Journal: Ecotoxicology

Supplementary Information (SI): Mercury concentrations in bats (Chiroptera) from a gold mining area in the Peruvian Amazon

Mónica Moreno-Brush1-2*, Alejandro Portillo3*, Stefan Dominik Brändel4-5, Ilse Storch1, Marco

Tschapka4-5, and Harald Biester2

1Chair of Wildlife Ecology and Management, University of Freiburg, Freiburg, Germany.

2Institut für Geoökologie, Abt. Umweltgeochemie, Technische Universität Braunschweig,

Braunschweig, Germany.

3Museo de Historia Natural, Universidad Nacional San Antonio Abad del Cusco, Cusco, Perú.

4Institute of Evolutionary Ecology and Conservation Genomics, University of Ulm, Ulm, Germany.

5Smithsonian Tropical Research Institute, Balboa, Panama.

*Corresponding author [email protected] · Phone: +49-531-391-5626 [email protected] · Phone: +51 974373550

1 SI- Table 1. Mercury concentrations (µg g-1) in the fur of bats collected in 2014 and 2015 in southeastern Peru. For each species, number of total samples (N) and samples below the analytical limit of quantification (N

Family Subfamilies and Species N N

Rhinophyllinae Rhinophylla pumilio 2 -- SAN, TAH 0.19 ; 0.45 0.32 0.19 Omnivorous

Carolliinae Carolia benkeithi 2 50 SAN, MAL < 0.16 ; 0.32 0.32 0.08 0.24 Frugivorous Carolia brevicauda 13 -- SAN, MAL, TAH 0.24 – 3.95 0.95 0.99 0.55 Frugivorous Carolia perspicillata 65 3 SAN, MAL, TAH < 0.55 ᵃ – 2.91 0.88 0.59 0.66 Frugivorous (continued)

2 SI-Table 1. (continued)

Family Subfamilies and Species N N

Desmodontinae Desmodus rotundus 1 100 SAN < 0.15 ᵃ Hematophagous

Phyllostominae Lophostoma silvicolum 6 17 SAN, MAL, TAH < 0.17 – 3.93 0.96 1.35 0.40 Animalivorous Micronycteris hirsuta 1 -- MAL 0.3 Animalivorous Phyllostomus discolor 2 -- MAL 1.02 ; 1.74 1.38 0.51 1.38 Omnivorous Phyllostomus elongatus 16 -- SAN, MAL, TAH 0.63 – 8.67 2.53 2.06 1.90 Omnivorous Phyllostomus hastatus 1 -- SAN 1.67 Omnivorous Tonatia saurophila 3 -- SAN, MAL 0.28 – 1.81 0.80 0.87 0.32 Animalivorous Trachops cirrhosus 1 -- MAL 0.86 Animalivorous Vampyrum spectrum 1 -- TAH 0.79 Animalivorous

Emballonuridae Cormura brevirostris 1 -- SAN 6.61 Insectivorous

Molossidae Molossus molossus 1 100 TAH < 0.33 ᵃ Insectivorous

Vespertilionidae Myotis riparus 1 -- TAH 3.66 Insectivorous ᵃ Maximum LOQ ᵇ Mean of values above the max LOQ c Based on Carrera, 2003; Fleming, 1988; Kalko et al., 1996; Moya et al., 2008.

SI-Table 2. P -values for pairwise comparisons of Hg concentrations in the fur of bats from genera Artibeus (A. planirostris) , Carollia (C. perspicillata , C. brevicauda) , and Phyllostomus (P. elongatus) . Data were analyzed using the Kruskal-Wallis ANOVA test (0.05 significance level). In case of a significant effect detected, pairwise comparisons were done using the Mann-Whitney test (*, **, and *** indicate p < 0.05, < 0.01, and < 0.001, respectively).

KW-ANOVA Pairwise comparison Significant Location p p difference Santa Rita (ASGM-affected) 2.8E-04 Carollia vs Phyllostomus 0.0024 ** Artibeus vs Carollia 0.0048 ** Artibeus vs Phyllostomus 0.028 *

Malinowski (ASGM-affected) 1.0E-11 Artibeus vs Carollia 7.4E-11 *** Artibeus vs Phyllostomus 1.2E-07 *** Carollia vs Phyllostomus 2.7E-04 ***

Tahuamanu (unaffected) 0.0076 Artibeus vs Carollia 0.019 * Carollia vs Phyllostomus 0.022 * Artibeus vs Phyllostomus 0.028 *

4 SI-Table 3. Estimated sample sizes that would have been required to achieve statistical power levels of 0.8 and 0.95 in pairwise comparisons of bats with different sex. Based on α = 0.05, using two-sample t-test (Cohen, 1988)

nᵃ mean stand.dev Power level 0.8 0.95 Carollia perspicillata (all data) females 28 1.09 0.66 38 62 male 37 0.70 0.49

C. perspicillata (MAL) females 12 0.61 0.36 9,529 15,776 male 13 0.63 0.45

Phyllostomus elongatus (all data) females 6 1.89 1.03 90 149 male 13 2.59 2.14

P. elongatus (SAN) females 4 2.22 1.07 10 16 male 3 5.23 2.98

P. elongatus (MAL) females 2 1.21 0.68 17 27 male 7 2.24 1.27 ᵃ Number of samples available in this study

5 SI-Table 4. Estimated sample sizes that would have been required of Phyllostomus elongatus and Carollia brevicauda to achieve statistical power levels of 0.8 and 0.95 in pairwise comparisons tests to evaluate differences in mercury concentrations at different study locations. Based on α = 0.05, using two-sample t- test (Cohen, 1988)

nᵃ mean stand.dev Power level 0.8 0.95 Phyllostomus elongatus SAN 6 3.82 2.56 26 42 MAL 7 2.19 1.33

MAL 7 2.19 1.33 8 13 TAH 3 0.76 0.13

Carollia brevicauda SAN 4 1.69 1.61 23 38 MAL 7 0.71 0.29

MAL 7 0.71 0.29 12 19 TAH 2 0.30 0.09

SAN 4 1.69 1.61 6 9 TAH 2 0.30 0.09 ᵃ Number of samples available in this study

SI-Fig. 1. Fur mercury (Hg) concentrations in Carollia perspicillata and Phyllostomus elongatus according to sex in southeastern Peru (sampling locations in Fig. 1). Data are presented as raw data points (black diamonds) and grouped box plots. Similar letters indicate that the populations were not significantly different according to the non-parametric post hoc Mann-Whitney U test at a 0.05 significance level.

7 SI-Fig. 2. Fur mercury (Hg) concentrations in Carollia perspicillata and Phyllostomus elongatus according to age in southeastern Peru (sampling locations in Fig. 1). Age was determined based on the length of the fourth metacarpal-phalangeal joint (Kunz and Anthony, 1982). Data are presented as raw data points (black diamonds) and grouped box plots. Similar letters indicate that the populations were not

8 significantly different according to the non-parametric post hoc Mann-Whitney U test at a 0.05 significance level.

REFERENCES Carrera, J.P., 2003. Distribución de murciélagos (Chiroptera) a través de un gradiente altitudinal en las estribaciones orientales de los Andes Ecuatorianos. (Thesis (Licentiate)). Pontificia Universidad Católica del Ecuador, Escuela de Biología. Quito.

Cohen, J., 1988. Statistical power analysis for the behavioral sciences, Second. ed. Lawrence Erlbaum Associates, Hillsdale, New Jersey.

Fleming, T.H., 1988. The short-tailed fruit bat: a study in plant-animal interactions. University of Chicago Press, Chicago.

Helsel, D.R., 2012. Statistics for censored environmental data using Minitab(R) and R, Second edi. ed. John Wiley & Sons, Inc., Hoboken.

Kalko, E.K. V., Handley, C.O., Handley, D., 1996. Organization, diversity, and long-term dynamics of a Neotropical bat community, in: Cody, M., Smallwood, J. (Eds.), Longterm Studies in Vertebrate Communities. Academic Press, Los Angeles, pp. 503–553.

Kunz, T.H., Anthony, E.L.P., 1982. Age estimation and post-natal growth in the bat Myotis lucifugusitle. J. Mammal. 63, 23–32.

Moya, M.I., Montaño-Centellas, F., Aguirre, L.F., Tordoya, J., Martínez, J., Galarza, M.I., 2008. Variación temporal de la quiropterofauna en un bosque de yungas en Bolivia. Mastozoología Neotrop. 15, 349–357.