Recovery and quantification of mycobacterium immunogenum DNA from metalworking fluids using dual-labeled probes

Mycobacteria in metalworking fluids (MWF) are associated with hypersensitivity pneumonitis but are difficult to recover using culture. Quantitative PCR is a promising approach to quantify mycobacteria, but three challenges exist: mycobacterial cell lysis, high-yield DNA extraction, and removal of PCR inhibitors. We used Mycobacterium spp. primers to amplify polymorphic regions of 16S-rDNA flanked with highly conserved regions. A standard curve was constructed by cloning M. immunogenum amplification product. We developed single tube DNA extraction employing mixer mill cell disruption, enzymatic digestions (lysozyme, proteinase K) followed by a mechanical disruption, and column purification. MWF was spiked with M. immunogenum, and DNA was successfully extracted. Mycobacterial 16S-RNA genes were quantified by comparing PCR amplification detection (Cycle Threshold) from our samples with that obtained from the standard curve. Recovery and quantification of mycobacterial DNA from spiked samples approached 100 %. A rapid method for quantification of mycobacteria in MWF was demonstrated

Psychosocial work factors and social inequalities in psychological distress: a population-based study.

BACKGROUND: Mental health problems (MHP) are the leading cause of disability worldwide. The inverse association between socioeconomic position (SEP) and MHP has been well documented. There is prospective evidence that factors from the work environment, including adverse psychosocial work factors, could contribute to the development of MHP including psychological distress. However, the contribution of psychosocial work factors to social inequalities in MHP remains unclear. This study evaluates the contribution of psychosocial work factors from two highly supported models, the Demand-Control-Support (DCS) and the Effort-Reward Imbalance (ERI) models to SEP inequalities of psychological distress in men and women from a population-based sample of Quebec workers. METHODS: Data were collected during a survey on working conditions, health and safety at work. SEP was evaluated using education, occupation and household income. Psychosocial work factors and psychological distress were assessed using validated instruments. Mean differences (MD) in the score of psychological distress were estimated separately for men and women. RESULTS: Low education level and low household income were associated with psychological distress among men (MD, 0.56 (95% CI 0.06; 1.05) and 1.26 (95% CI 0.79; 1.73) respectively). In men, the contribution of psychosocial work factors from the DCS and the ERI models to the association between household income and psychological distress ranged from 9% to 24%. No clear inequalities were observed among women. CONCLUSIONS: These results suggest that psychosocial work factors from the DCS and the ERI models contribute to explain a part of social inequalities in psychological distress among men. Psychosocial factors at work are frequent and modifiable. The present study supports the relevance of targeting these factors for the primary prevention of MHP and for health policies aiming to reduce social inequalities in mental health

Health Effects of Airborne Exposures from Concentrated Animal Feeding Operations

Toxic gases, vapors, and particles are emitted from concentrated animal feeding operations (CAFOs) into the general environment. These include ammonia, hydrogen sulfide, carbon dioxide, malodorous vapors, and particles contaminated with a wide range of microorganisms. Little is known about the health risks of exposure to these agents for people living in the surrounding areas. Malodor is one of the predominant concerns, and there is evidence that psychophysiologic changes may occur as a result of exposure to malodorous compounds. There is a paucity of data regarding community adverse health effects related to low-level gas and particulate emissions. Most information comes from studies among workers in CAFO installations. Research over the last decades has shown that microbial exposures, especially endotoxin exposure, are related to deleterious respiratory health effects, of which cross-shift lung function decline and accelerated decline over time are the most pronounced effects. Studies in naïve subjects and workers have shown respiratory inflammatory responses related to the microbial load. This working group, which was part of the Conference on Environmental Health Impacts of Concentrated Animal Feeding Operations: Anticipating Hazards—Searching for Solutions, concluded that there is a great need to evaluate health effects from exposures to the toxic gases, vapors, and particles emitted into the general environment by CAFOs. Research should focus not only on nuisance and odors but also on potential health effects from microbial exposures, concentrating on susceptible subgroups, especially asthmatic children and the elderly, since these exposures have been shown to be related to respiratory health effects among workers in CAFOs

Contraintes psychosociales au travail, télomères, marqueurs d'inflammation et fonction cognitive globale : une étude longitudinale de 24 ans

Contexte : La proportion de personnes âgées de 65 ans et plus est en constante augmentation, entrainant un accroissement des problèmes de santé fréquents dans cette tranche d'âge tels que les troubles neurocognitifs (TNC) mineurs et majeurs (démence). Face à cette problématique, l'identification de facteurs modifiables, permettant d'accroitre la réserve cognitive et ainsi, de retarder l'apparition des TNC, est une priorité pour la santé publique. Certains facteurs psychosociaux au travail, tels que la complexité intellectuelle, ont été suggérés comme facteurs ayant le potentiel de maintenir une bonne réserve cognitive à long terme. Ces facteurs, aussi appelés contraintes psychosociales au travail, sont reconnus pour leurs effets néfastes sur la santé, ayant été associés avec l'augmentation du risque de maladies cardiovasculaires et de problèmes de santé mentale. L'exposition chronique à des contraintes psychosociales peut entrainer une augmentation de l'inflammation et du stress oxydatif, deux mécanismes biologiques associés avec le risque de TNC. Objectif : L'objectif de cette thèse était d'évaluer l'effet de l'exposition à des contraintes psychosociales au travail sur 1) la fonction cognitive globale, 2) des biomarqueurs d'inflammation, soit la protéine-C réactive et l'interleukine-6, et un biomarqueur d'inflammation, d'oxydation et de vieillissement cellulaire, soit la longueur des télomères, ainsi que 3) l'effet intermédiaire potentiel de ces trois biomarqueurs dans la relation entre les contraintes psychosociales au travail et la fonction cognitive globale. Méthode : Cette thèse a été réalisée à partir des données de l'Étude prospective québécoise sur le travail et la santé (PROQ), une étude de cohorte occupationnelle prospective comprenant 9188 travailleurs cols-blancs de la région de Québec suivis depuis 24 ans. Un échantillon aléatoire de 3411 participants de l'Étude PROQ a été sélectionné pour faire partie de l'Étude sur les biomarqueurs comprenant un échantillon sanguin pour la mesure des biomarqueurs. Les contraintes psychosociales au travail ont été mesurées par questionnaire auto-rapporté validé. La fonction cognitive a été évaluée à l'aide du « Montreal Cognitive Assessment » (MoCA), un test de fonction cognitive globale validé. La protéine C-réactive, l'interleukine-6 et la longueur des télomères ont été mesurées selon des protocoles standardisés. Des mesures d'association ont été calculées à l'aide d'équations d'estimation généralisées ajustées pour plusieurs facteurs potentiellement confondants, dont des facteurs socio-démographiques, des habitudes de vie et des comorbidités. Une imputation multiple a été réalisée pour les données manquantes et une pondération par probabilité inverse de censure a été réalisée pour corriger pour le potentiel biais de sélection lié aux pertes au suivi. Résultats : Les résultats suggèrent que l'exposition répétée à un travail passif comportant une faible latitude décisionnelle et une faible demande psychologique, pourrait être associée à une moins bonne performance cognitive à long terme; que l'exposition à une combinaison de plusieurs contraintes psychosociales au travail, telles qu'une demande psychologique élevée, une latitude décisionnelle faible et un soutien social au travail faible, pourrait être associée avec des concentrations élevées de biomarqueurs d'inflammation; et que l'exposition à certaines contraintes psychosociales au travail, telles qu'un travail passif ou une faible latitude décisionnelle, pourrait être associée à des télomères plus courts. Par contre, l'effet intermédiaire de ces trois biomarqueurs dans la relation entre les contraintes psychosociales au travail et la fonction cognitive globale n'a pas été observé. Finalement, plusieurs différences ont été observées dans ces associations selon le sexe. Conclusion : Ces résultats apportent un appui supplémentaire à l'importance de considérer les expositions psychosociales au travail lors de l'introduction d'interventions permettant d'accroitre la santé cognitive et de maintenir la réserve cognitive. Ces résultats apportent également un appui à l'importance de ces expositions délétères en montrant l'impact qu'elles peuvent avoir sur des mécanismes biologiques d'inflammation, d'oxydation et de vieillissement cellulaire prématuré, associés au risque de plusieurs maladies chroniques. Les contraintes psychosociales au travail sont à considérer afin d'avoir une vision globale de l'ensemble des facteurs modifiables permettant de prévenir ces maladies et de contribuer au vieillissement en santé.Background: Due to the aging of the population, the burden of mild and major (dementia) neurocognitive disorders (NCD) has considerably increased in the past decades. The identification of modifiable factors that could maintain cognitive function is thus emerging as a public health priority. Some work-related psychosocial factors such as intellectual complexity, have been suggested to have the potential to support a good cognitive reserve in the long run. However, work-related psychosocial stressors are also recognized for their harmful effects on health, having been associated with an increased risk of cardiovascular disease and mental health problems. Chronic exposure to psychosocial stressors can lead to increased inflammation and oxidative stress, two biological mechanisms associated with the risk of NCD. Objective: The objective of this thesis was to evaluate the effect of work-related psychosocial factors exposure on 1) global cognitive function, 2) inflammatory biomarkers, namely C-reactive protein and interleukin 6, and telomere length, as well as 3) the potential mediating effect of these three biomarkers in the relationship between work-related psychosocial factors and global cognitive function. Method: This thesis was carried out using data from the Prospective Quebec (PROQ) study on work and health, an occupational cohort study including 9188 white-collar workers from Quebec City followed over 24 years. A random sample of 3411 participants from the PROQ Study was selected to be part of the Biomarker study, which included a blood sample for the measurement of biomarkers. Work-related psychosocial factors were evaluated using validated questionnaires. Global cognitive function was evaluated with the Montreal Cognitive Assessment, a validated test. C-reactive protein, interleukin-6 and telomere length were measured according to standardized protocols Associations were measured using generalized estimating equation adjusted for several potential confounders, including socio-demographic factors, lifestyle habits and comorbidities. Multiple imputation was performed for missing data and inverse probability of censoring weighting was performed to correct for potential selection bias related to losses to follow-up. Results: Analyses suggest that repeated exposure to passive work characterized by low decision latitude and low psychological demand may be associated with poorer long-term cognitive performance; that exposure to a combination of several psychosocial constraints at work, such as high psychological demand, low decision latitude and low social support at work, are associated with high concentrations of inflammatory biomarkers; and that exposure to certain psychosocial constraints at work, such as passive work or low decision latitude, are associated with shorter telomeres. However, biomarkers were not found to mediate the relationship between work-related psychosocial factors and global cognitive function. Finally, several differences were observed according to sex. Conclusion: The results of this thesis provide additional support for the importance of considering psychosocial exposures at work when introducing interventions to promote cognitive health. These results also support the importance of the deleterious effect of these exposures by showing the impact they can have on the biological mechanisms of inflammation, oxidative stress, and premature cellular aging, that are all associated with the risk of chronic disease. Work-related psychosocial factors must be considered to have a global vision of all the modifiable factors that make it possible to prevent disease development and contribute to healthy aging

Assessment of bacterial endospore viability with fluorescent dyes

Aim: To validate three fluorescence viability assays designed primarily for vegetative cells on pure Bacillus endospores. Methods and Results: Purified fresh and gamma-irradiated Bacillus endospores (Bacillus cereus, B. coagulans and two strains of B. subtilis) were used. The viability assays were: 5-cyano-2,3-diotolyl tetrazolium chloride (CTC) to test respiratory activity and early germination, DiBAC4(3) and Live/Dead BacLight to measure membrane energization and permeabilization, respectively. Gamma irradiation treatment completely eliminated spore culturability and was used as negative control. The untreated spores showed respiratory activity after 1 h of incubation and this was characteristic of almost 100% of spores after 24 h. The membrane potential assessment gave no answer about spore viability. A lower proportion of untreated spores had permeabilized membrane compared with gamma-irradiated spores using Live/Dead BacLight (P < 0·02). Conclusion: It is possible to use CTC and Live/Dead BacLight to rapidly test endospore viability and evaluate the proportion of spores in a preparation that could not be recovered with plate count. Significance and Impact of the Study: This study shows that fluorescence tests could be applied to assess viability in potentially pathogenic Bacillus spore preparations within 1 h

Assessment of particulates and bioaerosols in eastern Canadian sawmills

The purpose of this study was to quantify and identify the airborne contamination in eastern Canadian sawmills. Seventeen sawmills were chosen to cover a wide range of size, geographic distribution, and wood species processed. Within each sawmill different work sites (debarking, sawing, sorting, or planing) were studied separately. Area sampling was performed for exposure assessment. Microbial contaminants were assessed with all-glass impingers 30 and six-stage Andersen microbial samplers; appropriate selective media and culture conditions for bacteria, thermophilic actinomycetes, molds, and yeasts were used. Inhalable dust, endotoxins, temperature, and humidity also were measured. Penicillium species were the most predominant molds with up to 40 different Penicillium species identified. Debarking was the working site most highly contaminated by molds, bacteria, and endotoxins (p=0.0001). At this working site mold levels reached a maximum of 1.5×10 6 CFU/m 3 , whereas the median values for culturable bacteria and endotoxin were 21,620 CFU/m 3 and 1081 endotoxin units/m 3 , respectively. Planing sites were the most highly dust contaminated (median: 3.0 mg/m 3 ) (p <0.05). Sawmills of eastern Canada contain airborne biological contaminants that vary between working sites, and their microflora is different from that previously described in European sawmil

Six month tracking of microbial growth in a metalworking fluid after system cleaning and recharging

Large volumes of metalworking fluids (MWFs) are used in manufacturing industries for cooling and lubrication of metal pieces and tools during machining. MWFs accumulate microbial growth through continuous recirculation and reuse. We studied the progression of microbial contamination for 6 months after dumping, cleaning and recharging (DCR) of a large semi-synthetic MWF system managed with several biocides. Fresh, uncontaminated fluid was added to the system after extensive cleaning. The following samples were collected and analyzed: pre-DCR fluid (before system cleaning); neat fluid diluted to 6% with water; in use MWF 12 h and 1, 3 and 6 months post-DCR. Samples were analyzed for total microorganism concentrations by direct counting using fluorescence microscopy and by plate counting on various media (R2A, BHI, Middlebrooks and rose bengal under aerobic conditions). In addition, PCR was performed for the detection of mycobacteria. There was a rapid progression in the total bacterial counts as determined by fluorescence microscopy: 5·7 × 107 cells/ml in the pre-DCR used fluid, no measurable bacteria in the neat fluid, 6·9 × 106 cells/ml after 12 h and 2·2 × 106, 3·6 × 108 and 6·1 × 108 cells/ml after 1, 3 and 6 months, respectively. On average, only 0·2% of the direct count organisms were quantified on R2A cultures. PCR showed the presence of mycobacteria in the used MWF at 3 and 6 months. Mycobacteria were also identified from cultures on Middlebrooks and R2A. This study demonstrates that standard methods for cleaning MWF systems are inadequate since residual bacteria in the system can rapidly repopulate the newly charged MWF

Real-time PCR quantification of mycobacterium immunogenum in used metalworking fluids

Rapid detection and quantification of Mycobacterium immunogenum in field samples of metalworking fluids (MWFs) is important for factory fluid surveillance programs. The applicability of the developed DNA extraction and quantitative real-time PCR (qPCR) methods to detect and quantify M. immunogenum in used MWFs was evaluated. Total DNA from these samples was extracted, and M. immunogenum measured by qPCR by comparison with a standard curve derived from plasmid vectors. PCR counts were compared with bacterial culture counts. PCR counts of M. immunogenum varied from 1.42 × 103 to 3.68 × 106 cells/mL of MWFs. Recovery of M. immunogenum by bacterial culture varied from 2.5% to 70% of qPCR count in corresponding samples. Quantitative PCR could be used to measure M. immunogenum load in MWF samples with greater sensitivity and shorter processing time than the classic bacterial culture-based approach. The proposed qPCR approach could be routinely used in real-time PCR-equipped laboratories to provide early detection of M. immunogenum and to control proliferation that probably leads to hypersensitivity pneumonitis in exposed workers