Endotoxin exposure is associated with wheeze and asthma morbidity, while early life exposure may reduce risk of allergy and asthma. Unfortunately, it is difficult to compare endotoxin results from different laboratories and environments. We undertook this study to determine if lipopolysaccharide (LPS) extraction efficiency could account for differences among laboratories. We generated and collected aerosols from chicken and swine barns, and corn processing. We randomly allocated side-by-side filter samples to five laboratories for Limulus assay of endotoxin. Lyophilized aliquots of filter extracts were analyzed for 3-hydroxy fatty acids (3-OHFAs) as a marker of LPS using gas chromatography-mass spectrometry. There were significant differences in endotoxin assay and GC-MS (LPS) results between laboratories for all dust types (p < 0.01). Patterns of differences between labs varied by dust type. Relationships between assay and GC/MS results also depended on dust type. The percentages of individual 3-OHFA chain lengths varied across labs (p < 0.0001) suggesting that each lab recovered a different fraction of the LPS available. The presence of large amounts of particle associated LPS and absence of a freezing thawing cycle were associated with lower correlations between LPS and bioactivity, consistent with an absence of Limulus response to cell-bound endotoxin. These data suggest that extraction methods affect endotoxin measurements. The LAL methods may be most suitable when comparing exposures within similar environments; GC-MS offers additional information helpful in optimizing sample treatment and extraction. GC-MS may be of use when comparing across heterogeneous environments and should be considered for inclusion in future studies of human health outcomes.

Endotoxins from gram-negative bacteria pose a significant respiratory hazard. Establishing dose-response relationships is problematic because there are no standard procedures for sampling and analysis. The goal of this study was to compare endotoxin analyses in six laboratories using Limulus-based assays for analysis of organic dusts from three agricultural environments: chicken barns, swine barns, and corn processing facilities. For each dust generation experiment 14 side-by-side air samples were collected on 37-mm glass fiber filters at flows of 1.8 L/min. Each laboratory was randomly allocated two filters from each of seven experiments per dust type. Three laboratories used the QCL-1000 endpoint assay, and three used the kinetic-QCL method. To eliminate variability among different lots, a single lot of Limulus amebocyte lysate for endpoint assays and one similar lot for kinetic assays was provided. Precision of assays performed within labs was very good, with pooled coefficients of variation for replicate samples ranging from 1 to 11% over all labs and all dust types. There were significant differences between laboratories for all three dust types (p < 0.01). The pattern of differences between labs varied by dust type. For chicken dust, labs using the endpoint method reported higher results than those using kinetic methods. For swine and corn dusts, labs using the kinetic method reported the highest endotoxin values. For chicken dust, results from all labs except A and B were highly correlated (r = 0.86-1.00). For swine dust, only labs B and E, and C and D were correlated. For corn, A, B, and D were significantly correlated with most other labs. In conclusion, statistical differences in performance between laboratories were apparent and may be related to the extraction and analytical methods. The results of this study will be useful for standardization of sampling and analysis of airborne endotoxin in agriculture.

SUMMARYA total of 79 Australian isolates of beta-haemolytic Escherichia coli from cases of porcine postweaning diarrhoea (PWD), and 18 isolates of serotype O 149:K91 :K88 (F4) from unweaned pigs from Australia, Indonesia and Denmark, were examined by multilocus enzyme electrophoresis. These were divided into 57 electrophoretic types (ETs), with an overall mean genetic diversity per enzyme locus of 0-466. This value closely resembled that previously recorded for the whole species. Not only was the collection diverse, but there was considerable genetic heterogeneity amongst PWD isolates of the same serogroup.Isolates from serogroups 0 8 and 0 138 were most varied, whilst many from serogroups 0 141 and 0 149 were more closely related. In contrast, the isolates from the unweaned pigs all belonged to only one ET.

SUMMARY Approximately 23 viruses were isolated from healthy pigs, pigs with encephalitis, and in cases of reproductive failure. Five viruses were identified as enteroviruses and a total of 10 isolates were shown to cross‐react serologically to varying degrees. Twenty viruses were neutralised by a reference antiserum of serotype 8 porcine enterovirus. Intracerebral inoculation of colostrum‐deprived piglets with 2 of the characterised viruses caused lesions of encephalomyelitis which were not induced by oral infection. Intrafoetal inoculation of 2 sows with one characterised faecal isolate caused foetal death and abortion, but no adverse effects followed oral dosage.

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