R. Steven Wagner

Conflicting patterns of genetic structure produced by nuclear and mitochondrial markers in the Oregon slender salamander (Batrachoseps wrighti): Implications for conservation efforts and species management

Endemic to Oregon in the northwestern US, the Oregon slender salamander (Batrachoseps wrighti) is a terrestrial plethodontid found associated with late successional mesic forests. Consequently, forest management practices such as timber harvesting may impact their persistence. Therefore, to infer possible future effects of these practices on population structure and differentiation, we used mitochondrial DNA sequences (cytochrome b) and RAPD markers to analyze 22 populations across their range. Phylogenetic analyses of sequence data (774 bp) revealed two historical lineages corresponding to northern and southern-distributed populations. Relationships among haplotypes and haplotype diversity within lineages suggested that the northern region may have more recently been colonized compared to the southern region. In contrast to the mitochondrial data, analyses of 46 RAPD loci suggested an overall pattern of isolation-by-distance in the set of populations examined and no particularly strong clustering of populations based on genetic distances. We propose two non-exclusive hypotheses to account for discrepancies between mitochondrial and nuclear data sets. First, our data may reflect an overall ancestral pattern of isolation-by-distance that has subsequently been influenced by vicariance. Alternately, our analyses may suggest that male-mediated gene flow and female philopatry are important contributors to the pattern of genetic diversity. We discuss the importance of distinguishing between these two hypotheses for the purposes of identifying conservation units and note that, regardless of the relative contribution of each mechanism towards the observed pattern of diversity, protection of habitat will likely prove critical for the long-term persistence of this species.

Sequences of Tibetan Macaque (Macaca thibetana) and Tourist Behaviors at Mt. Huangshan, China

Abstract: Previous research on Tibetan macaques (Macaca thibetana) at Mt. Huangshan, China, suggested that ecotourism can have detrimental consequences. This study identified sequences of behaviors that typically occur in macaque-tourist interactions to examine whether particular tourist behaviors precipitate monkey responses. Focal sampling was used to record relevant behaviors from tourists and 10 macaques over 28 data collection sessions in August 2006. Data collectors recorded whether each behavior occurred as part of a sequence. Sequences were defined as two or more behaviors in which each behavior occurred within five seconds of the previous behavior. Of 3,129 total behaviors, 2,539 (81.1%) were from tourists and 590 (18.9%) were from monkeys. Tourists initiated significantly more sequences than did macaques (412 [84.6%] versus 75 [15.4%]). Tourist pointing, rail slapping, fleeing, and rock showing occurred significantly more than expected in tourist-macaque sequences. Points were also among the most common tourist behaviors preceding macaque threats. By discouraging tourists from engaging in these behaviors, macaque threats could be reduced, thereby improving macaque-tourist interactions. These results may aid in the management of other macaque tourist sites to minimize stress-inducing interactions.

Toxic Effects of the Herbicide Roundup® Regular on Pacific Northwestern Amphibians

Abstract One of the most widely used herbicides for commercial and home use is glyphosate, the active ingredient in Roundup® Regular. We examined toxicity of the herbicide Roundup® on 6 amphibian species: Ambystoma gracile, Ambystoma macrodactylum, Anaxyrus [Bufo] boreas, Pseudacris regilla, Rana cascadae, and Rana luteiventris. Larvae were exposed to 6 different Roundup® Regular treatments (0 (control), 0.1, 0.5, 1.0, 2.0, and 5.0 mg AI/L dilutions of glyphosate) and monitored for 16 d. Estimated acute lethal concentrations at 24 h (LC50) varied significantly among species (ANOVA, F(3, 56)  =  3.54, p < 0.0202), with concentrations ranging from 0.43 mg AI/L of Roundup® for P. regilla to 2.66 mg AI/L for A. boreas. Bufonid and ambystomatid larvae were less sensitive than Ranid and Pseudacrid species tested, with no salamander larval mortality occurring at 24 h. Mean time-to-death varied from 1 d for P. regilla to 8.3 d for A. gracile, respectively (ANOVA, F(5, 971)  =  108, p < 0.0001). For exposure times longer than 24 h, the A. boreas was not significantly different than the salamanders for time-to-death, based on Tukey-Kramer comparisons. Results suggest Roundup® Regular is highly toxic to the amphibians at levels below EPA standards for drinking water and at concentrations they may be exposed to during overspray. We recommend the use of less toxic glyphosate-based herbicides in aquatic systems, if applications are necessary, or made during times of year when amphibian larvae are not present.

Tourist impact on Tibetan macaques.

Abstract Ecotourism is a growing sector of the tourism industry, but few studies to date have quantified its impacts on local people, tourists and wildlife. We present a preliminary study on threat and affiliative behaviors of two groups of free-ranging Tibetan macaques (Macaca thibetana) as a function of habituation and tourist presence. Data indicate that the less habituated group spent less time within sight of tourists compared with the more habituated group. The more habituated group engaged in frequent affiliative behaviors while within sight of humans, whereas affiliative behavior was not observed in the less habituated group. The general pattern of threats consisted of adults primarily threatening juveniles and juveniles primarily threatening humans, possibly due to redirection. No clear pattern of threats as a function of tourist density emerged. Tourist feeding, although discouraged, potentially provided a catalyst for some aggression. Future research will focus on clarifying which human behaviors evoke specific threat responses from monkeys. These data will be used to refine the existing management plan for this monkey population.

Amphibian Chytridiomycosis in the Oregon Spotted Frog (RANA PRETIOSA) IN WASHINGTON STATE, USA

In recent years Batrachochytrium dendrobatidis (Bd), a chytridiomycete fungus pathogenic to amphibians (Longcore and others 1999), has been implicated as the proximate cause of amphibian declines around the world (Berger and others 1998; Daszak and others 2003; Muths and others 2003; Pounds and others 2006). Despite the insidious nature of Bd (Green and others 2002), few published data exist addressing its occurrence in the Pacific Northwest (PNW). When Pearl and others (2007) opportunistically examined 7 PNW amphibian species, they found Bd most often (57% of 21 individuals from 14 sampled populations) in the highly aquatic Oregon Spotted Frog (Rana pretiosa), an Endangered Species in Washington State (WDFW 2009). However, all R. pretiosa they sampled were from Oregon, and Bd was not detected in the Rana cascadae (Cascades Frog) and Anaxyrus boreas (Western Toad) specimens they sampled from Washington. Here, we report the detection of Bd in R. pretiosa from Washington. Since 1997 and 1998, respectively, we have been monitoring R. pretiosa populations at the Trout Lake Natural Area Preserve (TLNAP: UTM Zone 10, 610857-612950E, 5095880-5097574N, WGS84; elev. 594 to 599 m) and Conboy Lake National Wildlife Refuge (CLNWR: UTM Zone 10, 625223-635180E, 5086652-5095491N, WGS84; elev. 552 to 576 m), in Klickitat County, Washington. These sites represent 2 of only 3 areas where R. pretiosa is known to occur in Washington (McAllister and Leonard 1997). At both sites, monitoring included egg mass surveys during the annual breeding season following snowmelt (late February to mid-March at CLNWR; midMarch to early April at TLNAP). Surveys involved area-specific counts of individual egg masses and egg mass groups, each of which was marked with flags and geo-referenced using a Geographic Positioning System (GPS) to avoid double-counting. Sampling at CLNWR addressed 4 hydrologically distinct units, which were surveyed repeatedly until no new egg masses were found; at TLNAP, 3 units consistently used for breeding were sampled. We inferred a 1:1 correspondence between egg mass numbers and number of breeding females based on a combination of our direct observations of oviposition (n 5 13), the recapture of non-gravid females known to have laid eggs (n 5 84), and the relatively short interval over which the laying of new egg masses occurs (about 3 wk). Collectively, these data indicate that females lay only 1 clutch annually, and that egg mass numbers reflect the effective population numbers of adult females. Decline in egg mass numbers in 3 of the 4 surveyed units at CLNWR from 2004 to 2005 (Fig. 1) and 2 of the 3 units at TLNAP over the same period (Fig. 2), coupled with the increasing recognition of Bd as a cause of amphibian declines, motivated us to collect dead frogs found during 2006 and test them for Bd. Five dead adult R. pretiosa were tested for Bd; 1 from CLNWR and 4 from TLNAP. The CLNWR specimen collected on 13 March 2006 displayed feeble vital signs, minimal response to touch, and righting response was lacking. The frog died within 20 min of discovery, was preserved in 10% formalin, and then stored in 70% ethanol before histological examination using a standard wet-mount preparation of its epidermal tissue (Berger and others 1999), which was sloughing extensively as multiple epidermal layers. A dead adult female R. GENERAL NOTES

The vocal repertoire of Tibetan macaques (Macaca thibetana): A quantitative classification.

Vocal repertoires are basic and essential components for describing vocal communication in animals. Studying the entire suite of vocal signals aids investigations on the variation of acoustic structure across social contexts, comparisons on the complexity of communication systems across taxa, and in exploration of the evolutionary origins of species‐specific vocalizations. Here, we describe the vocal repertoire of the largest species in the macaque genus, Macaca thibetana. We extracted thirty acoustic parameters from call recordings. Post hoc validation through quantitative analyses of the a priori repertoire classified eleven call types: coo, squawk, squeal, noisy scream, growl, bark, compound squeak, leap coo, weeping, modulated tonal scream, and pant. In comparison to the rest of the genus, Tibetan macaques uttered a wider array of vocalizations in the context of copulations. Previous reports did not include modulated tonal screams and pants during harassment of copulatory dyads. Furthermore, in comparison to the rest of the genus, Tibetan macaque females emit acoustically distinct copulation calls. The vocal repertoire of Tibetan macaques contributes to the literature on the emergence of species‐specific calls in the genus Macaca with potential insights from social, reproductive, and ecological comparisons across species. Am. J. Primatol. 78:937–949, 2016.

Complex interactive effects of water mold, herbicide, and the fungus Batrachochytrium dendrobatidis on Pacific treefrog Hyliola regilla hosts

Infectious diseases pose a serious threat to global biodiversity. However, their ecological impacts are not independent of environmental conditions. For example, the pathogenic fungus Batrachochytrium dendrobatidis (Bd), which has contributed to population declines and extinctions in many amphibian species, interacts with several environmental factors to influence its hosts, but potential interactions with other pathogens and environmental contaminants are understudied. We examined the combined effects of Bd, a water mold (Achlya sp.), and the herbicide Roundup® Regular (hereafter, Roundup®) on larval Pacific treefrog Hyliola regilla hosts. We employed a 2 wk, fully factorial laboratory experiment with 3 ecologically realistic levels (0, 1, and 2 mg l-1 of active ingredient) of field-formulated Roundup®, 2 Achlya treatments (present and absent), and 2 Bd treatments (present and absent). Our results were consistent with sublethal interactive effects involving all 3 experimental factors. When Roundup® was absent, the proportion of Bd-exposed larvae infected with Bd was elevated in the presence of Achlya, consistent with Achlya acting as a synergistic cofactor that facilitated the establishment of Bd infection. However, this Achlya effect became nonsignificant at 1 mg l-1 of the active ingredient of Roundup® and disappeared at the highest Roundup® concentration. In addition, Roundup® decreased Bd loads among Bd-exposed larvae. Our study suggests complex interactive effects of a water mold and a contaminant on Bd infection in amphibian hosts. Achlya and Roundup® were both correlated with altered patterns of Bd infection, but in different ways, and Roundup® appeared to remove the influence of Achlya on Bd.