ABSTRACT
Introduction: The applications of fumigation and the challenges that high-containment facilities face in achieving effective large volume decontamination are well understood. The Biosecurity Research Institute at Kansas State University sought to evaluate a novel system within their biosafety level 3 (BSL-3) and animal biosafety level 3 agriculture (ABSL-3Ag) facility. Methods: The system chosen for this study is the CURIS® Hybrid Hydrogen PeroxideTM (HHPTM) system, comprising a mobile 36-pound (16 kg) device delivering a proprietary 7% hydrogen peroxide (H2O2) solution. To examine the system's efficacy in multiple laboratory settings, two BSL-3 laboratories (2,281 [65 m3] and 4,668 ft3 [132 m3]) with dropped ceiling interstitial spaces and an ABSL-3Ag necropsy suite (44,212 ft3 [1,252 m3]) with 21-foot (6.4 m) ceilings were selected. Biological indicators (BIs) of Geobacillus stearothermophilus (1.7 × 106 organisms) on steel spore carriers and H2O2 chemical indicators (CIs) were used to provide validation. Results: After cycle optimization, the smaller laboratory had a total of 60 BIs over two treatments that demonstrated a greater than 6-log reduction of bacterial spores. The larger laboratory (192 BIs) and the necropsy suite (206 BIs) had no BIs positive for spore growth when incubated at 60°C for 24 h per manufacturer's specifications. Conclusion: Overall successful results through multiple components of this study demonstrate that the HHP device, paired with the pulsed 7% H2O2 solution, achieved efficacy regardless of variables in laboratory size and layout. Perceived challenges such as 21-ft (6.4 m) ceiling heights, active equipment, and difficult to access ceiling interstitial spaces proved unfounded. Given the successful sterilization of all challenged BIs, the HHP system presents a useful alternative for high level decontamination within BSL-3 and ABSL-3Ag facilities.
ABSTRACT
BACKGROUND: Several alternatively-spliced mRNA transcripts of the follicle stimulating hormone receptor (FSHR) have been identified in sheep, including FSHR-1 (G protein-coupled form), FSHR-2 (dominant negative form), and FSHR-3 (growth factor type-1 form). Our objective was to determine which of these variants is predominantly expressed in follicles collected from ewes at various times after estrus. METHODS: Suffolk-cross ewes (n = 8) were allowed to come into estrus naturally and were euthanized 24 (n = 3), 36 (n = 3), or 48 (n = 2) hours after the onset of estrus. All visible follicles were measured, aspirated and pooled according to follicular diameter: small (<= 2.0 mm), medium (2.1-4.0 mm), large (4.1-6.0 mm), and preovulatory (> = 6.1 mm). Aspirated cells were separated from follicular fluid by centrifugation. Total RNA was extracted from cell pellets and reverse transcribed. The resulting cDNA was subjected to qPCR, using primer sets designed to amplify each variant specifically. Gene expression was normalized to that of beta-actin within samples, and compared by analysis of variance with the level of significant differences set at p < .05. RESULTS: Relative expression of FSHR-3 exceeded that of both FSHR-1 and FSHR-2 in medium follicles, and tended to be higher in small follicles (p = .09) regardless of time after onset of estrus, and thus results from different time points were pooled. Expression of FSHR-3 was greater than that of FSHR-2 and luteinizing hormone receptor (LHR) in small and medium follicles. Expression of LHR was greatest in preovulatory follicles. CONCLUSIONS: These experiments show that in addition to the well characterized G protein-coupled form of the FSHR, alternatively spliced variants of the FSHR may participate in follicular dynamics during follicular waves of the sheep estrous cycle. Furthermore, these results indicate that an "alternatively" spliced form of the FSHR (FSHR-3) is the predominant form of the FSHR in the sheep.
