Validation of method to monitor methaemoglobinaemia in birds using small (5 µL) blood samples in New Zealand bird fauna.

A minimally invasive blood sampling technique was produced to allow data to be generated from birds as small as 10 g in body mass. <br> These are data were used to validate the alternative and non-lethal method to determine the susceptibility of 9 birds to a methaemoglobin-forming agent. Data collection was typically at T0, 1 h, 3h, 24h, 48h, 72h with additional samples taken to monitor welfare states if required. <br> Data were collected from domestic chicken pullets (Hyline strain), pekin ducks (<em>Anas platyrhynchos</em>) and Japanese quail (<em>Coturnix japonica</em>), black-backed gulls (<em>Larus dominicanus</em>), pukekos (<em>Porphyrio melanotus</em>), North Island brown kiwi (<em>Apteryx mantelli</em>), weka and eastern rosellas (<em>Platycercus eximius)</em>. <br> Absorbance (630 nm and 575 nm), lactate, and haemoglobin data were generated by different instruments and summarised in this spreadsheet before formal analysis. Detailed clinical notes and data were recorded on a paper record (not included but archived at Manaaki Whenua - Landcare Research). <br> Data are used for the calculation of a methamoglobin absorbance index (MIA) that is denoted in these data as Eq 2 that was validated against a decline in haemoglobin (HbFe2+) using 5 micro L blood samples. <br> Haemoglobin (Hb) is normally maintained in the ferrous valence (HbFe2+) enabling it to form oxyhaemoglobin (HbO) for the transport of oxygen (O2). A methaemoglobinaemia absorbance index (MAI = absorbance 575 nm – absorbance 630 nm) was strongly and significantly associated with dose-dependent declines in HbFe2+in 9 bird species. Absorbance data were generated over the full range of the spectrometer (350 – 1000 nm) from 3 µL blood samples pipetted into a disposable cuvette (EKF Diagnostic: Cardiff, United Kingdom). The cuvette window was placed in the spectrometer’s light path using a custom-built holder that was 3D printed from opaque black polylactic acid (PLA) and inserted into a machined aluminium block housing that accepted the SMA 905 connectors from the optical fibre terminators(Ocean Optics: Largo, USA). Lactate (LAC) was measured using the Lactate Scout® (EKF diagnostics, Germany) by pipetting between 0.5-0.8 µL of whole blood onto the biosensor and reading the result as mmol L-1. A further 1 µL of whole blood was pipetted to the biosensor of the Cera-Chek total haemoglobin (Hbt) and packed cell volume (PCV) point-of-care monitor and read as g Hb dL-1and % of the cell fraction in the sample respectively. <br> <br> <br>