Ecological Impact of the Yellow Crazy ( gracilipes Smith, 1857; : Formicidae) on Ground Dwelling Assemblages in Dryevergreen Forest at Sakaerat Biosphere Reserve

Ms. Sasitorn Hasin Department of Forest Biology Faculty of Forestry, Kasetsart University Introduction

An is a non-native species whose introduction does or is likely to cause economic or environmental harm or harm to human, , or plant health . -- by the Definitions Subcommittee of the Invasive Species Advisory Committee; ISAC(2006).

Invasive ant species are introduced or alien ant species. They can cause a decline and change in diversity, community and population of native invertebrates, vertebrates and plants by their invasion and displacement -- By International Union for Conservation of Nature (IUCN) 5 species of the World’s Worst Invasive Ant Species

 Little fire ant 

 Big-headed ant  5 species of the World’s Worst Invasive Ant Species

 Argentine ant    Little fire ant  Red imported fire ant

 Big-headed ant  Yellow crazy ant Invasive ant species in Thailand

Group 1 : Invasive alien ant species; Harmful invasive ant species in Thailand. They has been considered to decrease diversity and population of native fauna in ecosystems.

Anoplolepis gracillipes (Yellow crazy ant) Solenopsis geminata (Fire ant)

Group 3: Alien ant species in Thailand, but they have been reported that they are invasive alien ant species in the other country. megacephala (Big head ) Tapinoma melanocephalum (Ghost ant)

Source by; Office of Natural Resources Environmental Policy and Planning; ONPA (2009). The yellow crazy ant and its impacts on ecosystems? Bird on the island invasion at A chick before and after being swarmed by yellow crazy ants. Note the damage around Pacific region the eyes and to the beak.

Chick after being swarmed by yellow crazy ants

Seabirds nest on the ground, leaving them vulnerable to the swarms of yellow-crazy ants. The birds and chicks have visibly suffered from attacks by the ants. Healthy chick

Source by U.S. Fish&Wildlife Service (http://usfwspacific.tumblr.com) Impact on Ecosystems by Yellow Crazy Ants

1. Absence of Yellow crazy ant

Photo source: Director of National Parks/Parks Conceptual diagram source: http://lib.znate.ru 2. Present the yellow crazy ant supercolonies

Source: http://www.stephenbelcher.net Photo source: Dr. Christopher Boland Ground dwelling are an important component of natural ecosystems. Recently research suggest that many arthropod groups have affected by infestation of Yellow crazy ant. Yellow crazy ant was indicated to harmful invasive ant species in Thailand. However, the Yellow crazy ant has never been recorded to cause the ecological impacts as seen on forest ecosystem, i.e. the decrease of arthropod.

Objectives

To assess the impact of the yellow crazy ant on ground dwelling arthropods Methodology

Study area

Sakaerat Environmental Research Station is situated in north eastern Thailand.  Three plots are invaded site where the Yellow crazy ant supercolonies formed approximately 3 year.

ID

Three plots were un-invaded site which sites are continuous areas of invaded site and Yellow crazy ant individuals and supercolonies were not presented. 40 m

40 m 40 m

40 m

A core area of Yellow crazy ant supercolonies is the centre of each plot. Arthropods sampling

ants will be collected by pitfall traps.

7.5 cm

10 cm The liquid - 95 % ethylene alcohol - Water - Detergent

6 cm 40 m

40 m

All individuals of arthropods were identified at least family by using systematic keys of Triplehorn and Johnson (2005). The number of family and individual of each family in each pitfall were counted. Data analysis

GLM univarate ANOVA to test the difference in number of individauls of arthropod groups, frequency and abundance among sites and between seasons, with site and season as fixed factors.

Whenever significant results (P < 0.05) occurred in ANOVA, a post- hoc test was done using Bonferroni pairwise comparisons.

Normality and homogeneity of data were confirmed prior to the analyses using Shapiro-Wilk’s and Levene’s tests, respectively.

Non-distributed variables were transformed through log10(x+1)

These statistical analyses were performed with SPSS ver. 20.0.0 for Windows. Results Arthropod composition

Total number of arthropods individauls 1/ Number of arthropods individauls (N per plots) Arthropods group Dry season Wet season Dry season Wet season UNI YCA UNI YCA UNI YCA UNI YCA Ant 1350 684 (356) 1728 995 (568) 450 ± 78 228 ± 23 576 ± 71 332 ± 11 Beetle 351 146 369 221 117 ± 17 49 ± 8.1 123 ± 14 74 ± 5 Bug - 1 - 10 - 1 ± 0 - 4 ± 3 Cricket 39 38 58 39 13 ± 4 13 ± 2 20 ± 4.2 13 ± 2 Crockroach 39 16 74 20 20 ± 10 6 ± 2 25 ± 4 7 ± 2b Dermaptera - 4 - 3 - 2 ± 1 - 1 ± 1 41 13 111 8 14 ± 8 5 ± 2 37 ± 22 3 ± 2 Centipede 6 2 24 3 2 ± 2 1 ± 1 8 ± 5 1 ± 1 Isopod 15 2 31 12 5 ± 3 1.7 ± 1 11 ± 8 4 ± 4 Millepede 16 - 15 2 6 ± 3 - 5 ± 3 1 ± 1 Opiliones - 37 - 32 - 13 ± 6 - 11 ± 7 Pseudoscorpion 15 6 23 7 5 ± 2 2 ± 2 8 ± 2 3 ± 2 Spider 48 38 83 32 16 ± 2 13 ± 4 27 ± 4 11 ± 2 Total 2310 1171 2943 1634 770 ± 76 391 ± 40.2 981 ± 60 545 ± 21 Frequency (%) Abundance (%) Arthropods group Dry season Wet season Dry season Wet season UNI YCA UNI YCA UNI YCA UNI YCA Ant 100 ± 0.0 100 ± 0.0 100 ± 0.0 97.9 ± 2.1 69.3 ± 3.9 69.2 ± 1.7 68.4 ± 5.2 72.0 ± 0.3 Beetle 95.8 ± 2.1 81.3 ± 9.5 89.6 ± 5.5 91.7 ± 5.5 15.8 ± 3.4 12.3 ± 1.2 12.6 ± 1.6 13.6 ± 1.0 Bug - 2.1 ± 2.1 - 6.3 ± 6.3 - 0.1 ± 0.1 - 0.7 ± 0.7 Cricket 39.6 ± 9.1 43.8 ± 3.6 52.1 ± 9.1 39.6 ± 9.1 1.6 ± 0.3 1.4 ± 0.4 2.4 ± 0.3 1.6 ± 0.4 Crockroach 27.1 ± 13.6 22.9 ± 4.2 68.8 ± 15.7 27.1 ± 4.2 1.8 ± 1.3 1.5 ± 0.6 3.0 ± 0.5 1.4 ± 0.4 Dermaptera - 6.3 ± 3.6 - 4.2 ± 4.2 - 0.4 ± 0.2 - 0.2 ± 0.2 Termites 16.7 ± 9.1 12.5 ± 6.3 33.3 ± 9.8 8.3 ± 5.5 1.9 ± 1.1 1.5 ± 0.9 4.4 ± 2.5 0.6 ± 0.3 Centipede 8.3 ± 8.3 4.2 ± 4.2 35.4 ± 21.7a 6.3 ± 3.6 0.3 ± 0.3 0.2 ± 0.2 1.0 ± 0.5 0.2 ± 0.1 Isopod 14.6 ± 9.1 2.1 ± 2.1 10.4 ± 5.5 2.1 ± 2.1 0.9 ± 0.7 0.2 ± 0.2 1.2 ± 0.9 0.9 ± 0.7 Millepede 16.7 ± 4.1 - 22.9 ± 10.4 2.1 ± 2.1 1.0 ± 0.7 - 0.6 ± 0.3 0.1 ± 0.1 Opiliones - 31.3 ± 15.7 - 27.1 ± 16.3 - 3.9 ± 2.1 - 2.2 ± 1.4 Pseudoscorpion 20.8 ± 5.5 10.4 ± 10.4 31.3 ± 9.5 10.4 ± 10.4 0.8 ± 0.2 0.7 ± 0.7 0.9 ± 0.3 0.5 ± 0.5 Spider 37.5 ± 3.6 39.6 ± 7.5 62.5 ± 14.4 35.4 ± 8.3 2.6 ± 0.4 3.7 ± 0.9 3.4 ± 0.7 2.3 ± 0.5 Total 100 100 100 100 Effect on ground dwelling arthropod

Dependent Variable Number of individauls Frequency (%) Abundance (%) (N per plots) d.f.n. d.f.d. F P d.f.n. d.f.d. F P d.f.n. d.f.d. F P Arthropods Arthropods group 11 52 22.3 0.000 11 52 18.2 0.000 11 52 29.7 0.000 season 1 52 6.2 0.02 1 52 4.2 0.05 1 52 0.01 0.94 sites 1 52 31. 7 0.000 1 52 21.6 0.000 1 52 1.1 0.31 season * Site 1 52 1.3 0.27 1 52 2.5 0.12 1 52 2.3 0.14 Arthropods group * season 11 52 0.7 0.75 11 52 0.6 0.83 11 52 0.6 0.81 Arthropods group * Site 8 52 1.9 0.09 8 52 2.2 0.05 8 52 1.8 0.11 Arthropods group * season * 7 52 0.4 0.92 7 52 0.9 0.52 7 52 0.5 0.81 Site Number of arthropods individauls(N per plots) Arthropods group Dry season Wet season UNI YCA UNI YCA Ant 450 ± 78a 228 ± 23b 576 ± 71a 332 ± 11b Beetle 117 ± 16.5a 49 ± 8.1b 123 ± 14.4 74 ± 5.0 Bug - 1 ± 0A' - 4 ± 3B' Cricket 13 ± 4 13 ± 2.7 20 ± 4.2 13 ± 2.6 Crockroach 20 ± 10a 6 ± 2b 25 ± 4a 7 ± 2b Dermaptera - 2 ± 1 - 1 ± 1 Termites 14 ± 8a 5 ± 2b 37 ± 22a 3 ± 2b Centipede 2 ± 2 1 ± 1 8 ± 5a 1 ± 1b Isopod 5 ± 3 1.7 ± 1 11 ± 8 4 ± 4 Millepede 6 ± 3 - 5 ± 3 1 ± 1 Opiliones - 13 ± 6 - 11 ± 7 Pseudoscorpion 5 ± 2 2 ± 2 8 ± 2 3 ± 2 Spider 16 ± 2 13 ± 4 27 ± 4a 11 ± 2b Total 770 ± 76 391 ± 40.2 981 ± 60 545 ± 21 Frequency (%) Abundance (%) Arthropods Dry season Wet season Dry season Wet season group UNI YCA UNI YCA UNI YCA UNI YCA

Ant 100 ± 0.0 100 ± 0.0 100 ± 0.0 97.9 ± 2.1 69.3 ± 3.9 69.2 ± 1.7 68.4 ± 5.2 72.0 ± 0.3

Beetle 95.8 ± 2.1 81.3 ± 9.5 89.6 ± 5.5 91.7 ± 5.5 15.8 ± 3.4 12.3 ± 1.2 12.6 ± 1.6 13.6 ± 1.0 Bug - 2.1 ± 2.1A' - 6.3 ± 6.3 B' - 0.1 ± 0.1 - 0.7 ± 0.7 Cricket 39.6 ± 9.1 43.8 ± 3.6 52.1 ± 9.1 39.6 ± 9.1 1.6 ± 0.3 1.4 ± 0.4 2.4 ± 0.3 1.6 ± 0.4 Crockroach 27.1 ± 13.6 22.9 ± 4.2 68.8 ± 15.7a 27.1 ± 4.2b 1.8 ± 1.3 1.5 ± 0.6 3.0 ± 0.5 1.4 ± 0.4 Dermaptera - 6.3 ± 3.6 - 4.2 ± 4.2 - 0.4 ± 0.2 - 0.2 ± 0.2 Termites 16.7 ± 9.1 12.5 ± 6.3 33.3 ± 9.8 a 8.3 ± 5.5b 1.9 ± 1.1 1.5 ± 0.9 4.4 ± 2.5a 0.6 ± 0.3b Centipede 8.3 ± 8.3 4.2 ± 4.2 35.4 ± 21.7a 6.3 ± 3.6b 0.3 ± 0.3 0.2 ± 0.2 1.0 ± 0.5 0.2 ± 0.1 Isopod 14.6 ± 9.1a 2.1 ± 2.1b 10.4 ± 5.5 2.1 ± 2.1 0.9 ± 0.7 0.2 ± 0.2 1.2 ± 0.9 0.9 ± 0.7 Millepede 16.7 ± 4.1 - 22.9 ± 10.4a 2.1 ± 2.1b 1.0 ± 0.7 - 0.6 ± 0.3 0.1 ± 0.1 Opiliones - 31.3 ± 15.7 - 27.1 ± 16.3 - 3.9 ± 2.1 - 2.2 ± 1.4 Pseudoscorpion 20.8 ± 5.5 10.4 ± 10.4 31.3 ± 9.5 10.4 ± 10.4 0.8 ± 0.2 0.7 ± 0.7 0.9 ± 0.3 0.5 ± 0.5 Spider 37.5 ± 3.6 39.6 ± 7.5 62.5 ± 14.4 a 35.4 ± 8.3 b 2.6 ± 0.4 3.7 ± 0.9 3.4 ± 0.7 2.3 ± 0.5 Total 100 100 100 100 70 900 ** * 60 800 700 50 600 40 500 400 30

300 (%) Frequency 20

Number of individaulsNumber 200 10 100 0 0 UNI YCA UNI YCA 100

80 Error bars represent standard error. 60 *, ** and (P < 0.05 and P < 0.01, respectively) indicate significant 40 differences between frequency in UNI-site and YCA-site within the (%) Abundance 20 arthropod group. 0 UNI YCA 1000 a' 70 a' UNI YCA 60 800 a 50 b'

600 b' 40 b 30

400 (%) Frequency 20

Number ofindividauls Number 200 10

0 0 Dry season Wet season Dry season Wet season

Black bar is un-invaded site and yellow bar is invaded site. Error bars represent standard error. Difference lower case letters indicate a significant difference between the UNI-site and YCA-site within the season. 120 UNI-site YCA-site 100

80 *** *** 60 ** 40 Frequency (%) Frequency * 20

0 CH IS PS AR BL CO FO OR TE Arthropod group

Error bars represent standard error. *, ** and *** (P < 0.05, P < 0.01 and P < 0.001, respectively) indicate significant differences between frequency in uninvaded site (UNI-site) and invaded site (YCA-site) within the arthropod group. Effect on dominance arthropod

Dependent Variable Number of individauls(N per plots) Frequency (%) Abundance (%) d.f.n. d.f.d. F P d.f.n. d.f.d. F P d.f.n. d.f.d. F P Coleoptera Families; beetle (COF) COF 8 34 7.86 0.000 8 34 4.38 0.000 8 34 7.73 0.000 season 1 34 1.64 0.21 1 34 0.69 0.41 1 34 0.01 0.92 Site 1 34 9.93 0.003 1 34 8.93 0.005 1 34 0.17 0.68 season * Site 1 34 0.03 0.86 1 34 0.34 0.57 1 34 0.02 0.88 COF * season 6 34 0.53 0.78 6 34 0.53 0.78 6 34 0.47 0.83 COF * Site 4 34 3.31 0.02 4 34 1.90 0.13 4 34 3.30 0.02 COF * season * Site 4 34 0.29 0.88 4 34 0.25 0.91 4 34 0.37 0.83 Orthoptera Families; Cricket (ORF) ORF 2 13 14.51 0.000 2 13 27.52 0.003 2 13 19.20 0.000 season 1 13 0.30 0.59 1 13 0.08 0.78 1 13 0.76 0.40 site 1 13 12.67 0.003 1 13 4.80 0.05 1 13 0.73 0.41 season * site 1 13 0.24 0.63 1 13 1.12 0.31 1 13 0.69 0.42 ORF * season 2 13 0.07 0.93 2 13 0.05 0.95 2 13 0.09 0.92 ORF * site 1 13 1.74 0.21 1 13 0.02 0.89 1 13 1.22 0.29 Formicidae; Ant season 1 8 4.49 0.67 1 8 1.00 0.35 1 8 0.06 0.81 site 1 8 18.51 0.003 1 8 1.00 1.35 1 8 0.20 0.67 season * site 1 8 0.04 0.84 1 8 1.00 2.35 1 8 0.20 0.67 Termitidae; Termites season 1 5 0.02 0.89 1 5 0.19 0.68 1 5 0.21 0.67 site 1 5 8.44 0.03 1 5 3.80 0.11 1 5 1.81 0.28 season * site 1 5 0.50 0.51 1 5 1.40 0.30 1 5 0.79 0.42 Spider season 1 8 0.89 0.38 1 8 0.57 0.48 1 8 0.20 0.67 site 1 8 8.41 0.02 1 8 1.67 0.23 1 8 0.02 0.91 season * site 1 8 2.10 0.19 1 8 1.85 0.21 1 8 2.63 0.14 Number of individuals, frequency and abundance of Coleoptera Families in 16 pitfall traps at un-invaded site (UNI) and invaded site (YCA). Values are averaged per site (Ave. ± SE) and n=3 for each site.

Total number of Number of individauls (N per plots) Frequency (%) Abundance (%) individauls Family Dry season Wet season Dry season Wet season Dry season Wet season Dry season Wet season UNI YCA UNI YCA UNI YCA UNI YCA UNI YCA UNI YCA UNI YCA UNI YCA Bostrichidae 35 45 68 53 12 ± 6 15 ± 6 23 ± 7 18 ± 8 37.5 ± 14.4 37.5 ± 9.5 64.6 ± 16.3 43.8 ± 10.8 1.7 ± 0.7 4.6 ± 1.7 2.8 ± 0.9 3.7 ± 1.6 Carabidae 14 11 44 27 5 ± 3 4 ± 3 15 ± 7 9 ± 3 22.9 ± 14.5 12.5 ± 9.5 32.3 ± 24.0 25 ± 7.2 0.7 ± 0.4 1.1 ± 0.9 1.9 ± 0.8 2.0 ± 0.8 Curculionidae 5 - - 4 2 ± 1 - - 2 ± 1 8.3 ± 5.5 - - 6.3 ± 3.6 0.3 ± 0.3 - - 0.3 ± 0.2 Elarteridae - 1 - 6 - 0 ± 0 2 ± 2 - 2.1 ± 2.1 - 6.3 ± 6.3 - 0.1 ± 0.1 - 0.4 ± 0.4 Nitituiidae 3 - - - 1 ± 1 - - - 2.1 ± 2.1 - - - 0.2 ± 0.2 - - - Scarabaeidae 9 4 5 7 3 ± 2 2 ± 1 2 ± 2 3 ± 1 12.5 ± 6.3 6.3 ± 6.3 8.3 ± 8.3 8.3 ± 5.5 0.6 ± 0.4 0.4 ± 0.4 0.2 ± 0.2 0.5 ± 0.3

Staphylinidae 167 68 120 91 56 ± 17a 23 ± 5b 40 ± 5a 31 ± 7b 70.8 ± 5.5 54.2 ± 15.0 66.7 ± 9.1 64.6 ± 13.7 9.8 ± 0.4a 6.7 ± 1.0b 4.7 ± 0.2a 6.5 ± 1.3b

Tenebrionidae 0 6 - 8 - 2 ± 2 - 3 ± 2 - 10.4 ± 10.4 - 12.5 ± 9.5 - 0.6 ± 0.6 - 0.6 ± 0.4 unknow 118 11 132 25 40 ± 10a 4 ± 3b 44 ± 7a 9 ± 6b 20.4 ± 11.6 12.5 ± 9.5 28.7 ± 9.5 20.8 ± 14.6 6.2 ± 1.3a 1.1 ± 0.8b 5.3 ± 0.9a 1.9 ± 1.5b Total 351 146 369 221 117 ± 16 49 ± 8 123 ± 14 74 ± 5 95.8 ± 2.1 81.3 ± 9.5 89.6 ± 5.5 91.7 ± 5.5 15.8 ± 3.4 12.3 ± 1.2 12.6 ± 1.6 13.6 ± 1.0

Pairwise comparison result was shown that number of individauls and abundance of two beetle groups, i.e. rove beetle and undescribed beetle, have significant higher value in un- invaded site than invaded site. DISCUSSION

Ant was the high number of individauls, frequency and abundance than the other arthropods group. This predominance can be explained by the diverse of species and their nesting habitat on terrestrial ecosystems, number of individauls per colony/nest and range of foraging behaviors of ant Ground-dwelling arthropods assemblage at un-invaded and invaded sites were strikingly difference. These results were unanticipated given the precedent for invasive ants to disrupt arthropod communities.

Beetle

Ant Spider Cricket

Five dominant arthropod groups, may be failed to coexist with the Yellow crazy ant super colonies. The impact on the composition of the arthropods assemblage may have resulted partly from much higher number of individauls of Yellow crazy ant. This finding might be suggests that Yellow crazy ant preyed on competed with arthropods differently than native ants. Although Yellow crazy ant appeared to present-day disrupt the communities of ground dwelling arthropods, the effects of Yellow crazy ant invasions in dryevergreen forest probably extend beyond the displacement of the native ant fauna. Such effects might have been difficult to detect in this study for several reasons.

First, pitfall traps largely fail to capture highly sedentary arthropods (e.g., larvae of ) or those that rarely occur on the ground (e.g. ).

Second, although the composition of arthropod based surveys s have numerous advantages, however, the effects of Yellow crazy ant on rare taxa/or key species would evade detection with the methods used here.

Third, the arthropod communities of dry evergreen forest experienced changes prior to invasion by Yellow crazy ant, and species sensitive to these disturbances may have already been lost.

Lastly, immigration of arthropods from un-invaded to a continuous areas of invaded sites could maintain populations of arthropods in invaded areas even if theYellow crazy ant prevent such species from reproducing successfully CONCLUSION

I anticipate that this work can be used as a base-line study from which future changes as a result of land utilization and climate change or the addition of further invasive species might be monitored.

Yellow crazy ant can be influenced by many factors such as the presence of competitors, food source and climates.

Subsequent to this survey, the Yellow Crazy ant has further spread in Dry evergreen forest at SERS, predominantly through human mediated dispersal. Funding sources

The Higher Education Research Promotion and National Research University Project of Thailand

Thesis Committee Thesis Advisor: Assist. Prof. Dr. Wattanachai Tasen Thesis Co-Advisor: Dr. Watana Sakchoowong Thank you very much for your attention

Photo by Yoshiaki Hashimoto

Table 1 Check lists of major invasive ant species of the world (modified from Holway et al., 2002)

Geographical Range Ant species Native Invasive species Anoplolepis gracilipes Africa , Tropical Africa, Asia, Australia, Caribbean, (Yellow crazy ant) Asia? Indian and Pacific Ocean

Linepithema humile Africa, Asia, North America, Atlantic South America (Argentine ants) and Pacific Ocean Africa Australia, North and South America, (Big-headed ant) Caribbean, Indian and Pacific Ocean Solenopsis invicta South America Caribbean, North America (Fire ant) Solenopsis geminata Central, North and Africa, Asia, Australia, Pacific and (Tropical fire ant) South America Indian Ocean Wasmannia auropunctata Central and South Africa, Caribbean, Pacific Ocean, (Little fire ant) America South and North America

Note: Red color species are found in Thailand. Origin of the yellow crazy ant is still unclear whether from tropical Asia or not. Steps to Ant invasion

Introductions are Non-native ant or alien ant species mostly due to human activity. Introduction

Survive but Do not Reproduce Survive and Reproduce

Establishment Non Invasive

Become part of the Spreads Rapidly natural fauna and flora Spread Invasive ant Naturalized Species species Methodology

1. Diversity and Distribution of invasive alien ant species in Thailand

2. Influence of environmental factors on distribution of the yellow crazy ant 2.1 Distribution of the yellow crazy ant along road construction 2.2 Distribution pattern of the yellow crazy ant from the road into two forest types

3. Interaction between the yellow crazy ant and the other arthropods 3.1 Effects of the yellow crazy ant super-colony on the nest abundance of other ant species 3.2 Impact of the yellow crazy ant on the other arthropods

4. To evaluate the impact of the yellow crazy ant on CO2 efflux in the dry evergreen forest Methodology

1. Diversity and Distribution of invasive alien ant species in Thailand

 I will survey ants species in - 32 areas including - 23 national parks, - 1 wildlife sanctuary, - 3 plantations, - 1agricultural land, - Sakaerat Environmental Research Station - Urban ecosystem (Bangkok).  Positions of these areas will be  Study areas recorded by GPS. Each study area will be conducted;

FA

UA

Forest Area or Public park Area in Urban Area (UA) Bangkok (FA) outside region

inside region

core region 2.1 Distribution of the yellow crazy ant along road construction

Research will be conducted along 4-km section of the main road constructed within the SERS

SV Main Road in SERS

20 m Main road number 304 number road Main Main road number 304 number road Main

4 KM

Ant sampling; survey point (SV) will be established at the edge of two sides of roadways, 20 m apart from each other, at least 400 survey points.