly aggressive plant pathogens and are often associated with the decline of many tree species, including oak and beech. Several fungi and bacteria species are known as potential antagonists usable as biological control agents. Phosphonate （H3PO3）, commonly branded as phosphite, has also been used in the past years to protect trees against invasive Phytophthora spp.. This study aimed at comparing the effects of selected antagonist microorganisms and phosphonate, when applied by microinjection or leaf treatment. Antagonistic species were first selected for their high inhibitory activity against problematic Phytophthora species, such as Phytophthora cactorum, P. quercina and P. plurivora attacking Quercus robur and Fagus sylvatica in Polish forests. Three endophytic species Trichoderma atroviride （two strains）, T. harzianum and Bacillus amyloliquefaciens showed a high control activity, and their efficacy was then assessed in comparison with a phosphonate treatment. Two application methods were experimented in this study： injection of a solution of spores or phosphonate into the sap vessels of beech or a foliar treatment on oak. Phosphonate and two strains of Trichoderma significantly reduced the necrotic area on oak leaves inoculated with P. plurivora and one strain of T. atroviride significantly reduced necrotic areas on beech branches. These results are therefore promising of a novel way to control Phytophthora spp. in forest stands and nurseries.展开更多
文摘In order to improve the spore yield of compound Bacillus spp. (B. amyloliquefaciens, B. laterosporus and B. megaterium), the effects of nutrient conditions including carbon source, nitrogen source, mineral salt and fermentation conditions including the inoculum age, inoculation amount, loading volume of liquid and initial pH on the spore yield were studied. The results indicated that the optimized medium was glucoses 20 g/L, soybean meal 30.0 g/L, K2HPO4 1.0 g/L;fermentation temperature is 37℃, the inoculum age 12 h, initial pH 7.0, 2% inoculation amount, loading volume of liquid 20 mL/250 mL. Under the optimized conditions of culture medium and fermentation for compound Bacillus spp., spore yield was 10.24 times more than the initial medium, and the spore formation rate reached more than 90%.
基金the National Key Technology R&D Program of China (2006BAD08A03)the Shandong Agricultural University Postdoctoral Foundation (200603065)
文摘The four effective antagonistic Bacillus strains, isolated from the rhizosphere soil of cucumber in a greenhouse, produced antifungal volatiles. These volatiles strongly inhibited the growth of the most tested pathogenic fungi with wide host plants, induced the mycelial morphological abnormalities, and decreased the sclerotoid production of Sclerotinia sclerotiorum in sealed plates. Spores of Botrytis cinerea exposed to these volatiles for 24-48 h in cavity slides cracked and the sporaceous inclusion became brown and effused to the suspension. An interesting phenomenon observed was that all the bacterial volatiles exhibited intense inhibitory activities against the pigment formation of tested pathogenic fungi, including Ascochyta citrullina, Alternaria solani, Alternaria brassicae, and so on. Interactions mediated by microbial volatiles could be widespread in soils, and volatiles may play an important role in reducing disease levels. A phylogenetic analysis based on 16S rDNA sequence placed the four bacteria in three species Paenibacillus polymyxa （BMP-11）, Bacillus subtilis （BL02）, and Bacillus pumilus （BSH-4 and ZB 13）. Through headspace sampling and GC-MS analysis, a rich profile was found from B. subtilis and overlapping volatile patterns could be found among the different species. Studies are under the way to find the possible action mechanisms and to seek the effective application of bacterial volatiles in greenhouse.
基金supported by Evonik Nutrition&Care GmbH,Germany and the United States Department of Agriculture(USDA)National Institute of Food and Agriculture,multistate project W400。
文摘Background:Previous research has shown that dietary supplementation of Bacillus spp.probiotics exerts beneficial effects on animals’growth.However,limited studies have evaluated the efficacy of Bacillus spp.on weaned pigs and their effects on host gut health and microbiome,and systemic immunity using a disease challenge model.The objective of this experiment was to investigate the effects of two Bacillus spp.strains(Bacillus subtilis DSM 32540 and Bacillus pumilus DSM 32539)on growth performance,diarrhea,intestinal health,microbiome,and systemic immunity of weaned pigs experimentally infected with an enterotoxigenic Escherichia coli(ETEC).Results:Pigs in PRO1(Bacillus subtilis DSM 32540)had greater(P<0.05)body weight on d 7 and 14 PI,greater(P<0.05)ADG from d 0 to 7 and d 7 to 14 PI,compared with pigs in CON(Control).Pigs in PRO1 had milder(P<0.05)diarrhea on d 2 and 3 PI compared with pigs in CON.However,no differences were observed in growth performance and diarrhea score between PRO2(Bacillus pumilus DSM 32539)and CON groups.Supplementation of PRO1 decreased(P<0.05)lymphocyte counts on d 7 and 14 PI,compared with CON.Supplementation of PRO1 and PRO2 both reduced(P<0.05)total coliforms in mesenteric lymph nodes on d 21 PI.Pigs in PRO2 had greater(P<0.05)goblet cell number and sulfomucin percentage in duodenal villi and greater(P<0.05)sialomucin percentage in jejunal villi than pigs in CON.Supplementation of PRO1 up-regulated(P<0.05)MUC2 gene expression in jejunal mucosa and reduced(P<0.05)PTGS-2 and IL1B gene expression in ileal mucosa on d 21 PI,compared with CON.Pigs in PRO1 had reduced(P<0.05)relative abundance of families Lachnospiraceae,Peptostreptococcaceae and Pasteurellaceae in the ileum.Conclusions:Supplementation of Bacillus subtilis DSM 32540 improved growth performance,alleviated diarrhea severity,enhanced gut health,and reduced systemic inflammation of weaned pigs infected with ETEC F18.Although Bacillus pumilus DSM 32539 was able to alleviate systemic inflammation,it had limited impacts on growth performance and severity of diarrhea of ETEC F18 challenged weaned pigs.
基金Sponsored by the National High Technology Research and Development Program of China(Grant No.2009AA062906)the"Twelfth Five-Year Plan"National Key Technology R&D Program of China(Grant No.2012BAD14B06-04)+2 种基金the National Natural Science Foundation of China(Grant No.51478140&51408200)State Key Laboratory of Urban Water Resource and Environment(Harbin Institute of Technology)(Grant No.2015DX06)Promising Youngsters Training Program of Heilongjiang University of Science and Technology(Grant No.Q20120201)
文摘A compound bioflocculant CBF,produced by mixed culture of Rhizobium radiobacter F2 and Bacillus sphaeicus F6, was investigated with regard to its production and flocculating properties. The optimization of the culture medium constituents including carbon source,nitrogen source and C / N ratio,metal ions and ionic strength on CBF production were studied. Flocculating properties of CBF were examined by a series of experiments and CBF had good flocculating activities in kaolin suspension with divalent cations and stable over wide range of p H. Studies of the flocculating properties revealed that the flocculation could be stimulated by cations Ca2+,Mg2+,Fe2+,Al3+and Fe3+. In addition,it was stable at 4-30 ℃ in the presence of Ca Cl2. It was found to be effective for flocculation of a kaolin suspension under neutral and weak alkaline conditions( p H 7. 0- 9. 0),and flocculating activities of higher than 95% were obtained when the CBF concentrations among 6- 14 mg / L at p H 8. 0. The results of this study indicate that CBF is a potential replacement of conventional synthetic flocculants and is widely applied in water treatment and downstream processing of food and fermentation industries.
基金Supported by Natural Science Foundation of Hebei Province( C2011201003)
文摘[ Objective] The paper was to screen the antagonistic Bacillus spp. strains against Phytophthora infestans （Mont.） de Bary and analyze its lipepeptide substances. [ Methods] Using flat isolation and culture method, the bacteria that could produce Bacillus spp. were isolated from rhizosphere soil, and carried out primary and secondary screening by confrontation culture. The lipopeptide substances of Y-3 strain with better antifungal effect were carried out I-IPLC analysis. [Result] Nine strains of bacteria had certain antagonistic effects against P. infestans in primary screening, Y-1 and Y-3 strains had significant inhibition effect against the growth of P. infestans in secondary screening, with inhibition rate higher than 80% ; the crude extract of lipepeptide produced by Y-3 strain had better antagonistic effect against P. infestans, with inhibition rate higher than 90%. HPLC analysis showed that the crude extract of lipepeptide mainly contained 8 components. [ Conclusion] The lipopeptide substances produced by Y-3 strain had significant inhibition effect, which had potential application value for biological control of potato late blight.
文摘The present study was taken up with a view to ascertain the possibility of introduction of alcohol resistant bacteria in vitro through the aseptically raised watermelon (Citrullus lanatus) seedlings in the backdrop of isolating such organisms from micropropagated watermelon stocks. Watermelon cv. Arka Manik seedlings grown in vitro from surface-sterilized seeds with the intact seed coat on MS medium appeared visibly clean largely, but upon subjecting them to tissue-indexing, the segments from the collar or root tissue showed bacterial colony growth on Nutrient Agar (NA) from 72% of such healthy seedlings and the cotyledon and hypocotyl tissue of 44% seedlings. The pooled colony growth from NA upon challenge with 90% alcohol yielded 10 distinct colony types, identified as B. pumilus (4×), B. subtilis (4×), B. cereus (1×) or B. safensis (1×) based on partial 16S rRNA sequence analysis. The shoot-tip tissue from the healthy index-negative seedlings cultured on watermelon proliferation medium partly turned index-positive within 2 - 4 sub-culture cycles while being apparently clean. On the other hand, those from the previously index-positive cultures tended to show obvious bacterial growth during subsequent in vitro culturing. The observations suggested the possibility of introduction of spore-forming Bacillus spp. through surface-sterilized seeds, their gradual emergence in vitro in visibly clean seedlings, possible transmittal of spores to the alcohol through tissue-culturing tools and the survival therein with the chances of unsuspected lateral spread. Seed coat removal followed by surface sterilization with sodium hypochlorite facilitated the raising of clean seedlings with no detectable bacterial association.
文摘Volatile organic compounds (VOCs) are harmful to human health and the environment. Recently, loess (Hwangtoh) was used as an eco-friendly interior paint formulation in Korea. It is used even more commonly as a filter carrier to remove VOCs. In this study, we isolated Bacillus strains from a loess filter. The strains that were tolerant to VOCs were labeled according to the series VOC01 to VOC35. Four strains—VOC03, VOC11, VOC18, and VOC30—were investigated for their ability to degrade cyclohexane and toluene. Strain VOC18 best degraded both VOCs, whereas VOC03 demonstrated no ability to degrade VOCs. In keeping with this, VOC18 grew best on cyclohexane or toluene as the sole carbon source. The strains were identified by their physiochemical and phylogenetic characteristics. Strain VOC18 was determined as a strain of Bacillus cereus;VOC11 and VOC30 were determined as differentiated strains of B. thuringiensis. Strain VOC03, which demonstrated high tolerance but no ability to degrade VOCs, was identified as a strain of B. megaterium.
文摘Phytophthora species are particularly aggressive plant pathogens and are often associated with the decline of many tree species, including oak and beech. Several fungi and bacteria species are known as potential antagonists usable as biological control agents. Phosphonate （H3PO3）, commonly branded as phosphite, has also been used in the past years to protect trees against invasive Phytophthora spp.. This study aimed at comparing the effects of selected antagonist microorganisms and phosphonate, when applied by microinjection or leaf treatment. Antagonistic species were first selected for their high inhibitory activity against problematic Phytophthora species, such as Phytophthora cactorum, P. quercina and P. plurivora attacking Quercus robur and Fagus sylvatica in Polish forests. Three endophytic species Trichoderma atroviride （two strains）, T. harzianum and Bacillus amyloliquefaciens showed a high control activity, and their efficacy was then assessed in comparison with a phosphonate treatment. Two application methods were experimented in this study： injection of a solution of spores or phosphonate into the sap vessels of beech or a foliar treatment on oak. Phosphonate and two strains of Trichoderma significantly reduced the necrotic area on oak leaves inoculated with P. plurivora and one strain of T. atroviride significantly reduced necrotic areas on beech branches. These results are therefore promising of a novel way to control Phytophthora spp. in forest stands and nurseries.
