Z16 microscope

Individuals preserved in 70% ethanol were cleared using KOH and acetic acid and mounted in slides with Canada balsam. The morphological identification of the specimens was done using a Leica Z16 microscope. We measured structures with taxonomic value and used the keys from Miller and Halbert (2014) (link) and Blackman and Eastop (2019) to identify species of Neophyllaphis. The photographs were taken with a Leica Z16 microscope, equipped with a CF500 camera and LAS 4.9 (Leica) image capture. Mounted specimens were deposited at the aphid collection of the Instituto de Biología Integrativa de Sistemas (Centro Mixto Universidad de Valencia-CSIC, Spain) and in the Centro de Investigación en Biología Celular y Molecular (CIBCM), Universidad de Costa Rica.

Three male specimens of H.rayi were examined and compared to male specimens of H.burnei (three specimens) and H.osburni (one specimen) borrowed from the
Florida Collection of Arthropods, Gainesville, Florida (FSCA).
The holotype and one paratypes of H.rayi MacGown & Hill, sp. nov. were deposited in the
Mississippi Entomological Museum (MEM)
and one paratype will be deposited in the Florida State Collection of Arthropods (FSCA). Genitalia were dissected and examined in 95% ethanol. Photomicrographs were captured using a Leica DFC 495 digital camera mounted on a Leica Z16 microscope with motorized Z-stepping, and image stacks were merged using Leica Application Suite v. 4.1.0 with Montage Module. All images were edited in Photoshop CS6. Measurements were made using a reticule placed in a 10× eyepiece of a Leica MZ16 stereomicroscope at a magnification of 10–100×.

Specimens from possible fire ant mounds reported by Kentucky citizens were collected by CAPS employees and placed in ethanol and/or hexane and sent to the MEM Invasive Insect Screening Center for identification and verification. Identifications were made using morphological methods and the keys found in [1 ]. When possible, identifications were verified using DNA analysis based on methods provided by Goodisman et al. [3 (link)] and using gas chromatography and mass spectrometer (GC/MS) methods to separate S. invicta, S. richteri, and their hybrid from one another based on different cuticular hydrocarbon and venom alkaloid profiles as described in Menzel et al. [2 ]. Specimens collected in ethanol were adequate for both DNA and morphological-based identifications, whereas samples stored in hexane were necessary for the GC/MS analyses. Photomicrographs were captured using a Leica DFC 495 digital camera mounted on a Leica Z16 Microscope with motorized Z-stepping, and image stacks were merged using Leica Application Suite V 4.1.0 with the Montage Module. All images were edited in Photoshop CS6. Voucher specimens were deposited in the MEM.