Antimicrobial activity of different proteins and their fragments from Bacillus thuringiensis parasporal crystals against clostridia and archaea

Yudina, T.G.; Bryukhanov, A. L.; Zalunin, I. A.; Revina, L. P.; Шестаков, А. И.; Voyushina, Nina E.; Chestukhina, G. G.; Netrusov, A. I.

doi:10.1016/j.anaerobe.2006.09.006

Cited by 20 publications

(17 citation statements)

References 20 publications

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“…Similarly, another in vitro study found that the active form of the Cry1Ab protein had no effect on a range of Grampositive and -negative bacterial strains (27). However, the concentration of Cry1Ab protein detected in the cecal digesta of pigs in the present study was 2.41 ng/ml, which was 90 times lower than that in the feed (55) and ϳ4,000 times lower than the concentrations used by Koskella and Stotzky (27) and Yudina et al (57) in their in vitro studies. This may account for the lack of an antibacterial response within the intestinal microbiota on feeding Bt maize.…”

Section: Discussioncontrasting

confidence: 50%

“…in vitro (57), the present study did not reveal any anticlostridial effects within the porcine cecum on administration of the Cry1Ab-containing Bt maize. Similarly, another in vitro study found that the active form of the Cry1Ab protein had no effect on a range of Grampositive and -negative bacterial strains (27).…”

Section: Discussionmentioning

confidence: 67%

“…An in vitro study has demonstrated antimicrobial activity of the Cry1Ab protein in both intact (130-kDa) and fragmented (ϳ60-kDa) forms against Clostridium butyricum, Clostridium acetobutylicum, and Methanosarcina barkeri (57). However, conflicting data were obtained by Koskella and Stotzky (27), who failed to observe activity in vitro against a range of Gram-positive and -negative bacteria, namely, Proteus spp., Pseudomonas aeruginosa, Enterobacter spp., Escherichia coli, Klebsiella, Agrobacterium radiobacter, Arthrobacter globiformis, and Bacillus spp.…”

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confidence: 78%

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High-Throughput Sequence-Based Analysis of the Intestinal Microbiota of Weanling Pigs Fed Genetically Modified MON810 Maize Expressing Bacillus thuringiensis Cry1Ab (Bt Maize) for 31 Days

Buzoianu

Walsh

Rea

et al. 2012

Appl Environ Microbiol

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The objective of this study was to investigate if feeding genetically modified (GM) MON810 maize expressing the Bacillus thuringiensis insecticidal protein (Bt maize) had any effects on the porcine intestinal microbiota. Eighteen pigs were weaned at ϳ28 days and, following a 6-day acclimatization period, were assigned to diets containing either GM (Bt MON810) maize or non-GM isogenic parent line maize for 31 days (n ‫؍‬ 9/treatment). Effects on the porcine intestinal microbiota were assessed through culture-dependent and -independent approaches. Fecal, cecal, and ileal counts of total anaerobes, Enterobacteriaceae, and Lactobacillus were not significantly different between pigs fed the isogenic or Bt maize-based diets. Furthermore, high-throughput 16S rRNA gene sequencing revealed few differences in the compositions of the cecal microbiotas. The only differences were that pigs fed the Bt maize diet had higher cecal abundance of Enterococcaceae (0.06 versus 0%; P < 0.05), Erysipelotrichaceae (1.28 versus 1.17%; P < 0.05), and Bifidobacterium (0.04 versus 0%; P < 0.05) and lower abundance of Blautia (0.23 versus 0.40%; P < 0.05) than pigs fed the isogenic maize diet. A lower enzyme-resistant starch content in the Bt maize, which is most likely a result of normal variation and not due to the genetic modification, may account for some of the differences observed within the cecal microbiotas. These results indicate that Bt maize is well tolerated by the porcine intestinal microbiota and provide additional data for safety assessment of Bt maize. Furthermore, these data can potentially be extrapolated to humans, considering the suitability of pigs as a human model.

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Section: Discussioncontrasting

confidence: 50%

Section: Discussionmentioning

confidence: 67%

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confidence: 78%

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High-Throughput Sequence-Based Analysis of the Intestinal Microbiota of Weanling Pigs Fed Genetically Modified MON810 Maize Expressing Bacillus thuringiensis Cry1Ab (Bt Maize) for 31 Days

Buzoianu

Walsh

Rea

et al. 2012

Appl Environ Microbiol

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“…Moreover, we were the first to employ highthroughput 16S rRNA gene sequencing to determine if consumption of GM food/feed influences intestinal microbial communities. Such studies are warranted, considering in vitro observations that the Cry1Ab protein is antimicrobial against intestinal bacteria, such as Clostridium, both in its intact and fragmented forms, at concentrations of 25 to 63 g/ml (15). Furthermore, the Cry1Ab protein is not completely degraded during intestinal transit when administered in feed at concentrations of 0.17 to 0.52 g/g feed and persists in the intestine at concentrations of 0.003 to 0.03 g/ml (8,9,16).…”

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confidence: 99%

Sequence-Based Analysis of the Intestinal Microbiota of Sows and Their Offspring Fed Genetically Modified Maize Expressing a Truncated Form of Bacillus thuringiensis Cry1Ab Protein (Bt Maize)

Buzoianu

Walsh

Rea

et al. 2013

Appl Environ Microbiol

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eThe aim was to investigate transgenerational effects of feeding genetically modified (GM) maize expressing a truncated form of Bacillus thuringiensis Cry1Ab protein (Bt maize) to sows and their offspring on maternal and offspring intestinal microbiota. Sows were assigned to either non-GM or GM maize dietary treatments during gestation and lactation. At weaning, offspring were assigned within sow treatment to non-GM or GM maize diets for 115 days, as follows: (i) non-GM maize-fed sow/non-GM maize-fed offspring (non-GM/non-GM), (ii) non-GM maize-fed sow/GM maize-fed offspring (non-GM/GM), (iii) GM maize-fed sow/non-GM maize-fed offspring (GM/non-GM), and (iv) GM maize-fed sow/GM maize-fed offspring (GM/GM). Offspring of GM maize-fed sows had higher counts of fecal total anaerobes and Enterobacteriaceae at days 70 and 100 postweaning, respectively. At day 115 postweaning, GM/non-GM offspring had lower ileal Enterobacteriaceae counts than non-GM/non-GM or GM/GM offspring and lower ileal total anaerobes than pigs on the other treatments. GM maize-fed offspring also had higher ileal total anaerobe counts than non-GM maize-fed offspring, and cecal total anaerobes were lower in non-GM/GM and GM/ non-GM offspring than in those from the non-GM/non-GM treatment. The only differences observed for major bacterial phyla using 16S rRNA gene sequencing were that fecal Proteobacteria were less abundant in GM maize-fed sows prior to farrowing and in offspring at weaning, with fecal Firmicutes more abundant in offspring. While other differences occurred, they were not observed consistently in offspring, were mostly encountered for low-abundance, low-frequency bacterial taxa, and were not associated with pathology. Therefore, their biological relevance is questionable. This confirms the lack of adverse effects of GM maize on the intestinal microbiota of pigs, even following transgenerational consumption.

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“…The effect of exogenous Cyt1Aa on Gram-negative bacteria has not been reported previously. Several d-endotoxins from B. thuringiensis have antibacterial effects on Gram-positive bacteria as well as on archaea, detected by zones of inhibition on agar and morphological changes in the treated bacteria (Revina et al, 2005;Yudina et al, 2003Yudina et al, , 2007. Here, we examined the effect of exogenous Cyt1Aa on both Gram-positive and Gram-negative bacteria.…”

Section: Introductionmentioning

confidence: 99%

Antibacterial activity of Cyt1Aa from Bacillus thuringiensis subsp. israelensis

2008

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Cyt1Aa is a d-endotoxin protein that is produced by Bacillus thuringiensis subsp. israelensis. It is a membrane pore-forming toxin that is lethal to insect larvae and is broadly cytolytic to vertebrate as well as invertebrate cells. Cyt1Aa is produced as a protoxin of 27 kDa. Proteolytic activation results in a reduction of the molecular mass to approximately 23-24 kDa and a threefold increase in activity. In this research, Cyt1Aa crystals were purified from B. thuringiensis IPS78/11 harbouring the expression vector pHT-cyAp20. The activity of the activated form of Cyt1Aa (23-24 kDa) was examined on a pathogenic strain of the Gram-negative Escherichia coli and the Gram-positive species Staphylococcus aureus. The Cyt1Aa minimal inhibitory concentration for E. coli and S. aureus was 1.25 and 5 mg ml "1 , respectively. Cyt1Aa was found to be bactericidal for E. coli, whereas it was bacteriostatic for S. aureus. Furthermore, Cyt1Aa increased the lethal effect when acting in combination with antibiotics. The association of Cyt1Aa with cells of these two bacteria was demonstrated by Western blot analysis using antibodies against the whole d-endotoxin crystal. Scanning electron microscopy displayed damage to Cyt1Aa-treated cells. Ion imbalance due to damage of the cell walls and membranes was confirmed by X-ray microanalysis. These experiments show that Cyt1Aa has an antibacterial effect on pathogenic species and demonstrate, apparently for the first time, that exogenous Cyt1Aa has a bactericidal effect upon Gram-negative bacteria.

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Antimicrobial activity of different proteins and their fragments from Bacillus thuringiensis parasporal crystals against clostridia and archaea

Cited by 20 publications

References 20 publications

High-Throughput Sequence-Based Analysis of the Intestinal Microbiota of Weanling Pigs Fed Genetically Modified MON810 Maize Expressing Bacillus thuringiensis Cry1Ab (Bt Maize) for 31 Days

High-Throughput Sequence-Based Analysis of the Intestinal Microbiota of Weanling Pigs Fed Genetically Modified MON810 Maize Expressing Bacillus thuringiensis Cry1Ab (Bt Maize) for 31 Days

Sequence-Based Analysis of the Intestinal Microbiota of Sows and Their Offspring Fed Genetically Modified Maize Expressing a Truncated Form of Bacillus thuringiensis Cry1Ab Protein (Bt Maize)

Antibacterial activity of Cyt1Aa from Bacillus thuringiensis subsp. israelensis

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