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PubMed2006

Geobacillus toebii subsp. decanicus subsp. nov., a hydrocarbon-degrading, heavy metal resistant bacterium from hot compost.

A thermophilic, spore-forming bacterial strain L1(T) was isolated from hot compost "Pomigliano Environment" s.p.a., Pomigliano, Naples, Italy. The strain was identified by using a polyphasic taxonomic approach. L1(T) resulted in an aerobic, gram-positive, rod-shaped, thermophilic with an optimum growth temperature of 68 degrees C chemorganotrophic bacterium which grew on hydrocarbons as unique carbon and energy sources and was resistant to heavy metals. The G+C DNA content was 43.5 mol%. Phylogenetic analysis of 16S rRNA gene sequence and Random Amplified Polymorphic DNA-PCR (RAPD-PCR) analysis of L1(T) and related strains showed that it forms within Geobacillus toebii, a separate cluster in the Geobacillus genus. The composition of cellular fatty acids analyses by Gas-Mass Spectroscopy differed from that typical for the genus Geobacillus in that it is lacking in iso-C15 fatty acid, while iso-C16 and iso-C17 were predominant. Isolates grew on a rich complex medium at temperatures between 55-75 degrees C and presented a doubling time (t(d)) of 2 h and 6 h using complex media and hydrocarbon media, respectively. Among hydrocarbons tested, n-decane (2%) was the more effective to support the growth (1 g/L of wet cells). The microorganism showed resistance to heavy metal tested during the growth. Furthermore, intracellular alpha-galactosidase and alpha-glucosidase enzymatic activities were detectable in the L1(T) strain. Based on phenotypic, phylogenetic, fatty acid analysis and results from DNA-DNA hybridization, we propose assigning a novel subspecies of Geobacillus toebii, to be named Geobacillus toebii subsp. decanicus subsp. nov., with the type strain L1(T) (=DSM 17041=ATCC BAA 1004).

Bacillaceae↗
PubMed2000

Effects of intramammary infection and parity on calf weaning weight and milk quality in beef cows.

The objectives of this study were to determine 1) the effect of intramammary infection on calf weaning weight, milk somatic cell count, and milk composition, and 2) the effect of parity on percentages of infected cows, infected quarters, and blind quarters. The number of infected quarters, milk somatic cell counts, milk components, and intramammary infection were studied at weaning in 164 beef cows. The percentage of infected cows ranged from 61.9% at first parity to 66.7% at fifth to ninth parities. Cows with three or four infected quarters had higher (P < .01) milk somatic cell counts than cows with zero, one, or two infected quarters. Among bacterial isolates, Staphylococcus aureus-infected quarters had the highest (P < .01) milk somatic cell count. Percentages of butterfat and lactose were lower (P < .01) in milk from infected quarters than from uninfected quarters. Infections by S. aureus and coagulase-negative staphylococci were the most common and accounted for 67 to 78% of the infections. Percentages of infected quarters and infections caused by S. aureus increased with parity (P < .01). Intramammary infections did not affect (P > .10) calf weaning weight. In conclusion, intramammary infection had no effect on calf weaning weight but increased milk somatic cell count and decreased the percentage of protein, lactose, solids-not-fat, and butterfat. The number of infected and blind mammary quarters increased with parity.

Animals↗
PubMed1997

Bacillus cereus necrotizing cellulitis mimicking clostridial myonecrosis: case report and review of the literature.

We describe a case of rapidly progressive necrotizing cellulitis in an immunocompromised farmer caused by Bacillus cereus, and review 15 additional cases of serious soft tissue infection due to this organism reported in the English language literature. These cases illustrate the potential for B. cereus to cause fulminant soft tissue disease indistinguishable from that caused by clostridia.

Bacillaceae Infections↗
PubMed1997

Effect of storage temperatures and ingredients on growth of Bacillus cereus in coffee creamers.

Growth of Bacillus cereus ATCC 33018 was evaluated in half and half (10.5% fat), whipping cream (30% fat), and nondairy creamer (7.5% fat). Samples were inoculated with approximately 10 vegetative cells/ml or 100 spores/ml and were subsequently stored at 4, 7, 23 and 32 degrees C. Within 9 h at 32 degrees C and 11 h at 23 degrees C, in both half and half and whipping cream, vegetative cells and spores reached population levels that can cause foodborne illness. No growth occurred in any product stored at 4 or 7 degrees C. Sodium stearoyl lactylate, a fatty acid derivative that is used as an emulsifier, inhibited growth of spores and vegetative cells in the nondairy creamers stored at either 32 or 23 degrees C.

Bacillaceae Infections↗
PubMed2005

Prevalence of enterotoxigenic Bacillus cereus and its enterotoxins in milk and milk products in and around Nairobi.

OBJECTIVES: To determine the prevalence of enterotoxigenic Bacillus cereus (B. cereus) and enterotoxins in milk and milk products. DESIGN: A random sampling of milk products was carried out. SETTING: Market milk and milk products were collected from retail shops in Nairobi and analysed for contamination with enterotoxigenic B. cereus and its enterotoxins using reverse passive latex agglutination and TECRA ELISA immunoassay tests. SUBJECTS: Ninety six milk samples including 36 raw milk, 42 pasteurised milk, 10 yogurt and eight fermented milk samples. Forty seven Bacillus cereus isolated from milk and milk products. MAIN OUTCOME MEASURES: Isolation of enterotoxigenic B. cereus from milk and milk products and detection of B. cereus hemolytic (hemolysin BL) and non-hemolytic enterotoxins in milk. RESULTS: Fifty seven percent of the samples were contaminated with B. cereus. Eighty one percent (38 out of 47) of the isolates produced non-hemolytic enterotoxins, while 25 (53.2%) of the isolates produced hemolysin BL. Eighteen (38.3%) of the isolates produced both hemolysin BL and non-hemolytic enterotoxins. About fourteen percent (14.3%) of the pasteurised milk samples tested positive for non-hemolytic enterotoxin. CONCLUSION: Enterotoxigenic B. cereus and enterotoxins occur in market milk and their presence poses a potential risk of causing food poisoning. The risk can be reduced if milk products undergo thorough quality control checks and are always kept at below 4 degrees C till consumption. Post pastuerisation contamination which is commonly blamed for spoilage of milk products by B. cereus is not necessarily the most important source of this organism.

Animals↗
PubMedDate not supplied

Neonatal intensive care unit: reservoirs of nosocomial pathogens.

Improvement in the care and treatment of neonates had contributed to their increased survival. Nosocomial infection remains an important problem in intensive care units. Hospital wards had been shown to act as reservoirs of pathogenic microorganisms associated with infection. To assess the prevalence of pathogenic organisms in the environment of the neonatal unit, 92 swabs were randomly collected from cots, incubators and various equipments in the unit and were cultured on Blood agar and MacConkey agar plates. Air contamination was detected by exposing the same types of agar plates for 3 hours in several areas of the unit. After 48 hours incubation, isolates were identified biochemically. There is marked congestion in the unit. Ninety one percent of swabs yielded growth, with coagulase negative Staphylococcus being the predominant organism (44%), followed by Bacillus species (20%), E. coli (12.5%), and Klebsiella (8.5%), Pseudomonas species (3.6%) and moulds (3.6%). Sedimentation plates had colony counts of from 10 - 100 per plate and the majority of the cultures were polymicrobial cultures. The presence of various Gram-negative bacili including known neonatal pathogens (like E. Coli and Pseudomonas) especially on ward equipment and congestion in the ward has the potential to cause nosocomial infection.

Air Microbiology↗
PubMed1998

Heat resistance of Alicyclobacillus acidoterrestris spores as affected by various pH values and organic acids.

Alicyclobacillus acidoterrestris, a thermoacidophilic sporeformer, has caused spoilage of fruit juices which had been treated with thermal processes intended to commercially sterilize the juice. The objective of this research was to document the effect of pH, acid, and temperature on the heat resistance of spores of three fruit-juice isolates of A. acidoterrestris. The thermal resistance of spores of A. acidoterrestris strains VF, WAC, and IP were studied in a model fruit-juice system composed of 12% glucose and 30 mM of either citric, malic, or tartaric acid, adjusted to selected pH values ranging from 2.8 to 4.0. Decimal reduction times (D values) and inactivation rates were determined. Spores of strains VF and WAC were similarly resistant to heat under acidic conditions, while strain IP spores were less resistant. In the range of pH 2.8 to 4.0, a statistically effect of hydrogen ion concentration on heat resistance was observed at lower temperatures, but not at the higher temperatures, but not at the higher temperatures. For examples, at 91 degrees C and pH 3.1 and 3.7, D values were 31.3 and 54.3 min, respectively, while at 97 degrees C D values at pH 3.1 and 3.7 were 7.9 and 8.8 min, respectively. The type of acid did not significantly affect the heat resistance. The zd values ranged from 5.9 to 10 degrees C, depending on the acid, pH, and the strain. The models generated from this research can be used to determine adequate thermal processes, accounting for the acid type, pH, and temperature, to destroy A. acidoterrestris spores in beverages, since this organism is able to survive the typical hot-fill and hold process (2 min at 88 to 96 degrees C) currently used to process fruit juice.

Bacillaceae↗
PubMed1999

Alicyclobacillus in orange juice: occurrence and heat resistance of spores.

Spore suspensions of a pure culture of Alicyclobacillus acidoterrestris DSM 2498 were submitted to different heat treatments (60 degrees C for 60 min, 60 degrees C for 30 min, 70 degrees C for 20 min, 80 degrees C for 5 min, 80 degrees C for 10 min, 80 degrees C for 30 min, and boiling for 5 min) to determine the best activation conditions in orange juice. The best treatment for spore activation was shown to be 70 degrees C/20 min. Seventy-five samples of concentrated orange juice from 11 different suppliers were examined for the presence of thermophilic acid-tolerant spore formers by the most probable number technique using Bacillus acidocaldarius medium (BAM broth) and incubation at 44 degrees C for 5 days after a prior spore activation. After incubation, isolation was carried out using BAM agar medium incubating at 44 degrees C for 5 days. Typical colonies were submitted to a microscopic examination, evaluation for the presence of spores, and various biochemical tests. Of the orange juice samples examined, 14.7% were found to be positive for Alicyclobacillus. The thermal death time open tube method was used to determine the heat resistance of the spores of strains confirmed as being Alicyclobacillus. The D-values determined were in the range from 60.8 to 94.5 min at 85 degrees C, 10.0 to 20.6 min at 90 degrees C, and 2.5 to 8.7 min at 95 degrees C. The z-values were between 7.2 degrees C and 11.3 degrees C. The results demonstrated the occurrence of Alicyclobacillus in orange juice and the high heat resistance of the spores that could survive the heat treatments normally applied in the processing of orange juice.

Bacillaceae↗
PubMed2000

Heat resistance of Alicyclobacillus acidocaldarius in water, various buffers, and orange juice.

The effect of the pH or the composition of the heating medium and of the sporulation temperature on the heat resistance of spores of a thermoacidophilic spore-forming microorganism isolated from a dairy beverage containing orange fruit concentrate was investigated. The species was identified as Alicyclobacillus acidocaldarius. The spores showed the same heat resistance in citrate-phosphate buffers of pH 4 and 7, in distilled water, and in orange juice at any of the temperatures tested (D120 degrees C = 0.1 min and z = 7 degrees C). A raise in 20 degrees C in the sporulation temperature (from 45 to 65 degrees C) increased the heat resistance eightfold (from D110 degrees C = 0.48 min when sporulated at 45 degrees C to 3.9 min when sporulated at 65 degrees C). The z-values remained constant for all sporulation temperatures. The spores of this strain of A. acidocaldarius were very heat resistant and could easily survive any heat treatment currently applied to pasteurize fruit juices.

Bacillaceae↗
PubMed2000

Efficacy of disinfectants in killing spores of Alicyclobacillus acidoterrestris and performance of media for supporting colony development by survivors.

Alicyclobacillus has recently emerged as a spoilage microorganism of concern in a wide range of pasteurized fruit products. The focus of this study was to determine the efficacy of chemical disinfectants in killing Alicyclobacillus acidoterrestris spores. Direct plating media were evaluated for their suitability to support germination and outgrowth of spores surviving exposure to these disinfectants. Significant (P < or = 0.05) reductions of about 2.2, 0.4, and 0.1 logs in the number of viable A. acidoterrestris spores in a five-strain mixture were achieved when spores were suspended in 200 ppm chlorine, 500 ppm acidified sodium chlorite, or 0.2% H2O2 solutions, respectively, for 10 min at 23 degrees C. When treated with either 1,000 ppm chlorine or 4% H2O2, the number of spores was reduced by more than 5 logs. Treatment with 8% trisodium phosphate or 80 ppm Tsunami did not significantly reduce numbers of viable spores. Spores of individual strains of A. acidoterrestris varied little in resistance to the same chemical treatment. K agar (pH 3.7) was judged best for recovering chemically treated spores, compared to orange serum agar (pH 5.0) and potato dextrose agar (pH 3.5). Experiments were done to determine the effectiveness of chemical treatments in killing a mixed-strain inoculum of A. acidoterrestris spores on the surface of apples. Treatment with 500 ppm chlorine or 1,200 ppm acidified sodium chlorite for 1 min significantly (P < or = 0.05) reduced the number of viable spores, but reductions were less than 1 log. Hydrogen peroxide (2%) was ineffective in killing spores remaining on the apple skin after treatment.

Agar↗
PubMed2003

Identification of essential histidines in cyclodextrin glycosyltransferase isoform 1 from Paenibacillus sp. A11.

The isoform 1 of cyclodextrin glycosyltransferase (CGTase, EC 2.4.1.19) from Paenibacillus sp. A11 was purified by a preparative gel electrophoresis. The importance of histidine, tryptophan, tyrosine, and carboxylic amino acids for isoform 1 activity is suggested by the modification of the isoform 1 with various group-specific reagents. Activity loss, when incubated with diethylpyrocarbonate (DEP), a histidine modifying reagent, could be protected by adding 25 mM methyl-beta-cyclodextrin substrate prior to the modification. Inactivation kinetics of isoform 1 with DEP resulted in second-order rate constants (k(inactivation)) of 29.5 M(-1)s(-1). The specificity of the DEP-modified reaction for the histidine residue was shown by the correlation between the loss of isoform activity and the increase in the absorbance at 246 nm of N-carbethoxyhistidine. The number of histidines that were modified by DEP in the absence and presence of a protective substrate was estimated from the increase in the absorbance using a specific extinction coefficient of N-carbethoxyhistidine of 3,200 M(-1)cm(-1). It was discovered that methyl-beta-CD protected per mole of isoform 1, two histidine residues from the modification by DEP. To localize essential histidines, the native, the DEP-modified, and the protected forms of isoform 1 were digested by trypsin. The resulting peptides were separated by HPLC. The peptides of interest were those with R(t) 11.34 and 40.93 min. The molecular masses of the two peptides were 5,732 and 2,540 daltons, respectively. When the data from the peptide analysis were checked with the sequence of CGTase, then His-140 and His-327 were identified as essential histidines in the active site of isoform 1.

Amino Acid Sequence↗
PubMed1978

[Restriction of washing and its effect to the normal human skin flora. Quantitative and qualitative investigations of the aerobic skin flora (author's transl)].

The behaviour of the aerobic skin flora of the flexor sides of the forearms, under a three-week restriction of washing, was investigated in twenty-four patients for its quantitative and qualitative aspects. The combined scrabbing-washing method was used as described by Burtenshaw with Ringer's solution. After a three-week restriction of washing, using the Wilcoxon Test, statistically no significant changes in the total number of microorganisms were found in areas investigated in the intermittent sampling of the skin flora. A three-week restriction of washing in the region of the flexor sides of the forearms does not induce significant changes in the composition of the normal aerobic skin flora. The percentage of differences in the total number of bacteria fluctuates up to the mark in wide limits; statements about standard deviations are not permitted because of the technical and experimental conditions. There was no shifting of the Resident flora to other groups of bacteria in the patients under study. Pathogenic microorganisms could not be found in higher numbers after the second sampling.

Actinomycetales↗
PubMed1999

[Bacillary endophthalmitis. Four case reports].

PURPOSE: Bacillary endophthalmitis occurring after penetrating ocular trauma with an intraocular foreign body is always associated with poor visual outcome. Bacilli cause fulminant infection associated with tissue damage in the intraocular structures. CASE REPORTS: Our series consisted of four patients with penetrating ocular trauma and endophthalmitis caused by B. cereus or B. licheniformis. Three eyes ultimately developed phtisis. Only on eye recovered good vision (2.5/10 P4). DISCUSSION: Recommended early treatment includes topical, subconjunctival, parenteral antibiotics. A review of the literature indicates that intravitreal antibiotic infusion is crucial for sufficient concentration to control infection. Early vitrectomy is recommended in the management of endophthalmitis. Vitreous and intraocular foreign bodies should be cultured to identify pathogens and ascertain antibiotic susceptibilities.

Administration, Topical↗