These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer‐reviewed by leading experts in the field, making this an essential research companion.

The mode of action of azole compounds implies a potential to affect endocrine systems of different organisms and is reason for environmental concern. The occurrence and fate of nine agricultural azole fungicides, some of them also used as biocides, and four azole pharmaceuticals were studied in wastewater treatment plants (WWTPs) and lakes in Switzerland. Two pharmaceuticals (fluconazole, clotrimazole, 10-110 ng L(-1)) and two biocides (propiconazole, tebuconazole, 1-30 ng L(-1)) were consistently observed in WWTP influents. Loads determined in untreated and treated wastewater indicated thatfluconazole, propiconazole, and tebuconazole were largely unaffected by wastewater treatment, but clotrimazole was effectively eliminated (> 80%). Incubation studies with activated sludge showed no degradation for fluconazole and clotrimazole within 24 h, but strong sorption of clotrimazole to activated sludge. Slow degradation and some sorption were observed for tebuconazole and propiconazole (degradation half-lives, 2-3 d). In lakes, fluconazole, propiconazole, and tebuconazole were detected at low nanogram-per-liter levels. Concentrations of the pharmaceutical fluconazole correlated with the expected contamination by domestic wastewater, but not those of the biocides. Per capita loads of propiconazole and tebuconazole in lakes suggested additional inputs; for example, from agricultural use or urban runoff rainwater.

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Although the use of hexachlorocyclohexane (HCH), one of the most popular insecticides after the Second World War, has been discontinued in many countries, problems remain from former production and waste sites. Despite the widespread occurrence of HCHs, the environmental fate of these compounds is not fully understood. In particular, environmental metabolites of the more persistent beta-HCH and delta-HCH have not been fully identified. Such knowledge, however, is important to follow degradation and environmental fate of the HCHs. In the present study, several hydroxy metabolites that formed during incubation of beta- and delta-HCH with the common soil microorganism Sphingobium indicum B90A were isolated, characterized, and stereochemically identified by gas chromatography-mass spectrometry (GC-MS) and nuclear magnetic resonance spectroscopy (NMR). The metabolites were identified as isomeric pentachlorocyclohexanols (B1, D1) and tetrachlorocyclohexane-1,4-diols (B2, D2); delta-HCH additionally formed a tetrachloro-2-cyclohexen-1-ol (D3) and a trichloro-2-cyclohexene-1,4-diol (D4), most likely by hydroxylation of delta-pentachlorocyclohexene (delta-PCCH), initially formed by dehydrochlorination. The dehydrochlorinase LinA was responsible for conversion of delta-HCH into delta-PCCH, and the haloalkane dehalogenase LinB was responsible for the transformation of beta-HCH and delta-HCH into B1 and D1, respectively, and subsequently into B2 and D2, respectively. LinB was also responsible for transforming delta-PCCH into D3 and subsequently into D4. These hydroxylations proceeded in accordance with SN2 type reactions with initial substitution of equatorial Cls and formation of axially hydroxylated stereoisomers. The apparently high reactivity of equatorial Cls in beta- and delta-HCH toward initial hydroxylation by LinB of Sphingobium indicum B90A is remarkable when considering the otherwise usually higher reactivity of axial Cls. Several of these metabolites were detected in groundwater from a former HCH production site in Switzerland. Their presence indicates that these reactions proceed under natural environmental conditions and that the metabolites are of environmental relevance.