Scientists took a rare chance to prove we can quantify biodiversity by ‘testing the water’

Recent study conducted at a UK fishery farm provides new evidence that DNA from water samples can accurately determine fish abundance and biomass

Organisms excrete DNA in their surroundings through metabolic waste, sloughed skin cells or gametes, and this genetic material is referred to as environmental DNA (eDNA).

As eDNA can be collected directly from water, soil or air, and analysed using molecular tools with no need to capture the organisms themselves, this genetic information can be used to report biodiversity in bulk. For instance, the presence of many fish species can be identified simultaneously by sampling and sequencing eDNA from water, while avoiding harmful capture methods, such as netting, trapping or electrofishing, currently used for fish monitoring.

While the eDNA approach has already been applied in a number of studies concerning fish diversity in different types of aquatic habitats: rivers, lakes and marine systems, its efficiency in quantifying species abundance (number of individuals per species) is yet to be determined. Even though previous studies, conducted in controlled aquatic systems, such as aquaria, experimental tanks and artificial ponds, have reported positive correlation between the DNA quantity found in the water and the species abundance, it remains unclear how the results would fare in natural environments.

However, a research team from the University of Hull together with the Environment Agency (United Kingdom), took the rare opportunity to use an invasive species eradication programme carried out in a UK fishery farm as the ultimate case study to evaluate the success rate of eDNA sampling in identifying species abundance in natural aquatic habitats. Their findings were published in the open-access, peer-reviewed journal Metabarcoding and Metagenomics.

“Investigating the quantitative power of eDNA in natural aquatic habitats is difficult, as there is no way to ascertain the real species abundance and biomass (weight) in aquatic systems, unless catching all target organisms out of water and counting/measuring them all,”

explains Cristina Di Muri, PhD student at the University of Hull.
Drained pond after fish translocation.
Photo by Dr. Watson H.V.

During the eradication, the original fish ponds were drained and all fish, except the problematic invasive species: the topmouth gudgeon, were placed in a new pond, while the original ponds were treated with a piscicide to remove the invasive fish. After the eradication, the fish were returned to their original ponds. In the meantime, all individuals were counted, identified and weighed from experts, allowing for the precise estimation of fish abundance and biomass.

“We then carried out our water sampling and ran genetic analysis to assess the diversity and abundance of fish genetic sequences, and compared the results with the manually collected data. We found strong positive correlations between the amount of fish eDNA and the actual fish species biomass and abundance, demonstrating the existence of a strong association between the amount of fish DNA sequences in water and the actual fish abundance in natural aquatic environments,”

reports Di Muri.
Environmental DNA sampling using water collection bottles
Photo by Dr. Peirson G.

The scientists successfully identified all fish species in the ponds: from the most abundant (i.e. 293 carps of 852 kg total weight) to the least abundant ones (i.e. one chub of 0.7 kg), indicating the high accuracy of the non-invasive approach.

“Furthermore, we used different methods of eDNA capture and eDNA storage, and found that results of the genetic analysis were comparable across different eDNA approaches. This consistency allows for a certain flexibility of eDNA protocols, which is fundamental to maintain results comparable across studies and, at the same time, choose the most suitable strategy, based on location surveyed or resources available,”

elaborates Di Muri.

“The opportunity of using eDNA analysis to accurately assess species diversity and abundance in natural environments will drive a step change in future species monitoring programmes, as this non-invasive, flexible tool is adaptable to all aquatic environments and it allows quantitative biodiversity surveillance without hampering the organisms’ welfare.”

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Original publication:

Di Muri C, Lawson Handley L, Bean CW, Li J, Peirson G, Sellers GS, Walsh K, Watson HV, Winfield IJ, Hänfling B (2020) Read counts from environmental DNA (eDNA) metabarcoding reflect fish abundance and biomass in drained ponds. Metabarcoding and Metagenomics 4: e56959. https://doi.org/10.3897/mbmg.4.56959

Scientists use forensic technology to genetically document infanticide in brown bears

Modern open-source software helped the researchers identify the male that killed a female and her two cubs

Scientists used a technology designed for the purposes of human forensics, to provide the first genetically documented case of infanticide in brown bears, following the murder of a female and her two cubs in Trentino, the Italian Alps, where a small re-introduced population has been genetically monitored for already 20 years.

The study, conducted and authored by Francesca Davoli, The Italian Institute for Environmental Protection and Research (ISPRA), Bologna, and her team, is published in the open access journal Nature Conservation.

To secure their own reproduction, males of some social mammalian species, such as lions and bears, exhibit infanticidal behaviour where they kill the offspring of their competitors, so that they can mate with the females which become fertile again soon after they lose their cubs. However, sometimes females are also killed while trying to protect their young, resulting in a survival threat to small populations and endangered species.

“In isolated populations with a small number of reproductive adults, sexually selected infanticide can negatively impact the long-term conservation of the species, especially in the case where the female is killed while protecting her cubs,” point out the researchers.

“Taking this into account, the genetic identification of the perpetrators could give concrete indications for the management of small populations, for example, placing radio-collars on infanticidal males to track them,” they add. “Nevertheless, genetic studies for identifying infanticidal males have received little attention.”

Thanks to a database containing the genotypes of all bears known to inhabit the study site and an open-source software used to analyse human forensic genetic profiles, the scientists were able to solve the case much like in a television crime series.

orsa occultata - leggeraUpon finding the three corpses, the researchers were certain that the animals had not been killed by a human. In the beginning, the suspects were all male brown bears reported from the area in 2015.

Hoping to isolate the DNA of the perpetrator, the researchers collected three samples of hairs and swabbed the female’s wounds in search for saliva. Dealing with a relatively small population, the scientists expected that the animals would share a genotype to an extent, meaning they needed plenty of samples.

However, while the DNA retrieved from the saliva swabs did point to an adult male, at first glance it seemed that it belonged to the cubs’ father. Later, the scientists puzzled out that the attacker must have injured the cubs and the mother alternately, thus spreading blood containing the inherited genetic material from the father bear. Previous knowledge also excluded the father, since there are no known cases of male bears killing their offspring. In fact, they seem to distinguish their own younglings, even though they most likely recognise the mother.

To successfully determine the attacker, the scientists had to use the very small amount of genetic material from the saliva swabs they managed to collect and conduct a highly sophisticated analysis, in order to obtain four genetic profiles largely overlapping with each other. Then, they compared them against each of the males reported from the area that year. Eventually, they narrowed down the options to an individual listed as M7.

“The monitoring of litters is a fundamental tool for the management of bear populations: it has allowed the authors to genetically confirm the existence of cases of infanticide and in the future may facilitate the retrieval of information necessary to assess the impact of SSI on demographic trends,” conclude the researchers.

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Original source:

Davoli F, Cozzo M, Angeli F, Groff C, Randi E (2018) Infanticide in brown bear: a case-study in the Italian Alps – Genetic identification of perpetrator and implications in small populations. Nature Conservation 25: 55-75. https://doi.org/10.3897/natureconservation.25.23776