Specimen collection and preparation methods

Coleopterans

The collections of beetles in the territory of the Kurgalsky Peninsula was carried out used different means such as the light and soil traps, sweep netting and shaking off branches with an entomological net, sifting of various substrates using an entomological sieve, as well as generally accepted manual collection methods. The cameral preparation of the material was carried out by generally accepted entomological methods. The collected specimens were mounted on entomological pins, labelled and placed in the drawers of the Coleoptera collection of the Zoological Institute in taxonomic order.

Lepidopterans

Since there are no generally accepted universal methods of conducting investigations for different groups of lepidopterans, a combined technique was used, including observation of flying and feeding individuals and their catching with a net, observation and catching of individuals sitting on trunks and branches of trees, mowing with a net from grass and from tree branches, search for open-living caterpillars on different parts of plants for breeding imagos from them. Species identification was carried out visually when capturing or observing sitting specimens, sometimes – for the largest and most characteristic species – in flight. If it was impossible to make an exact visual identification in the field, the material was collected for subsequent laboratory processing (visual comparison with the fund collections and atlases, preparation and study of the structure of the genitalia).

Blood-sucking mosquitoes

The collection of larvae and pupae of mosquitoes was carried out according to standard methods using a household sieve with a diameter of 20 cm, the larvae and pupae were caught and then transferred into a plastic container filled with water. From this container, the material was transported to the Zoological Institute for further work. Adult insects were caught using an exhauster.

Rearing, determination, and fixation of mosquitoes:

Individual and mass rearing of mosquitoes was conducted in the Zoological Institute. During individual rearing, 4th-instar larvae were placed individually in a 200-ml containers, which were covered with a mosquito net on top. For mass rearing, groups of larvae and pupae were placed in 1-liter containers. Larvae of the 2nd and 3rd instar were kept in 10-liter containers until reaching the 4^th instar. Containers with emerged imago were placed for 5 min into the freezer until the state of torpidity of an adult insect occurs, after which the imago was removed from a freezer, and its further determination was carried out under a binocular microscope. A similar procedure was carried out with wild captured adult females. Species determination of mosquitoes was carried out according to standard keys and tables developed by authors (Gutsevich et al., 1970; Khalin, 2009; Becker et al., 2010). Adults were placed in 5-ml tubes (25 specimens per tube). Labels were used to indicate species, sex, and place of collection of the preimaginal phase. Tubes with the material were stored in a special refrigerator, maintaining a temperature of -80°C.

References:

• Becker N., Petric D., Zgomba M., Boase C., Madon, M., Dahl C., Kaiser A. 2010. Mosquitoes and their control. Second Edition. Berlin, Heidelberg: Springer-Verlag. 608 p.
• Gutsevich, A.V., Monchadsky, A.S., Stackelberg, A.A. 1970. Fauna SSSR. Nasekomye dvukrylye. Volume 3. No. 4. Komary semeystva Culicidae. Leningrad: Nauka. 384 p. [in Russian]

Amphibians, reptiles

• Route accounting/walking routes for various types of habitats. Equipment: rubber boots, GPS-navigator, photo camera, notepad.
• Survey of reservoirs/catch with a net of coastal vegetation in reservoirs used by amphibians for reproduction. Equipment: herpetological net, photo camera.
• Accounting for the laying of amphibians for estimating the number of breeding individuals/visual accounting of the laying of frog roe in reservoirs in the selected area.

Equipment: rubber boots, photo camera.

• Tracking on the roads/pedestrian and road routes on the roads for the collection and registration of crushed animals. Equipment: motor transport, photo camera.
• Photofixing/photographing animals and biotopes. Equipment: photo camera.
• Specification/visual definition in the field and laboratory definition. Equipment: photo camera, binocular microscope.
• Data processing/data processing for accounting and distribution, work with maps.

Mammals

• For predatory mammals (Carnivora) and ungulates (Artiodactyla) route surveys are planned according to generally accepted methods. The length and location of the transects are determined by the representativeness of various types of biotopes.
• Counts of insectivores (Lipotyphla) and rodents (Rodentia) are carried out at several monitoring sites located in the most typical habitats. When extrapolating data, the so-called a typological approach, the essence of which is that the species composition of mammals in similar biotopes in a limited area will not be fundamentally different (naturally, such an approach can only be used to analyze the distribution of background and subdominant species).
• To search for bats (Chiroptera), route and point counting techniques using an ultrasonic detector are used.
• The search for rare species of mammals is planned to be carried out purposefully, based on the available literature and stock data, as well as taking into account the habitat preferences of each of them.
• The spatial distribution and abundance of invasive species are determined on routes in winter and spring.