Characterization of Streptococcus bovis from the rumen of the dromedary camel and Rusa deer

Aims: Isolation and characterization of Streptococcus bovis from the dromedary camel and Rusa deer. Methods and Results: Bacteria were isolated from the rumen contents of four camels and two deer fed lucerne hay by culturing on the semi-selective medium MRS agar. Based on Gram morphology and RFLP analysis seven isolates, MPR1, MPR2, MPR3, MPR4, MPR5, RD09 and RD11 were selected and putatively identified as Streptococcus. The identity of these isolates was later confirmed by comparative DNA sequence analysis of the 16S rRNA gene with the homologous sequence from S. bovis strains, JB1, C14b1, NCFB2476, SbR1, SbR7 and Sb5, from cattle and sheep, and the Streptococcus equinus strain NCD01037T. The percentage similarity amongst all strains was >99%, confirming the identification of the camel isolates as S. bovis. The strains were further characterized by their ability to utilize a range of carbohydrates, the production of volatile fatty acids (VFA) and lactate and the determination of the doubling time in basal medium 10 supplemented with glucose. All the isolates produced L-lactate as a major fermentation end product, while four of five camel isolates produced VFA. The range of carbohydrates utilized by all the strains tested, including those from cattle and sheep were identical, except that all camel isolates and the deer isolate RD11 were additionally able to utilize arabinose. Conclusions: Streptococcus bovis was successfully isolated from the rumen of camels and deer, and shown by molecular and biochemical characterization to be almost identical to S. bovis isolates from cattle and sheep. Significance and Impact of the Study: Streptococcus bovis is considered a key lactic acid producing bacterium from the gastrointestinal tract of ruminants, and has been implicated as a causative agent of lactic acidosis. This study is the first report of the isolation and characterization of S. bovis from the dromedary camel and Rusa deer, and suggests a major contributive role of this bacterium to fermentative acidosis.

Stable production of hyaluronic acid in Streptococcus zooppidemicus chemostats operated at high dilution rate

Hyaluronic acid is routinely produced through fermentation of both Group A and C streptococci. Despite significant production costs associated with short fermentations and removal of contaminating proteins released during entry into stationary phase, hyaluronic acid is typically produced in batch rather than continuous culture. The main reason is that hyaluronic acid synthesis has been found to be unstable in continuous culture except at very low dilution rates. Here, we investigated the mechanisms underlying this instability and developed a stable, high dilution rate (0.4 h(-1)) chemostat process for both chemically defined and complex media operating for more than 150 h of production. In chemically defined medium, the product yield was 25% higher in chemostat cultures than in conventional batch culture when arginine or glucose was the limiting substrate. In contrast, glutamine limitation resulted in higher ATP requirements and a yield similar to that observed in batch culture. In complex, glucose-limited medium, ATP requirements were greatly reduced but biomass synthesis was favored over hyaluronic acid and no improvement in hyaluronic acid yield was observed. The successful establishment of continuous culture at high dilution rate enables both commercial production at reduced cost and a more rational characterization and optimization of hyaluronic acid production in streptococci. (c) 2005 Wiley Periodicals, Inc.

Comparison of gelation profile of yoghurts during fermentation measured by RVA and ultrasonic spectroscopy

The gelation profile of yoghurts from conventionally treated (85 degrees C/30 min) and UHT treated (143 degrees C/6s) milks at 16, 18, and 20% total solids was analyzed during fermentation for 4 hrs using the invasive Rapid Visco Analyzer (RVA) and the non-invasive ultrasonic spectroscope. The viscosity measured by the RVA and the ultrasonic velocity measured by the ultrasonic spectroscope exhibited similar sigmoid trends with respect to fermentation time. The ultrasonic spectroscope detected the onset of gelation of yoghurt milk earlier (by an average of 52 min) than did the RVA, indicating a higher sensitivity of ultrasonic spectroscopy. The delay of gelation time of UHT-treated yoghurt milk as compared to conventionally treated yoghurt milk was detected by both techniques. A non-significant ( P > 0.05) effect of solids content in the yoghurt milks on their gelation time was also observed by both instruments.

Alkali hydroxide-induced gelation of pectin

The mechanism of pectin gelation depends on the degree of methoxylation. High methoxyl pectin gels due to hydrophobic interactions and hydrogen bonding between pectin molecules. Low methoxyl pectin forms gels in the presence of di- and polyvalent cations which cross link and neutralise the negative charges of the pectin molecule. Monovalent cations normally do not lead to gel formation with high methoxyl pectin solutions free of divalent cations, especially Ca. The present study found that alkali (NaOH or KOH) added to high methoxyl pectin leads to gel formation in a concentration-depended manner. It was also found that monovalent cations (Na and K) induce gelation of low methoxyl pectin and the time required for gel formation (setting time) depends on the cation concentration. The results indicate that a combined char-e neutralisation and ionic strength effect is responsible for the monovalent cation-induced gelation of pectin. (C) 2003 Elsevier Ltd. All rights reserved.

Concurrent microscopic observations and activity measurements of cellulose hydrolyzing and methanogenic populations during the batch anaerobic digestion of crystalline cellulose

This study compares process data with microscopic observations from an anaerobic digestion of organic particles. As the first part of the study, this article presents detailed observations of microbial biofilm architecture and structure in a 1.25-L batch digester where all particles are of an equal age. Microcrystalline cellulose was used as the sole carbon and energy source. The digestions were inoculated with either leachate from a 220-Lanaerobic municipal solid waste digester or strained rumen contents from a fistulated cow. The hydrolysis rate, when normalized by the amount of cellulose remaining in the reactor, was found to reach a constant value 1 day after inoculation with rumen fluid, and 3 days after inoculating with digester leachate. A constant value of a mass specific hydrolysis rate is argued to represent full colonization of the cellulose surface and first-order kinetics only apply after this point. Additionally, the first-order hydrolysis rate constant, once surfaces were saturated with biofilm, was found to be two times higher with a rumen inoculum, compared to a digester leachate inoculum. Images generated by fluorescence in situ hybridization (FISH) probing and confocal laser scanning microscopy show that the microbial communities involved in the anaerobic biodegradation process exist entirely within the biofilm. For the reactor conditions used in these experiments, the predominant methanogens exist in ball-shaped colonies within the biofilm. (C) 2005 Wiley Periodicals, Inc.

Properties of a unique mutant of Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus that exhibits a partial many polyhedra and few polyhedra phenotype on extended serial passaging in suspension cell cultures

Serial passaging of wild-type Helicoverpa armigera, single-nucleocapsid (HaSNPV) in H. zea (HzAMI) illsect Cell Cultures results ill rapid selection for the few polyhedra (FP) phenotype. A unique HaSNPV mutant (ppC19) was isolated through plaque purification that exhibited a partial many polyhedra (MP) and FP phenotype. Oil serial passaging in suspension cell cultures, ppC19 produced fivefold more polyhedra than a typical FP mutant (FP8AS) but threefold less polyhedra than the wild-type virus. Most importantly, the polyhedra of ppC19 exhibited MP-like virion occlusion. Furthermore, ppC19 produced the same amount of budded virus (BV) as the FP mutant, which was fivefold higher than that of the wild-type virus. This selective advantage was likely to explain its relative stability in polyhedra production for six passages when compared with the wild-type Virus. However, subsequent passaging of ppC19 resulted in a steel) decline in both BV and polyhedra yields, which was also experienced by FP8AS and the wild-type virus Lit high passage numbers. Genomic deoxyribonueleic Licid profiling of the latter suggested that defective interfering particles (DIPS) were implicated in this phenomenon and represented another Undesirable mutation during serial passaging of HaSNPV Hence, a strategy to isolate HaSNPV Clones that exhibited MP-like polyhedra production but FP-like BV production, coupled with low multiplicities of infection during scale-up to avoid accumulation of DIPS, could prove commerically invaluable.

Comparison of cellulose solubilisation rates in rumen and landfill leachate inoculated reactors

The aim of this study was to conduct a number of controlled digestions to obtain easily comparable cellulose solubilisation rates and to compare these rates to those found in the literature to see which operational differences were significant in affecting cellulose degradation during anaerobic digestion. The results suggested that differences in volumetric cellulose solubilisation rates were not indicative of the true performance of cellulose digestion systems. When cellulose solubilisation rates were normalised by the mass of cellulose in the reactor at each time step, the comparison of the rates became more meaningful. Cellulose solubilisation was surface area limited. Therefore, changes in the loading rate of cellulose to the reactor altered the volumetric solubilisation rate without changing the mass normalised rate. Comparison of mass normalised solubilisation rates from this study and the literature demonstrated that differences in reactor configuration and operational conditions did not significantly impact on the solubilisation rate whereas the difference in composition of the microbial communities showed a marked effect. This work highlights the importance of using appropriately normalised data when making comparisons between systems with differing operational conditions. (c) 2005 Elsevier Ltd. All rights reserved.

Lasers - An effective artificial source of radiation for the cultivation of anoxygenic photosynthetic bacteria

The laser diode (LD) is a unique light source that can efficiently produce all radiant energy within the narrow wavelength range used most effectively by a photosynthetic microorganism. We have investigated the use of a single type of LID for the cultivation of the well-studied anoxygenic photosynthetic bacterium, Rhodobacter capsulatus (Rb. capsulatus). An array of vertical-cavity surface-emitting lasers (VCSELs) was driven with a current of 25 mA, and delivered radiation at 860 nm with 0.4 nm linewidth. The emitted light was found to be a suitable source of radiant energy for the cultivation of Rb. capsulatus. The dependence of growth rate on incident irradiance was quantified. Despite the unusual nearly monochromatic light source used in these experiments, no significant changes in the pigment composition and in the distribution of bacteriochlorophyll between LHII and LHI-RC were detected in bacterial cells transferred from incandescent light to laser light. We were also able to show that to achieve a given growth rate in a light-limited culture, the VCSEL required only 30% of the electricity needed by an incandescent bulb, which is of great significance for the potential use of laser-devices in biotechnological applications and photobioreactor construction. (c) 2006 Wiley Periodicals, Inc.