Current Considerations and Practical Solutions for Overcoming Nanoparticle Interference with LAL Assays and Minimizing Endotoxin Contamination.

Monitoring endotoxin contamination in drugs and medical devices is required to avoid pyrogenic responses and septic shock in patients receiving these products. Endotoxin contamination of engineered nanomaterials and nanotechnology-based medical products represents a significant translational hurdle. Nanoparticles often interfere with an in vitro limulus amebocyte lysate (LAL) assay commonly used in the pharmaceutical industry for the detection and quantification of endotoxin. Such interference challenges the preclinical development of nanotechnology-formulated drugs and medical devices containing engineered nanomaterials. Protocols for the analysis of nanoparticles using LAL assays have been reported before. Here, we discuss considerations for selecting an LAL format and describe a few experimental approaches for overcoming nanoparticle interference with the LAL assays to obtain more accurate estimations of endotoxin contamination in nanotechnology-based products. The discussed approaches do not solve all types of nanoparticle interference with the LAL assays but could be used as a starting point to address the problem. This chapter also describes approaches to prevent endotoxin contamination in nanotechnology-formulated products.

A dye elution method for the quantification of insecticidal crystal proteins from Bacillus thuringiensis and its compatibility with the presence of agro-industrial raw materials and waste products.

The insecticidal crystal proteins produced by Bacillus thuringiensis during sporulation are active ingredients against lepidopteran, dipteran, and coleopteran insects. Several methods have been reported for their quantification, such as crystal counting, ELISA, and SDS-PAGE/densitometry. One of the major tasks in industrial processes is the analysis of raw material dependency and costs. Thus, the crystal protein quantification method is expected to be compatible with the presence of complex and inexpensive culture medium components. This work presents a revalidated elution-based method for the quantification of insecticidal crystal proteins produced by the native strain B. thuringiensis RT. To quantify proteins, a calibration curve was generated by varying the amount of BSA loaded into SDS-PAGE gels. First, SDS-PAGE was performed for quality control of the bioinsecticide. Then, the stained protein band was excised from 10% polyacrylamide gel and the protein-associated dye was eluted with an alcoholic solution of SDS (3% SDS in 50% isopropanol) during 45 min at 95°C. This protocol was a sensitive procedure to quantify proteins in the range of 2.0-10.0 µg. As proof of concept, proteins of samples obtained from a complex fermented broth were separated by SDS-PAGE. Then, Cry1 and Cry2 proteins were properly quantified.

Nanozyme with tailored selectivity and highly catalytic activity for efficient sensing of endotoxin and sourcing gram-negative bacteria.

Endotoxin detection is important for determining bacterial contamination and infection in fields of food, pharmaceutical and clinical disease diagnosis. The horseshoe crab deformed cell lysate analysis is regarded as the gold-standard method, but the endangered and high-cost horseshoe crab animals required in sensing process further raise animal ethical issues and hinder their applications. The colorimetric methods based on nanozymes are simple and economical, but the low selectivity and sensitivity are still the bottleneck for their further application. Herein, we successfully developed a phenylboronic acid functionalized iron-based nanozyme with higher selectivity and highly catalytic activity for endotoxin sensing. The as-prepared colorimetric sensor using the obtained nanozyme as sensing probes shows a good linear relationship for endotoxin sensing in the range of 1-20 µg mL-1, with a LOD = 0.42 µg mL-1, along with good selectivity and reproducibility. The sensor can also be well applied to detecting endotoxin in practical samples such as beer and serum. Moreover, the parameters including time and temperature which could affect the endotoxin release from E. coli were also studied and optimized, based on the relationship between endotoxin and Gram-negative bacteria, the as-prepared sensor achieves the qualitative and quantification of E. coli.

A cross-sectional study on occupational exposure to microorganisms, endotoxin, hydrogen sulfide, and dust during work at drilling waste treatment plants.

This cross-sectional study aims to obtain knowledge about workers' exposure to airborne dust, bacterial and fungal species, endotoxin, biofilm formation, and hydrogen sulfide (H2S) in drilling waste treatment plants. In total, 408 full-shift personal samples, 66 work areas, 40 drilling waste, and reference (outdoor air and seawater) samples were analyzed. Some workers were exposed to high levels of endotoxin (207 EU/m3), bacteria (3.8 × 104 colony forming units (CFU)/m3 and 9.8 × 104 DNA copies/m3), or fungi (1.4 × 107 CFU/m3 and 3,600 copies/m3). The exposure levels to endotoxin, bacteria, and peaks of H2S were dependent on the treatment technique. All types of drilling waste contained large concentrations of bacteria compared to the seawater references. Elevated concentrations of airborne bacteria were found close to drilling waste basins. In total, 116, 146, and 112 different bacterial species were found in workers' exposure, work areas, and the drilling waste, respectively. An overlap in bacterial species found in the drilling waste and air (personal and work area) samples was found. Of the bacterial species found, 49 are classified as human pathogens such as Escherichia coli, Enterobacter cloacae, and Klebsiella oxytoca. In total, 44 fungal species were found in the working environment, and 6 of these are classified as human pathogens such as Aspergillus fumigatus. In conclusion, across the drilling waste treatment plants, human pathogens were present in the drilling waste, and workers' exposure was affected by the drilling waste treated at the plants with elevated exposure to endotoxin and bacteria. Elevated exposure was related to working as apprentices or chemical engineers, and working with cleaning, or slop water, and working in the daytime.

Bacterial contamination of coffee and personal exposure to inhalable dust and endotoxin in primary coffee processing factories in Ethiopia.

INTRODUCTION AND OBJECTIVE: Endotoxins from gram-negative bacteria might be released when the coffee cherries are processed and may cause respiratory health problems among workers in the coffee industry. The relationship between bacterial contamination and occupational exposure to endotoxin levels has not been thoroughly explored previously in primary coffee processing factories in Ethiopia, or elsewhere. The aim of this study was to characterize the level of personal endotoxin exposure and its relations with bacterial contamination of coffee cherries in such factories in Ethiopia. MATERIAL AND METHODS: A cross-sectional study was conducted from March 2020 - February 2021 in 9 primary coffee processing factories in 3 regions in Ethiopia. A total of 180 personal air samples were collected to analyze workers' exposure to inhalable dust and endotoxin. Correlation tests were performed to assess the relationship between total bacteria and endotoxin levels and between inhalable dust and endotoxin levels. RESULTS: The geometric mean (GM) of personal inhalable dust exposure among machine room workers and hand pickers were 9.58 mg/m3 and 2.56 mg/m3, respectively. The overall GM of endotoxin exposure among machine room workers and hand pickers were 10,198 EU/m3 and 780 EU/m3, respectively. Gram-negative bacteria were found in all 54 coffee samples. The correlation between inhalable dust and endotoxin exposure was significant (r=0.80; P <0.01). CONCLUSIONS: About 92% of the samples from hand pickers and all samples from machine room workers exceeded the occupational exposure limit of 90 EU/m3 recommended by the Dutch Expert Committee on Occupational Standards. Prevention and control of bacterial contamination of the coffee in primary coffee processing are suggested to reduce endotoxin exposure that might cause respiratory health problems among coffee workers.

Production of pro-inflammatory mediators stimulated by exposure of poultry house workers to airborne dust particulates.

INTRODUCTION AND OBJECTIVE: Poultry house employees spend a significant part of their work shift being exposed to airborne particulate pollutants. The aim of this study was to assess their exposure at different stages of chicken production cycle, based on quantification of pro-inflammatory mediators (IL-1ß, IL-6, IL-8, and TNFα) in nasal lavage (NAL) samples. MATERIAL AND METHODS: The concentrations of airborne dust at 3 different stages of the production cycle (i.e. empty poultry house, with 7- and 42-day-old chickens) were stationary measured using Grimm spectrometer, as well as CIS and Button samplers. The dust collected by the latter 2 samplers was analyzed for endotoxin and (1→3)-ß-D-glucan content. NAL samples were collected from employees after their work shift to determine the pro-inflammatory mediator levels. RESULTS: The maximum particulate aerosol, endotoxin, and (1→3)-ß-D-glucan concentrations at workplaces reached the levels of 4.12 mg/m3, 45.21 ng/m3, and 56.54 ng/m3, respectively. The IL-1ß, IL-6, and IL-8 concentrations in NAL samples ranged between 0.62-18.12 pg/mL, <0.70-25.37 pg/mL, and <3.50-259.5 pg/mL, respectively. All TNFα levels were below 4 pg/mL. There were no significant differences between these cytokine concentrations in NAL samples collected at different stages of chicken breeding in either 'winter' or 'summer' seasons. CONCLUSIONS: Inhalation stimulation with poultry dust containing endotoxins and (1→3)-ß-D-glucans resulted in the production of pro-inflammatory mediators, which proves the course of immunological processes in the exposed employees that may lead to adverse effects. The use of nasal lavage fluid in the control of such exposure confirms that NAL analysis is a reliable laboratory tool for assessing the impact of poultry dust on exposed farm workers.

Indoor air quality in remote first nations communities in Ontario, Canada.

A recent study of the health of Indigenous children in four First Nations Communities in remote northwestern Ontario found that 21% of children had been admitted to hospital for respiratory infections before age 2 years. Here we report a detailed analysis of the housing conditions in these communities. We employed a variety of statistical methods, including linear regression, mixed models, and logistic regression, to assess the correlations between housing conditions and loadings of biocontaminants (dust mite allergens, fungal glucan, and endotoxin) and indoor concentrations of PM2.5, CO2, benzene, and formaldehyde. The houses (n = 101) were crowded with an average of approximately 7 people. Approximately 27% of the homes had sustained CO2 concentrations above 1500 ppm. Most homes had more than one smoker. Commercial tobacco smoking and the use of non-electric heating (e.g., wood, oil) were associated with increased fine particle concentrations. Over 90% of the homes lacked working Heat Recovery Ventilators (HRVs), which was associated with increased fine particle concentrations and higher CO2. Of the 101 homes, 12 had mold damage sufficient to increase the relative risk of respiratory disease. This resulted from roof leaks, through walls or around the windows due to construction defects or lack of maintenance. A similar percentage had mold resulting from condensation on windows. Endotoxin loadings were much higher than any previous study in Canada. This work provides evidence for the need for more effort to repair existing houses and to ensure the HRVs are properly installed and maintained.

Application of a novel chemical assay for the quantification of endotoxins in bacterial bioreactor samples.

A novel chemical assay, the so-called Kdo-DMB-liquid chromatography (LC) assay, was used for the accurate and cost-effective determination of the endotoxin content in supernatants of Gram-negative bacteria bioreactor samples. During mild acid hydrolysis, the endotoxin-specific sugar acid 3-deoxy-D-manno-oct-2-ulsonic acid (Kdo) is quantitatively released. Kdo is reacted with 1,2-diamino-4,5-methylenedioxybenzene (DMB) to obtain the highly fluorescent derivate Kdo-DMB. It is separated from the reaction mixture by reversed phase-(U)HPLC and detected by fluorescence. From the Kdo content the endotoxin content of the sample is calculated. For three batch cultivations of Escherichia coli K12 and a fed-batch cultivation of Pseudomonas putida KT2440, the evolution of the endotoxin content in dependence on the cultivation time was monitored. Under optimal, constant cultivation conditions a linear correlation between the endotoxin content and the easy-to-access bioreactor parameters optical density at 600 nm and dry cell weight was found for both endotoxin kinds. Under stress cultivation conditions the E. coli K12 cultivation showed a stronger increase of the endotoxin content at harvest in comparison to optimal conditions. Optical density and dry cell weight may be used for production reactors as an economic real-time estimation tool to determine the endotoxin content at different cultivation time points and conditions. The optical density can further be used to establish straightforward sample dilution schemes for endotoxin quantification in samples of unknown endotoxin content. The endotoxin content [ng mL-1] measured by the Kdo-DMB-LC assay and the endotoxin activity [EU mL-1] obtained by the compendial Limulus Amoebocyte Lysate assay show a high correlation for the bacterial bioreactor samples tested.

Establishment of monoclonal antibody and scFv immuno-based assay for Cry2Aa toxin in spiked grain samples.

Bacillus thuringiensis (Bt) Cry toxins have been widely used in the development of genetically modified organisms (GMOs) for pest control. This work aimed to establish more cost effective methods for used Cry2Aa toxins. Three immunoassay methods (IC-ELISA, DAS-ELISA, and CLEIA) were successfully developed in this work. The mAb was used as the detecting antibody, for the IC-ELISA, the range of IC20 to IC80 was 1.11 µg/mL - 60.70 µg/mL, and an IC50 of 10.65 µg/mL. For the DAS-ELISA, the limit of detection (LOD) and limit of quantitation (LOQ) were 10.76 ng/mL and 20.70 ng/mL, respectively. For the CLEIA, the LOD and LOQ were 6.17 ng/mL and 7.40 ng/mL, respectively. The scFv-based detections were the most sensitive for detecting Cry2Aa. The LOD and LOQ for the DAS-ELISA were 118.75 ng/mL and 633.48 ng/mL, respectively. The LOD and LOQ for the CLEIA, read as 37.47 ng/mL and 70.23 ng/mL, respectively. The fact that Cry2Aa toxin was recovered in spiked grain samples further demonstrated that the approaches might be used to identify field samples. These methods provided good sensitivity, stability, and applicability for detecting Cry2Aa toxin, promising ultrasensitive monitoring and references for Cry toxins risk assessment.

Exposure levels of animal allergens, endotoxin, and ß-(1,3)-glucan on a university campus of veterinary medicine.

OBJECTIVES: The study aimed to determine the allergen, endotoxin and ß-(1,3)-glucan concentrations at various areas on a university campus of veterinary medicine. METHODS: Dust samples were collected four times a year for three years using electrostatic dust collectors (EDC) at 25 different locations on a campus of veterinary medicine and in laboratories of inorganic chemistry as a control area representing animal-free environment. Major animal allergens from dog, cat, horse, cattle and mouse, domestic mite (DM) allergens, and ß-(1,3)-glucan were measured using enzyme immunoassays and endotoxin using the limulus amoebocyte lysate (LAL) assay. Seasonal, annual and local influences on exposure levels were analyzed using Bayesian mixed models. RESULTS: With the exception of mouse allergens, all other determinants were found in almost all locations on the campus and in the control area, but in up to 10.000-fold variable concentrations. By far the highest levels of feline, canine, equine and bovine allergens were detected in buildings where the respective species were examined. The highest levels of mouse and DM allergens, ß-(1,3)-glucan and endotoxin occurred together and were associated with locations where large animals were present. In buildings without animals, allergen levels were considerably lower but still elevated at several locations compared to the control area, especially for dog and horse allergens, and ß-(1,3)-glucan. Significant seasonal effects were observed for dog, cat, horse and DM allergens, and ß-(1,3)-glucan. Variations between years were less apparent than between seasons (except for ß-(1,3)-glucan). CONCLUSIONS: The strongest influencing factor on the concentration of mammalian allergens was the presence of the corresponding animal at the collection site. Seasonal influence on allergen concentrations was observed, while the overall exposure remained constant over the years. At locations with horses, elevated levels of mite allergens, endotoxin, and ß-(1,3)-glucan can be expected, probably due to passive transfer from stable environment.

Indoor Airborne Microbiome and Endotoxin: Meteorological Events and Occupant Characteristics Are Important Determinants.

Airborne bacteria and endotoxin may affect asthma and allergies. However, there is limited understanding of the environmental determinants that influence them. This study investigated the airborne microbiomes in the homes of 1038 participants from five cities in Northern Europe: Aarhus, Bergen, Reykjavik, Tartu, and Uppsala. Airborne dust particles were sampled with electrostatic dust fall collectors (EDCs) from the participants' bedrooms. The dust washed from the EDCs' clothes was used to extract DNA and endotoxin. The DNA extracts were used for quantitative polymerase chain (qPCR) measurement and 16S rRNA gene sequencing, while endotoxin was measured using the kinetic chromogenic limulus amoebocyte lysate (LAL) assay. The results showed that households in Tartu and Aarhus had a higher bacterial load and diversity than those in Bergen and Reykjavik, possibly due to elevated concentrations of outdoor bacterial taxa associated with low precipitation and high wind speeds. Bergen-Tartu had the highest difference (ANOSIM R = 0.203) in ß diversity. Multivariate regression models showed that α diversity indices and bacterial and endotoxin loads were positively associated with the occupants' age, number of occupants, cleaning frequency, presence of dogs, and age of the house. Further studies are needed to understand how meteorological factors influence the indoor bacterial community in light of climate change.

Electrochemical Approach for the Synthesis of Ultrasmall Cu13 Clusters and Their Application in the Detection of Endotoxin.

Ultrasmall metal nanoclusters are attractive for their size-dependent optical and electrochemical properties. Here, blue-emitting copper clusters stabilized with cetyltrimethylammonium bromide (CTAB) are synthesized using an electrochemical approach. The electrospray ionization (ESI) analysis reveals that the cluster has 13 copper atoms in the core. The clusters are then used for the electrochemical detection of endotoxin─a bacterial toxin in Gram-negative bacteria. Differential pulse voltammetry (DPV) is used to detect endotoxins with high selectivity and sensitivity. It shows a lower detection limit of 100 ag mL-1 with a linear range of 100 ag mL-1 to 10 ng mL-1. The sensor is efficient for detecting endotoxins from human blood serum samples.

Health symptoms, inflammation, and bioaerosol exposure in workers at biowaste pretreatment plants.

Biowaste pretreatment plants have been built within the last years in Denmark in order to recycle pre-sorted biowaste from houses, restaurants, and industry. We investigated the association between exposure and health at six biowaste pretreatment plants (visited twice) across Denmark. We measured the personal bioaerosol exposure, took blood samples, and administered a questionnaire. Thirty-one persons participated, 17 of them twice, resulting in 45 bioaerosol samples, 40 blood samples, and questionnaire answers from 21 persons. We measured exposure to bacteria, fungi, dust, and endotoxin, the total inflammatory potential of the exposures, and serum levels of the inflammatory markers serum amyloid A (SAA), high sensitivity C-reactive protein (hsCRP), and human club cell protein (CC16). Higher exposures to fungi and endotoxin were found for workers with tasks inside the production area compared to workers with main tasks in the office area. A positive association was found between the concentration of anaerobic bacteria and hsCRP and SAA, whereas bacteria and endotoxin were inversely associated with hsCRP and SAA. A positive association between hsCRP and the fungal species Penicillium digitatum and P. camemberti were found, whereas an inverse association between hsCRP and Aspergillus niger and P. italicum were found. Staff with tasks inside the production area reported more symptoms of the nose than those working in the office area. To conclude, our results indicate that workers with tasks inside the production area are exposed to elevated levels of bioaerosols, and that this may affect workers' health negatively.

Work in nursing homes and occupational exposure to endotoxin and bacterial and fungal species.

Indoor microbial exposure may cause negative health effects. Only little is known about the occupational microbial exposure in nursing homes and the factors that influence the exposure. The exposure in nursing homes may be increased due to close contact with elderly persons who may carry infectious or antimicrobial-resistant microorganisms and due to handling of laundry, such as used clothing and bed linen. We investigated the microbial exposure in 5 nursing homes in Denmark, by use of personal bioaerosol samples from different groups of staff members taken during a typical working day, stationary bioaerosol measurements taken during various work tasks, sedimented dust samples, environmental surface swabs, and swabs from staff members' hands. From the samples, we explored bacterial and fungal concentrations and species composition, endotoxin levels, and antimicrobial resistance in Aspergillus fumigatus isolates. Microbial concentrations from personal exposure samples differed among professions, and geometric means (GM) were 2,159 cfu/m3 (84 to 1.5 × 105) for bacteria incubated on nutrient agar, 1,745 cfu/m3 (82 to 2.0 × 104) for bacteria cultivated on a Staphylococcus selective agar, and 16 cfu/m3 air for potential pathogenic fungi incubated at 37 °C (below detection limit to 257). Bacterial exposures were elevated during bed making. On surfaces, the highest bacterial concentrations were found on bed railings. The majority of bacterial species found were related to the human skin microflora, such as different Staphylococcus and Corynebacterium species. Endotoxin levels ranged from 0.02 to 59.0 EU/m3, with a GM of 1.5 EU/m3. Of 40 tested A. fumigatus isolates, we found one multiresistant isolate, which was resistant towards both itraconazole and voriconazole, and one isolate resistant towards amphotericin B. In conclusion, we give an overview of the general microbial exposure in nursing homes and show that microbial exposures are higher for staff with more care and nursing tasks compared with administrative staff.

Exposure levels of dust, endotoxin, and microorganisms in the Danish recycling industry.

INTRODUCTION: Recycling of domestic waste and a number of employees in the recycling industry is expected to increase. This study aims to quantify current exposure levels of inhalable dust, endotoxin, and microorganisms and to identify determinants of exposure among recycling workers. METHODS: This cross-sectional study included 170 full-shift measurements from 88 production workers and 14 administrative workers from 12 recycling companies in Denmark. The companies recycle domestic waste (sorting, shredding, and extracting materials from waste). We collected inhalable dust with personal samplers that were analysed for endotoxin (n = 170) and microorganisms (n = 101). Exposure levels of inhalable dust, endotoxin, and microorganisms and potential determinants of exposure were explored by mixed-effects models. RESULTS: The production workers were 7-fold or higher exposed to inhalable dust, endotoxin, bacteria, and fungi than the administrative workers. Among production workers recycling domestic waste, the geometric mean exposure level was 0.6 mg/m3 for inhalable dust, 10.7 endotoxin unit (EU)/m3 for endotoxin, 1.6 × 104 colony forming units (CFU)/m³ of bacteria, 4.4 × 104 CFU/m³ of fungi (25 °C), and 1.0 × 103 CFU/m³ of fungi (37 °C). Workers handling paper or cardboard had higher exposure levels than workers handling other waste fractions. The temperature did not affect exposure levels, although there was a tendency toward increased exposure to bacteria and fungi with higher temperatures. For inhalable dust and endotoxin, exposure levels during outdoor work were low compared to indoor work. For bacteria and fungi, indoor ventilation decreased exposure. The work task, waste fraction, temperature, location, mechanical ventilation, and the company size explained around half of the variance of levels of inhalable dust, endotoxin, bacteria, and fungi. CONCLUSION: The production workers of the Danish recycling industry participating in this study had higher exposure levels of inhalable dust, endotoxin, bacteria, and fungi than the administrative workers. Exposure levels of inhalable dust and endotoxin among recycling workers in Denmark were generally below established or suggested occupational exposure limits (OEL). However, 43% to 58% of the individual measurements of bacteria and fungi were above the suggested OEL. The waste fraction was the most influential determinant for exposure, and the highest exposure levels were seen during handling paper or cardboard. Future studies should examine the relationship between exposure levels and health effects among workers recycling domestic waste.

Panorama da qualidade biológica da água destinada ao tratamento dialítico em pacientes sob cuidados críticos em Unidades de Terapia Intensiva / Overview of the biological quality of water intended for dialysis treatment in patients under critical care in Intensive Care Units

A incidência da lesão renal aguda caracteriza-se como evento frequente em pacientes críticos internados em Unidades de Terapia Intensiva e está associada ao aumento de mortalidade, causando grande impacto à Saúde Pública. As intercorrências clínicas são minimizadas com intervenções dialíticas, acarretando a exposição do paciente a volumes expressivos de água tratada durante a terapia renal em leito. As análises microbiológicas e de determinação de endotoxinas bacterianas em amostras de água tratada e em soluções de dialisato foram executadas em dois hospitais públicos do município de São Paulo, seguindo metodologias analíticas preconizadas em compêndios oficiais. A avaliação demonstrou que a porcentagem de resultados satisfatórios no período de 2010 a 2022 variou entre 35,2 a 100% e de 40 a 100% para as unidades hospitalares I e II para a água tratada, respectivamente; e, 100% para as soluções de dialisato para a unidade hospitalar I. A eficácia de ações delineadas pelas equipes técnicas das unidades hospitalares, na adequação da água destinada à terapia dialítica, aponta para a importância em estimular outras instituições hospitalares na padronização e implantação de melhoria contínua de seus sistemas de tratamento de água para uso em procedimento dialítico, prevenindo riscos adicionais aos pacientes expostos à terapia renal.

The incidence of acute kidney is high among critically ill patients admitted to Intensive Care Units and is associated with increased mortality, having a major impact on public health. Clinical complications are minimized with dialysis interventions, which expose patients to significant volumes of treated water during in-bed renal therapy. Microbiological analyzes and determination of bacterial endotoxins were performed on treated water samples and dialysate solutions in two public hospitals in São Paulo city, using analytical methodologies recommended in official compendia. The evaluation showed that the percentage of satisfactory results for treated water ranged from 35.2% to 100% in Hospital Unit I and from 40% to 100% in Hospital Unit II between 2010 and 2022. For dialysate solutions in Hospital Unit I, the percentage of satisfactory results was 100% during the same period. The effectiveness of actions implemented by the technical hospital teams, in adapting water for dialysis therapy, points to the importance of encouraging other hospital institutions to standardize and implement a program of continuous improvement for their water treatment systems used in dialysis procedures. This will help to prevent additional risks to patients exposed to renal therapy.

An algorithm for quantitatively estimating occupational endotoxin exposure in the biomarkers of exposure and effect in agriculture study: II. Application to the study population.

BACKGROUND: We developed an algorithm to quantitatively estimate endotoxin exposure for farmers in the Biomarkers of Exposure and Effect in Agriculture (BEEA) Study. METHODS: The algorithm combined task intensity estimates derived from published data with questionnaire responses on activity duration to estimate task-specific cumulative endotoxin exposures for 13 tasks during four time windows, ranging from "past 12 months" to "yesterday/today." We applied the algorithm to 1681 participants in Iowa and North Carolina. We examined correlations in endotoxin metrics within- and between-task. We also compared these metrics to prior day full-shift inhalable endotoxin concentrations from 32 farmers. RESULTS: The highest median task-specific cumulative exposures were observed for swine confinement, poultry confinement, and grind feed. Inter-quartile ranges showed substantial between-subject variability for most tasks. Time window-specific metrics of the same task were moderately-highly correlated. Between-task correlation was variable, with moderately-high correlations observed for similar tasks (e.g., between animal-related tasks). Prior day endotoxin concentration increased with the total metric and with task metrics for swine confinement, clean other animal facilities, and clean grain bins. SIGNIFICANCE: This study provides insight into the variability and sources of endotoxin exposure among farmers in the BEEA study and summarizes exposure estimates for future investigations in this population.

An algorithm for quantitatively estimating occupational endotoxin exposure in the Biomarkers of Exposure and Effect in Agriculture (BEEA) study: I. Development of task-specific exposure levels from published data.

BACKGROUND/OBJECTIVE: Farmers conduct numerous tasks with potential for endotoxin exposure. As a first step to characterize endotoxin exposure for farmers in the Biomarkers of Exposure and Effect in Agriculture (BEEA) Study, we used published data to estimate task-specific endotoxin concentrations. METHODS: We extracted published data on task-specific, personal, inhalable endotoxin concentrations for agricultural tasks queried in the study questionnaire. The data, usually abstracted as summary measures, were evaluated using meta-regression models that weighted each geometric mean (GM, natural-log transformed) by the inverse of its within-study variance to obtain task-specific predicted GMs. RESULTS: We extracted 90 endotoxin summary statistics from 26 studies for 9 animal-related tasks, 30 summary statistics from 6 studies for 3 crop-related tasks, and 10 summary statistics from 5 studies for 4 stored grain-related tasks. Work in poultry and swine confinement facilities, grinding feed, veterinarian services, and cleaning grain bins had predicted GMs > 1000 EU/m3 . In contrast, harvesting or hauling grain and other crop-related tasks had predicted GMs below 100 EU/m3 . SIGNIFICANCE: These task-specific endotoxin GMs demonstrated exposure variability across common agricultural tasks. These estimates will be used in conjunction with questionnaire responses on task duration to quantitatively estimate endotoxin exposure for study participants, described in a companion paper.

Ophthalmic viscoelastics commonly used in cataract surgery: A microbiota investigation.

PURPOSE: To survey commonly used, sterile ophthalmic viscoelastic materials used during routine cataract surgery for the presence of bacterial DNA and/or viable bacteria and endotoxin quantification. METHODS: Samples from three different ophthalmic viscoelastic manufacturers and three different production lots per manufacturer were collected for 16 S ribosomal ribonucleic acid (rRNA) sequencing and conventional aerobic and capnophilic bacterial culture. Other samples of viscoelastic material from the same three manufacturers were collected for endotoxin quantification using a commercially available Limulus amebocyte lysate (LAL) assay. Statistical analysis was performed using Sigma Plot 14.0, and R v4.0.2.0. Differences (p ≤ .05) between sample collection sites in total DNA concentration, microbial richness, mean intra-group distances, and endotoxin quantification alongside reagent controls were evaluated. RESULTS: Culture yielded two isolates, identified as Staphylococcus epidermidis and Bacillus megaterium. 16 S rRNA sequencing revealed no differences between brands in richness or overall composition. The most common bacterial DNA detected across all brands was Staphylococcus sp., Cutibacterium sp., Flavobacterium sp., and Lactobacillus sp. A significant difference was found between the median endotoxin concentration between Anvision and Hyvisc® viscoelastic (Anvision: 0.171 EU/mL, Hyvisc®: 0.03 EU/mL; p < .001). CONCLUSIONS: No brand-specific differences in bacterial DNA were detected in the viscoelastic materials. Staphylococcus, Cutibacterium, Flavobacterium, and Lactobacillus were the dominant contributors to the bacterial DNA detected. Although Anvision viscoelastic samples contained significantly more endotoxin than Hyvisc® viscoelastic samples, endotoxin concentrations were below the FDA limit of 0.2 EU/mL for both manufacturers. These data further the understanding of inflammatory outcomes following cataract surgery.

Exposure Characterization of Wood Dust Particulate, Endotoxins, and (1-3)-ß-d-Glucans, and Their Determinants in Mozambiquan Wood Processing Workers.

OBJECTIVES: Dust generated from wood processing comprises a heterogeneous mixture of inorganic and organic particles, including wood fragments, microorganisms, endotoxins, (1-3)-ß-d-glucans, and allergens. This study characterized exposure to wood dust and its determinants in the Mozambiquan wood processing industry. METHODS: A total of 124 personal inhalable samples, collected from a stratified random sample of 30 workers, were analysed for dust particulate, endotoxins, and (1-3)-ß-d-glucans. Mixed-effects models were developed to investigate significant exposure determinants. RESULTS: The geometric mean (GM) inhalable dust particulate concentrations were 3.29 mg m-3, 98 endotoxin units (EU) m-3, and 123 ng m-3 for (1-3)-ß-d-glucans. Significant predictors for higher particulate levels included machinery (GMR = 1.93), sawing (GMR = 2.80), carpentry (GMR = 2.77), or painting (GMR = 3.03) tasks. Lebombo-ironwood species was associated with higher dust particulate levels (GMR = 1.97). Determinants of endotoxin concentrations included working with dry wood and damp cleaning methods, which were associated with lower levels. Working in closed buildings (GMR = 3.10) and dry sweeping methods were associated with higher (1-3)-ß-d-glucan concentrations (GMR = 1.99). CONCLUSIONS: Work tasks in certain exposure groups (machinery, sawing, carpentry, painting), processing certain wood species (Lebombo-ironwood) and working in closed buildings were associated with higher exposures, whilst using dry wood and damp cleaning practices reduced exposure levels.