Amphibian reptilian 8

Published 2014 * in the United States of America VOLUME • NUMBER AMPHIBIAN & REPTILE CONSERWION COLOMBIA amphibian-reptile-conservation.org ISSN: 1083-446X elSSN: 1525-9153 Board of Directors Howard Craig Hassapakis ARC: Editor, Publisher, ARC: and Chairman of the Board Treasurer Clark, Jr USA Garcia and Associates, Franco Andreone Bruce Waldman of Directors, Museo Regionale di Scienze Naturali, ITAEY University of Peradeniya, SRI EANKA Virginia EGYPT Ted R Kahn KOREA Neotropical Conservation Foundation, USA Indraneil Das Peter Uetz USA Commonwealth University, Michael Hutchins Chair, Department of Conservation and Science, American Zoo and Aquarium Association [1990-2005] and former Executive Director/CEO, The Wildlife Society [2005-2012], USA former DirectorAVihiam Suez University, USA Seoul National University, Madhava Meegaskumbura Adel Ibrahim & Associate Editor; MAEAYSIA Universiti Malaysia Sarawak, Walter R Erdelen Conway former Assistant Director-General for Natural Sciences of the United Nations Educational, Scientihc and Cultural Organization (UNESCO); FRANCE Editor Craig Hassapakis USA Utah Valley University, Africa Issue Branch Museum, SOUTH AFRICA Bill Port Elizabeth Associate Editors Howard Erik Wild BRAZIL Belo Horizonte, Clark, Raul Diaz Jr Garcia and Associates, USA Mayra Oyervides La Sierra University, USA Branch Port Elizabeth Museum, SOUTH AFRICA Bill The University of Texas-Pan American, USA Copy Editor Ruthe Smith California, USA Editorial Board C Kenneth Dodd, University of Florida, USA Museo Regionale Commonwealth University, di Scienze Naturali, ITAEY USA Universiti Malaysia Sarawak, Gunther Kohler Senckenberg Forschungsinstitut und Naturmuseum, GERMANY MAEAYSIA Madhava Meegaskumbura University of Peradeniya, SRI EANKA Aurelio Ramirez-Bautista Javier Sunyer Universidad Nacional Autonoma de Nicaragua-Eeon, MEXICO NICARAGUA Larry David Wilson Centro Zamorano de Biodiversidad, Manuel Acevedo Universidad de San Carlos de Guatemala, HONDURAS GUATEMAEA Melissa Medina-Flores Universidad Nacional Autonoma de Honduras, Ulisses HONDURAS Janeiro, Rafaqat Masroor Museum of Natural History, PAKISTAN Zoological Federal Rio de BRAZIE Roman Nazarov Museum, Moscow State University of Nis, SERBIA Jianping Jiang Chengdu Institute of Biology, Chinese Academy of Sciences, CHINA University, SOUTH KOREA Javier Alvar ado-Diaz Universidad Michoacana de San Nicolas de Hidalgo, MEXICO Rdmulo Romeu da Nobrega Alves Universidade Estadual da Paraiba, BRAZIE Antonio Salas Environment and Sustainable Development, Cesar Aguilar Universidad de San Marcos, Port Elizabeth Jelka Crnobrnja Isailovic EGYPT Kangwon National University, RUSSIA Adel Ibrahim Suez University, Caramaschi Museu Nacional, Universidade SOUTH KOREA Daesik Park Universidad Autonoma del Estado de Hidalgo, Pakistan Seoul National University, Indraneil Das Peter Uetz Virginia Bruce Waldman Franco Andreone Jr PERU PERU Bill Branch Museum, SOUTH AFRICA Santiago J Sanchez-Pacheco University of Toronto, CANADA Oxyrhopus petolarius, Yotoco, Reserva Amphib Reptile Conserv forestal, 2012, Fdo Castro December 2014 Volume | | Number e87 | Official journal website: amphibian-reptile-conservation.org 8(2) Amphibian & Reptiie Conservation [Special Section]: 1-18; S1-S24 (e87) Conservation status of the herpetofauna, protected areas, and current problems in Valle del Cauca, Colombia ^Alejandro Valencia-Zuleta, Andres Felipe Jaramillo-Martmez, Andrea Echeverry-Bocanegra, Ronald Viafara-Vega, Oscar Hernandez-Cordoba, Victoria E Cardona-Botero, Jaime Gutierrez-Zuhiga, and Fernando Castro-Herrera Universidad del Valle, Grupo Laboratorio de Herpetologia, Departamento de Biologia, Cali, COLOMBIA — Abstract In this study, we present an analysis of the conservation status of amphibian and reptile species by associating the natural protected areas and municipalities with the distribution of richness in Valle del Cauca We establish the percentage of species of amphibians and reptiles in each of the lUCN (International Union for Conservation of Nature) threat categories and assign local conservation categories to all species, construct distribution maps for the records of species in relation to their threat status, and analyze the endemic and total number of species in each of the protected areas We found that nearly 50% of the species in the Valle del Cauca are under some degree of risk or threat, that the largest percentage are in the Vulnerable (VU) category, whereas 37% of the fauna is not threatened (Least Concern [LC] and Near Threatened [NT]), and 13% is categorized as Data Deficient (DD) Although the distribution of species is scattered throughout the territory, patterns are maintained within the various regions, with areas of greater richness found in the Pacific region and the cordilleras; the municipalities with the largest number of species under some level of threat are Buenaventura, Darien, El Cairo, Dagua, Cali, La Cumbre, and Yotoco The types of protected areas with the largest number of species are the Reserves Forestales Protectoras Nacionales (RFPN) 37%, followed by the Parques Nacionales Naturales (PNN) 18%, the Reserves Forestales Protectoras Regionales (RFPR) 10%, and the Parques Naturales Regionales (PNR) 7.5%; 17% (~ 57 spp.) of the species in the Valle del Cauca have not been recorded in any of the protected areas, and more than 65% of these are under some type of threat We consider this study a starting point for evaluating conservation priorities for the herpetofauna of Valle del Cauca Key words Amphibians, Resumen reptiles, distribution, lUCN, population declines, threats —En este trabajo presentamos un analisis del estado de conservacion de las especies de anfibios y reptiles relacionando las areas naturales protegidas y los municipios la distribucion de riqueza en Valle del Cauca Establecemos los porcentajes de especies de anfibios y reptiles en cada categoria de amenaza establecida por UlCN (Union Internacional para la Conservacion de la Naturaleza) y asignamos categorias de conservacion local a todas las especies, se realizo mapas de distribucion de los registros de las especies en relacion a los estados de amenaza, y analizamos el numero de especies totales y endemicas en cada area protegida Encontramos que cerca del 50% de las especies en el Valle del Cauca presentan algun grade de riesgo o amenaza, que la mayor proporcion se encuentra en la categoria vulnerable (VU), mientras que el 37% de la herpetofauna no se encuentra en riesgo (preocupacion menor [LC] y casi amenazado [NT]) y el 13% esta categorizada en datos deficientes (DD) Aunque la distribucion es diferencial a lo largo del territorio, se conservan patrones a lo largo de las regiones, sitios de mayor riqueza en la region pacifica y las cordilleras; y los municipios mayor numero de especies algun grado de amenaza son Buenaventura, Darien, El Cairo, Dagua, Cali, La Cumbre y Yotoco Los tipos de areas protegidas mayor numero de especies son las Reservas Forestales Protectoras Nacionales (RFPN) 37%, seguidas por los Parques Nacionales Naturales (PNN) 18%, Reservas Forestales Protectoras Regionales (RFPR) 10%, y los Parques Naturales Regionales (PNR) 7.5%; el 17% (~ Correspondence Email: ^alejandwvalencia08@ gmail.com (Corresponding author, Alejandro Valencia-Zuleta) Amphib Reptile Conserv December 2014 Volume Number e87 | Valencia-Zuleta et al 57 spp.) de las especies del Valle del Cauca no se han registrado en ningun tipo de area protegida y mas del 65% de ellas presents algun tipo de amenaza Consideramos este trabajo un punto de partida para evaluar prioridades en la conservacion de la herpetofauna vallecaucana Palabras claves Anfibios, reptiles, distribucion, UICN, declive poblacional, amenazas Citation: Valencia-Zuleta A, Jaramillo-Martmez AF, Echeverry-Bocanegra A, Viafara-Vega R, Hernandez-Cordoba O, Cardona-Botero VE, GutierrezZuniga J, Castro-Herrera F 2014 Conservation status of the herpetofauna, protected areas, and current problems in Valle del Cauca, Colombia Amphibian & Reptiie Conservation 8(2) [Special Section]: 1-18; S1-S24 (e87) Copyright: © 2014 Valencia-Zuleta et al This an open-access is Commons Attribution-NonCom- under the terms of the Creative article distributed official any medium, and authorized official journal website mercial-NoDerivatives 4.0 International License, which permits unrestricted use for non-commercial and education purposes only, provided the original author and the publication credit sources, which will and authorized publication sources are recognized and properly credited The be duly enforced, are as follows: official journal title Amphibian & Reptile Conservation, official in Received: 12 March 2014; Accepted: 24 November 2014; Published: 09 December 2014 ferent levels can assign to protected areas, however, Introduction still have not been implemented in the country (Vasquez and Many cline populations of amphibians and reptiles are in de- (Mendelson et al 2006; Bohm et al SeiTano 2009) Currently, 197 reserves of 2013), primar- and species, diseases, 2001; Stuart et al 2007; Rovito illegal trafficking 2004; Mendelson et al 2009; Bohm types exist in the Valle del Cauca; three natural national parks are the of habitat loss, climate change, introduced ily as a result all most important because of their large size and location in (Young et al areas of high herpetofaunal diversity, in the dpto and in 2006; Wake the country et al et al 2013) Estimates — the “Cordillera Occidental” (= the Western Cordillera) and the 15-36% of the world’s species of reptiles threatened (Bohm et al 2013), and according to Stu- “Regibn Pacifica” (= Pacific Region) indicate that (Cardona et are are present in these areas, as their biological patrimony art et al (2004) 22.5% of the species evaluated by lacked sufficient information to evaluate their though the lUCN lUCN status 2013); environmental problems, however, al has not been fully elucidated (Patino 2010) Al- Valle del standardized the use of categories that Cauca is one of the dptos with the greatest have not been evaluated or lack the necessary amount of herpetofaunal species richness (333 species), which represents 24% of the amphibian and 25% of the reptile species recorded from the country (Cardona et al 2013) We are unaware, however, of the number of information for conducting an assessment; in the case of threatened species in the dptos, or plans for their con- can be applied to any taxon and has attempted to catalogue the majority of species cies still reptiles, 59% (lUCN 2012), many spe- of the species have not been assessed book of amphibians, Castro-H and Bohvar-G (2010) included 68 species under some type of servation In a red In Colombia, in addition to the above mention factors that threaten populations of amphibians and reptiles threat, (Rueda 1999; Ruiz and Rueda-A 2008; Velasquez et al 2008; Isaacs and Urbina 2011; Urbina 2011; Urbina et al of amphibians in Valle de Cauca provided by CoiTedor et al 2011; Vargas and Amezquita 2013), the social prob- lem associated with the planting and eradication of and along with an action plan for the conservation (2010); these publications are considered pioneer ef- programs usu- forts in conseiwation; in general, research ally are illicit crops threatens the fauna because of the destruction of developed separately and independently The objective of this paper is to present an analysis primary forests and the use of pesticides such as Glifo- of the conservation status of the species of amphibians sato (Arroyo and reptiles by associating the natural protected areas and A and Lynch 2009; Brain and Solomon 2009) mining crisis also has developed in the country, where municipalities with the distribution of richness in Valle people for economic pur- del Cauca, as a starting point for evaluating conservation mining permits are granted to poses while the long-term impact on the environment caused by these res 2006; activities is priorities for the herpetofauna of this region ignored (Mancera and Alva- UPME 2007; Hernandez et al 2013) In response to these problems, early in the 1930s Materials and Methods “areas naturales protegidas” (= natural protected areas) were designated in the country, which led to the Study area fomia- tion of “zonas forestales protectoras” (= protected forest areas) in the department (dpto= a territorial division Colombia m autonomy in the administration of replanning, and the promotion of economic that has gional issues, and social development within its territory Cauca is a dpto in southwestern Colombia that of 42 municipalities (Eig 1) with a total surface Valle del consists area of 22,142 km^; under the including very it contains a diversity of landscapes, humid warm terms established by the Constitution) of Valle del Cauca vial forests in (decree 1393/40) Regulations for deteimining the exact forests, management that range in elevation categories that competent authorities at dif- Amphib Reptile Conserv tropical forests, transition, dry and lowland montane premontane plu- and very dry tropical montane forests to 4,000 m This to pluvial from sea level December 201 Volume Number e87 I I I Conservation status of the herpetofauna in Colombia were scored from dpto has allocated 233,889 for parks and natural re- areas All these criteria which because of their ecological importance and abundance of natural resources have been established in various strategic ecosystems and protected areas (Gomez means no risk in the particular serves, et al and is unknown where high risk, (see supplemental material at am- criteria, phibian-reptile-eonservation.org for the definition of the score in each criteria) 2007) for each species, Sources to 4, of information it Based on the data obtained calculated the weighted average for the different natural groups (amphibians, lizards, and snakes), and assigned a pereentage in the In updating their Cauca, Cardona on the herpetofauna of Valle del score to each criteria according to the natural group, (2013) considered the following: because the same list et al ral • Geographic data: es, field notes, Obtained from bibliographic sourc- some of and online databases from the follows: at the With the weighted average of each species, these were assigned to Naturales (ICN), and the National does not affect each natu- for the percentage assigned in each criteria) and biological collections of amphibInstitute criteria group in the same way (see supplemental material ians and reptiles at the Universidad del Valle (UV-C), History final de Ciencias Museum of Natural 3.0, Smithsonian Institution (USNM) the categories proposed LC 0-1.4, NT CR 1-3.3, DD tified 5-2.0, by the lUCN, VU 1-2.6, EN 2.7- 3.4^.0 Each category was jus- lUCN aecording to the appendix of the (2012), Ad- especially considering the threats to each species • Threat category (species recorded from the dpto were catalogued using the following in speeies, deaths ditionally, the threat status for traffic or lUCN Red by humans, distribution within the dpto (eco-regions, localities, life zones), tats, books of amphibians and rep- Colombia (Castano-M 2002; Rueda-A et al 2004), and the red book of amphibians from Valle del Cauca (Castro-H and Bohvar-G 2010) occurrence in disturbed habi- tiles in publications, and the presence of species in protected map of Valle del Cauca (Colombia) North: List of Threatened Species (http://www iuenredlist.org/), the red frequency of observation, number of citations in Fig Political each species reported from the dpto was examined by searching through the criteria): trafifieking caused by vehicular as El Aguila (AGE), El Cairo (CR), Ansermanuevo (ASN), Argelia (ARG), Cartago (CTG), Ulloa (ULA), Alcala (ACL), Toro (TR), Versalles (VRSL), Obando (OBD), La Union (UN), El Dovio (DV), Rolda- (RDNL), La Victoria (VTR), Zarzal (ZRZ), Bolivar (BLV); East: Sevilla (SVL), Caicedonia (CDN); Middle: Bugalagrande (BGG), Tmjillo (TIE), Andalucia (ADL), Rio Erio (RE), Tulua (TL), San Pedro (S/PD), Yotoco (YTC), Darien (DR), Buga (BG), Guacari (GCR), Ginebra (GNB), Vijes (VJ), Restrepo (RTP), Cumbre (CMB), El Cerrito (CRT); South: Palmira (PMR), Yumbo (YMB), Cali (CL), Candelaria (CDR), Pradera (PDR), Elorida (ERD), Jamundi (JMD); West: Buenaventura (B/tura), Dagua (DG) nillo Amphib Reptile Conserv December 2014 Volume I | Number e87 | Valencia-Zuleta et • Protected areas: Each species to geographic location was recorded according al Manejo Integrado (DM1) is a geographical space where landscapes and ecosys- Distrito de • and the use of bibliographic resources on protected areas within the dpto, consid- tems retain ering the important areas with a wide extension and though with the ability to hold a great diversity of herpeto- whose The definition of protected areas were defined based on the Decree-Law 622 of 1977 and 2372 of 2010 of the National Government (in parentheses the areas that were chosen in this work): set to fauna • Parques Nacionales Naturales (PNN) is composition and function, their structure natural al- have been modified and and cultural associated values are reach the human population who allocated their sustainable use, preservation, restoration, knowledge, and enjoyment (La Plata and Enclave Subxerofitico Atuncela) an area • of great extent permitted ecological autoregulation their its and whose ecosystems in general have not Each species was recorded based on documented geographic location within the mu- Munieipalities: nicipalities of the dpto been substantially altered by human exploitation Analysis of the Data or occupation, where plant and animal species, geomorphological resorts, historical or cultural The species distribution model for each threat category was performed using all the records collected from the events have scientific, educational, aesthetic and recreational value and their perpetuation is management regime Las Hermosas, Uramba- (Farallones de Cali, Earth 7.1.2.2014; these models were constructed in Santuario de Flora y Fauna (SFF) is dedicated to preserving wildlife species or plant conununities to (Isla we used is Mal- a regional geographic area where landscapes and strate- gic ecosystems, maintain their structure, The position and function values are associated with com- natural and cultural human for preservation, restoration, disposition knowledge, and Reservas Forestales Protectoras (RFP) is graphical area where forest ecosystems main- and composition have been modified and associated natural values are accessible to the population to who into aeeount The percentages of amphibian and reptile species for each threat status was determined, and through histograms indicate the endemic number of species and total number of species in each of the protected areas a geo- tain their function, although their structure 2006) For this work, the 19 climate layers of the above 0.5 taken enjoyment (La Sierpe and Paramo del Duende) • al WorldClim project (www.worldclim.org, spatial resolution of 30 arc second or ~ km^) To evaluate the predictive ability of the models generated, the Area Under the Curve (AUG) score was taken into account The AUG is a ranked approaeh for assessing model fit, which determines the probability that a presence loeation will be ranked higher than a random background location (Phillips et al 2006) The predietion models generated by MAXENT were mapped in ArcGIS 10.1 (ESRI 2013), with only the detection probabilities pelo) Parque Natural Regional (PNR) 3.3.3a data were available (Phillips et preserve genetic resources of native flora and fauna (Decreto 622 de 1977), Max- The software generated models ustheory of maximum entropy only when presence Ent Version ing the • using Google different Bahia Malaga, Tatama) • museums and georeferenced subjected to an appropriate Results human Status of threats to the herpetofauna allocated their preservation, sustainable use, restoration, knowledge, and en- Approximately (RFPN) (Amaime, Anchicaya, San Cipriano and Escalerete rivers, Bosque de some degree of risk or threat The majority of amphibians forests, national Yotoco, Dagua, Cali, Tulua, Sonso-Guabas, (Bitaco and Frayle-Desbaratado) protection Reserva Natural (RN) is of the species in the dpto showed (60%) are in one of the threat categories, with the Vulnerable (VU) containing the most species (59), followed by the Critically Endangered (CR) and Endangered (EN), each with 29, and 27 in the Near Threatened (NT) (Fig A) categories Conversely, more than one-third of the Cerro Dapa-Carisucio) and regional (RFPR) • 51% joyment In this type of protected area are the an area in which undis- show some degree of risk, with those in the NT and VU containing the largest number of species (38 in each), followed by the EN (14), and a few (six) in the CR (Fig 2B) categories Of the remaining herpetofaunal species in the dpto, 17% show no risk (EC), and reptile species turbed conditions exist or have undergone mini- mal human disturbanee of flora, fauna, and soil, and it is intended for conseiwation, research, and study of its natural wealth (Laguna de Sonso) 14% are Data Deficient (DD; Amphib Reptile Conserv see supplemental material) December 2014 Volume Number e87 | | | Conservation status of the herpetofauna in Colombia VU EN CR DD ® LC m NT = Amphibians, (B) = Fig Threat status of the percentage of herpetofaunal species in Valle de Cauca: (A) Eighty percent of the amphibian families contain species under some level of threat, in the family Craugastoridae in ries In general, the NT with 40% dillera and in the Interandean Valley species are found in of the species Reptiles two important (Fig 3C) EN The areas, the Pacific re- gion and the western Cordillera in the northern part of the one of the threat catego- species are represented mostly dpto; in the central Cordillera, a few representatives are and in the families Craugastoridae, Centrolenidae, Hylidae, found in the high elevation areas of Dendrobatidae, and Leptodactylidae In addition, more Buga, to the north, and Palmira, Pradera, and Florida, to than one-half of the VU species are in the family Crau- gastoridae; in particular, the the VU and EN the south (Fig 3D) The distribution of the same pattern and include the families Craugastoridae, tral and western Cordilleras and groups of Pristimantis in highland areas of the western CR species are grouped in the Craugas- in addition, the centrolenids toridae, Bufonidae, and Centrolenidae, families with the in the western Cordillera 70% of the Significantly, the of the family Hemiphractidae, which are restricted to the dpto, but EN categories (Fig 3F) and CR As with the amphibians, most families of (84%) contain species under some level of threat In particular, most of the threatened species are in the families Colubridae, Dactyloidae, Dipsadidae, and Gymnophthalmidae, with most in the NT and VU categories Over 30% of the NT species are in the family Colubridae, followed by the Dipsadidae and Dactyloidae, whereas the VU species are mostly in the Dactyloidae and Dipsadidae The majority of EN species are in the families Colubridae, Dipsadidae, and Gymnophthalmidae Eurthermore, the CR species are represented by one species In particular, reptiles Herpetofaunal richness and the Pacific region (Fig 3E) similar patterns to species in the threat 90% of the municipalities in Valle del est La Cumbre (19), and Yotoco (11) contain the greatnumber of species under some level of threat (Fig 4) A (42), similar pattern was found in these municipalities, where the majority of species fall into the VU category, followed by the EN, and last by the CR, with the only exceptions in the municipalities of Cali and La Cumbre The municipalities of Buenaventura (six amphibians, five reptiles), El Cairo (10, zero), and Darien (eight, one) contain the largest number of CR species The modeling of the maps present an AUC of 0.7540.83, indicating a better performance than the random et al 2001) and dendrobatids are found Cauca contain one species in at least one of the threat categories, whereas 62% of the municipalities contain more than two species The municipalities of Buenaventura (82 species), Darien (61), El Cairo (51), Dagua (45), Cali each family, except for the Dactyloidae models (Manel show and central Cordilleras; DD species are distributed all along the greatest risk of losing species, along with representatives in CR species is important, based on the presence of Atelopus in the cen- species follow Centrolenidae, Dendrobatidae, and Hylidae Significantly, Sevilla, Tulua, is scattered throughout Valle del Cauca, but the areas (see Cardona-B et al The herpetofauna and protected areas [2013] to define ecoregions in the Valle amount of richness are the Cordilleras (Fig A) The NT spe- del Cauca) with the greatest Pacific region cies and the show a wide numfollowed by Protected areas in Valle del Cauca with the greatest RFNP (228 species), RFPR (60), PNR (45), DMI ber of species are the distribution along the western Cordil- and the PNN (120), cover a large area along the Interandean Valley and the RN (21), and the area with the lera (specifically in the northern Pacific (Fig 3B) foothills The VU and central area), species are found along the (three species of reptiles) and northern and central portions of the western the number The majority of the RFPN and the PNN are in the VU, and the SSF is species in and in the remain- ing areas most of the species are in EC, except for the Cordillera, but are less represented in the central Cor- Amphib Reptile Conserv least (34), December 2014 Volume I | Number e87 | Valencia-Zuleta et Fig Distribution species, (C) SEE maps VU species, (Eig 5) The most documented areas for the richness of herpetofauna in the (D) EN species, (E) CR species, areas that protect the largest al and (E) number of in Valle del Cauca (A) richness, (B) NT DD species Endemic species REPN (162 species), and the PNR (25), and the species in a threat category are the the PNN (84), the REPR (37), areas that protect the least SEE dpto, (13, four, 17% and numbers are the three, respectively) Nineteen endemic species are found in the dpto, which DMI, RN, and represents only Throughout the represent the largest (~ 57 spp.) of the species are not found in a more than 65% of those fall into one of the threat categories (NT = four, VU =10, EN = seven, and CR = 15) In addition, information is not available 31% The protected of these species (DD) the largest number of species is (183 species), followed by the the PNN REN and VU number of species DD With with 75% in 15% are categorized as regard to the threat categories for reptiles, three species are in the area with (13), CR (six species), EN (three), (two); the remaining CR, and DD the other three are because they lacked sufficient information for an assess- de Anchicaya ment Earallones de Cali (Eig 6) The endemic species are distributed in four types of protected areas, the REPN of the rivers San Cipriano and Escalerete and the REPR de Bitaco (49) contain a high (84), ing that the PNN and the REPN, which number of species, and it is worth notPNR el Paramo del Duende and the SPP Isla (90), the Amphib Reptile Conserv of the species diversity Amphibians one of the threat categories: protected area, and for 6% December 2014 Volume | | Number e87 | Conservation status of the herpetofauna Colombia in BVU DEN bCR Fig Municipalities in Valle del Cauca with (C) Cali, (D) Darien, (E) El Cairo, and (E) the greatest number of species in the threat categories: (A) Buenaventura, (B) Dagua, La Cumbre we show de Malpelo are the types of areas with the greatest num- and Bolivar-G (2010), ber of endemic species Only four species (Pristimantis in the diaphonus, Anomaloglossus atopoglossus, Nymphargus (12), armatus, and Geophis betaniensis) are not found in any disappearance has increased in certain species, which of these areas troublesome Discussion The lack of a threat status among the reptiles results from insufficient basic ecological information and the number of and CR species in the NT (one) a significant increase EN (11), VU (10), categories, indicating that the risk of is actual distribution of their populations (Urbina-Cardona Conservation status of amphibians and reptiies 2008), for which an evaluation of the threat status has focused on specific species or groups The need to recognize (e.g., the red book of reptiles in Colombia), and thus has become a problem the status of a species in a specific For area should be considered baseline information for devel- for planning conservation strategies oping studies and management plans for its conservation status of populations of reptile species in a given area has The lUCN been proposed categorizations are generally applied globally as a mechanism to this reason, the change and attitudes for each taxon to determine the status of a species at the generate interest in preserving these organisms (Dodd and are considered advanced 2001), the protection and restoration of large areas these local or regional levels, on organism inhabit (Roe and Bolfvar-G 2010); information studies (Castro-H et al 2004; Franga and Araujo certain species (e.g., population status, natural his- 2006), species-specific information, field studies, de- tory) is necessary to elicit an approximate categorization mographics, natural history, and possible threats (Cagle Records for the DD species (13% of the species in this 2008; Fifes (some are only known from tions), and thus it is species (60%) in in Valle del Cauca, so that their original descrip- resents nearly one-half of the total herpetofauna of the in the dpto; in show a moderate history allow medium first to assess many cases, them affect their many to avoid these conditions cause the current status of abundance characteristics of their natural The conservation of snakes remains term (VU) In comparison with the results of Castro-H Amphib Reptile Conserv a local effective strategies can the is might make them vulnerable and one of the threat categories, which rep- risk of extinction or population decline over the is species of reptiles, and in spite of their low density threats a high proportion of amphibian dpto, and the majority of these species more be accomplished This study not possible to determine their status The dpto contains 2013) Significantly, this study proposal that easily addresses the status of reptile species study) are not well represented in herpetological collections et al many subjective, be- species remains un- December 2014 Volume I | Number e87 | Amphibian & Reptiie Conservation Official journal website: 8(2) [General Section]: 30-35 amphibian-reptile-conservation.org Short Communication surveys in Western Panama indicate populations of Atelopus varius frogs are persisting in regions where Batrachochytrium dendrobatidis is now enzootic Field ^Rachel Perez, ^’^Corinne L Richards-Zawacki, ^Alexander R Krohn, ^Matthew Robak, "^Edgardo J Griffith, "^Heidi Ross, ^Brian Gratwicke, ^Roberto Ibanez, and *Jamie Voyles ^ ^Department of Biology, New Mexico Institute of Mining and Technology, Socorro, New Mexico 87801, USA ^Department of Ecology and Evolutionary Biology, Tulane University, New Orleans, Louisiana 70118, USA ^Department of Environmental Science Policy and Management, University of California, Berkeley, Berkeley, California 94720, Conservation Biology Institute, Washington D.C., Key words USA USA '^El Valle Amphibian Conservation ^Smithsonian Tropical Research Institute, Center, El Valle, PANAMA ^Smithsonian PO Box 0843-03092, Balboa, Ancon, PANAMA Chytridiomycosis, Central America, Harlequin frog, golden frog, threatened species, emerging infec- tious disease, chytridiomycosis, Batrachochytrium dendrobatidis Citation: Perez R, Richards-Zawacki CL, Krohn Panama AR, Robak M, Griffith EJ, indicate populations of Atelopus varius frogs are persisting Reptile Conservation 8(2) [General Section]: 30-35 in Ross in Western Amphibian & H, Gratwicke B, Ibanez R, Voyles J 2014 Field surveys regions where Batrachochytrium dendrobatidis is now enzootic (e85) Copyright: © 2014 Perez et al This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercialNoDerivatives 4.0 International License, which permits unrestricted use for non-commercial and education purposes only, in any medium, provided the original author and the official and authorized publication sources are recognized and properly credited The official and authorized publication credit sources, which will be duly enforced, are as follows: official journal title Amphibian & Reptile Conservation; official journal website Received: 18 June 2014; Accepted: 25 August 2014; Published: 11 November 2014 document Bd invasion and characterize 5J-related amphibian losses (Brem and Lips 2008) Second, A zeteki has been repeatedly tested in controlled laboratory infection experiments and found to be highly The stunning Harlequin frogs of the genus Atelopus, once common, are now among the most imperiled of all amphibian species (La Marca et al 2005; Zippel et al 2006) Of 88 and thus helped described Atelopus species in Central and South America, 65 (74%) are Critically Endangered (La Marca et al 2005) The most pressing threat to these frogs is ch54ridiomycosis, a fungal disease caused susceptible to lethal ch)4ridiomycosis (Bustamente et 2010; DiRenzo by Ba- adaptive sociated with die-offs of amphibians around the world lison et 1998; Longcore et al al et al 2014) Third, immune al responses are suppressed by 2014) Thus, Atelopus species have dynamics and have also provided motivation for progressive conservation action (Gagliardo et Kilbum Knowles 2007) and we established new study we In October 2012, al 2008) revisited study sites that were surveyed fox Atelopus from 2001 to 2004 (Richards and 2010), including declines in t\wQQ Atelopus species: A varius, A zeteki, become wave spreading to east caused et al (El- important focal species in the study of ch)4ridiomycosis mass mortality events, resulting in catastrophic losses in amphibian diversity (Lips et al 2006; Brem and Lips 2008; Woodhams et al 2008; from west Bd al 1999; Kilpatrick et 2010) In Western Panama, an epidemic 2014; Ellison et al al recent immunogenetics research suggests that A zeteki trachochytrium dendrobatidis (hereafter “5 0.999) from a linear regression of log (plasmids) vs log (zoospores) (t4 by running = 210.6, P < 0.0001) that we obtained the plasmid standard set alongside a series of standards containing known quantities of zoospores (obtained from Alex Hyatt, Australian Laboratory) If one of three replicate positive, we checked Animal Health wells turned up Cycle Threshold (Ct) value to de- termine whether non-amplification in two of three wells could have been caused by a low-level infection (near the detection threshold) and verified that the qPCR was not inhibited (IPC amplified normally) In cases of inhibition or Ct values far from the detection threshold, and considered them positive if we re-ran Bd was detected in any of the three re-run wells We surveyed 16 which were field sites from 2012 to 2013, 10 of where Atelopus species were found in 2004 (Richards and Knowles 2007), prior to the chytridiomycosis epidemic The remaining six sites were chosen based on other biologists’ sightings of Atelopus Figure sites Figure A female Harlequin frog, Atelopus varius This spe- Endangered by lUCN, has been the mountains of Western Panama cies, classified as Critically found in small numbers in A pair of Atelopus varius in amplexus, found in the mountains of Western Panama Amphib Reptile Conserv 31 November 2014 Volume Number e85 | | | Atelopus varius populations persisting after chytridiomycosis Table Summary of infection where Atelopus varius m) (735 Prevalence N Lower 95% Upper 95% Atelopus varius 0% 0/1 0.975 Colostethus flotator 0% 0/1 0.975 Craugastor bransfordii 0% 0/1 0.975 Craugastor crassidigitus 0% 0/1 0.975 Craugastor fitzingeri 0% 0/1 0.975 Lithobates warszewitschii 60% 3/5 0.147 0.947 Pristimantis cerasinus 100% 1/1 0.025 Pristimantis cruentus 0% 0/2 0.842 Pristimantis museosus 0% 0/1 0.975 Pristimantis ridens 0% 0/1 0.975 25% 2/8 0.0715 0.591 Terahyla spinosa 0% 0/1 0.975 Glass frog metamorph 0% 0/1 0.975 Atelopus varius 0% 0/6 0.459 Colostethus flotator 0% 0/7 0.41 25% 1/4 0.073 0.524 0/1 0/1 0.975 42% 1/5 0.057 0.437 Silverstoneia flotator 33% 1/3 0.008 0.906 Smilisca spp 100% 3/3 0.292 Atelopus varius 0% 0/1 0.975 Craugastor bransfordii 0% 0/2 0.842 Craugastor crassidigitus 0% 0/1 0.975 Craugastor longirostris 0% 0/1 0.975 Craugastor fitzingeri 0% 0/2 0.842 Dendrobates auratus 0% 0/3 0.708 Dendrobates minnutus 0% 0/1 0.975 Diasperous spp 0% 0/3 0.708 Pristimantis caryophyllaceus 0% 0/1 0.975 Rhinella alata 0% 0/10 0.308 Silverstoneia flotator 0% 0/1 0.975 Sachatamia albomaculata Highland (521 m) Colostethus panamensis Pristimantis ridens Rhaebo Lowland Hertz (e.g., (0 m) et al haemititicus 2012) or predicted habitat in species distribution models We (Fig 1, Table 1) At one varius, five adult males, found persisting pop- amplexus site, we found one juvenile males each of the other two respective We (Fig 2) have intentionally only provided general mation, rather than precise risk sites site (Table A sites confirmed that Bd is cause other species (e.g., al om vary in elevation from 45 to 750 areas 1) These m and all three al 2006) and concerning be- at least A zeteki is 2014; Ellison et al known to be al 2010; DiRenzo et 2014) found that A varius is ma Furthermore, our positive persisting in multiple sites qPCR results suggest that these populations have survived despite the presence of Bd Prior to this study Hertz sites et al (2012) was the only study to document sightings of A varius in the wild in are in Panama where fungal epidemics were associated with mas- Amphib Reptile Conserv is still following a chytridiomycosis outbreak in western Pana- Lithobates warszewitschii and Sach- (Table We did we know that they previously had restricted ranges We present in two of these three Bd positive on these transects 2010) et al et al efforts for these species are fection experiments (Bustamente et atamia albomaculatd), but none of the Atelopus samples were 2006 (Lips highly susceptible to chytridiomycosis in laboratory in- coordinates, due to the If Although suiwey (Zipple et 1) site infor- populations based on detection of Bd on skin swabs Lips 2008; Kilbum underway, the absence of these species of illegal animal collections We Brem and to not detects, zeteki ox A chiriquiensis at any of our study We found individual adult in amphibian declines from 2004 2006; and two adult females, including one pair at sive suitability varius at three of 16 (18.7%) field sites ulations of at three sites persist Species Site (Elevation) Highland still prevalence of Batrachochytrium dendrobatidis from amphibians found 32 since 2006 Those observations were made in November 2014 Volume Number e85 | | | Perez 2009 at a site in tions may changes America Proceedings of the National Academy of Sciences of the United States of America 95(15): Santa Fe National Park These popula- be persisting for a wide range of biotic in host behavioral, innate or acquired tral (e.g., immune 9031-9036 Brem FM, responses, anti-5(i microbial communities) or abiotic environmental/thermal conditions) reasons (e.g., How- Lips KR 2008 Batrachochytrium dendroba- among Panamanian amphib- tidis infection patterns because there have been few coordinated efforts to ian species, habitats and elevations during epizootic new populations, resurvey historical localities, or for Bd infections, the question of how these popu- and enzootic stages Diseases of Aquatic Organisms 81(3): 189-202 ever, locate test et al —and whether any other Atelopus populations have survived—remains be unraveled We lations Panamanian Golden Frog infected with a pathogenic chytrid fungus Integrative Zoology 5(2): 143-153 DiRenzo GV, Langhammer PF, Zamudio KR, Lips KR 2014 Fungal Infection Intensity and Zoospore Output of Atelopus zeteki, a Potential Acute Chytrid Supershedder PloS One 9(3): e93356 Ellison AR, Savage AE, DiRenzo GV, Langhammer P, Lips KR, Zamudio KR 2014 Fighting a Losing Battle: Vigorous Immune Response Countered by Pathogen Suppression of Host Defenses in the Chytridiomycosis-Susceptible Vrog Atelopus zeteki G3: Genes believe that the lack of post-decline survey effort has not likely a consequence of the enormity of the challenge of monitoring these species while simultaneously establishing conservation pro- grams of chytridiomycosis to entire to abate the threat amphibian communities Post-decline surveys are critical for conservation of Atelopus species, as well as for other neotropical am- Documenting rediscovered species is critically important for informing conservation and management phibians initiatives (Minteer et al 2014) and, in this case, could Genomes be accomplished with photographs, rather than collecting the individuals Moreover, understanding the variables that permit some populations to persist and A being bred in captivity zeteki) with the expectation for P, Griffith Hertz A, Lotzkat reit S, Carrizo A, Ponce amphibian species importance of con- in the central the wild,” even after long periods without 6(2): any sightings Worldwide Wildlife Fund, Association of Zoos and Aquariums (AZA) Con- ci JP, (e46) Green in anuran AZA Amphibian Taxon AdviLatin American Studies, DM fimgus, chytrid for using the Kilpatrick We thank We thank Maggie Unkefer, Clif- 2010 Ubiquity of the pathogenic Batrachochytrium communities in AM, La Marca E, Lips KR, P 2010 The ecology Letters S, Puschendorf R, Ibanez work was conducted under lACUC NMT: 2013-1, lACUC Tulane: 0453, STRI lACUC: strophic their support This Young BE 2005 Catapopulation declines and extinctions in Neo- tropical harlequin frogs (Bufonidae: Atelopus) Bio- SE/AH-1-13, tropica SE/AH-1-12, and SE/AH-4-13 Lips 3>1 {2): KR, Brem Voyles J, 190-201 F, Brenes R, Reeve ID, Alford RA, Collins JP 2006 Emerging infectious dis- ease and the loss of biodiversity in a Neotropical Literature Cited Berger L, Speare R, Daszak AA, Goggin CL, 7(4): 109-118 R, Rueda-Almonacid JV, SE/AH-4-12, dendrobatidis, Panama EcoHealth Briggs CJ, Daszak Medina, Karina Klonoski, and Simone Des Roches for ANAM List and impact of chytridiomycosis: an emerging disease of amphibians Trends in Ecolog}’ & Evolution 25(2): ford Richardson, Kelly Cruz and the Cruz family, Daniel 2012-0901-2015, lUCN Red 537-548 the Smithsonian Tropical Research Institute for allowing facilities Reptile Consei^ation March 2014] Kilbum VL, Ibanez R, Sanjur O, Bermingham E, Sura- Conservation Fund, the Chicago Board of Trade Endan- access to their & mountain range of www.iucm-edlist.org [Accessed: 09 Endowment Fund, Riverbanks Conservation Support Fund, Bay and Paul Foundations, Minnesota Zoo Ulysses S Seal Conservation Grant, Maryland Zoo this research St- categories and criteria Available: servation and Project Golden Frog funded 9-30 lUCN 2013 Guidelines —Disney M, Kohler G, B 2012 Field notes on findings of threatened western Panama Amphibian sory Group, Stone Center for Ross Zoo Yearbook as an example International tinued monitoring for species presumed to be “extinct in gered Species Fund, The J, 42(1): 125-135 of one day return- them to the wild (Gagliardo et al 2008; Zippel et al 2011) Our discovery of extant populations of A varius Acknowledgments Mendelson E, amphibian conservation, using the programmes in Panama A varius ing in 5(i-enzootic areas underscores the Crump H, Zippel K 2008 The principles of rapid response while others die (e.g., Genetics: g3-114 Gagliardo R, out will be critical to conservation, especially since several species are Effects of tem- perature and hydric environment on survival of the to been so much an oversight, but HM, Livo LJ, Carey C 2010 Bustamante have persisted P, amphibian community Proceedings of the National Academy of Sciences of the United States of America Green DE, Cunningham 103(9): 3165-3170 Parkes H 1998 Chytridiomycosis Longcore JE, Pessier AP, Nichols DK 1999 Batracho- causes amphibian mortality associated with population declines in the rain forests Amphib Reptile Conserv of Australia and Cen- chytrium dendrobatidis gen 33 et sp nov., a chytrid November 2014 Volume Number e85 | | | Atelopus varius populations persisting after chytridiomycosis pathogenic to amphibians Mycologia 91(2): 219-227 disease in changing climates: strategies of an amphib- Mendelson JR, Lips KR, Gagliardo RW, Rabb GB, Collins JP, Diffendorfer JE, Brodie ED 2006 Biodiversity-Confronting amphibian declines and extinctions ian pathogen Ecology 89(6): 1627-1639 KC, Ibanez R, Eindquist ED, Richards CE, Jaramillo CA, Griffith EJ 2006 Implicaciones en la con- Zipell servacion de las ranas doradas de Panama, asociadas Science 313(5783): 48 Minteer BA, Collins Eove KE, Puschendorf R 2014 Avoiding (Re) extinction Science 344(6181): 260- Available: http://erevistas.saber.ula.ve/index.php/her- 261 petotropicos/article/view/643 [Accessed: 25 October su revisidn taxonomica Herpetotropicos JP, Richards CE, Knowles EE 2007 Tests of phenotypic and genetic concordance and 2014] Zippel K, Johnson K, Gagliardo R, Gibson R, their application to the The Amphibian Ark: Molecular Ecology 16(15): 3119-3133 Woodhams DC, Alford RA, Briggs CJ, Johnson M, RolfomddLQ) at A global community for ex situ conservation of amphibians Herpetological Conser- lins-Smith EA 2008 Eife-history trade-offs influence is McFad- den M, Browne R, Martinez C, Townsend E 2011 conservation of Panamanian golden frogs (Anura, Bu- Rachel Perez 3(1) vation and Biology a M.S student in the department of biology at 6(3): 340-352 New Mexieo Teeh She reeeived her B.S University of California, Riverside Her main researeh interests are in the areas of epidemiology, eeoim- munology, and amphibian eonservation She is eurrently investigating innate defenses and mierohabitat eonditions of neotropieal amphibians Corinne Richards-Zawacki is a professor in the Eeology and Evolutionary Biology department at Tulane University Her researeh lies at the interseetion of eeology and evolutionary biology in that she approaehes questions about how ehanges in elimate and habitat shape population and eommunity proeesses in a way that explieitly eonsiders their evolutionary implieations The questions she asks address (1) the effeets of elimate and landseape ehanges on speeies distributions and diversity, (2) how reproduetive isolation evolves during speeiation, and (3) how elimate and host/pathogen evolution shape the dynamies of wildlife diseases She is passionate about amphibian eonservation and has 12 years of experienee working on eonservation-oriented projeets in Panama Mueh of her work in Panama has foeused on the Critieally Endangered Panamanian golden Ifogs Her lab has also published studies related to the eaptive management of amphibian speeies threatened by Bd Alexander Krohn is a eurrent Ph.D eandidate at the University of California, Berkeley, in the department of Environmental, Seienee, Poliey and Management His dissertation foeuses on the eonvergent evolution of melanism in desert reptiles, but he has been interested in herpetology, tropieal eeology, and eonservation sinee he was in middle sehool Matthew Robak is broadly interested in amphibian eonservation Tulane University where he is researehing how He is eurrently a Ph.D eandidate at differenees in temperature affeet amphibians’ immune responses to Batrachochytrium dendrobatidis exposure Edgardo J Griffith is a world-renowned leader in amphibian eonservation in founders of the El Valle Amphibian Conservation Center (EVACC) Edgardo proven that multi-speeies ex situ amphibian eonservation the dramatie amphibian deelines in is the only option Panama His work has been featured Panama and one of the one of the pioneers is we have that has moment to fight at the in several eonservation books, doeumentaries, and peer-reviewed seientifie papers Edgardo has 14 years working with the Panamanian amphibians, ineluding the Panamanian golden frog in situ and breeding them ex situ Amphibian Conservation Center (EVACC) Heidi and her husband were awarded the San Diego Medal of International Conservation in 2012 for their work at EVACC Heidi has 10 years of experienee ex situ experienee with Panamanian amphibians leading to the breeding of nine of the 12 priority speeies in the EVACC eolleetion, ineluding the Panamanian golden frog Heidi Ross Amphib Reptile Conserv is the projeet direetor at the El Valle 34 November 2014 Volume Number e85 | | | Perez Brian Gratwicke al is et al a conservation biologist that leads the amphibian eonservation program at the Nation- Zoo (Washington, DC, USA) Brian also leads the Previous eonservation experienee ineludes in the Caribbean, and Panama Amphibian Reseue and Conservation Projeet work on freshwater eeology tiger eonservation efforts in Asia He in Afriea, tropieal marine eeology has published more than 25 peer-reviewed papers and book ehapters and was a eontributor to Hotspots Revisited Brian reeeived a Ph.D in zoology from Oxford University He obtained his baehelor’s and master’s degrees respeetively, at the versidad de zoology and fisheries eeology, from the University of Zimbabwe Roberto Ibanez based in is the in-eountry Direetor of the Smithsonian Tropieal Researeh Panama (1996—2014), and versity (2003—2013) He Panama Amphibian Reseue and Conservation Institute Projeet, He has been part-time Assoeiate Professor at the Uni- professor during de Panama Field Study Semester of MeGill Uni- reeeived his B.S in Biology from the Universidad de Panama, and his Master’s degree and Ph.D in Zoology from the Eeology and Evolutionary Biology Department at the University of Conneetieut lished He has studied the amphibians and reptiles of Panama for more than 30 years more than 30 peer-reviewed papers, more than 20 lowlands of eentral Panama Jamie Voyles is a and a book guide to the has pub- amphibians of the a distinguished researeher of the national researeh system of Panama New Mexieo Teeh working on emerging infeetious She eonduets eh54ridiomyeosis researeh in Central Ameriea and in California She member of multiple working groups involved in eonservation to amphibianreseue.org Amphib Reptile Conserv is eurrently an Assistant Professor at diseases in wildlife is He notes, He initiatives, investigating disease-related amphibian deelines She is aetively sueh as Amphibian Reseue and Conservation Projeet, and eontributes and AmphibiaWeb 35 November 2014 Volume Number e85 | | | Amphibian & Reptiie Conservation Official journal website: 8(2) [General Section]: 36-43 amphibian-reptile-conservation.org Confirmation of introduced Louisiana pinesnakes, Pituophis ruthveni, in Fiorida based on molecular analyses ^’*Kenneth L Krysko, ^Dustin C Smith, ^Michael R Rochford, "^Guy N Kieckhefer and ^Florida Museum of Natural History, ^’^Leroy Division of Herpetology, 1659 P III, Nunez Museum Road, University of Florida, Gainesville, Florida 32611, USA ^Zoo Miami, Florida 33177, USA ^University of Florida, Fort Lauderdale Research and Education Center, 3205 College Avenue, Fort Lauderdale, Florida 33314-7719, USA ‘^1113 Wyoming Street, Belgrade, Montana 59714, USA Miami, 12400 Southwest 152nd Abstract —As Street, more wide-range phylogenetic studies are compare DNAfrom these data sets available, the opportunity arises to to suspected introduced individuals in order to confirm species and determine their geographic origins Two recently collected Pituophis specimens in Miami-Dade County, Florida, were examined using molecular analyses Maximum likelihood and Bayesian inference methods place our specimens within the P catenifer sayi I P ruthveni clade Additional morphological evidence support their identification as the Louisiana pinesnake, Pituophis ruthveni StuW 1929, a species indigenous to a small area in western Louisiana and eastern Texas and candidate for listing by the U.S Fish and Wildlife Service Although P ruthveni is viewed as a distinct species from P catenifer sayi based on allopatry and differences in color pattern, no molecular evidence was found supporting the recognition of P ruthveni as a separate species However, adding other mtDNA and nuclear DNA genes might provide needed data for distinguishing between these two named taxa identification Key words DNA, exotics, ND4, mitochondrial, MR, Kieckhefer based on molecular analyses Amphibian & Citation: Krysko KL, Smith DC, Rochford veni, in Florida mtDNA, III nonnative, phylogenetics, Squamata, species GN, Nunez LR 2014 Confirmation of introduced Louisiana pinesnakes, Pituophis ruth- Reptile Conservation 8(2) [General Section]: 36-43 (e86) Copyright: © 2014 Krysko et al This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercialNoDerivatives 4.0 International License, which permits unrestricted use for non-commercial and education purposes only, in any medium, provided the original author and the official and authorized publication sources are recognized and properly credited The official and authorized publication credit sources, which will be duly enforced, are as follows: official journal title Amphibian & Reptile Conservation; official journal website Received: 19 August 2014; Accepted: 27 October 2014; Published: 19 November 2014 est to highest Introduction numbers) biological control, zoos, cargo/ plant shipments, and the pet trade stages 2-5 after Colautti and Pinesnakes, bullsnakes, and gophersnakes {Pituophis Macisaae 2004) are those transferred from their native range into a new nonindigenous area Over the past cen- Holbrook 1842) are large (up to 254 em total length) eonstrictors native to North Ameriea, charaeterized by Introduced species (e.g., has become increasingly clear how disruptive disproportionately small heads, four prefrontal seales, human-eaused biological introductions have been to the planet While not all introduced species eause obvious harm, some introdueed speeies can eventually become upwards between the intemasals (Conant and Collins 1991) Based primarily on moleeular data using Parsimony and Maximum eeonomic Likelihood analyses with 893 base pairs (bp) of the nieo- tury it threats and a large and lead to serious conservation prob- As of 2005, was rostral plate that extends estimat- tinamide adenine dinucleotide dehydrogenase subunit ed that the eost of environmental damages, losses, and lems (Simberloff et al 1997) it control due to introdueed speeies exeeeded $120 billion (ND4) region (Rodriguez-Robles and De Jesus-Eseobar 2000), the P melanoleucus species complex contains per year in the United States alone (Pimentel et al 2005) three Prior to 2011, the state of Florida had 137 doeumented species; melanoleucus P (sensu strieto; Pinesnakes; with three subspecies introdueed reptile and amphibian taxa (56 being estab- lodingi, which ranks highest in the world (Krysko et al 2011a, 2012) Invasion pathways in Florida include (few- P m melanoleucus, P m mugitus), P P c author, affinis, P c annectens, P catenifer, c P c m cateni- fer (gophersnakes and bullsnakes; with six subspecies lished), Correspondence Email: * kenneyk@ufl.edu (Corresponding ^gnkieck@gmail com fherizinosaur@yahoo com eurrently recognized P deserti- KLK); ^dustsmi@miamidade.gov; ^ mikerochford@hotmail.com', ; Amphib Reptile Conserv 36 November 2014 Volume Number e86 | | | Krysko Figure Map et al of Zoo Miami bounded in green Note that major roadways, residential areas, and undeveloped proteeted lands surround zoo property Dots represents loeations of Pituophis found on zoo property; yellow female) and red cola, P c = UF-Herpetology 163092 pumilis, P c sayi), and P ruthveni (Louisi- ell et al al 1976) Although New eontaining only Jersey south to extreme southern peninsular Florida (i.e., species because Miami-Dade County; Krysko et al 2011b) and west to Kentueky and southeastern Louisiana (Rodriguez- is Thomas 1998; Reiehling 1995; Stull 1929; occurs in the eastern United States from southern De (gravid (adult male) ana pinesnake) Pituophis melanoleucus (Daudin 1803) Robles and = UF-Herpetology 157954 P ruthveni P c sayi, it is it is et nested within a elade recognized as a separate oceurs in ahopatric populations and somewhat diagnosable using eolor pattern charaeters (Collins 1991; Knight 1986; Reiehling 1995; Rodriguez- De Jesus-Escobar 2000; Thomas Robles and Jesus-Eseobar 2000) This species lacks et al 1976) a dark line from the eye to the angle of the jaw, has a Pituophis ruthveni dorsal pattern either absent (uniform blaek), obseure, or imperiled species by the U.S Fish and Wildlife Serviee whitish to brownish with 23-30 distinct dark dorsal body (2013) blotches that are elearly separated from each other both The anteriorly and posteriorly along the body and tail (Knight Thomas et al Pituophis catenifer oceurs from the Paeifie Ocean 1986; Powell et 1976) treme al bar 2000; Powell et al 1998) De southern Pituophis melanoleucus from ex- peninsular Florida (UF-Herpetology Dade County, and because of ongoing dense and Texas, and from Canada south to Mexieo (Rodriguez-Robles and known also a eandidate for listing as an 45970) was eohected in 1980 in a disturbed pineland (with Casuarina and Schinus) in Cutler Ridge, Miami- 1998; Reiehling 1995; east to Wiseonsin, Illinois, last is Jesus-Eseo- tion this speeies is believed to be extirpated along the Atlantie Coast Ridge (Krysko et This speeies typically has urbaniza- al 2011b) In 2010, two a dark line from the eye to the angle of the jaw, and a yel- Pituophis were eohected on the Atlantie Coast Ridge at low or cream-eolored dorsal pattern with 41-79 distinet dark dorsal blotehes that are clearly separated from eaeh other both anteriorly and posteriorly along the body and Zoo Miami, Miami-Dade County; one was found tail (Knight 1986; Powell Thomas et al 1976) et al et al on color pattern 1998; Reiehling 1995; introduced sometimes laeks a dark line et al 1998) from the eye brown 2011a) This species to the angle P eies alone, these snakes were suspected to ruthveni and reported to represent the known vouehers Pituophis ruthveni occurs in allo- 2004; Powell an undeveloped area and another near publie aeeess Based patric populations in western-central Louisiana to eastern Texas (Ealy in first for this speeies in Florida (Krysko et Many doeumented be al introduetions categorize spe- based on sometimes vague superficial morphology, such as color patterns, which may or may not be arbitrary of human 28-38 dark dorsal blotches; near the head the blotches obscure However, as more wide-range phylogenetie studies are conducted and published, the oppor- the ground coloration, whereas near the tunity arises for other researchers to the jaw, and has a pale tinctly separated dorsal pattern with tail they are dis- known from eaeh other (Knight 1986; Pow- Amphib Reptile Conserv 37 constructs eompare DNA from data sets to suspected introdueed individuals in November 2014 Volume Number e86 | | | Verification of introduced Pituophis ruthveni in Florida Figure Well-dQYQlopQd Pituophis embryo (UF-Herpetology 164295) oviposited from wild colleeted gravid female (UF-Herpe- tology 157954) in Miami, Miami-Dade County, Florida tember 2010 and revealed a well-developed embryo (UF- order to confirm species identification as well as deter- mine we conduct Herpetology 164295; Fig molecular analyses of Pituophis in a coalescent frame- On 25 December 2010 their geographic origins In this paper, 1,425 tory, Zoo Miami is situated at 12400 SW Street, Miami, Miami-Dade County, Florida, USA (Fig 25.61 1926°N, 80.398042°W, Datum WGS84, elev m) The property 152* Museum Memphis Zoo, and the male (UF-Herpetology 163092) is housed at Zoo Miami of which are undeveloped is currently housed at the Laboratory techniques managed lands, predominantly of pine rockland habitat Zoo Miami property is surrounded by a mixture of natural areas, disturbed areas, and a county park, followed by rification Kit dense urbanization Sciences, LLC) Using total cellular On 16 May 2010 at female, 1,302 1645 h, an didult DNA isolations were obtained using QIAquick PCR Pu- Pituophis (gravid and DNeasy Blood and Tissue Kit (Qiagen DNA as a template and Polymerase Chain Reaction (PCR) methodology mm gy 157954; see of Natural His- University of Florida The female (UF-Herpetology 157954) ; ha, 106 another adult (male, km southwest of the first snake sion of Herpetology, Florida and specimen acquisition 300 h, The well-developed embryo, shed skins from the two adults, and digital images were deposited in the Divi- and Methods consists of ca 1215 mm lake and 0.26 Site description at SVL, 1,635 mm TL) Pituophis (UF-Herpetology 163092) was collected in an undeveloped area (25.60304°N, 80.40295°W), across a large man-made work to confirm species identity and phylogenetic placement of our two specimens, followed by more detailed examination of morphology and color pattern Material 2) SVL, 1,486 mm TL; UF-HerpetoloFig 86 in Krysko et al 2011) was col- (Saiki et al 1988), mitochondrial amplified and sequenced for the DNA (mtDNA) was ND4 region follow- lected in a service area behind a large animal exhibit ing Rodriguez-Robles and (25.60395°N, 80.4006°W) This snake was observed ND4 region includes a section of the end of the ND4 gene, and two subsequent transfer ribonucleic ac- by zoo staff the previous day along an adjacent public which were sequenced using the primers ND4 and Leu (Arevalo et al 1994) PCR was conducted in 25 pi reactions: 9.5 pi H^O, 12.5 pi GoTaq® Master Mix (Promega Corp, Madison, Wisconsin, USA), 1.0 pi each primer (10 pM), and 1.0 pi DNA template PCR parameters included initial denaturing at 94 °C for three min, followed by 35 cycles of amplification: de- on 28 June 2010; all three eggs contained a dark blood spot, but only one egg had an obvious network of veins developing The first two eggs failed to develop and were discarded on 06 July 2010 The third egg had an unpleasant odor and was frozen on 14 September 2010; it was dissected on 20 Sep- Amphib Reptile Conserv ’ ids (tRNA”‘®, tRNA^®^), and oviposited three eggs on 22 June 2010 The eggs were viewed with a Jesus-Escobar (2000) The walkway, but was not captured This snake was retained in captivity De light 38 November 2014 Volume Number e86 | | | Krysko et al CA_SanBenito_Lamprope/f/5 9efi//a_HWG1485 98 TX_Culberson_Bogerfop/?/s suboct//ans_CME116 CA_Riverside_Anzo^s elegans_W^Z^ OHOttawaPanfPerophis — 100 U'U/p/>JUS_CAS1 84362 FL_Hi llsborough_P/fiiop/7/s melanoletJCLis mugitus_W\yZ^ 3063 - FL^Waku na^Pituophis melanoleucus mugitusJJSN M2 1 452 \^C_Brur\5'Mck_Pituophis melanoleucus mela^oleucus_W\VZ^502^9 98 •“I NJ_Cumberland_P/fuop/i/s meianoleucus melanoieucus_MVZ225b20 - NJ_Burlington_P/f/;op/7/s meianoleucus melanoleucus_W\WZ22b52) — Pituophis meianoleucus lod^nglJH\NG2Q5^ I L Pituophis meianoleucus lodlngl_HV\lG2652 q: Pituophis deppei jani 94 93 CD Mexico_Michoacan_P/^yop/7is deppei deppei_2 100 Mexico_Durango_P/fL/op/i/s deppei b'eppe/_1 ^0 ar/5_12 CA_SanDiego_P/ft/oppys catenifer annectens_M^/Z^ 50206 84l- CA_SanDiego_P/fL/op/7/s catenifer annecteas_MVZ149983 CASantaBarbaraPyfyop/r/s catenifer pumilus_29 CA_SantaBarbara_P/fi;op/7/s catenifer pumllus_30 0.02 Figure Maximum Likelihood phylogeny for Pituophis (Squamata: Colubridae) snakes, ineluding the two known P ruthveni Miami-Dade County, Florida Note that values (> Inset photograph of UF-Herpetology 157954 by Dustin C Smith (highlighted in yellow, UF-Herpetology 157954 and 163092) eolleeted in Miami, 50%) above nodes represent bootstrap support (UF-Herpetology 157954 and 163092) are KJ938643 naturing at 94 °C for one min, annealing at 52 °C for one min, and extension at 72 °C for one min, followed and KJ938644, respectively A total of 48 specimens with 875 base pairs (bp) of se- Three pi of each PCR product were electrophoresed on a 1% agarose by °C a final extension at 72 gel, visualized with Hayward, California, standard Sequence for seven GelRed™ haplotypes were estimated using both staining (Biotium Inc., USA), and compared with a files quence data were analyzed Relationships among mtDNA with for Bio- technology Research, University of Florida) were as- by Biomatters Available from —DNA sequence data were downloaded from GenBank for 46 snakes, including 42 Pituophis, and one of each Lampropeltis getula, Pantherophis vulpinus, Bogertophis subocularis, and Arizona elegans incorporating the original data set from Rodriguez-Robles and De Jesus-Escobar (2000) and current taxonomy after Pyron and Burbrink (2009) GenBank Accession numbers for our two Pituophis specimens Amphib Reptile Conserv distributed rate heterogeneity (GTR + F) BlackBox (Stamatakis 2006; Stamatakis et al 2008) from the CIPRES Science Gateway (Miller et al 2010) BI was conducted using BEAST (ver 1.8; Drummond and Rambaut 2007) from the UF-HPC Galaxy instance http:// www.geneious.com) Phylogenetic analyses gamma and 1,000 nonparametric bootstrap replicates (Felsenstein 2004) to assess node support using RAxML-HPC sembled and edited as necessary with Geneious software (ver 6.1, created Likeli- hood (ML) and Bayesian Inference (BI) methods ML was conducted with the General Time Reversible model DNA from the automated sequencer (Genomics Division, Interdisciplinary Center Maximum (http://hpc.ufl.edu; al 2005; Goecks Blankenberg et al 2010) et al 2010; Giardine et The Bayesian Information Criterion in jModelTest (ver 2.1.4; Darriba et al 2012; Guindon and Gascuel 2003) determined the best-fit nucleotide substitution model to be Hasegawa, Kishino, and Yano with a proportion of invariant sites and gamma distributed rate heterogeneity 39 (HKY + I + T) A relaxed November 2014 Volume Number e86 | | | Verification of introduced Pituophis ruthveni in Florida CA_SanBeii1fl_LaffipiTope!fe9eliy/a_HWG1485 CARiverside_Artzofiseiega;is_HVZ137665 TX_Culberson_Sogdop/i/s sui5«ci/)0fi5_CMEI 0N_0tl3wa_PaafteropWs Ki/lpiii[;s_CAS184362 Pifyop/jiscfeppe/^ani 99 Meaco_MichDacan_fl'(i/opljis [teppef rfeppa_2 99| deppei P Meaco_Durarigo_filuflp/?(s deppei cteppe/_1 99 GualeTOla_P('t[jlwMS_MVZ150219 NJ_C jmbeHand_Pft:;!)p/jis m »7;e/a/!o/ei/cus_MVZ225520 NJ_Burlinglon_Pituophis_rr_rrelanoleu(!us_MVZ225521 Pifiypp/:is ;ne/a™/e(;c!;s /t>dingf_HWG2651 P^uophk mknolsucus Wng[_HIWG2652 CA_Alameda_Pwwp/;is ca/en/fe; cafefflfer_JAR77 CA_Napa_PtftJopPis cafeiiife^ cafemfer CA_MerdocinD_Pm.^fe cateiffer cafenffer_CAS20l258 NV Mineral PifuopA/s Mte/i;fefdeseriia)Ja_MVZ137577 CAKemPdmphis catew/fef dssert'coteRSR1 15 UT_Ulah_P[i'iropft*s' caimkr desejl'ico/a Mexico_BajaCal_Rli;!)p/)(5 calmer lymans_IO CA_Monterey_Pi7t/(3pfjJS cafe/?ter cafenffe,''_JAR75 Mexico_BajaCal_fl'lrjt)pn(s rafenifer Mgmfus Mexico_BajaCal_RK;op/)(5 cafepder lxmans_l2 Prfuop/iis calender toa;;s_1 CA_SanDiego_Pflw>pfiis ralenife'aifleciefJs_HVZ15CI206 CA_SanDifigo_Pf!uophis t;atenite'3iinecitefls_HVZ1499B3 CA_SantaBaitara_Pfltjopft,ficafeni/efpOT/ius_2& CA_SantaBaitara_Ptopft,isiafe.ri//efpum/lusJ MeBCO_BajaCal_PfW&pP(S i«/telrelis_38 Me)dco_BajaCal_fl'l[jt)pP(s rafemfer !xmans_1 Meaco_BajaCal Rlijoprts i«rteljraI('s_JAR78 Prfvop/iisverfebra/IsJO AZ_Mana)pa_P/f(/(!pf]ys :a!e;!ffer affiffl5_WVZ1 62369 P- AZ_Cochise_P/!(/opf?ls calaiifer 3]^nis_MVZ137697 catenifer CO_6arfield_PJ!'i/op/;fsc3teajferdese.^'coia_UVZ150216 TX_Js1fDavis_P/fuopf}rsc3te,r!/fersayi_MVZ15D218 NM_Ljna_Pikipi)fs c ca?ei'w'fera;i?f]fs_H8S151 MO_SaintLouis_P/luoptecaIer:/'fersay(_MVZ226247 LA,_Bienville_Pt[/opft/s -c s futhveniJ2 a; OK_C!evelard_P(‘f[/op^fs catepifersayi_37 LA,_8ienwille_PfT[/opf]/'s LA,_Bienwille_PrT[/opfi/s I' rutoiJS to CO_Jefferson_R1[jopb(sca!a,7ifer53ji_35 a; FL_MmiDade_UF163092 FL MiamiOade UF157954 Figure Bayesian Inference phylogeny for Pituophis (Squamata: Colubridae) snakes, including the two known P ruthveni (high- UF-Herpetology 157954 and 163092) collected in Miami, Miami-Dade County, Florida Note that values (> 95%) above nodes represent posterior probabilities Inset photograph of UF-Herpetology 163092 by Dustin C Smith lighted in yellow, phylogenetics method was used without relying on a potentially arbitrary molecular clock (Zuckerkandl pothesis with posterior probabilities and FigTree (Drummond et al 2006) An un- correlated lognormal relaxed clock with coalescent con- 95% (Kingman 1982), estimated base stant population size (ver 1.4) The most credible inferences of phylogenetic relationships were confined to nodes where nonparametric bootstrap values > 70% and posterior probability (Pp) was > Pauling 1965) that might incorporate uncertainty in the tree estimation process was created using randomly generated starting tree, and normal distribution for the ucld.mean parameter priors were used Two independent runs were performed consisting of three heated and one cold Markov Chain Monte Carlo (MCMC) estimated for 40 million generations, with every 1,000th sample being retained Both runs were analyzed independently (to confirm chains were converging and not sampling local optima) using Tracer (Hillis and Bull 1993; Felsenstein 2004) frequencies, Morphology and color pattern We determined sex, number of MCMC (ver 1.6) for ESS length, ventrals, subcaudals, supralabials, infralabi- sal scale rows; and color pattern of dorsum and venter compared these data to those found in the literature values >200, as well as for a split stan- dard deviation less than 0.005 for -InL tree values chains that indicate parameter stationarity among Results was achieved —Both MF and BI Trees sampled prior to stationarity were discarded as Phylogenetic analyses bum-in, which occurred prior to five million generations produced identical phylogenetic groupings Trees from both independent MCMC mns were com- bined and burn-in was removed using LogCombiner 1.8), tail preoculars, postoculars, temporals, loreals, and dor- als, We snout-vent length (SVF), the best statistically supported tree clade credibility tree) with using TreeAnnotator mean (ver 1.8), Amphib Reptile Conserv and (Figs Although some of these clades are organized differ- monophyly of Pituocongment with the find- ently in relation to one another the (ver Maximum phis is was obtained ings by Pyron and Burbrink (2009), though (i.e heights 4) methods and a phylogenetic hy- well supported, which is the latter study used only single samples for each species Both of 40 November 2014 Volume Number e86 | | | Krysko our two Pituophis specimens have the same lotype, mtDNAhap- currently uncertain if and both phylogenetic methods place them within the P catenifer sayi Morphological data for P UF -Herpetology 157954 wild tail and heavily patterned 163092 include 212 blotches, parietal stripe present, Data for UF-Herpetology Oscar Rodriguez for collecting specimens; Steve Reich- 27-30-24 dorsal venter scale rows, 57 subcaudals, 8/8 ventrals, postoculars, temporals, 1/1 loreals, 32 body blotches, and David L Reed and Gustav Paulaboratory space; and R Alexander Pyron and ling for providing data; (left/ lay for right) supralabials, 11/11 infralabials, 1/1 preoculars, 7/7 scale rows, — Acknowledgments We are thankful to everyone who assisted in this study: Ryan Zach, Kevin Kopf, and loreals, 34 body blotches, established in the vast suggesting reproduction might have taken place in the in- abials, 11/11 infiralabials, 1/1 preoculars, 7/7 postoculars, 1/1 is cess areas, an adult male and gravid female were found clude 226 ventrals, 55 subcaudals, 8/8 (left/right) supral- temporals, ruthveni protected undeveloped habitats surrounding public ac- ruthveni clade / P et al 27-31-23 dorsal Craig Hassapakis for reviewing this paper 11 tail blotches Literature Cited Discussion Arevalo E, Davis SK, Sites Jr JW 1994 Mitochondrial DNA sequence divergence and phylogenetic relation- ML Our mam and BI phylogenies produced identical phylogenetic groupings (Figs and 4) as those found in the ML analysis by Rodriguez-Robles and De Escobar (2000) However, we found no among chromosome races of the Sceloporus grammicus complex (Plirynosomatidae) in central Mexico Systematic Biology 387^18 Blankenberg D, Von Kuster G, Coraor N, Ananda G, ships Jesus- some provided on the support for eight by Rodriguez-Robles and De JesusEscobar (2000) Our two Pituophis specimens were Mangan M, Nekrutenko A, Taylor J 2010 Galaxy: A web-based genome analysis tool for experimentalists Current Protocols in Mo- placed within a well-supported lecular Biology Chapter relationships, original ML and no support values are Lazarus R, tree P catenifer sayi / P ruth- 10.1 002/047 1 42727 same group of specimens (except for our Florida specimens) uncovered by Rodriguez-Robles and veni clade, the De P combined mtDNA and single nuclear (nDNA) (Pyron and Burbrink 2009) and phenetic moiphological similar(Reichling 1995) analyses Nonetheless, Collins JT 1991 Viewpoint: no molecular support for the recognition as a separate species One of the Rodriguez-Robles and lecular studies gion), is De ment we found tiles of P ruthveni limitations of our Jesus-Escobar ’s (2000) the use of only a single locus for Herpetological Review 22: mo- JT 1991 A arrangerep- 42^3 Field Guide to Reptiles pany, Boston, Massachusetts, re- USA 363 p Darriba D, Taboada GL, Doallo R, Posada D 2012 genes might provide needed data for distinguishing be- jModelTest tween these two named lel taxa Pituophis ruthveni is curit A new taxonomic and Amphibians ofEastern and Central North America Third Edition, Expanded Houghton Mifflin Com- and adding additional mtDNA and unlinked nDNA rently recognized as a separate species because Distributions some North American amphibians and Conant R, Collins and (ND4 and 135-141 10: a doi: A neutral terminology to define ‘invasive’species Diversity ruthveni were also found to be sister taxa based on 19.10.1-21 mb 0s89 Colautti RI, Macisaac HJ 2004 Jesus-Escobar (2000) Pituophis catenifer sayi and ity 19:Unit More models, new heuristics and paral- computing Nature Methods Dorcas occurs 2: ME, 9: Willson JD, Reed RN, 772 Snow RW, Rochford able using color pattern characters, the most diagnostic MR, Miller MA, Meshaka WE et al 2012 Severe ma mm al declines coincide with proliferation of inva- being 28-38 dark dorsal body blotches and the blotches sive obscuring the ground coloration anteriorly (Collins 1991 Proceedings of the National Academy of Sciences of the United States ofAmerica 109: 2418-2422 in allopatric populations and is believed to be diagnos- Reichling 1995; Rodriguez-Robles and De Jesus-Esco- in Everglades National Park Drummond AJ, Ho SYW, Phillips MJ, Rambaut A bar 2000) Our two Pituophis specimens exhibit these we categorized them as P Bumiese pythons 2006 ruthveni Relaxed phylogenetics and dating with confidence Before our specimens were found, Pituophis ruthveni Public Library ofScience Biology A: e88 doi: 10 1371/ three characters, thus was not known species is to be kept at Zoo Miami, journal.pbio 0040088 therefore this Drummond not representative of a zoo-mediated introduc- pathway and was “BEAST: Bayesian by sampling trees.” Biomed AJ, Rambaut A 2007 by an outside per- evolutionary analysis son Other species such as the Reticulated python, Ma- Central Evolutionary Biology tion likely released layopython reticulatus (see Kaiser et al et al 2014), and Pacific Coast giant patterns of the Louisiana pine snake {Pituophis rmth- Stauro- eastern Texas Texas Journal of Science 56: typus salvinii, are other examples of reptile species that ven'i) in on zoo property, the latter possibly established (Smith et al 2011) Although we are 383-394 have been illegally released Amphib Reptile Conserv 214 Ealy MJ, Fleet RR, Rudolph DC 2004 Diel activity 2013; Reynolds musk turtle, 7: Ernst 41 CH, Ernst EM 2003 Snakes of the United States November 2014 Volume Number e86 | | | Verification of introduced Pituophis ruthveni in Florida and Canada Smithsonian Books, Washington D.C., USA and London, United Kingdom 668 p Felsenstein cal sociates, Sunderland, Massachusetts, USA 664 p and taxonomic tini Genome Research 15: 1,451-1,455 Goecks J, Nekmtenko A, Taylor J, The Galaxy Team porting A and reproducible, Am- p stability in the snake tribe Lampropel- (Serpentes: Colubridae) Molecular Phylogenetics and Evolution 52: 524-529 Reichling SB 1995 The Taxonomic status of the Louisiana Pine Snake {Pituophis melanoleucus ruthveni) comprehensive approach for sup- accessible, to Pyron RA, Burbrink FT 2009 Neogene diversification P, 2010 Galaxy: USA 150 Kansas, Zhang Y, Blankenberg D, Albert I, Taylor J, Miller W, Kent WJ, Nekmtenko A 2005 Galaxy: A platform for interactive large-scale genome analysis Shah ED 1998 A Key Jr phibians and Reptiles of the Continental United States and Canada University Press of Kansas, Lawrence, Giardine B, Riemer C, Hardison RC, Burhans R, Elnitski L, 52: 273-288 Powell R, Collins JT, Hooper 2004 Inferring Phylogenies Sinauer As- J Economics and transparent its relevance to the evolutionary species concept Genome Journal of Herpetology 29\ 186-198 Reynolds RG, Niemiller ML, Revell LJ 2014 Toward Guindon S, Gascuel O 2003 A simple, fast and accurate method to estimate large phylogenies by maximum- a Tree-of-Life for the boas and pythons: Multilocus computational research in the Biology 1 : sciences life R86 species-level phylogeny with unprecedented taxon sampling Molecular Phylogenetics and Evolution 71 likelihood Systematic Biology 52: 696-704 DM, Hillis Bull JJ 1993 An empirical test of bootstrap- 201-213 ping as a method for assessing confidence in phylogenetic analysis Systematic Biology 42: 182-192 lecular systematics of CMR, Luiselli L, O’Shea M, Schleip WD, Wiister W 2013 Best Kaiser H, Crother BI, Kelly Ota H, Passes P, practices: In the 21st century, taxonomic decisions herpetology are acceptable only when Rochford MR, Somma EA, Wasilewski JA, Kieckhefer III RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA 1988 Primer-directed enzymatic amplification of DNA with thermostable DNA polymerases Science 239 (4839): 487^91 Strangers CR, CueSmith DC, Gillette The GN, Granatosky MC, vinii Rochford MR, Enge KM, Cueva D 2012 Taxonomic Gillette zotti FJ 68 by Krysko et al IRCF Reptiles & Amphibians Brien ML, Cherkiss MS, Wilkins L, Maz- Dietary habits of the Burmese python Py- ¥\ondSi Herpetological Bulletin 101: 5-7 Stamatakis A 2006 RAxML-VI-HPC: Maximum likeli- hood based phylogenetic analyses with thousands of taxa and mixed models Bioinformatics 22: 2,688- CR, 2,690 Stamatakis A, Hoover corrections of from Florida (2011) Zootaxa 3199: 66- P, Rougemont J 2008 A fast boot- strapping algorithm for the RAxML Web-Servers Systematic Biology 57: 758-771 Stebbins RC 1985 A Field Guide to Western Amphibians Miller New Non-indige- thon molurus bivittatus, in Everglades National Park certain verified non-indigenous reptiles reported Gray 1864 (Kinostemidae), a Snow RW, Agreement 08013 Florida Fish and Wildlife Conservation Commission, Tallahassee, Florida, USA Somma LA, Musk Turtle, Staurotypus sal- 18:55-56 ect 524 p Krysko KL, Pacific Coast Giant nous Species in Florida Report, Proj- in Florida Final in of Nonindigenous Species in Florida Island Press, Washington, D.C., USA 479 p Smith DC, Krysko KL, Sorensen TA, Sider MN 2011 reptiles in Florida phibians and Reptiles DC, Brown TC (Editors) 1997 Paradise: Impact and Management Simberloff D, Schmitz from 1863 through 2010: Outlining the invasion process and identifying invasion pathways and stages Zootaxa 3028: 1-64 Krysko KL, Enge KM, Moler PE 2011b Atlas of Amand and Saiki Nielsen SV 2011a Verified non-indigenous amphibians and bull, Evolution 14: 35-50 in supported by a Stabile JE, World gopher, species complex Molecular Phylogenetics tal Colubridae) Journal of Herpetology 20: 77-79 JP, New pinesnakes {Pituophis: Colubridae), a transcontinen- body of evidence and published via peer-review Herpetological Review 44: 8-23 Kingman JFC 1982 The Coalescent Stochastic Processes and their Applications 13: 235-248 Knight JL 1986 Variation in snout morphology in North American snake Pituophis melanoleucus (Serpentes: Krysko KL, Burgess va D, Enge KM, Mo- Rodriguez-Robles JA, de Jesus-Escobar JM 2000 MA, CIPRES Pfeiffer W, Schwartz T and Reptiles Peterson Field Guide Series Houghton Mifflin Company, Boston, Massachusetts, USA 448 2010 Creating the Science Gateway for inference of large phy- logenetic trees Pp 1-8 In: Proceedings of the Gate- P- way Computing Environments Workshop (GCE), 14 November 2010, New Orleans, Louisiana, USA 115 Stull OG 1929 The description of a new subspecies of Pituophis melanoleucus from Eouisiana Occassional enviromnental and economic costs associated with Papers of the Museum ofZoology, University ofMichigan No 205 Thomas RA, Davis BJ, Culbertson MR 1976 Notes on alien-invasive species in the United States Ecologi- variation and range of the Louisiana pine snake, Pituo- P- Pimentel D, Zuniga R, Morrison D 2005 Update on the Amphib Reptile Conserv 42 November 2014 Volume Number e86 | | | Krysko et al Zuckerkandl E, Pauling L 1965 Evolutionary diver- phis melanoleucus ruthveni Stull (Reptilia, Serpentes, gence and convergence in proteins Pp 97-177 Colubridae) Journal of Herpetology 10: 252-254 Evolving Genes and Proteins Editors, Bryson V, Vo- U.S Fish and Wildlife Service 2013 U.S Fish and Wildlife Service Species Assessment and Listing gel HJ Form Available: http://ecos.fws.gov/docs/candidate/ assessments/20 13/r4/C02C_V01 pdf [Accessed: In: Academic Press, New York, New York, USA 629 p 25 November 2013] Kenneth L Krysko is Museum a senior herpetologist at the Florida of Natural History, University of Florida (UF), where he has helped eurate the international researeh and teaehing colleetions for 15 years He studies ecology more than and molecular systematics on both native and normative herps and produced Florida’s first Atlas of amphibians and reptiles His research on introduced species has made UF-Herpe- tology the information center for invasive species in Florida documenting species introductions as back as 1863 and far He recently published a large illustrated that Florida has the monograph worst invasive problem in the entire world He received his B.S in Biological Sciences from Florida State UniverM.S in Biological Sciences from Florida International University, and Ph.D in Wildlife Ecology and reptile sity, Conservation from the University of Florida Dustin C Smith is a conservation and research specialist at Zoo Miami He graduated with a B.A from the University of South Florida He has been involved with research projects 2001 Panama, on projects in Puerto Rico and Belize Most current research involves native and non-native amphibians and reptiles in southern Florida and throughout the southeastern US, and of his in Costa Rica, in is currently working Michael R Rochford is the Invasive Species Coordinator at the University of Florida’s Fort Lauderdale Research and Education Center, where he has worked for the last eight years He received his B.S in Biology from Kansas State University in 2004 He has worked extensively on radio-telemetry and diet studies of Burmese pythons in Elorida as well as on projects to assess populations of American alligators and American crocodiles In addition to his research and conservation interests, he also has strong interest in finding and photographing herpetofauna of the United States and Mexico Trey “Guy” Kieckhefer graduated with a B.S in Biology from Eckerd College in Saint Petersburg, Elorida He worked for the University of Elorida studying invasive Burmese pythons in the Everglades, and currently works at the Tavernier Science Center in the Elorida Keys for the National Audubon Society He conducts surveys for prey fishes in the Everglades and submerged aquatic vegetation He is interested in Ichthyology, Ornithology, exotic species, and the overall health of the Everglades ecosystem Leroy a P Nunez is a collections assistant within the Division of Herpetology at the Elorida Natural History, University of Elorida (UE) Amphib Reptile Conserv He Ecology at the of obtained an Associate’s Degree in Zoology from Santa Ee College and a Bachelor’s Degree in Biology from UE Interdisciplinary Museum He is currently working School of Natural Resources and Environment on at his Master’s Degree in UF His current research involves studying phytogenies of invasive squamates in Florida and his future research projects will focus on phylogenetics and developmental evolutionary biology 43 November 2014 Volume Number e86 I I I CONTENTS Special Section Alejandro Valencia-Zuleta, Andres Felipe Jaramillo-Martinez, Andrea Echeverry-Bocanegra, Ronald Viafara-Vega, Oscar Hernandez- Cordoba, Victoria E Cardona-Botero, Jaime GutierrezZuNiGA, AND Fernando Castro -Herrera Conservation status of the herpetofauna, proteeted areas, and — eurrent problems in Valle del Cauea, Colombia Alejandro Valencia-Zuleta et al — Supplemental Material SI General Section Gernot Vogel, Patrick David, and Irvan Sidik — On Trimeresurus sumatranus ignation of a neotype and the description of a new (Raffles, 1822), with the des- species of pitviper from Sumatra (Squamata: Viperidae: Crotalinae) Richards-Zawacki, Alexander R Krohn, Matthew Robak, Edgardo J Griffith, Heidi Ross, Brian Gratwicke, Roberto Ibanez, and Jamie Voyles Field surveys in Western Panama indicate populations of Atelopus varius frogs are persisting in regions where Batrachochytrium Rachel Perez, Corinne dendrobatidis Kenneth nez L is L — now enzootic 30 Krysko, Dustin C Smith, Michael R Rochford, Guy — Confirmation of introduced Louisiana pinesnakes, Pituophis N Kieckhefer III, and Leroy ruthveni, in Florida based P Nu- on molecular 36 analyses Back cover Table of Contents Cover: Bothrocophias colombianus, by Jorge Alberto Zuniga Baos and Luis Enrique Vera Perez; Allobates juanii, by Alvaro Velasquez; Bolitoglossa savagei and Podocnemis lewyana, by Juan David Jimenez Bolano; Caecilia thompsoni, by Esteban Alzate; Anolis macrolepis, by Fernando Castro Instructions for Authors: Located at the Amphibian & Reptile Conservation website: http://amphibian-reptile-conservation.org/submissions.html Copyright: © 2014 Craig Hassapakis/Tw/?/?z/?/^i« & Reptile Conservation volume 2014 NUMBER ... December 2014 Volume | | Number e87 | Official journal website: amphibian- reptile-conservation.org 8( 2) Amphibian & Reptiie Conservation [Special Section]: 1- 18; S1-S24 (e87) Conservation status of... I | Number e87 | Official journal website: amphibian- reptile-conservation.org 8( 2) Amphibian & Reptiie Conservation [Special Section]: 1- 18; S1-S24 (e87) Supplemental Material Conservation status... Burrowes, 1 989 ) CR Centrolene buckleyi (Boulenger, 188 2) VU CR, B2ab(iii) la VU VU, Cal, Cai, Tu, B2ab(iii) Tr VU VU CR CR EN, VU VU EN, CR, Alace Espada, 187 2) Centrolene heloderma (Duellman, 1 981 )