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Super-flies and parasites

2 Posts tagged with the forensic_entomology tag
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Hello Super-flies and Parasites fans!

 

We are back with all things nasty from the Parasites and Vectors division here at the Museum. There have been some exciting developments in the New Year, most importantly the launch of the Museum’s brand new website!

 

This is another ‘Forever Flies’ series of blog posts, bringing you news from the Museum'sforensic entomologygroup.

green-bottle fly.jpg

Forever Flies is our forensic entomology blog series. This image shows a carrion-eating greenbottle blowfly.

 


Forensic Entomology

You will remember from my previous Forever Flies post that forensic entomology is the study of the insects and arthropods found at a crime scene. The most common role for Museum forensic entomologists is establishing a minimum time since death in suspicious cases, by analysing the carrion insects on the body.

 

2014-10-16 Gross maggots with adult.jpg   issue2forensic3_large.jpg

Blowflies use the bodies of dead animals to grow and develop. The rate at which they do this, going from egg to larva to pupa to adult fly, is pretty consistent and depends largely on ambient temperature. Forensic entomologists use this to determine the minimum post-mortem interval (PMImin), which helps crime scene investigators determine approximate time-of-death.


Thanks to entomological expertise (Greek – entomo = insect, logos = knowledge) scientists can collect insects from a corpse and/or crime scene, determine what stage in their life cycle the insects have reached and, using their knowledge on the duration of each stage of the insects’ life cycle, determine how long ago the parent insect laid her eggs on the corpse.

 

This gives an incredibly useful estimate of the minimum amount of time this body has been dead (minimum post-mortem interval - PMImin), which helps crime scene investigators determine approximate time-of-death. The more accurate this minimum post-mortem interval is, the more accurate the time of death can be. Knowing time of death can focus the police investigation and suggest the likelihood of a suspect’s involvement.

 

Scientists can also use these insects to determine if the body has been moved since death and how long a body was exposed above ground before burial.

 

Metamorphosis in pupae


Flies spend over 50% of their developmental life in the pupae stage, protectively encased inside a hard shell (called a puparium) where they slowly transform from a maggot into a fly in a process called metamorphosis (Greek again - Meta = change, morphe = form).

 

A puparium looks quite bland and boring but underneath there are all sorts of wonderful things going on. Scientists can remove the shell and, using traditional microscopy, take a look at the fascinating changes of metamorphosis. But this process does destroy the pupa sample, making it difficult to work out how long it takes for the pupa to go through the different stages of metamorphosis.

 

Scientists know that the length of time metamorphosis takes to complete really depends on temperature, the question is can we use our knowledge of the process to pinpoint a more accurate estimate of PMImin?  What forensic scientists need is a standardised method to work out:

  1. At what stage in the metamorphosis process is the pupa
  2. how long did it take to reach this stage

 

If these two points can be determined then scientists can provide a far more accurate PMImin.

 

The ‘MORPHIC’ project

 

Dr Daniel Martin-Vega, a forensic entomologist, has joined the Museum from the University of Alcalá in Spain to research carrion fly pupae and to develop a standardised protocol for aging pupae (as in determining their age) that can be used by forensic scientists. This project is called MORPHIC and is funded by the European Commission through a Marie-Curie fellowship.

 

It sounds all neat, logical and tidy but there is A LOT of work and dedication involved!

 

For this projectDaniel is raising two species of the carrion-loving blowflies, the greenbottle blowfly Lucilia sericata and the bluebottle blowfly Calliphora vicina. The flies live in netting covered cages, where they feed and reproduce whilst he monitors them.

 

Daniel feeding flies_resized2.jpgDaniel showing me the Diptera (insect) culture room. Each netting-covered box has a species of carrion blowfly in it. He is researching the pupae of these flies to see if he can improve the estimate of  PMImin and thus improve the information given to crime scene investigators.


He also has to collect the post-feeding maggots and place them in a box with some nice clean soil for them to happily grow until they are ready to start the metamorphosis process. These boxes are then placed in a cabinet kept at a specific temperature. Since the rate of metamorphosis largely depends on temperature it is very important the Daniel can control this environmental factor in order to document the rate of change at different temperatures.

 

where do you keep the maggots_resized2.jpg

The maggot house! This is a comfy box with soil where maggots crawl around and prepare to pupate. When the maggots start pupating Daniel has to come in every 6 hours or so to monitor and collect them for his research

 

puppa&maggots_resized.jpg

Blowfly maggots and pupae.

 

 

Once the maggots start to pupate Daniel has to collect the pupae:

 

I come in every 6 hours when the maggots start to pupariate in order to collect blowfly pupae at 6-hour intervals during the first 48 hours after puparium formation (the period when the greatest morphological changes of metamorphosis occur). Luckily, I only do this from time to time. After that, the collection of pupae is just daily until the adult flies’ emergence.

picking out pupae_resized.jpg

Daniel sieving out the pupae from the box.

 

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Maggots and pupae, oh my!

 

Watch those maggots wriggle about!

 

He then has to sieve out the pupae from the soil and carefully place them in a petridish labelled with the blowfly species name, the date collected and the time collected. These petridishes are also placed in the special temperature-control cabinet.

 

 

Fly puppa sorted_resized.jpg

Daniel has separated out the pupae of different species of blowfly. Each petridish with pupae has the species name, the date collected and the time collected.

 

where do you keep the maggots_resized.jpg

The petridishes are kept at a specific temperature. Since the rate of metamorphosis largely depends on temperature it is very important the Daniel can control this environmental factor.

 

Daniel uses the Museum’s wonderful micro-computed tomography (micro-CT) scanner to take detailed images of the inside of the pupae without destroying them. A micro-CT scanner is a type of X-ray scanner that produces 3D images, much like a hospital CAT scanner, but at a much smaller scale and a higher resolution. The results are like 3D microscope images! 

 

Thomas Simonsen and Daniel Martin-Vega analysing CT images of 5 pupae.jpg Thomas Simonsen and Daniel Martin-Vega operating Micro-CT scanner.jpg

Daniel with colleague Dr Thomas Simonsen using the Museum’s micro-CT scanner to look at 3D images of blow-fly pupae. The micro-CT scanner uses x-ray technology to produce 3D 'microscopy' images at high resolution without damaging the sample.

 

By using the Museum’s micro-CT scanner Daniel can take these detailed images at specific time points of the metamorphosis process.  He will then have a catalogue of images of the blow fly pupal development at specific temperatures. This catalogue of images will be used to develop a standardised tool to determine the age of blow fly pupae. Then when pupae are collected from a crime scene, they can be compared to this catalogue and scientists will be able to determine how long the fly has been in its pupal stage. Giving scientists a more accurate estimate of PMImin! Ta daaaaa!

 

C_vicina-48h.jpgC_vicina-216h.jpg

Micro-CT scanner images of a bluebottle blowfly Calliphora vicina pupa. The one on the left is at 48 hours, the one on the right at 216 hours. You can see the difference in development between the two pupa images.

 

Micro-CT scan of blowfly pupa.jpg

Dorsal micro-CT scanner image of a blowfly pupa.

 

I hope you enjoyed this post. If you fancy a stab at a bit of CSI work why not check out the Museum's Crime Scene Live After Hours events.

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Hello Super-flies & Parasites fans!

 

This time we are departing from the familiar world of blood flukes and having a look at something new and exciting: Welcome to the ‘Forever Flies’series of blog posts. I’ve really enjoyed writing this bit as it’s totally new to me and I’m learning so much from our wonderful scientists in the Forensic Entomology group of the Parasites & Vectors division here at the Museum.

 

About forensic entomology


I think I mentioned forensic entomology way back in the first ever Super-flies and Parasites post. But to refresh our memories and delve a bit deeper here’s an explanation taken from the group’s website:

'Forensic entomology is the study of insects and other arthropods (ie spiders, mites) in a situation where a crime may have been committed. The insects recovered from a crime scene can provide vital information for the investigating team. The most common role for Museum forensic entomologists is establishing a minimum time since death in suspicious cases, by analysing the carrion insects on the body.'

 

Flies use the bodies of dead animals to grow and develop, fulfilling a vital nutrient recycling role in an ecosystem. They can turn one vertebrate body into thousands or millions of flies, which are then fed on by other animals - insects, frogs and birds for example. The rate at which they do this, going from eggs to larvae to pupa to adult fly, is pretty consistent.

 

Knowing enough about the species of flies we can exploit this information when they develop on corpses. By determining how long the insects have been feeding on the tissues of the corpse, we can determine the length of time elapsed since flies found the body; thereby providing crime scene investigators with a minimum post-mortem interval.

 

Some flies however don’t hang about and wait for death, they prefer feeding on live animal tissues, and can cause a horrible disease called myiasis, a major economic and animal welfare problem.


2014-10-16 Gross maggots with adult.jpg

Female Bluebottle blow fly, Calliphora vicina, and maggots feeding on a dead pig.

 

Dr Martin Hall and colleagues within the Museum and around the world work together on the taxonomy and biology of flies that develop as larvae on living or dead vertebrate animals. Their expertise and research greatly contributes to the control of a painful and damaging disease, myiasis, but also to the field of forensic entomology, helping CSIs determine crucial information from a crime scene.

 

So there you have it, some real life crime scene investigation stuff! These are the guys CSIs turn to for help! (Cue CSI theme song!)

 

The beauty of maggots lies in their mouthparts

 

I asked Martin for a little blurb to get the Forever Flies series rolling and he sent me some surprisingly beautiful photos of maggots! Here’s what he has to say about them:

To most people maggots are repulsive creatures; they all look much the same and have zero redeeming features.

 

2014-10-16 Gross maggots 1-1.jpg2014-10-16 Gross maggots 2.jpg

Maggots, thought of as repulsive creatures with zero redeeming features. Read on to see how they are transformed by confocal microscopy into beautiful works of art.


However, viewed under a microscope they become much more interesting, with a range of characters that can be used in discrimination, especially when you look at the business end of the maggot, its mouthparts!

 

It’s not so easy to ask a tiny 2mm long newly hatched maggot to 'open wide' to view the teeth, and traditionally we have been limited to viewing slide-mounted specimens by light microscopy. Scanning electron microscopy has its merits, but only for the external features. In normal light microscopy, imaging of these mouthpart structures is limited by problems of resolution, illumination and depth of field.


Lucilia sericata L1 - Light microscope high power.jpg

Greenbottle blowfly, Lucilia sericata light microscope high power image. With normal light microscopy the relationship of the sclerites of the cephaloskeleton (mouthparts) to each other is unclear.

 

At the Museum we have been using a laser confocal microscope for the first time to look inside these maggots to view the mouthparts, the so-called cephaloskeleton, in three dimensions. The mouthparts are crucial to the maggots in establishing themselves on their food source, be it a live animal or a decomposing corpse. The images produced by the confocal microscope rely on the autofluorescence of structures of the cephaloskeleton.


Lucilia sericata LI - Confocal microscope low power.jpg

Greenbottle blowfly, Lucilia sericata confocal microscope low power image - relationships are still unclear but, with the autofluorescence under laser light, the structures look so much more beautiful!

 

We were especially interested in the relationships of the small sclerites to each other and the so called 'hump', present in newly hatched larvae of Lucilia greenbottle blowflies but absent in Calliphora bluebottles.


Lucilia sericata L1 - Confocal microscope high powerWITH ARROW.jpg

Greenbottle blowfly, Lucilia sericata confocal microscope high power image: The structure relationships are becoming clearer (the 'hump' is arrowed) and this is finalised in the next image.

 

The 177 optical sections scanned by the confocal microscope enabled us to rotate and view this structure in three dimensions (see green false-coloured sclerite in image below) and see clearly for the first time how it relates to other structures.

 

In addition to their academic and practical value in identification, the images are also things of beauty in their own right and would not look out of place in an art gallery!


Lucilia sericata - Confocal high power - false colour sclerites named.jpg

Greenbottle blowfly, Lucilia sericata confocal microscope high power image and false colour sclerites. We have rotated the original to give three dimensionality. The red wavelength was selected, as this is the wavelength that gave most autofluorescence of the cephaloskeleton, to enable us to discard other structures and here false-colour has been added to show the different structures, including the 'hump' (in green) or epistomal sclerite.

 

The work was done at the Museum and involved Drs Andrzej Grzywacz (a visitor under the EC-funded SYNTHESIS project) and Krzysztof Szpila from the Nicolaus Copernicus University in Torun, Poland, collaborating with Tomasz Góral and Martin Hall. We used the Museum’s Nikon A1-Si Confocal Microscope.

 

For more information see the article published online in Parasitology Research on 19 September 2014.


By Dr Martin Hall

 

I hope you enjoyed the first 'Forever Flies' post. For more on flies head over to Erica McAlister's Diptera blog.

 

There will be more coming soon but just to let you know there may be a couple of weeks break whilst some important maintenance work is done to the site and my life

 

See you soon