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Validating Morphometrics with DNA Barcoding to Reliably Separate Three Cryptic Species of Bombus Cresson (Hymenoptera: Apidae)

1
Department of Environmental Conservation, University of Massachusetts Amherst, 160 Holdsworth Way, Amherst, MA 01003, USA
2
Private Practice, Eau Claire, WI 54701, USA
*
Author to whom correspondence should be addressed.
Currently retired.
Insects 2020, 11(10), 669; https://doi.org/10.3390/insects11100669
Received: 14 September 2020 / Revised: 21 September 2020 / Accepted: 27 September 2020 / Published: 30 September 2020
(This article belongs to the Section Insect Ecology, Diversity and Conservation)
Evidence of bumble bee population declines has led to an increase in conservation efforts to protect these important pollinators. However, effective conservation requires accurate species identification. We provide quantitative methods to accurately identify three cryptic species of bumble bees using morphometric measurements of the cheek length and width, and antennal segments. We validated the accuracy of our methods with DNA analysis. We predicted that these methods would reliably identify both the queens and worker bees of Bombus vagans and B. sandersoni. We expanded these methods to include an uncommon form of Bombus perplexus with all light hair on its thorax, rather than the more common light on top and dark below, that can mistakenly be identified as B. vagans or B. sandersoni. Although the species we consider here, Bombus vagans, B. sandersoni and B. perplexus, are not currently listed as species of concern in North America, there is uncertainty of their population status, some of which is due to difficulty in species identification, which we have resolved. Recent history informs us that some bumble bee species experience rapid declines within a few decades. Our methods to correctly identify these cryptic species is key to monitoring their status and population trends.
Despite their large size and striking markings, the identification of bumble bees (Bombus spp.) is surprisingly difficult. This is particularly true for three North American sympatric species in the subgenus Pyrobombus that are often misidentified: B. sandersoni Franklin, B. vagans Smith B. perplexus Cresson. Traditionally, the identification of these cryptic species was based on observations of differences in hair coloration and pattern and qualitative comparisons of morphological characters including malar length. Unfortunately, these characteristics do not reliably separate these species. We present quantitative morphometric methods to separate these species based on the malar length to width ratio (MRL) and the ratios of the malar length to flagellar segments 1 (MR1) and 3 (MR3) for queens and workers, and validated our determinations based on DNA barcoding. All three measurements discriminated queens of B. sandersoni and B. vagans with 100% accuracy. For workers, we achieved 99% accuracy by combining both MR1 and MR3 measurements, and 100% accuracy differentiating workers using MRL. Moreover, measurements were highly repeatable within and among both experienced and inexperienced observers. Our results, validated by genetic evidence, demonstrate that malar measurements provide accurate identifications of B. vagans and B. sandersoni. There was considerable overlap in the measurements between B. perplexus and B. sandersoni. However, these species can usually be reliably separated by combining malar ratio measurements with other morphological features like hair color. The ability to identify bumble bees is key to monitoring the status and trends of their populations, and the methods we present here advance these efforts. View Full-Text
Keywords: Apoidea; Bombini; malar ratio; Bombus vagans; Bombus sandersoni; Bombus perplexus Apoidea; Bombini; malar ratio; Bombus vagans; Bombus sandersoni; Bombus perplexus
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MDPI and ACS Style

Milam, J.; Johnson, D.E.; Andersen, J.C.; Fassler, A.B.; Narango, D.L.; Elkinton, J.S. Validating Morphometrics with DNA Barcoding to Reliably Separate Three Cryptic Species of Bombus Cresson (Hymenoptera: Apidae). Insects 2020, 11, 669.

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