Production and characterization of pureClostridiumspore suspensions

Abstract

Aims: A general protocol was derived for optimizing the production of pure, high concentration Clostridium endospore suspensions. Methods and Results: Two sporulation methods were developed that yielded high concentrations of notably pure Clostridium sporogenes, C. hungatei and C. GSA‐1 (Greenland ice core isolate) spore suspensions (10 ml of 109 spores ml−1 with >99% purity each). Each method was derived by evaluating combinations of three sporulation conditions, including freeze drying of inocula, heat shock treatment of cultures, and subsequent incubation at suboptimal temperatures that yielded the highest percentage of sporulation. Pure spore suspensions were characterized in terms of dipicolinic acid content, culturability, decimal reduction time (D) value for heat inactivation (100°C) and hydrophobicity. Conclusions: While some Clostridium species produce a high percentage of spores with heat shock treatment and suboptimal temperature incubation, other species require the additional step of freeze drying the inocula to achieve a high percentage of sporulation. Significance and Impact of the Study: Pure Clostridium spore suspensions are required for investigating species of medical and environmental importance. Defining the conditions for optimal spore production also provides insight into the underlying mechanisms of Clostridium sporulation.

U(VI) reduction by spores of Clostridium acetobutylicum

“…Other sporulation approaches were also tested but did not result in a high yield of spores with the ability to reduce U(VI). Specifically, cells that were heatshocked for 15 min at 80 C and incubated at 23 C according to a protocol by Yang et al (2009) produced spores that did not reproducibly reduce U(VI). Additionally, cells allowed to sporulate naturally gave a low spore yield, but those spores had the ability to reduce U(VI).…”

Padronização de um modelo de infecção por Clostridium difficile em hamsters sírios Mesocricetus auratus

“…Os esporos de C. diffi cile foram produzidos de acordo com YANG et al (2009). As estirpes de C. diffi cile foram inoculadas em erlenmeyer contendo 250mL de BHI (Difco Laboratories, EUA), seguindo de incubação a 37°C por 72 horas em ambiente de anaerobiose.…”

<i>Bacillus pumilus</i>: Possible Model for the Bioweapon <i>Bacillus anthracis</i>

“…sporogenes was nearly half that at 36%. Yang reported [19] that the viability of Cl. sporogenes cultured on T-soy was 30%.…”