Lymphocytic choriomeningitis virus in employees and mice at multipremises feeder-rodent operation, United States, 2012.

We investigated the extent of lymphocytic choriomeningitis virus (LCMV) infection in employees and rodents at 3 commercial breeding facilities. Of 97 employees tested, 31 (32%) had IgM and/or IgG to LCMV, and aseptic meningitis was diagnosed in 4 employees. Of 1,820 rodents tested in 1 facility, 382 (21%) mice (Mus musculus) had detectable IgG, and 13 (0.7%) were positive by reverse transcription PCR; LCMV was isolated from 8. Rats (Rattus norvegicus) were not found to be infected. S-segment RNA sequence was similar to strains previously isolated in North America. Contact by wild mice with colony mice was the likely source for LCMV, and shipments of infected mice among facilities spread the infection. The breeding colonies were depopulated to prevent further human infections. Future outbreaks can be prevented with monitoring and management, and employees should be made aware of LCMV risks and prevention.

Development of a quantitative method for assessment of dose in in vitro evaluations using a VITROCELL® VC10® smoke exposure system.

The assessment of potential cytotoxicity or genotoxicity of combustible tobacco products has historically been performed using partitioned exposures (i.e. total particulate matter [TPM], gas vapor phase [GVP]) rather than whole smoke. The VITROCELL® VC10® smoke exposure system offers multiple platforms for air liquid interface (ALI) or air agar interface (AAI) exposure to mimic in vivo-like conditions for assessing the toxicological impact of whole smoke using in vitro assays (e.g. cytotoxicity, mutagenicity and DNA modifications). The aims of this study were to investigate dosimetry during whole smoke exposure in the VITROCELL® VC10® smoking robot using quartz crystal microbalances (QCMs) and to support the use of photometers for concurrent assessment of 'dose' during whole smoke exposures. QCM results showed consistent deposition across different exposure chambers, between dilution bars, experiments and modules. Higher levels of variation were noted at higher airflows (i.e., >8 L/min). Dosimetry assessed using photometers also showed a high level of consistency between experiments, with no notable impact on deposition on the QCM when the photometers were placed 'in-line' between the dilution bar and the exposure module. However, the use of photometers alone may be not be sufficient to estimate deposition; the predictability of the data-generated equation was poor. Further development of dosimetry methodology and information for use in validated in vitro biological test methods is needed to facilitate on-going aerosol-based research and relative assessment.

Development, qualification, validation and application of the Ames test using a VITROCELL® VC10® smoke exposure system.

The Ames test has established use in the assessment of potential mutagenicity of tobacco products but has generally been performed using partitioned exposures (e.g. total particulate matter [TPM], gas vapor phase [GVP]) rather than whole smoke (WS). The VITROCELL®VC10® smoke exposure system offers multiple platforms for air liquid interface (ALI), or air agar interface (AAI) in the case of the Ames test exposure to mimic in vivo-like conditions for assessing the toxicological impact of fresh WS in in vitro assays. The goals of this study were to 1) qualify the VITROCELL®VC10® to demonstrate functionality of the system, 2) develop and validate the Ames test following WS exposure with the VITROCELL®VC10® and 3) assess the ability of the Ames test to differentiate between a reference combustible product (3R4F Kentucky reference cigarette) and a primarily tobacco heating product (Eclipse). Based on critical function assessments, the VITROCELL®VC10® was demonstrated to be fit for the purpose of consistent generation of WS. Assay validation was conducted for 5 bacterial strains (TA97, TA98, TA100, TA1535 and TA102) and reproducible exposure-related changes in revertants were observed for TA98 and TA100 in the presence of rat liver S-9 following exposure to 3R4F WS. In the comparative studies, exposure-related changes in in vitro mutagenicity following exposure of TA98 and TA100 in the presence of S9 to both 3R4F and Eclipse WS were observed, with the response for Eclipse being significantly less than that for 3R4F (p < 0.001) which is consistent with the fewer chemical constituents liberated by primarily-heating the product.

Development, qualification, validation and application of the neutral red uptake assay in Chinese Hamster Ovary (CHO) cells using a VITROCELL® VC10® smoke exposure system.

Cytotoxicity assessment of combustible tobacco products by neutral red uptake (NRU) has historically used total particulate matter (TPM) or solvent captured gas vapor phase (GVP), rather than fresh whole smoke. Here, the development, validation and application of the NRU assay in Chinese Hamster Ovary (CHO) cells, following exposure to fresh whole smoke generated with the VITROCELL® VC10® system is described. Whole smoke exposure is particularly important as both particulate and vapor phases of tobacco smoke show cytotoxicity in vitro. The VITROCELL® VC10® system provides exposure at the air liquid interface (ALI) to mimic in vivo conditions for assessing the toxicological impact of smoke in vitro. Instrument and assay validations are crucial for comparative analyses. GOALS OF THIS STUDY: 1) demonstrate functionality of the VITROCELL® VC10® system by installation, operational and performance qualification, 2) develop and validate a cellular system for assessing cytotoxicity following whole smoke exposure and 3) assess the whole smoke NRU assay sensitivity for statistical differentiation between a reference combustible cigarette (3R4F) and a primarily "heat-not-burn" cigarette (Eclipse). RESULTS: The VITROCELL® VC10® provided consistent generation and delivery of whole smoke; exposure-related changes in in vitro cytotoxicity were observed with reproducible IC50 values; comparative analysis showed that the heat-not-burn cigarette was significantly (P<0.001) less cytotoxic than the 3R4F combustible cigarette, consistent with the lower levels of chemical constituents liberated by primarily-heating the cigarette versus burning.