Disappearing Butterflies

HOW TO SOLVE A BIOLOGICAL MYSTERY USING MUSEUM COLLECTIONS AND DNA TECHNOLOGY


By Rebecca Whitla, PhD student at Oxford Brookes University


The Black-veined white butterfly (Aporia crataegi) was a large, charismatic butterfly with distinctive black venation on its wings. Once commonly found in the UK, the species unfortunately went extinct here in around 1925, with the last British specimens collected from Herne Bay in Kent. It isn’t fully understood why the species disappeared from the UK, but climate change, predation, parasites, and disease have all been suggested to have caused its disappearance — perhaps with several of these factors contributing to its decline. Central to solving the mystery of the disappearance of the Black-veined white will be the collections of butterflies that are stored in museums like OUMNH.

Butterflies tend to be well-represented in museum collections, and the Black-veined white is no exception. While the species has now been extinct in the UK for around 100 years, Lepidoptera enthusiasts from previous centuries often captured wild Black-veined white specimens for their personal collections. The abundance of Black-veined white butterflies in museum collections, like the collections at OUMNH, serve as a valuable repository for scientific research — including my own!

Black-veined white butterflies in the collections at OUMNH

Between June and December 2021, I undertook a research project using OUMNH’s Black-veined white butterflies. My task was to extract enough DNA from the butterflies to use for ‘whole genome sequencing’ — in other words, I was attempting to extract DNA from butterfly specimens to decode their complete DNA sequence. Getting DNA sequences from the historical specimens that are kept in Museums is no easy task, as DNA degrades over time. Nonetheless, animal specimens from natural history museums have successfully been used for whole genome sequencing and genetic analysis in the past, including species as diverse as longhorn beetles and least Weasels.

In order to work out how to extract DNA from the specimens, I had to try a variety of methods. This included experimenting to find out whether butterfly legs or abdomen fragments yielded more DNA, and whether non-destructive methods of DNA extraction were as effective as destructive methods. An example of a non-destructive method of DNA extraction would be a process like soaking a sample overnight and using the leftover liquid for DNA extraction, whereas a destructive method might involve mashing a whole leg or abdomen segment to use as a DNA source.

Preparing a DNA sample

Overall, I found that destructively sampling the legs of the butterflies gave the most reliable results, and also had the added benefit of not destroying the wings or abdomen of the specimens. Keeping the wings and abdomens of the butterflies intact will likely prove useful for conducting morphological studies in future.

Now that I have a reliable DNA extraction method, the next step in my research will be to analyse more Black-veined white specimens from a span of different time periods leading up to the species’ disappearance. I will then compare samples collected from each time period to calculate the genetic diversity of the species at each point in time, leading up to its disappearance. If I find a steady decline in the species’ genetic diversity over time, this may indicate a gradual extinction of the species. This is because we expect that, as numbers of a species decrease, inbreeding will become common, resulting in less diversity in the species’ DNA. However, if the populations of Black-veined white butterflies went extinct very suddenly, the decline in genetic diversity will probably be less pronounced. Learning more about the fate of the Black-veined White could not only help us unlock the historical mystery of the species’ decline in Britain, but will also help us understand more about the species’ decline in other parts of the world.

Re-Collections: Jane Willis Kirkaldy

By Evie Granat, Project Officer Trainee with the Freshwater Habitats Trust and Museum volunteer


The Museum is lucky enough to house several specimens presented by Jane Willis Kirkaldy (1867/9 – 1932). They serve as a reminder of a passionate and dedicated tutor, and of a key figure behind the development of women’s education at Oxford University.


Jane Willis Kirkaldy was born somewhere between 1867 and 1869, and spent her youth in London with her parents and five siblings. After completing her secondary education at Wimbledon High School, Kirkaldy gained entry to Somerville College (Oxford) on an exhibition scholarship in 1887. She finished her degree in 1891, becoming one of the first women to achieve a First Class Hons in Natural Sciences (Zoology). However, since the University didn’t award women degrees in the nineteenth century, it wasn’t until 1920 that Kirkaldy received her MA.

Upon completing her undergraduate studies, Kirkaldy worked for a short period as a private tutor in Castle Howard before returning to Oxford in 1894. Whilst researching at the University, she produced two papers for the Quarterly Journal of Microscopical Science, including an article entitled “On the Head Kidney of Myxine”. This study of the renal systems of hagfish was written with the aid of experimental work carried out by renowned zoologist Walter Weldon at his UCL laboratory. She also studied lancelets under the Oxford Linacre Professor of Zoology, publishing “A Revision of the Genera and Species of Branchiostomdae” in 1895.

Kirkaldy’s achievements are especially noteworthy given how few women studied Natural Sciences at Oxford during the nineteenth century. In addition to her contributions to the scientific field, she also helped advance women’s education at Oxford University. In 1894, The Association of the Education of Women named Kirkaldy a tutor to female students in the School of Natural Sciences. The following year she ceased all research to concentrate fully on teaching, co-authoring ‘Text Book of Zoology’ with Miss E.C. Pollard in 1896, and Introduction to the Study of Biology with I. M. Drummond in 1907. She eventually became a tutor or lecturer at all of Oxford’s Women’s Societies, and a Director of Studies at all five of the women’s colleges. Amongst the many female scientists that came under her care was the Nobel Prize-winning chemist Dorothy Crowfoot Hodgkin.

Left: Page from one of our donations books listing Jane Willis Kirkaldy as the donor of a series of Middle Devonian fossils (from the Eifel) to the Museum in October 1901. Right: Chromite from East Africa, also donated to the Museum by Kirkaldy.

Beyond the Department of Natural Sciences, Kirkaldy was an important figure at Oxford — she served as a member of the Council of St. Hugh’s College for 14 years, and was made an honorary fellow of Somerville College in 1929. At the Museum of Natural History, she presented beetles from New Guinea (1890), Devonian Fossils from the Eiffel (1901), and Chromite from near Beira, Mozambique (1924).

Kirkaldy retired from the University in 1930 due to ill health, before passing away in a London care home in 1932. Oxford University subsequently dedicated the junior and senior ‘Jane Willis Kirkakdy Prizes’ in her memory, which still exist to this day.


References

https://www.firstwomenatoxford.ox.ac.uk/article/principals-and-tutors

https://archive.org/details/internationalwom00hain/page/160/mode/2up

https://www.ias.ac.in/article/fulltext/reso/022/06/0517-0524

http://wimbledonhighschool.daisy.websds.net/Filename.ashx?tableName=ta_publications&columnName=filename&recordId=72

http://wimbledonhighschool.daisy.websds.net/Filename.ashx?tableName=ta_publications&columnName=filename&recordId=71

https://archive.org/details/internationalwom00hain/page/160/mode/2up

Dorothy Crowfoot Hodgkin: Patterns, Proteins and Peace: A Life in Science, by Georgina Ferry

Quarterly Journal of Microscopical Science

Presenting… Christmas Island

By Eileen Westwig, Collections Manager in the Museum’s Life Collections.

About 320 km south of Java in the Indian Ocean lies Christmas Island. Although discovered and named on Christmas Day in 1643, the island remained unexplored until its first settlement in 1888, a development which had dire consequences for some of its native species.

Christmas Island is home to a variety of endemic animals such as rats, land crabs, butterflies and many birds. The accumulation of bird droppings over thousands of years made the island rich in phosphate, and the commercial potential of these deposits brought many expeditions to the island. With the ships’ cargo came black rats.

Two species of endemic rats, Maclear’s Rat (Rattus macleari) and the Bulldog Rat (Rattus nativitatis) went extinct within 20 years of settlement, despite having been previously very numerous on the island.

One of the skins of Maclear’s Rat (Rattus macleari) collected by H.E. Durham, and now held in the Life Collections of the Oxford University Museum of Natural History.

Maclear’s Rat, seen at the top of the page in an illustration from an 1887 publication, was described as chestnut brown above, with a partly white, long tail. It was once the most numerous mammal on the island ‘occurring in swarms’. The Bulldog Rat had a much shorter tail and a layer of subcutaneous fat up to 2 centimetres thick, the function of which is unknown to this day.

The likely cause of their extinction was the introduction of diseases by the ship rats, to which the Christmas Island rodents had no immunity. The disappearance of the native rats also had a knock-on effect: the parasitic Christmas Island Flea (Xenopsylla nesiotes) depended on the rats as hosts, and so the fleas became extinct with the rats’ demise.

In 1901 Dr. Herbert E. Durham, a British parasitologist investigating the origins of beriberi disease, led an expedition to Christmas Island. During his visit he collected several specimens of Maclear’s Rat, but was unable to find any Bulldog Rats, despite a lengthy search and the offer of a reward. Two of the nine Maclear’s Rats Durham obtained showed abundant parasites, trypanosomes, in their blood.

Christmas Island possesses quite a number of peculiar species in its fauna, and it is regrettable that observations were not made before animals had been imported to this isolated station, as well as that my own notes are so incomplete.

Dr. Herbert E. Durham

Durham also found blood parasites in the native fruit bats (Pteropus melanotus) but noted that these were unlikely to have been introduced, instead were “an old standing native occurrence.” These bats still inhabit various islands in the Indian Ocean, including Christmas Island, where they are critically endangered.

Original letter by H.E. Durham offering his Christmas Island specimens to the Museum in 1938.

The Museum holds a range of material from Christmas Island, including six skins and three skulls of Rattus macleari, which were collected by H. E. Durham in 1901-02, and donated in 1938.

Visit the Museum’s Presenting… case between now and 6 March to see Christmas Island specimens from the collections.

Mammoth tusks and cocktail sticks

By Pete Brown, Move Project Assistant

As part of the Museum of Natural History Move Project Team I have helped move and repackage tens of thousands of specimens since 2017, removing boxes filled at any time over the last 150 years from their old storage location in a deconsecrated church building near Oxford.

At our new facility we have been documenting and repacking the contents in new, clean containers and placing them in environmentally stable, safe warehouses specially adapted for museum storage.

Some objects are trickier to store than others. Things that are long, heavy, curvy and fragile are tricky. Mammoth tusks are long, heavy, curvy, and fragile. This means:

  1. They’re not going to fit in a normal box.
  2. They’re going to be difficult to move around.
  3. That beautiful curve will mean that all the weight of the tusk may be bearing down on just two small contact points where the tusk meets the storage surface.
  4. Because those points are fragile, they’re likely to get damaged.

A lot of weight can rest on small areas of the tusk, putting strain on the specimen and potentially causing damage

The tusk in this article is a prime example. The area nearest the camera in the photo above provided just a tiny point of contact with the floor and was very loose, almost to the point of detaching. It needed to be repaired, and stored in such a way that it wouldn’t get damaged again.

Pete Brown carries out delicate conservation work on the mammoth tusk

I filled some of the missing areas around the fragile area with an easily removable fine acrylic putty to prevent further movement and loss of the original material. A cotton tape sling helped to suspend the fragment in place during the work.

Thick plastazote provided a sturdy, slightly yielding bed for the tusk to lie on in storage, but to prevent the tusk from getting damaged again more needed be done to reduce the pressure on the points of contact.

The dark grey foam material, plastazote, is often used as a cushioned support for museum objects

I cut depressions into the plastazote where the tusk naturally lay to increase the total surface area supporting the weight of the tusk, and fixed plastazote wedges and supports in place with cocktail sticks to again increase the contact area and prevent movement. Cotton fabric ties, fed through slits in the plastazote, also helped to guard against unwanted movement.

Cocktail sticks: not just for cheese and pineapple

The repaired end of the tusk is now only supporting a fraction of the weight it used to, and once the tusk and the plastazote bed are placed into their new custom-made crate it will be ready for long-term, safe, damage-free storage!

The end of the tusk after treatment

To keep up with all the move project action, follow the museum hashtag #storiesfromthestore on Twitter @morethanadodo.

 

Understanding beeswax

By Tuuli Kasso, PhD in Science Fellow at the Natural History Museum of Denmark, University of Copenhagen. Tuuli is a visiting researcher, who has used the Museum’s collection to help her understanding of beeswax. 

When working on the dissertation for my MSc in Archaeological Science last year, I explored the medieval craftsmanship of sealing wax. I was interested in the way the medieval wax seals had flaked, as the beeswax dried out. Drawing on my previous education in conservation techniques, I began a close investigation of the prestigious material, beeswax.

Medieval craftsmen used a range of dangerous materials to make sealing wax. The red pigment cinnabar, a mercury (II) sulphide, and red lead, are now known to be extremely poisonous.

Although some of the ingredients of sealing wax are very hazardous, there is nothing dangerous in beeswax… except the bees! Produced by honey bees, Apis mellifera, honey and beeswax were important commodities in the Middle Ages. Beekeeping was a skilful profession, housing colonies in woven hives, known as skeps. Colonies were carefully selected to overwinter for the next season.

Manuscript illuminations provide detailed information on the types and construction of beehives in the Middle Ages.England, 13th century. British Library Royal 12 C XIX f. 45.

Beeswax was also important in the Middle Ages for lighting, and beeswax candles were preferred for their pleasant smell. After the Protestant Reformation in the 16th and 17th centuries, the religious use of candles decreased, so demand for beeswax declined.

Even today, the Catholic and Orthodox Churches still require the candles they use to contain a proportion of beeswax.

On my quest to understand the degradation of beeswax in sealing wax and write my disseration, I was very lucky to use some samples from the entomological collections from the Oxford University Museum of Natural History. After some early mornings spent amongst the Westwood collection, I found the perfect specimens of natural honeycombs, from the 19th century. The old hand-written labels were also a lovely encounter when exploring the historical collections.

I compared the samples to modern beeswax and medieval seal samples, and learned that the degradation of beeswax is caused by multiple factors, triggered also by storage conditions. The composition of beeswax is very complex, and there are differences caused by the age of the bee in addition to geographical provenance.

A selection of bee specimens from the Museum’s collection.

The recent catastrophic decline of bee populations has drawn focus to save the bees, and in my PhD research (University of Copenhagen and University of Cambridge) I will explore the recovery of ancient DNA and proteins of bees from beeswax, to cast light on the health of bee populations over time.

From pin to paper

Katherine Child, image technician in the Museum’s Life collections, doesn’t just use photography to capture the beauty of specimens. She is also an artist and has been trying out innovative techniques for her paintings. You may remember her amazing moth illustrations created with deposits of verdigris on pinned insects and she’s now using that technique to explore Museum staff’s favourite insect specimens.

Verdigris is a green corrosion often found on old pins within entomology collections (as well as elsewhere, on things like statues and copper pipes). Last year, after learning that the substance was once used as a pigment, I decided to try and make my own paint.

A clearwing moth before conservation, showing verdigris spreading where metal reacts with insect fats, or lipids.

Verdigris forms when copper or a copper alloy reacts with water, oxygen, carbon dioxide or sulphur. While a beautiful shade of green, the substance is damaging in natural history collections, where it can actually develop inside specimens and if left, split them irreversibly. So as part of the conservation of the Hope Entomological Collections, verdigris is removed.

I started to collect up the substance as it was cleaned from specimens and after about three years (you only get a little bit per pin) I was ready to make my paint! After my first moth project, the only question was, what to paint next…?

Attelabid_small
Byctiscus populi or ‘The Attelabid that changed my life’, chosen by Zoë (collections manager) who said ‘I saw a pink version of this species in the Natural History Museum in London and that’s when I decided I wanted to study entomology’.

With an estimated 6 million insects and arachnids in the entomology collections, it’s very easy to feel overwhelmed. You can pull open any one of thousands of draws and find astonishing specimens. While I have favourites, my first inclinations as to what to paint still felt a little arbitrary. After mulling over various possibilities, I decided to get help!

Chosen by DPhil student Leonidas, Agalmatium bilobum is a little bug which lays its eggs on tree bark, then covers them with mud to protect them.

I asked my co-workers what their favourite insects were, then opened the question out to regular volunteers and visitors of the Life collections. I loved finding out why people chose the things they did. Answers varied from ‘It was the first spider I ever looked at under a microscope aged 12’ to ‘Because they’re cool’ to ‘Because they have an ingenious way of manipulating spiders!’

Nuctenea_small
One of arachnologist Russell’s favourite spiders: Nuctenea umbratica. Though common in the UK, umbratica is Latin for “living in the shadows”, and it often hides away during the day. The slight transparency of the paint lends itself to a spider’s glittering eyes.

 

Painting this live African Mantis Sphodromantis lineola (chosen by conservator Jackie) was made slightly more challenging by the fact that the subject thought Katherine’s pencil might be tasty.

Most of the subjects I painted were based on specimens from the Museum’s collections or specimens individuals had brought in from their own collections, but one favourite was a live African Mantis, housed in the department to help with education and outreach. When I began to draw her she was intrigued by the movement of my pencil and came to the front of the tank, to follow every mark I made with her intimidating gaze.

A detail from the final painting

Attelabid that...
Katherine’s fabulous finished painting, which will be framed and displayed in the Life collections department.

Though time consuming, the painting was loads of fun to research and do. It’s fantastic to be surrounded not only by extremely knowledgeable people, but also by people with a genuine passion for what they do and a love for the insects (and spiders) they study.