Material and Methods

Sampling sites
The Mt. Malindang Range Natural Park is located at the core of the Province of Misamis
Occidental that lies at 8º17’ N and 8º29’ N latitude and between 123º32’ E and 123º46’ E
longitude. It has a total land area of 53,028 hectares that covers the forest lands of the Cities of
Oroquieta, Ozamis and Tangub; and the municipalities of Sapang Dalaga, Concepcion, Don
Victoriano, Calamba, Aloran, Panaon, Lopez Jaena, Jimenez, Tudela Sinacaban, Clarin and
Bonifacio.
Ten sampling sites will be established representing ten of the major river catchments in the
province of Misamis Occidental. Site 1 will be established in Bagumbang river in the municipality
of Bonifacio, site 2 in Migcanaway River in Tangub City, sites 3 and 4 will be established in Labo
and Clarin rivers in Ozamiz City and Clarin, respectively. Sites 5 and 6 will be located in Palilan
and Aloran rivers in the municipalities of Jimenez and Aloran. Site 7 will be established in
Layawan river in Oroquieta city, site 8 in Daisog river in Lopez Jaena, sites 9 and 10 in Langaran
river and Dioyo river in Plaridel and Baliangao, respectively. In each sampling site, three sub-sites
will be established representing the upstream, midstream and downstream of the river.

Figure 1. Map of the sampling sites in Mt. Malindang Range Natural Park.

Sample Collection and Identification

Opportunistic sampling method will be utilized using a standard entomological hand net.
Sampling collection will be done between 8 a.m. and 5 p.m. for 1 hour in each sampling site
covering all possible habitats in river banks, streams, sandy areas, shrubs and moist rocks.
Collected samples will be preserved in 96% ethanol and will be examined using a
stereomicroscope. Species identification will be done by comparing morphological characters
using existing taxonomic keys and published works.

Measurement of the Morphological Traits

Measurement of the morphological traits for species identification will be done following
the methods of Jaskula et al., (2020). The following body parameters right mandible length (RML),
length of head (LH), width of head (WH), length of pronotum (LP), width of pronotum (WP),
length of elytra (LE), maximum elytra width (MEW) and total body length (TBL) will be
measured in both male and female collected samples.

Determination of Habitat Preferences

Habitat preferences will be classified based on vegetation type in the area, climatic and soil
parameters. Air temperature, soil parameters such as temperature, soil pH and salinity will be
measured and recorded. In every sampling site, soil pH and soil humidity (%) will be measured in
three places where tiger beetles will be observed and average values of those measurements will
be noted. Soil structure will also be determined following the methods of Jaskuła et al., (2019).
Three sub-samples of soil will be collected (in total 150 ml of volume) in the same places of the
sampling site where pH and humidity of soil will be measured for laboratory analysis.

Laboratory analysis
To check the soil structure, all samples will be dried separately on Petri dishes in an electronic
drier. Next, every sample will be weighed, and sifted on electronic sieves. All received parts of
soil particles gravel (>2 mm), sand (0.0632 mm), silt (0.063–0.002 mm), and clay (≤0.002 mm)
will be weighed. To estimate the proportion of particular soil particles in each sample, the values
of their weight will be compared with the total weight of the entire soil sample.
To check soil salinity, in the case of every sample, a volume of two ml of soil will be dissolved in
100 ml of distilled water. Then, using the WTW Multi 350i probe, electrical conductivity of soil-
water solution will be measured (three measurements will be done to note the average value used
in further analysis).

Statistical methods
Statistical analysis concerns 112 samples of 11 Cicindelidae taxa (presence/absence
data): Calomera aulica aulica (Caa), C. fischeri fischeri (Cff), C. littoralis
nemoralis (Cln), Cephalota besseri besseri (Cbb), C. chiloleuca (Cch), C. circumdata
circumdata (Ccc), Cicindela monticola rumelica (Cmr), Cylindera germanica
germanica (Cgg), C. trisignata hellenica (Cth), C. trisignata trisignata (Ctt), and Myriochila
melancholica (Mm). Species Cicindela maritima kirgisica (Cmk) and samples AL-02 and UA-03
were excluded from macrohabitat analysis due to the fact that they were outliers in the analysed
communities. This material was supplemented by data on: 1/five microhabitat environmental
parameters—altitude, soil pH, soil humidity, soil salinity, and soil sediment granulometry
(percentage share of gravel, sand, silt, and clay), 2/five macrohabitats types—saltmarshes, banks
of rivers, banks of lakes, sandy sea beaches, sandy-stony sea beaches, and 3/four climatic zones:
arid-steppe-cold (Bsk), temperate with dry, hot summer (Csa), temperate with no dry season and
hot summer (Cfb), and cold without any dry season and with warm summer (Dfb).

Multivariate statistics were calculated for biotic and environmental data. Principal Component
Analysis (PCA) on normalised data was conducted for the environmental ordination of sites
investigated, divided into two geographical regions—A (Black Sea Basin) and B (Mediterranean
Sea Basin). According to Non-metric Multidimensional Scaling (calculated using
presence/absence transformed data, the Bray–Curtis similarity index, and 50 restarts, excluding
outlier samples UA-03 and AL-02), tiger beetle samples were divided into four climatic zones:
Bsk, Csa, Cfb, and Dfb. Taxa characteristic of each of four zones and dissimilarity between those
community types were obtained using the SIMPER analysis with Bray–Curtis similarity and cut-
off for low contributions 100%. Detrended Canonical Correspondence Analysis (DCCA) was
implemented to recognise data distribution (linear or unimodal) with detrending by segments. As
the length of DCCA gradient was 1.5 SD units for the first axis and 1.58 for the second DCCA
axis, redundancy analysis (RDA) with scaling focused on inter-species correlation and species
scores divided by standard deviation was conducted to recognise the main environmental factors
determining species occurrence. To test the significance of environment-species relation, the
unrestricted Monte Carlo Permutation Test was applied with automatic selection under the full
model for all environmental variables. Statistical analyses were performed using PRIMER 6 and
Canoco 4.5 software (Clark & Gorley, 2001; Ter Braak & Šmilauer, 2002).

Statistical Analysis
Statistical comparison of the climatic and soil parameters of each sampling site will be
done using Principal Component Analysis (PCA) of the SPSS Statistics software.

Literature cited
Acal, D. A. P., Wiesner, J., Nuñeza, O. M., & Jaskuła, R. (2021). Tiger beetles (Coleoptera,
Cicindelidae) of Northern Mindanao region (Philippines): checklist, distributional maps,
and habitats. ZooKeys, 1017, 37.
Anichtchenko, A., & Medina, M. N. (2019). A new Neocollyris (Heterocollyris) subspecies from
Mindanao, Philippines (Coleoptera, Carabidae, Cicindelinae). Acta Biologica
Universitatis Daugavpiliensis, 19(1), 13-15.

Cabras, A. A., & Wiesner, J. (2016). Tiger beetles (Coleoptera: Carabidae: Cicindelinae) of
Mainit Hotspring and Mati Protected Landscape, Mindanao with notes on their ecology
and threats. International Research Journal of Biological Sciences, 5(9), 1-6.

Jaskuła, R., Płóciennik, M., & Schwerk, A. (2019). From climate zone to microhabitat—
environmental factors affecting the coastal distribution of tiger beetles (Coleoptera:
Cicindelidae) in the south-eastern European biodiversity hotspot. PeerJ, 7, e6676.

Jaskuła, R., Schwerk, A., & Płóciennik, M. (2021). Morphological variability in Lophyra
 flexuosa (Fabricius, 1787)(Coleoptera, Cicindelidae) in desert countries is affected by
sexual dimorphism and geographic aspect. Ecology and evolution, 11(23), 17527-17536.

Pearson DL, Vogler AP. 2001. Tiger beetles: the evolution, ecology and diversity of the
cicindelids. Ithaca/London: Cornell University Press. 333

Pepito, M. J., Torrejos, C., Cabras, A., Medina, M. N., & Cudera, R. (2019). Preliminary List of
Carabidae and Cicindelidae (Coleoptera) Fauna in Lake Holon, T’boli, South Cotabato,
Philippines. Hand, 14, 18.

Medina, M. N. D., Cabras, A. A., & Wiesner, J. (2019). Thopeutica petertaylori, a new tiger
beetle species (Coleoptera: Cicindelidae) from Mindanao, Philippines.

Zettel, H., & Wiesner, J. (2018). Cylindera (Conidera) mindoroana sp. n.(Coleoptera:
Cicindelidae), a new tiger beetle species from the Philippines.