Effect of visitors on the behaviour of three Asian small‐clawed otters Aonyx cinereus

The Asian small‐clawed otter Aonyx cinereus is classified as Vulnerable by the International Union for Conservation of Nature Red List. The aim of this study was to evaluate the effects of the presence of visitors on the behavioural repertoire of three Asian small‐clawed otters at Cattolica Aquarium, Italy. Observational data sampling took place over two periods of 11 days each. The first period was in August when the Aquarium was open and there were visitors present (open period = OP). The second period was between September and October, on days when the Aquarium was closed to visitors (closed period = CP). Focal sampling was integrated with behaviour sampling during data collection. To evaluate the significant differences in frequency and duration of behaviours during the OP and CP periods, a Wilcoxon Signed Rank test was performed. The results of this study showed that the absence of visitors stimulated significant differences in frequency and duration of activity (such as ‘locomotion’ and ‘waiting’) and social behaviours (such as ‘play’). The otters also spent significantly more time visible than ‘out of sight’ when there were no visitors present. This preliminary research provides information that may influence the way zoos and aquatic parks manage their otters, and contributes to the improvement of the welfare of animals in human care

Real-time molecular optical micro-imaging of EGFR mutations using a fluorescent erlotinib based tracer

Abstract Background EGFR mutations are routinely explored in lung adenocarcinoma by sequencing tumoral DNA. The aim of this study was to evaluate a fluorescent-labelled erlotinib based theranostic agent for the molecular imaging of mutated EGFR tumours in vitro and ex vivo using a mice xenograft model and fibred confocal fluorescence microscopy (FCFM). Methods The fluorescent tracer was synthesized in our laboratory by addition of fluorescein to an erlotinib molecule. Three human adenocarcinoma cell lines with mutated EGFR (HCC827, H1975 and H1650) and one with wild-type EGFR (A549) were xenografted on 35 Nude mice. MTT viability assay was performed after exposure to our tracer. In vitro imaging was performed at 1 μM tracer solution, and ex vivo imaging was performed on fresh tumours excised from mice and exposed to a 1 μM tracer solution in PBS for 1 h. Real-time molecular imaging was performed using FCFM and median fluorescence intensity (MFI) was recorded for each experiment. Results MTT viability assay confirmed that addition of fluorescein to erlotinib did not suppress the cytotoxic of erlotinib on tumoral cells. In vitro FCFM imaging showed that our tracer was able to distinguish cell lines with mutated EGFR from those lines with wild-type EGFR (p < 0.001). Ex vivo FCFM imaging of xenografts with mutated EGFR had a significantly higher MFI than wild-type (p < 0.001). At a cut-off value of 354 Arbitrary Units, MFI of our tracer had a sensitivity of 100% and a specificity of 96.3% for identifying mutated EGFR tumours. Conclusion Real time molecular imaging using fluorescent erlotinib is able to identify ex vivo tumours with EGFR mutations