Pipetting for dummies

By Keafon Jumbam

PracticingI’m back in the lab once again to hone my skills on faecal hormonal assays. The unsuccessful ending to my first lab visit didn’t deter me from trying again. If anything, the lessons I gathered from that experience made me more determined to succeed in my analyses this time around. Besides, I’m surrounded by several lab experts who are always happy to offer support and guidance; starting with this – relax and develop a pipetting rhythm that works for you.

And on that note, I plunged into the fundamentals of hormonal assays.

Back to basics

The prerequisite for me to obtaining a good assay was to master the steps and types of reagents needed as well as brushing up on my pipetting techniques. So I set aside a couple of days for these tasks but the monotony of the mundane exercise – and in particular, pipetting – started to get to me by day two. I needed to persist through it for two main reasons: To get a ‘feel’ of the different assay liquids and subsequently handle them better and secondly, to develop that all too important pipetting rhythm!

Finally tackling the real deal – hormonal assay

By day three, I had a firm grasp of the basics and was confident in my abilities to pull off a good assay. The assay involves preparing a standard curve from a serial dilution of different volumes of liquids and reagents, which are then pipetted into a “plate” of 96 tiny wells. The wells are closely packed and certainly require your utmost concentration during pipetting. In fact, it is not uncommon to completely ignore visitors when busy with a plate; unless of course it’s an emergency.

pipettes

Three’s a charm

You would think that with all those days of practise I’ve had, surely I would nail the first assay but it turned out to be a disaster – and the second one too – both plates had HUGE outliers spanning the length of the standard curves. Was I already re-establishing my previous pattern of disappointment? Self-doubt was slowly setting in…But I thought to approach this non-academically and perhaps a bit more like a sportswoman. This is a physical feat, and after all, expert runners benefit from having somebody critiquing their technique.

The moment of truth... my hard work about to be interpreted...
The moment of truth… my hard work about to be interpreted…

So I solicited the help of an expert to invigilate my every step and it paid off – she quickly identified the mistake – yes, it was my pipetting technique! Another bout of training later and I was geared up for a third assay plate. I could hardly contain my excitement when it came out a success! The standard curve was on point as were the rest of my results. Practise does make perfect. I know it’s too soon to claim total victory but as I head back to the field to collect more faecal samples, I’m more confident in my abilities to analyse them in the nearest future. Watch this space…

A thing of beauty -- not a single outlier on my standard curve!
A thing of beauty — not a single outlier on my standard curve!

Rabies babies

By Keafon Jumbam
“Batties, batties! – we’ve just come across another dead batty.” For the past couple of months, this has been an almost daily radio call. In the beginning it was mostly unknown foxes on the growing obituary list, but our own have also joined the ranks… Ernie was the first to depart, followed by Bentley and then Bertha – the one and only project mom (gasp; there goes my existing link to maternal care studies!). To say we have been under immense stress would be an understatement. We live in constant fear of who will be next.

Initially, we couldn’t make sense of why batties were dropping dead like flies all over the reserve. An onsite vet dissected the carcasses but the cause of death remained unclear. Samples of the brain, lungs and heart were sent to laboratories for further investigation and the results came back positive for rabies. Then our panic really intensified because of the extremely high risk to ourselves. Rabies is often fatal in humans too, and I hadn’t taken any rabies shots yet, thanks to the public hospital policy in Phuthaditjhaba that wouldn’t administer vaccines unless the patient had been bitten and tested positive for rabies. I panicked even more as I reflected on a few instances in the past when I had been nipped in the leg by naughty batties demanding for more raisins. Was I already infected?

It turns out I wasn’t alone; some of my team-mates were also losing their minds over the outbreak and needed rabies shots and boosters. Orders for vaccines were hurriedly put through and although they arrived within a few days, it felt like months of waiting.

Newly collared and vaccinated foxes
Newly collared and vaccinated foxes

The vaccines weren’t our only concern; we needed to attend to the batties too. It is the start of breeding season and we need to keep track of them as they disperse in search of mates. Thanks to issues with our suppliers, most of our batties lacked radio collars. What is more, the mate-search and pairing up process comes with fierce scuffles and territorial fights that further increase the spread of rabies in our population. And batties are so cute you don’t realize they can be deadly. Baine for instance, had wandered off to a neighboring farm in search of a mate, but was sent packing with bloodshot eyes and half of his head mauled to a pulp. The vets couldn’t have come at a better time – they attended to their injuries, vaccinated them against rabies and then collared them.

Alas it was too late for Bentley who was found dead upon the vets’ arrival. His badly injured brother Baine was by his side on the night he was found dead. The deceased’s partner – Catelyn – was in no mood to play the grieving wife; she had already found solace in the paws of another fox.

His brother's keeper: Ben and Baine used to be inseparable
His brother’s keeper: Ben and Baine used to be inseparable

And so we took our cue from Cat, stopped moping, and ended the stressful weeks with a party themed Rabies Babies, in honour of the lives we’d lost. Dressed up as doctors and nurses in white lab coats and dissecting aprons, we danced our sorrows away. And you know the party wouldn’t be complete without some shots right? 🙂