A comparison of passive and active dust sampling methods for measuring airborne methicillin-resistant Staphylococcus aureus in pig farms.

Methicillin-resistant strains of Staphylococcus aureus (MRSA) are resistant to most ß-lactam antibiotics. Pigs are an important reservoir of livestock-associated MRSA (LA-MRSA), which is genetically distinct from both hospital and community-acquired MRSA. Occupational exposure to pigs on farms can lead to LA-MRSA carriage by workers. There is a growing body of research on MRSA found in the farm environment, the airborne route of transmission, and its implication on human health. This study aims to directly compare two sampling methods used to measure airborne MRSA in the farm environment; passive dust sampling with electrostatic dust fall collectors (EDCs), and active inhalable dust sampling using stationary air pumps with Gesamtstaubprobenahme (GSP) sampling heads containing Teflon filters. Paired dust samples using EDCs and GSP samplers, totaling 87 samples, were taken from 7 Dutch pig farms, in multiple compartments housing pigs of varying ages. Total nucleic acids of both types of dust samples were extracted and targets indicating MRSA (femA, nuc, mecA) and total bacterial count (16S rRNA) were quantified using quantitative real-time PCRs. MRSA could be measured from all GSP samples and in 94% of the EDCs, additionally MRSA was present on every farm sampled. There was a strong positive relationship between the paired MRSA levels found in EDCs and those measured on filters (Normalized by 16S rRNA; Pearson's correlation coefficient r = 0.94, Not Normalized; Pearson's correlation coefficient r = 0.84). This study suggests that EDCs can be used as an affordable and easily standardized method for quantifying airborne MRSA levels in the pig farm setting.

Occupational and environmental exposure to SARS-CoV-2 in and around infected mink farms.

OBJECTIVE: Unprecedented SARS-CoV-2 infections in farmed minks raised immediate concerns regarding transmission to humans and initiated intensive environmental investigations to assess occupational and environmental exposure. METHODS: Air sampling was performed at infected Dutch mink farms, at farm premises and at nearby residential sites. A range of other environmental samples were collected from minks' housing units, including bedding materials. SARS-CoV-2 RNA was analysed in all samples by quantitative PCR. RESULTS: Inside the farms, considerable levels of SARS-CoV-2 RNA were found in airborne dust, especially in personal inhalable dust samples (approximately 1000-10 000 copies/m3). Most of the settling dust samples tested positive for SARS-CoV-2 RNA (82%, 75 of 92). SARS-CoV-2 RNA was not detected in outdoor air samples, except for those collected near the entrance of the most recently infected farm. Many samples of minks' housing units and surfaces contained SARS-CoV-2 RNA. CONCLUSIONS: Infected mink farms can be highly contaminated with SARS-CoV-2 RNA. This warns of occupational exposure, which was substantiated by considerable SARS-CoV-2 RNA concentrations in personal air samples. Dispersion of SARS-CoV-2 to outdoor air was found to be limited and SARS-CoV-2 RNA was not detected in air samples collected beyond farm premises, implying a negligible risk of environmental exposure to nearby communities. Our occupational and environmental risk assessment is in line with whole genome sequencing analyses showing mink-to-human transmission among farm workers, but no indications of direct zoonotic transmission events to nearby communities.

Spatial Variation of Endotoxin Concentrations Measured in Ambient PM10 in a Livestock-Dense Area: Implementation of a Land-Use Regression Approach.

BACKGROUND: Results from studies on residential health effects of livestock farming are inconsistent, potentially due to simple exposure proxies used (e.g., livestock density). Accuracy of these proxies compared with measured exposure concentrations is unknown. OBJECTIVES: We aimed to assess spatial variation of endotoxin in PM10 (particulate matter ≤10µm) at residential level in a livestock-dense area, compare simple livestock exposure proxies to measured endotoxin concentrations, and evaluate whether land-use regression (LUR) can be used to explain spatial variation of endotoxin. METHODS: The study area (3,000 km2) was located in Netherlands. Ambient PM10 was collected at 61 residential sites representing a variety of surrounding livestock-related characteristics. Three to four 2-wk averaged samples were collected at each site. A local reference site was used for temporal variation adjustment. Samples were analyzed for PM10 mass by weighing and for endotoxin by using the limulus amebocyte lysate assay. Three LUR models were developed, first a model based on general livestock-related GIS predictors only, followed by models that also considered species-specific predictors and farm type-specific predictors. RESULTS: Variation in concentrations measured between sites was substantial for endotoxin and more limited for PM10 (coefficient of variation: 43%, 8%, respectively); spatial patterns differed considerably. Simple exposure proxies were associated with endotoxin concentrations although spatial variation explained was modest (R2<26%). LUR models using a combination of animal-specific livestock-related characteristics performed markedly better, with up to 64% explained spatial variation. CONCLUSION: The considerable spatial variation of ambient endotoxin concentrations measured in a livestock-dense area can largely be explained by LUR modeling based on livestock-related characteristics. Application of endotoxin LUR models seems promising for residential exposure estimation within health studies. https://doi.org/10.1289/EHP2252.

Dynamics of MRSA carriage in veal calves: a longitudinal field study.

Colonization of Methicillin-Resistant Staphylococcus aureus (MRSA) in food producing animals has public health implications, but intervention targets have not yet been identified. In this field study occurrence and dynamics of MRSA in veal calves were investigated longitudinally on three farms. Determinants generally associated with MRSA carriage, such as environmental exposure and antimicrobial use, were explored. In addition, the reliability and reproducibility of MRSA detection in nasal samples from veal calves were investigated as well as the additional value of rectal samples to establish MRSA status of an individual animal. On these three farms, MRSA prevalence and MRSA air loads in stables rapidly increased during the production cycle, especially after releasing calves from their individual houses, but not simultaneously with or directly after treatment with antimicrobials. These observations constitute the hypothesis that antimicrobial use may not necessarily be the only condition for MRSA transmission in veal calves, but indicate that other factors may contribute to transmission as well. MRSA in calves was present both nasally and rectally. The reproducibility and repeatability of the nasal samples were moderate. The results of this study give a better understanding of the dynamics of MRSA in a field situation.

Evaluation of isolation procedures and chromogenic agar media for the detection of MRSA in nasal swabs from pigs and veal calves.

Since the emergence of MRSA in livestock, screening of animals for the detection of MRSA is widely practised. Different procedures are published for animal samples but a systematic comparison of methods has not been performed. The objective of this study was to compare three available commonly used procedures and three chromogenic agars for detecting MRSA in nasal swabs from pigs (n=70) and veal calves (n=100). Procedures 1 and 2 used a pre-enrichment comprising Mueller Hinton broth with 6.5% NaCl followed by selective enrichment with 4 microg/ml oxacillin+75 microg/ml aztreonam (procedure 1) and 5 microg/ml ceftizoxime+75 microg/ml aztreonam (procedure 2) respectively. Procedure 3 used a selective enrichment broth only, containing 4% NaCl, 5 microg/ml ceftizoxime+50 microg/ml aztreonam. After selective enrichment, media were streaked on to three different chromogenic agars. Significantly more MRSA were found for pig as well as for veal calf samples with procedures 1 and 2. No significant differences were found between procedures 1 and 2. For nasal swabs from pigs significantly more MRSA-positive samples were found when MRSA Screen (Oxoid) or MRSASelect (Bio-Rad) agars were used compared to MSRA ID (bioMérieux). For calf samples no significant differences between the different agars were found. In conclusion, the results of this study show that procedures 1 and 2, both using additional high salt pre-enrichment are superior and should be recommended for MRSA detection in nasal swabs from pigs and veal calves. The preferred choice of chromogenic agar depends on the sample matrix.