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SpectralWorks Ltd.. (11/7/13). "Press Release: AnalyzerPro Goes to the Dogs – »Scentomics« to Protect Endangered African Wild Dogs".

Organisation Organisation SpectralWorks Ltd.
Products Product AnalyzerPro® mass spectrometry data mining software
  Product 2 GC/MS (gas chromatography/mass spectrometry)
     


AnalyzerPro is being used in groundbreaking research by the Paul G Allen Family Foundation Laboratory for Wildlife Chemistry in Maun, Botswana to search for and identify the active components of African wild dog scent marks.

African wild dogs are intensely social large predators that live in packs of up to about 30 adults and a litter of pups that they all care for. Also known as the painted dog, the African wild dog is one of Africa's most endangered carnivores, and with less than 6000 remaining, and self-sustaining populations in only 3 countries, plausible population models predict that wild dogs will be extinct in the wild in 30-40 years unless new ways are found to protect them. Finding those new ways is the mission of the Bioboundary® project, the brainchild of Dr John "Tico" McNutt of the Botswana Predator Conservation Trust (BPCT), an independent research organization that has been working on the large predators of Botswana's world-renowned Okavango Delta region since 1989.

Botswana is one of a handful of countries whose protected wildlife areas are large enough to sustain wild dog populations, but even here the dogs' propensity to roam over huge distances takes them out of protected areas and into livestock areas where they come into fatal conflict with humans. Predators in livestock areas threaten peoples' livelihoods, and the major cause of wild dog deaths is humans, so keeping the dogs and humans apart will benefit both sides in the conflict.

Bioboundaries will tap into the chemical signals that wild dog packs use to inform other dogs that an area is occupied and to stop packs from trespassing into one another's home ranges. Artificial territorial boundaries that mimic the effects of natural pack territory markers will stop wild dogs inside protected areas from trespassing into livestock areas.

Developing artificial scent marks begins with deformulating the natural scents, and with funding from the Paul G Allen Family Foundation the BPCT set up a dedicated gas chromatography - mass spectrometry laboratory to identify the active signalling constituents of African wild dog scent marks collected from the BPCT's habituated study packs. All mammal secretions and excretions are bewilderingly complex and it would be utterly impractical to replicate wild dog scent marks by identifying everything in them and blindly reconstructing the mixture; an artificial scent mark needs to contain only the compounds that contribute to the chemical signal. The challenge then becomes to sift out which of the hundreds of compounds in natural scents marks are actually sending chemical messages. Differences in composition that correlate with signalling function, and compounds that fit the design criteria for a territorial mark; stability, robustness to widely varying temperature and humidity, persistence, and specificity to wild dogs are the clues that will lead to the formula for an artificial "Keep Out" signal.

Comparisons of results from wild dogs with those from other mammals show that none of the compounds identified so far in wild dog urine are unique to wild dogs, and the major peaks on a total-ion chromatogram are common metabolites that occur in the urine and faeces of other mammals, so they are not likely to be signalling compounds. All the interesting things are happening among the minor peaks, produced by pico- to nanomolar gas phase concentrations which are easily within the range of a wild dogs' sense of smell but close to the limits of detection for GC-MS analysis.

In theory, a 30 m capillary gas chromatography columns can separate 300 - 400 compounds, but in natural materials the component's retention times are randomly scattered, and this causes dozens of overlapped and co-eluting peaks. On a typical chromatogram from a mammal odour least 20 % of the peaks overlap, and as the detection limit falls, the complexity rises; on a dog mark chromatogram there is no clean baseline. Overlapped peaks cannot be properly quantified or identified, and most failures to find an MS library match are due to mixed spectra from co-elutions, only a minority are due to new, interesting, compounds. Deconvolution with AnalyzerPro unravels the complexity of the minor peaks of the dog scent chromatograms.

The challenges posed by deformulating African wild dog scent messages are very similar to the challenges posed in the rapidly developing field of metabolomics - both need to find small differences in composition between very complex chemical mixtures, and identify the compounds responsible. But wild dog semiochemistry is metabolomics with a twist. Unlike the samples from uniform lab animals living under controlled conditions, the samples from free-ranging large predators include huge amounts of biological noise. Total peak area varies by more than four orders of magnitude, and of hundreds of components only a dozen or less are found in every sample.
Dr Peter Apps, who runs the laboratory commented, "To be any use for African wild dog semiochemistry, deconvolution software has to be flexible enough to detect and integrate minor peaks when the whole sample composition changes from sample to sample and there are no reliable landmark peaks that can be used to align retention times between chromatograms. I torture-tested the trial version of AnalyzerPro against chromatograms where I already knew that there were tricky multiple co-elutions and retention shifts. AnalyzerPro found all of the hidden components that I knew were there, and several that I missed because they were completely obscured by major peaks".

"AnalyzerPro has features that are especially useful for unravelling the complexities of wild dog scent signals. The MatrixAnalyzer™ generates a list of all the components of a set of samples, and reports their peak areas for each sample, and by selecting settings in the MatrixAnalyzer method it is possible to search for target components even when the chromatograms were run under completely different conditions, or are from very different materials. Search parameters can be set differently for individual components - so the retention time window can be set wider for a compound that is susceptible to variable retention. The deconvolution software interfaces seamlessly with searches of the NIST spectral library, and the options to export results and raw data in various formats make it easy to transfer data for statistical analysis."

"AnalyzerPro has already converted 65 chromatograms of wild dog urine volatiles into a set of comparable data on 862 components that we are now exploring with multivariate statistics, and we are running pairwise comparisons of wild dogs with leopards, lions, spotted hyaenas and cheetahs. The components that correlate with signalling will be identified and formulated into prototype artificial scents that the BPCT field team will test on our study packs".

For more on the BPCT's research programmes visit www.bpctrust.org


About SpectralWorks Limited

SpectralWorks Ltd is a leading UK based software development company.

We are dedicated to providing innovative solutions targeted for markets within the life sciences industry and have strong working relationships with a number of the major instrument manufacturers. Coupled with their collaborations with academia and industry, they have the right balance between scientific and software development expertise to provide the best scientific solutions for the end user.

In addition to a wide range of mass spectrometry software products, they provide highly respected consultancy services. These services are able to cover the complete software development cycle or specific steps within a project life cycle. These services are frequently utilized by instrument vendors and end users that demand the best for their laboratories. Visit www.spectralworks.com

A list of SpectralWorks articles and contact details on LaboratoryTalk is available here:
www.laboratorytalk.com/news/slw/slw000.html

   
Record changed: 2016-01-10

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