Green and Golden Bell Frog Plan of Management

Transcription

1 Green and Golden Bell Frog Plan of Management Arncliffe Prepared for Roads and Maritime Service May 2016

2 DOCUMENT TRACKING Item Detail Project Name Green and Golden Bell Frog Plan of Management Arncliffe Project Number 14SYD-349 Meredith Henderson Project Manager (02) Level Sussex Street Sydney NSW 2000 Prepared by Matthew Dowle Reviewed by Meredith Henderson Approved by Mark Adams Status Final Version Number 17 Last saved on 13 May 2016 Cover photo Green and Golden Bell Frog (Litoria aurea) hiding among reeds (Eco Logical Australia). This report should be cited as Eco Logical Australia Green and Golden Bell Frog Plan of Management - Arncliffe. Prepared for NSW Roads and Maritime Service. ACKNOWLEDGEMENTS This document has been prepared by Eco Logical Australia Pty Ltd with support from Roads and Maritime Services, Dr Arthur White and Prof Michael Mahony. Disclaimer This document may only be used for the purpose for which it was commissioned and in accordance with the contract between Eco Logical Australia Pty Ltd and Roads and Maritime Service. The scope of services was defined in consultation with Roads and Maritime Service, by time and budgetary constraints imposed by the client, and the availability of reports and other data on the subject area. Changes to available information, legislation and schedules are made on an ongoing basis and readers should obtain up to date information. Eco Logical Australia Pty Ltd accepts no liability or responsibility whatsoever for or in respect of any use of or reliance upon this report and its supporting material by any third party. Information provided is not intended to be a substitute for site specific assessment or legal advice in relation to any matter. Unauthorised use of this report in any form is prohibited. Template 08/05/2014 ECO LOGICAL AUSTRALIA PTY LTD ii

3 Contents 1 Introduction Background Purpose of the Plan of Management Objective of plan Legislative context and related documents Threatened Species Conservation Act Environment Protection and Biodiversity Conservation Act Secretary s Environmental Assessment Requirements Guidelines and related documents Green and Golden Bell Frog Description Habitat Species status and distribution Sydney Key Populations Arncliffe population Key threatening processes Impacts, mitigation and management measures Description of impacts Current management Proposed management Project specific mitigation measures Construction mitigation measures Habitat enhancement and management within adjacent habitat including the RTA ponds Habitat creation at Marsh St and the establishment of a captive breeding population Monitoring and Reporting Monitoring implementation of mitigation strategies Monitoring effectiveness of management and mitigation measures during construction Population monitoring during operation Population performance criteria during construction and operation Goals and performance indicators for mitigation measures Contingency measures and corrective actions for declines in the frog population Monitor densities, distribution and habitat use Evidence of consultation with OEH Review of Plan of Management Annual reporting of monitoring ECO LOGICAL AUSTRALIA PTY LTD iii

4 References Appendix A Unanticipated find procedure Appendix B Consultation with OEH Appendix C Hygiene protocol Murray et al Appendix D Consultation with DP&E ECO LOGICAL AUSTRALIA PTY LTD iv

5 List of figures Figure 1: Proposed impact area and Green and Golden Bell Frog habitat... 3 Figure 2: Maximum known number of adults between 2002/03 and 2014/15 at the RTA ponds (White unpubl. data) Figure 3: Frog exclusion fence design Figure 4: Approximate location of frog underpass under M5 East Figure 5: Frog underpass design concept Figure 6: Area proposed for habitat at Marsh Street List of tables Table 1: Proportion of male and female Green and Golden Bell Frogs from 2002/03 to 2014/15 (White unpubl. data) Table 2: Construction mitigation and management measures within the construction zone Table 3: Habitat enhancement measures Table 4: Monitoring requirements to assess effectiveness of mitigation measures Table 5: Population monitoring and corrective actions for Green and Golden Bell Frog at Arncliffe Abbreviations Abbreviation CEMP DotE DP&E ELA EPA Act EPBC Act LEP LGA MNES Description Construction and Environmental Management Plan Commonwealth Department of the Environment NSW Department of Planning and Environment Eco Logical Australia Pty Ltd Environmental Planning and Assessment Act 1979 (NSW) Environmental Protection and Biodiversity Conservation Act 1999 (Commonwealth) Local Environmental Plan Local Government Area Matters of National Environmental Significance ECO LOGICAL AUSTRALIA PTY LTD v

6 OEH Roads and Maritime SEARs TSC Act NSW Office of Environment and heritage NSW Roads and Maritime Services Secretary s Environmental Assessment Requirements Threatened Species Conservation Act 1995 (NSW) ECO LOGICAL AUSTRALIA PTY LTD vi

7 1 Introduction 1.1 Background Roads and Maritime Services (Roads and Maritime) has gained approval to construct and operate the New M5 (the project), which comprises a new, tolled multi-lane road link between the existing M5 East Motorway, east of King Georges Road, and St Peters. The project also includes an interchange at St Peters and connections to the existing road network. The project is declared to be State Significant Infrastructure (SSI) and approval was sought under Part 5.1 of the NSW Environmental Planning and Assessment Act 1979 (EP&A Act). In addition to State approval, the project is a controlled action under the Environment Protection and Biodiversity Conservation Act 1999 (EPBC Act). Construction activities associated with the project would affect habitat of Litoria aurea (Green and Golden Bell Frog) at the Arncliffe surface works area. There are expected to be direct and indirect impacts. Permanent road facilities are proposed on land owned by Roads and Maritime, adjacent to existing purpose built breeding ponds. The impacts to the breeding ponds relate to impacts arising from construction within approximately 32 metres of the ponds and will not arise from direct physical removal of habitat. Direct impacts do involve removal of around 7.82 hectares of foraging, sheltering and dispersal habitat. Green and Golden Bell Frogs at this location form the Arncliffe key population, which is covered in The Green and Golden Bell Frog Key Population of the Lower Cooks River Management Plan (DECC 2008a). The plan addresses threats and issues affecting the conservation of the species in the Lower Cooks River, in accordance with the draft species Recovery Plan (DEC 2005). Green and Golden Bell Frogs have been continuously recorded at the Arncliffe location for over 20 years (White 2015). The species has been breeding and foraging at this site, which contains both suitable terrestrial and aquatic habitat (White 2015). Purpose built breeding ponds for the species are currently located on Roads and Maritime land adjacent to Marsh Street and the Arncliffe surface works area. These breeding ponds are regularly managed through the manipulation of water levels to control vegetation and predators, including the Plague Minnow (Gambusia holbrooki), and Chytrid fungus (a specific frog fungal disease) through salt water flushing. The ponds are known as the RTA ponds and referred to as the RTA ponds hereon. The local population centres on the artificially created habitat at the RTA ponds. The RTA ponds are considered to be the key source for adult frogs for the local population, which disperse across the Kogarah Golf Course. It is unlikely that other ponds within the golf course provide suitable significant breeding habitat as they contain Plague Minnow. However, occasional breeding events in the golf course ponds have been recorded (Dr Arthur White pers. comm 2015). 1.2 Purpose of the Plan of Management This Plan of Management has been prepared to support the Commonwealth and State environmental approvals process. The Plan has also been developed to provide a framework for the construction team to incorporate Green and Golden Bell Frog management actions in their Construction Environmental Management Plans (CEMP), and to ensure that any actions are consistent with relevant Roads and Maritime guidelines and the impact assessment process. In particular, this Plan of Management outlines mitigation and management measures to be implemented prior to construction and in the event the species is found in the construction zone during ECO LOGICAL AUSTRALIA PTY LTD 1

8 the proposed works. The Plan of Management also outlines management measures to enhance habitat adjacent to the RTA ponds within the Kogarah Golf Course for the duration of the construction activities. The project has attempted to avoid potential adverse impacts to the Green and Golden Bell Frog population. There are two main foci for management: provision of new habitat and an insurance population. The Plan of Management sets out strategies to create new artificial habitat on Roads and Maritime land at Marsh Street supported by a captive breeding program. These strategies would be further detailed in a Habitat Creation and Captive Breeding Plan which would be prepared with the advice of independent expert ecologists prior to the commencement of any works in the vicinity of the RTA ponds. This strategy demonstrates that significant effort is being directed toward minimising risk to the population as a result of potential indirect impacts. The Plan of Management has been considered in relation to available management guidelines and policies outlined in Section Objective of plan This plan has the following objectives: Minimise or eliminate all avoidable construction impacts by removing and excluding frogs from the construction zone and implementing strict ongoing construction protocols and exclusions. Compensate for unavoidable construction impacts by augmenting existing foraging habitat Insure against stochastic impacts on RTA ponds by establishing a captive breeding colony and managing non construction related threats known to adversely impact the RTA ponds. At least double the availability of suitable habitat in the vicinity by creating new breeding habitat at Marsh Street wetlands and re-instating habitat within Kogarah Golf Course post construction. Together these objectives are designed to increase the security of the species at Arncliffe which is the ultimate aim of this management plan. Biologically meaningful key performance indicators (KPIs) have been developed to assess and monitor the effectiveness of all actions. These include the responses to not meeting KPIs (thresholds), the personnel responsible and reporting mechanisms. This plan will be subject to expert review by independent experts, with knowledge of the biology and ecology of Green and Golden Bell Frog, as well as the relevant agencies. ECO LOGICAL AUSTRALIA PTY LTD 2

9 Green and Golden Bell Frog Plan of Management Arncliffe M5E frog underpass Figure 1: Proposed impact area and Green and Golden Bell Frog habitat ECO LOGICAL AUSTRALIA PTY LTD 3

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11 2 Legislative context and related documents 2.1 Threatened Species Conservation Act 1995 The Green and Golden Bell Frog is listed as Endangered under the NSW Threatened Species Conservation Act 1995 (TSC Act). The TSC Act requires that a public authority take appropriate measures to implement actions included in a Recovery Plan for which they have agreed to be responsible. In addition, the TSC Act specifies that public authorities must not make decisions that are inconsistent with the provisions of the draft species Recovery Plan (DEC 2005). Construction activities associated with the project, would affect habitat of the Green and Golden Bell Frog. The management of potential habitat and mitigation measures for the project is to be undertaken in a manner consistent with the TSC Act requirements and the draft species Recovery Plan. The project is consistent with the draft Recovery Plan because it provides for fully offset impacts. This is consistent with the objective of no net loss of habitat. 2.2 Environment Protection and Biodiversity Conservation Act 1999 The Green and Golden Bell Frog is listed as Vulnerable under the Commonwealth Environment Protection and Biodiversity Conservation Act 1999 (EPBC Act). The EPBC Act significant impact guidelines (DEWHA 2009) for this species consider that a significant impact is possible if actions result in the removal or degradation of terrestrial habitat within 200 metres of known habitat. The current project footprint proposes to remove known Green and Golden Bell Frog habitat as part of the construction activities for permanent and temporary facilities. The project has been referred to the Commonwealth for approval under the EPBC Act and has been determined to be a controlled action on the basis of impacts to Green and Golden Bell Frog. 2.3 Secretary s Environmental Assessment Requirements The NSW Secretary s Environmental Assessment Requirements (SEARs) for the project were issued on 5 March 2015 and a revised SEARs issued on 26 August 2015 and included a requirement to undertake an assessment of potential impacts of the project on biodiversity values. In addition, matters for further consideration were provided by the NSW Office of Environment and Heritage (OEH), which included specific consideration of the Green and Golden Bell Frog. In anticipation that additional conservation measures are likely to be required for impacts on this species, this plan sets out a targeted and detailed management strategy for this species at this location. 2.4 Guidelines and related documents This Plan of Management has been considered in relation to available management guidelines and policies including: Draft Recovery Plan for the Green and Golden Bell Frog (DEC 2005). Plan of Management Green and Golden Bell Frog Key Population of the Lower Cooks River (DECC 2008a). EPBC Act Policy Statement Significant Impact Guidelines for the vulnerable green and golden bell frog Litoria aurea (DEWHA 2009). Best Practice Green and Golden Bell Frog Habitat Guide (DECC 2008b). Protecting and restoring Green and Golden Bell Frog habitat (DECC 2008c). ECO LOGICAL AUSTRALIA PTY LTD 5

12 Hygiene protocol for the control of disease in frogs (DECC 2008d). Environmental Impact Assessment Guidelines: Green and Golden Bell Frog (NPWS 2003). Species expert reports and annual monitoring at Kogarah Golf Course (White 2015). Biodiversity Guidelines protecting and managing biodiversity on RTA projects (NSW Roads and Traffic Authority 2011). In addition, this plan has been prepared with reference to: The Commonwealth EPBC Act Policy Statement Translocation of listed threatened species assessment under chapter 4 of the EPBC Act (SEWPaC, 2013). Policy and Procedure Statement No 9 Policy for the translocation of threatened fauna in NSW (NPWS, 2001). Approvals required implementing this plan In anticipation that implementation of this plan, including the proposals to undertake habitat creation and captive breeding activities, would be part of the conditions of the planning approval under the EP&A Act, separate TSC Act licensing of these activities would not be required. The program and plan requires Animal Ethics Committee approval according to the Code of practice for the care and use of animals in research in Australia (National Health and Medical Research Council and Commonwealth Scientific and Industrial Research Organisation). ECO LOGICAL AUSTRALIA PTY LTD 6

13 3 Green and Golden Bell Frog 3.1 Description The Green and Golden Bell Frog is a relatively large dull olive to bright emerald green frog that can range in size from around 45 millimetres to 100 millimetres snout to vent length (Cogger 2000, OEH 2015). Its distinctive characteristics are a gold or creamish white stripe running along the side, extending from the upper eyelids almost to the groin, with a narrow dark brown stripe beneath it, from nostril to eye. It also has a blue or bluish-green colour on the inside of the thighs (OEH 2015). The Green and Golden Bell Frog can be distinguished from similar species by its wart-free skin, expanded finger and toe pads, and lack of spotting or marbling on the hind side of the thigh. Tadpoles of the species are relatively large ( millimetres at limb bud development stage) and juvenile frogs are smaller versions of the adults that metamorphose at around millimetres snout to vent length (DEC 2005). It is active by day and usually breeds in summer when conditions are warm. However, the breeding has been recorded from September to February, with a peak breeding period following heavy rains in the warmer January to February months. Breeding patterns are influenced by geography with southerly and higher altitude populations having a narrower window of opportunity for breeding than more northerly and lower altitude populations (DEC 2005, DotE 2015). The species is known to be highly mobile, and may move among breeding sites with large distances travelled in a single day/night or up to kilometres (Pyke and White 2001). Male frogs call while ECO LOGICAL AUSTRALIA PTY LTD 7

14 floating in water and amongst fringing vegetation and females produce a raft of eggs that initially float before settling to the bottom of the water body (DEC 2005). Tadpoles are known to feed on algae and other plant-matter within the water body, while adult frogs are known to eat mainly insects, but may also eat other frogs. 3.2 Habitat Green and Golden Bell Frogs can occupy a broad range of habitats, including natural, artificial and disturbed habitats, and breed in ephemeral ponds (Pyke & White 1996, DEC 2005). They have been recorded associated with coastal swamps, marshes, dune swales, lagoons, lakes and other estuarine wetlands as well as riverine floodplain wetlands and billabongs and constructed water bodies such as storm water detention basins, farm dams, bunded areas, drains and ditches (DEC 2005). Green and Golden Bell Frog need various habitats for different aspects of their life cycle including foraging, breeding, sheltering, over-wintering and dispersal. They will also use different habitats or habitat components on a temporal or seasonal basis (DotE 2015). The species has been found in a wide range of water bodies except fast flowing streams (Pyke & White 1996) and has been associated with habitats such as marshes, dams and stream-sides, particularly those containing Typha spp. (Bullrushes) or Eleocharis spp. (Spikerushes). Breeding habitat consists of water bodies that are still, shallow, ephemeral, unpolluted, unshaded, with aquatic plants present and free of Plague Minnow (Gambusia holbrooki) and other predatory fish. Breeding habitats also occur near terrestrial habitats containing grassy areas and vegetation no taller than woodlands for foraging and dispersal, and a range of diurnal shelter sites, such as rocks, logs, tussock forming vegetation and other cover for refuge (Pyke & White 1996, DotE 2015). 3.3 Species status and distribution The Green and Golden Bell Frog is listed as Endangered under the NSW TSC Act and as Vulnerable under the Commonwealth EPBC Act. The NSW Scientific Committee, when producing the original schedules for listing of the species as endangered stated that: [the] Population [was] severely reduced over entire range; [and] severe threatening processes [operate]. The Green and Golden Bell Frog has declined from a status where it was regarded as an extremely abundant species, with a widespread and almost continuous distribution between the north coast near Brunswick Heads, south along the coast to Victoria, to one where it now has only a fragmented distribution throughout this former range. It is currently considered to be absent from at least 90% of its former distribution (White and Pyke 1996; DEC 2005). The Green and Golden Bell Frog occurs mainly along the coastal lowland areas of eastern NSW and Victoria. Its distribution now extends from Yuraygir National Park near Grafton on the North Coast of NSW, to the vicinity of Lake Wellington, just west of Lakes Entrance in south-eastern Victoria. The furthest inland record of the species is near Hoskinstown in the Southern Tablelands, just outside the ACT (DotE 2015). At the time of the Recovery Plan production, there were 43 populations described as key populations, known or considered likely to persist throughout the species range within NSW. ECO LOGICAL AUSTRALIA PTY LTD 8

15 3.3.1 Sydney Key Populations Sydney still contains some of the largest but also most disturbed and isolated populations of the Green and Golden Bell Frog, as a result of development and other human disturbances rather than a naturally patchy distribution. Eight key populations exist within the greater Sydney Region, with other transient sites believed to also exist, consisting of small populations of migrating individuals. The Sydney based eight key populations are at: Kurnell Homebush Bay (Sydney Olympic Park lands) Greenacre Clyde/Rosehill (wetlands at the confluence of the Parramatta and Duck Rivers) Merrylands (Holroyd Gardens estate) Arncliffe (Marsh Street Wetlands) this location St Marys Hammondville Arncliffe population As part of the original M5 East project opened in 2001, Roads and Maritime provided breeding ponds for the Green and Golden Bell Frog on Roads and Maritime owned land occupied by Kogarah Golf Club in Arncliffe. The new frog habitats are known as the RTA ponds. This site is in the immediate vicinity of planned construction activity for the New M5. The RTA Ponds were purpose built and while they are permanent, they have the capability of being periodically emptied. Water levels are managed in accordance with the plan of management as part of the M5 East approvals and in response to particular triggers. The two ponds were planted with emerging and fringing aquatic vegetation such as Eleocharis sp and Schoenoplectus sp. These species are rushes and sedges which are used by the frogs for basking. The areas surrounding the ponds were planted with native grasses such as Rytidosperma sp. (Wallaby Grass) and a few low shrubs. No trees were planted as these shade the ponds, which is less favourable for the frogs. A boulder field was constructed to provide basking or sheltering habitat. The entire site was fenced with cyclone mesh fencing to prevent access to the ponds. Vegetation is managed by Roads and Maritime with the assistance of a frog and tadpole community group. Formal monitoring of the frogs in the area started in November Monitoring was initially confined to the remaining areas of the Marsh Street wetland and Eve Street wetlands, however, with the construction of the two frog ponds at Arncliffe and the partial loss of the Marsh Street wetland, monitoring focussed almost entirely on the new frog habitats, the Kogarah Golf Course and the remaining portion of the Marsh Street wetland. Monitoring gradually declined then ceased at Marsh Street and Eve Street wetlands due to lack of frogs present. Monitoring has been ongoing with most survey work being carried out during the warmer months of the year (from August to May). Surveys carried out over the summers of and 2000/01 showed a progressive increase in the number of adult Green and Golden Bell Frogs found in the two frog ponds at Arncliffe and a decrease in the number of frogs in the Marsh Street wetlands. A graph showing the results of the monitoring effort between 2002/03 and 2014/15 is shown below (Figure 2). This graph demonstrates presumed recent declines in the size of the population. While no ECO LOGICAL AUSTRALIA PTY LTD 9

16 specific study of the reasons for decline has been undertaken, Dr Arthur White believes this could be due to a range of factors including: excessive plant growth overshadowing of the existing frog ponds more extensive mowing of grassed areas on the Kogarah Golf Course increasing the risk of predation to frogs foraging on the golf course. Roads and Maritime (M5 operations) has undertaken works within the frog ponds over summer 2015/16 which has removed the excessive plant growth in the existing frog ponds Figure 2: Maximum known number of adults between 2002/03 and 2014/15 at the RTA ponds (White unpubl. data) The surveys have estimated the maximum known number of adults based on the Petersen-Lincoln index. Note that this estimate for 2013/14 could not be made due to the low numbers of frogs captured on the two successive nights surveyed in February The zero does not mean there were no frogs present; it was that the estimate could not be reliably performed. No errors or confidence limits were calculated for any of these data. Sex ratios are important to understand in populations of Green and Golden Bell Frog. This is because this species relies on a fast growth rates and rapid maturation to maintain the population (Pickett et al. 2014). These lifecycle attributes increase the importance of breeding events to enable replacement of adults in subsequent generations as adults generally do not live beyond two years (Bower et al. 2014). To enable successful breeding and to improve genetic diversity, there would need to be enough males and females to breed. The table below describes the proportion of females and males tagged per survey season for the area that includes the RTA ponds, Kogarah Golf Course and the Marsh Street wetlands (Table 1). ECO LOGICAL AUSTRALIA PTY LTD 10

17 Population age structures were not provided in any of the RTA pond monitoring reports from White (White ). Age structure is important to understand if there are enough adults of suitable breeding age to sustain the population. Table 1: Proportion of male and female Green and Golden Bell Frogs from 2002/03 to 2014/15 (White unpubl. data) Survey season Female Male Unknown* 2002/ / / / / / / / / / / / / * denotes that the individual captured would have been too young to determine sex. Proportions have been rounded to the nearest whole number. 3.4 Key threatening processes A number of factors associated with direct and indirect consequences of human activity have contributed to the decline of Green and Golden Bell Frogs, including (extracted from DEC 2005): Habitat loss, modification and disturbance. The removal of and disturbances to habitat has occurred across large areas as a result of development, and is considered the most significance key threatening process. This includes the reduction of wetlands and poorly drained coastal flood plain land that formerly constituted prime habitat, which has been drained, in-filled or developed. Habitat fragmentation. This has historically occurred over wide areas as a result of developments or through construction of significant barriers to natural movement (e.g. major roads). In some cases, this has prevented connections within a population, effectively limiting gene flow and dispersal. Predation by introduced fish. Predation occurs on the eggs and tadpoles of frogs by the introduced Plague Minnow (Gambusia holbrooki). Other introduced fish are likely to be the European Carp (Cyprinus carpio) and Gold Fish (Carassius auratus). Disease Chytrid fungus. The Chytrid fungus (Batrachochytrium dendrobatidis) has been implicated in the decline of frogs across the world, and is thought to be a significant contributor to the decline of frogs in Australia. ECO LOGICAL AUSTRALIA PTY LTD 11

18 Water quality and pollutant issues. Developments and other activities occurring within a catchment have consequences for downstream areas and may include altered flow regimes, increased nutrient loads, weed infestation, other contaminants and rubbish. Deteriorating runoff water quality and increased soil erosion and sedimentation reduces the area s suitability for frogs. Other threats. Other possible threats indicated by anecdotal evidence includes predation by the foxes, cats, dogs and rats, road mortality, mowing near breeding and foraging habitat, predator/prey interactions with Cane Toads, artificial and natural opening of coastal lagoon estuaries, changes to flow regimes of streams and associated wetlands and excessive grazing and trampling of habitat. ECO LOGICAL AUSTRALIA PTY LTD 12

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20 4 Impacts, mitigation and management measures 4.1 Description of impacts Construction activities of the project are likely to result in direct and indirect impacts to Green and Golden Bell Frog habitat. The activities of the project include permanent facilities to be established on land adjacent to the existing RTA ponds. However, a construction area and temporary facilities are also proposed around 32 metres from the RTA ponds, which would extend onto land owned by Rockdale City Council (Figure 1). This construction area is known as the Arncliffe surface works area. Unless mitigated, the proposed temporary and permanent activities are likely to impact on Green and Golden Bell Frogs within the site area, shown on Figure 1, resulting in: Potential direct impacts: Removal of around 7.82 hectares of foraging, sheltering and dispersal habitat Potential mortality of frogs from heavy machinery movements within the construction zone Removal of one ephemeral pond that has previously been recorded as containing a breeding event. Potential indirect impacts: To the RTA ponds (leading to reduction in the capacity of the ponds to function as habitat) by: o Increase in shading by construction exclusion fence o Increase in shading from the permanent facilities during winter o Increase in dust from heavy vehicle movements o Increase in noise by heavy vehicle movements and tunnel boring o Increase in light from 24 hour construction operation o Increase in vibration from heavy vehicle movements, tunnel boring and stockpiling o Reduction of water quality through sedimentation and contaminants originating from construction zone o Accidental introduction of predatory fish o Introduction of frog pathogen by construction personnel and construction equipment and machinery. A reduction of habitat connectivity to other areas within the golf course Limiting foraging habitat Reduction in breeding success Potential mortality of individuals as a result of habitat modification. Temporary impacts for up to four years represent the bulk of the impacts to Green and Golden Bell Frog habitat at the Arncliffe surface works area. The habitat temporarily affected will be reinstated, to the current levels of habitat, at the completion of the project. 4.2 Current management Current management of Green and Golden Bell Frog habitat within the RTA ponds includes regular manipulation of water levels and drainage of breeding ponds. This creates habitat preferred for breeding by the frog and enables the flushing of salt water and periods of dryness for the management of: Predators, including Plague Minnow (Gambusia holbrooki) ECO LOGICAL AUSTRALIA PTY LTD 14

21 Chytrid fungus, a disease that affects all frog species, including the Green and Golden Bell Frog, which is present within this population, although it has not been tested since Ponds on the Kogarah Golf Course are also artificial, but are not purpose built for frog habitat and water levels are not manipulated to manage threats to the frogs or to provide suitable habitat. Many of the golf course ponds contain fringing vegetation such as Typha spp. (Cumbungi) and Juncus spp., suitable for a diversity of frog species, including the Green and Golden Bell Frog. Water quality in many of the golf course ponds is low as a result of nutrient run-off from management activities on the golf course along with saline influences. In addition, adjacent vegetation exists as exotic grasses that are regularly mown for the purposes of golf course maintenance and use as fairways. Habitat types within the golf course are described as follows (extracted from Management Plan Green and Golden Bell Frogs Lower Cooks River Key Populations, DECC 2008a): Breeding habitat: primarily consists of the purpose built artificial ponds (RTA ponds). These ponds were built as a requirement of a previous development approval. Other breeding habitat (ephemeral) also exists and includes golf course water hazards, although breeding in these ponds is occasional. Foraging habitat: Includes grassed areas (native or exotic), tussock vegetation and emergent sedges and reeds bordering water features and ponds. The drainage channel and reed beds that border the southern extremity of the golf course may also provide foraging and dispersal habitat. Sheltering habitat: includes similar vegetation to that used as foraging areas that contain rock piles, fallen timber, tussock grasses and other artificial sheltering sites. Sheltering habitat is present around the RTA ponds. Dispersal habitat: typically includes wet areas such as creek lines, drains, stormwater canals, connecting vegetation, and other easements and depressions. However, in the golf course, fairways currently provide movement habitat between the RTA ponds and foraging habitat. An artificial frog passage was built underneath the M5 to facilitate movement between the golf course and habitat to the west and south (Marsh Street Wetlands and Old Spring Creek Wetland site). However, this passage is not used because of the relatively hostile environment across which frogs would need to travel (White A., pers. comm. 2015). The underpass consisted of a circular culvert 60 centimetres in diameter that ran the entire length of the passage. The total length of the culvert was 38 metres. Near the mid-way point, the culvert sloped upwards and an open skylight was installed in the space in the median strip in the motorway. Travelling southwards from the centre section, the culvert dipped quickly downwards so that it could pass under the southern section of the M5 East and emerged at the edge of the stormwater basin that had formed the Marsh Street wetland. The centre skylight was the only light point in the culvert. The northern entrance to the culvert was only accessible by maintaining a cleared vegetation area on the southern side of the SWSOOS. The frog use of the underpass was monitored for the first two years only. It became clear that the frogs were not using the underpass. Either the underpass was too narrow, too dark or too long for the frogs to use. This did not mean that frogs did not move between the northern and southern areas either side of the M5 East. Occasionally Bell frogs were captured beneath the M5 near the cycleway underpass. This area is 300 meters to the east of the frog culvert and much more exposed. Few frogs were found making the passage beneath the M5. The impoverished condition of the Marsh Street wetland after the installation of the M5 resulted in few Bell frogs remaining in this area (and hence few frogs moving beneath the M5). Frog monitoring since 2000 failed to detect any Bell frogs in the Marsh Street wetland after ECO LOGICAL AUSTRALIA PTY LTD 15

22 Over-wintering habitat: Boulder piles were constructed surrounding the RTA ponds to provide over-wintering habitat. However it is unclear whether the frogs actually use the boulders as over-wintering habitat. 4.3 Proposed management Roads and Maritime is seeking to manage impacts to Green and Golden Bell Frog habitat. The actions include the implementation of mitigation and management measures, and enhancement of habitat. These are to be delivered by those undertaking the construction works and Roads and Maritime. This plan along with the Habitat Creation and Captive Breeding Plan, required by this plan, is to be used by the successful tender of the project for detailed design and construction. These plans are to be implemented as part of the works through a Construction Environmental Management Plan (CEMP). The mitigation and management measures outlined in Section 4.4 of this Plan of Management must be implemented to minimise potential impacts to Green and Golden Bell Frog individuals and known habitat. The framework for implementation of the project specific mitigation and management actions are to be provided in the CEMP and should require all elements associated with the design, construction, cost and responsibilities. 4.4 Project specific mitigation measures All measures are to be incorporated in a CEMP. They are to follow an adaptive approach that seeks for continued improvement of this Plan of Management and its mitigation measures. The actions and measures are outlined in the tables below: Construction related activities within the construction zone (Section 4.4.1) Habitat enhancement and management within adjacent habitat including the RTA ponds (Section 4.4.2) Habitat creation at Marsh Street and captive breeding (Section 4.4.3) Construction mitigation measures Management measures relating to construction activities within the construction zone are outlined in Table 2. These measures should be considered as a minimum requirement and would be the responsibility of the construction environmental manager. More specific construction measures to address impacts on frogs are contained within the CEMP, including induction ECO LOGICAL AUSTRALIA PTY LTD 16

23 Table 2: Construction mitigation and management measures within the construction zone Mitigation measure Description Who Timing Clear delineation of the construction boundary. Areas to be cleared should be marked and checked with surveyor s pegs and equipment to ensure that the minimum area of take is adopted. Define the construction clearing areas Clearing should only occur within these areas. Once areas are cleared, the area of take should be calculated to ensure that no additional areas have been cleared. Contractor - CDS Project ecologist Pre-construction The distance between the RTA ponds and the edge of the clearing required for the construction zone is expected to be at least 32 metres. Establishment of a physical barrier, using frog exclusion fencing between all construction works, existing RTA Ponds and reminder of the golf course. This frog fencing should be designed in consultation with a person who has had at least five years experience in the management of Green and Golden Bell Frogs. Frog-exclusion fences have been used routinely on construction sites and other hazardous areas where threatened frog species occur. Frog exclusion fences consists of a continuous curtain of impervious material (usually shade cloth fabric) strung between support posts. The fence is at least 1 metre high and has an overhanging lip. The current fence design has a 25 cm horizontal lip and then another 30 hanging vertical lip at the top of the fence. The overhanging lip is designed to prevent frogs Establish a Frog exclusion zone climbing over the fence (see Figure 3) The base of the fence is buried (to stop frogs from digging under the fence. If there are gates in the fence, the base section of the gate can still maintain a seal with the ground by being weighed down with length of flexible chain. Contractor - CDS Project ecologist Pre-construction There should be a section of fence directly adjacent to the RTA ponds (marked in Figure 1 as noise wall fence and yellow dashes) which will: Reduce sound and dust Not exclude daylight Exclude frogs Exclude construction activities to clearly separate frog habitat to be retained from construction zone. This section of fence is to be inspected daily. Any breaches of the fence are to be raised with the ECO LOGICAL AUSTRALIA PTY LTD 17

24 Mitigation measure Description Who Timing Contractor for remediation. The remainder of the construction zone should be fenced to clearly separate frog habitat from the construction zone (marked in Figure 1 as frog exclusion fence and black dots). The frog noise wall will be constructed between the RTA pond site and the adjoining works compound. The wall is intended to reduce the amount of noise and dust that might otherwise reach the RTA ponds. To be effective the wall needs to be 4 metres high and 28 metres long. Because the wall is so high and is located on the NW side of the RTA ponds, it risks overshadowing the ponds. To alleviate this, the top 2 m of the wall will be constructed of transparent plastic. The noise wall will be continuous with perimeter frog fences and will form part of the barrier between the RTA ponds and the frog habitat in the Frog Enhancement Area and the works site. This fence should: Exclude humans from entering the construction zone Exclude frogs from the construction zone. The remaining section of fence is to be inspected weekly. Any breaches of the fence are to be raised with the Contractor for remediation. Conduct a pre-clearance survey within the construction zone immediately prior to constructions works being undertaken. An ecologist with a minimum of five years experience in the management of frogs is to conduct the pre-clearing survey. The survey should include two diurnal and two nocturnal surveys, with the last nocturnal survey Undertake pre-clearance survey and salvage activities conducted the night prior to works being undertaken. Winter to spring frog encounters: If Green and Golden Bell Frogs are encountered sheltering underneath rock, rubble or wood they need Contractor - CDS Project ecologist Pre-construction and construction to be assessed for an over wintering position or torpor. Then the frogs are to be collected in accordance with the following protocol: Placed in a clean, plastic holding container with a small amount of purified water Frogs should be micro-chipped if not already tagged Adult frogs should be sexed, snout-vent length measured, weight recorded, condition of the frog, date and location of collection ECO LOGICAL AUSTRALIA PTY LTD 18

25 Mitigation measure Description Who Timing Transported to a suitable over-winter location in consultation with the project ecologist and based on the advice of an independent expert If frogs are injured, they are to be taken to a vet or suitably experienced frog keeper and euthanased. If frogs are not in torpor, the procedure for spring to autumn encounters applies. Spring to autumn frog encounters: If active frogs are encountered during the pre-clearance surveys or daily checks, then they are to be collected in accordance with the following protocol: Placed in a clean, plastic holding container with a small amount of purified water Frogs should be microchipped if not already tagged Adult frogs should be sexed, snout-vent length measured, weight recorded, condition of the frog, date and location of collection Relocated to Symbio Zoo, the artificial habitat created at Marsh Street, or the RTA ponds based on the advice of the project ecologist Prior to works commencing, a number of water bodies within the construction zone will need to be decommissioned. Dam decommissioning needs to be done in the presence of a suitably qualified and experienced ecologist. Any frogs encountered will need to be collected as per above. Pre-clearance surveys should include searching for tadpoles. Tadpoles should be netted and then identified using Anstis (2013). Green and Golden Bell Frog tadpoles should be collected and handled as above, apart from tagging. Tadpoles encountered between autumn and spring should not be released, but should be kept in a suitable over-wintering facility. Tadpoles encountered from spring to autumn should be released into the RTA Ponds, artificial habitat at Marsh Street or relocated to Symbio, based on the advice of the project ecologist. If tadpoles are not Green and Golden Bell Frogs, these should be released into ponds other than the RTA Ponds on the golf course. Site inductions Site inductions should contain a relevant section on the Green and Golden Bell Frog. The induction should incorporate: What to do in the event of unexpected finds of frogs within the construction zone. All persons working in the construction zone Construction ECO LOGICAL AUSTRALIA PTY LTD 19

26 Mitigation measure Description Who Timing Highlighting the enhanced frog habitat area and why this is a no-go zone. Construction Environmental Manager All persons working Stop work procedure Implement a stop work or unanticipated find procedure for when Green and Golden Bell Frogs are observed within the construction zone (Appendix A). The procedure will include a process to notify the construction environmental manager and suitably qualified ecologist, a relocation procedure and when it is okay to re-commence works. in the construction zone Construction Environmental Manager Construction Sediment and erosion control Establish appropriate sediment and erosion control to prevent silt, sediments, spills and other contaminants from reducing water quality in frog habitat. These controls should be regularly inspected, particularly after heavy rain events. Contractor - CDS Construction Environmental Manager Pre-construction and Construction Light spill management Directional lighting should be used in the vicinity of the transparent frog exclusion fence. Directional lighting should aim to reduce night time light spill onto the RTA ponds. Contractor - CDS Construction Dust from heavy vehicle haulage, dumping and storing of spoil and general vehicle movements will Dust suppression need to be minimised. Dust may reduce water quality in the RTA ponds. Bulk water carriers and sprayers should apply town water only to reduce dust. Slurry run-off should be managed in accordance with the sediment and erosion control measures. Contractor - CDS Construction Contaminated lands management Develop appropriate procedures to manage contaminated fill that may occur in surrounding soils during the construction works and any habitat enhancement, if applicable. Contractor - CDS Construction Develop appropriate procedures to manage acid sulphate soils during construction and operation, if applicable. Management of acid sulphate soils should be carried out in accordance with the Roads and Acid sulphate soils management Maritime guideline or approved procedure. Relevant documents include: RTA Guidelines for the Management of Acid Sulphate Materials, April 2005 NSW Acid Sulphate Soils Manual (Stone et al 1998) NSW EPA publication Assessing and Managing Acid Sulphate Soils. Contractor - CDS Construction ECO LOGICAL AUSTRALIA PTY LTD 20

27 Mitigation measure Description Who Timing Use of herbicides and other chemicals Herbicides should not be used near the RTA ponds and within the enhanced frog habitat area. If herbicides are to be used within the construction zone, spray drift must not be able to reach aquatic habitat. This applies to herbicides in solution in surface water run-off. Contractor - CDS Construction Re-instate all habitats that was temporarily impacted from the construction activities within the Arncliffe Habitat re-instatement surface works area. Habitat reinstatement should be conducted in accordance with relevant guidelines and policies and be conducted in consultation with the Kogarah Golf Course and the Rockdale City Council. Contractor - CDS Post construction ECO LOGICAL AUSTRALIA PTY LTD 21

28 Figure 3: Frog exclusion fence design ECO LOGICAL AUSTRALIA PTY LTD 22

29 4.4.2 Habitat enhancement and management within adjacent habitat including the RTA ponds To compensate for loss of foraging habitat and to encourage frog movement to other areas within the Kogarah Golf Course, habitat adjacent to the RTA ponds and between the existing M5 East Motorway and the construction zone will be enhanced. These measures are shown in Table 3. These measures should be considered as a minimum requirement. The detailed design and construction of the habitat management works, associated costs and implementation should be outlined in the CEMP. The implementation of the measures will be the responsibility of the construction environmental manager. Table 3: Habitat enhancement measures Mitigation measure Description Where Who Timing Enhanced frog habitat will consist of the following: Enhance habitat adjacent to RTA frog ponds Improved areas of foraging habitat consisting of tussocky grasslands and swales Areas of vegetation and other structures, such as logs, suitable for sheltering At least three wet areas that will act as stepping stones to encourage frogs to move between the RTA ponds and the remainder of the golf course. Establishment and enhancement of frog habitat is to be conducted in accordance with: Adjacent habitat Contractor - CDS Project ecologist Frog expert Construction Best Practice Green and Golden Bell Frog Habitat Guide (DECC 2008b). Protecting and restoring Green and Golden Bell Frog habitat (DECC 2008c). To be designed by person with at least five years experience in the design of frog fencing or by a frog expert. Hygiene protocol Develop a hygiene protocol for persons working outside the construction zone and within the Kogarah Golf Course. This is to reduce the risk of the introduction and spread of Chytrid Fungus. The hygiene protocol is to be developed in accordance with the following: Adjacent habitat Any entering enhanced habitat person the frog Pre-construction and maintenance ECO LOGICAL AUSTRALIA PTY LTD 23

30 Mitigation measure Description Where Who Timing Hygiene protocol for the control of disease in frogs (DECC Any person 2008d). Environmental Impact Assessment Guidelines: Green responsible for and Golden Bell Frog (NPWS 2003). daily checks of frog RTA Biodiversity Guidelines protecting and managing biodiversity on RTA projects (NSW Roads and Traffic Authority 2011). fencing The hygiene protocol should follow Murray et al (2011), which is appended to this plan. The RTA Ponds would continue to be managed by Roads and Maritime in accordance with the M5 East approval. The Plan of Management (Biosphere 2000) outlines the management actions required but would include maintaining the RTA ponds and enhancing the frog habitat zone. This includes: Maintain the existing RTA ponds Water supply systems Water level management Salt water supply Water quality monitoring Water supply: Develop sustainable water supply plans to supply fresh water for the existing breeding ponds and any enhanced frog habitat ponds. Considerations to include stormwater harvesting, reuse from the groundwater treatment plant and groundwater bores. RTA ponds and adjacent habitat Roads and Maritime Project ecologist (CDS) Frog expert Construction Water level management: the ponds were constructed with sumps and drainage valves. If the ponds needed to be emptied, the valve is manually raised and water drains into an open drain along the northern boundary of the site. Ponds are drained when it is deemed that the ponds are either becoming too overgrown with emergent vegetation or the pond has become infested with fish. If excess vegetation is the reason, the pond is drained in winter and the excess vegetation manually harvested and removed. The pond is then refilled in early spring. ECO LOGICAL AUSTRALIA PTY LTD 24

31 Mitigation measure Description Where Who Timing Salt water supply: Develop arrangements for regular (six monthly) saline flushing of breeding and sheltering ponds for Chytrid fungus control. Maintenance of the surrounding vegetation: The area immediately surrounding the ponds is to be maintained as low sedges, to a distance of 0.5 to 1 metres from the top of the pond. Beyond this a mowed grass strip is to be maintained. The grass strip is 1-2m metres wide. Outside of the mowed strip is a zone of taller shrubs and grasses that extends to the boundary fence. These vegetation zones are maintained through voluntary working bees (usually by Frog and Tadpole Study Group volunteers). Large works, such as tree removal and large area weed removal is done by contractors engaged by Roads and Maritime Services. Use of herbicides: Herbicides are not to be used near the RTA ponds or areas of aquatic habitat or in such a way that could impact upon aquatic habitat. Water quality: The value of water quality (WQ) monitoring is uncertain. The RTA Ponds were only monitored between 2000 and 2005 for basic parameters such as turbidity, dissolved oxygen content, oxidationreduction potential, ph, temperature, conductivity and salinity. The two independent reviewers as well as Dr White indicated that although WQ could be monitored, the benchmarks for WQ are not clear for this species. The approach that has been suggested is to drain ponds of water and refill. This is the action that is currently taken at the RTA Ponds. However the triggers are not necessarily related to WQ, but rather other thresholds. These thresholds would include: Presence of Plague Minnow Reduction in frog health attributable to chytrid Increase in shading as a result of vegetation colonisation and growth in the ponds. All of these thresholds are included in the monitoring program at the RTA ECO LOGICAL AUSTRALIA PTY LTD 25

32 Mitigation measure Description Where Who Timing Ponds. The presence of these threats triggers a response, which ranges from flushing with saline water to emptying the ponds and refilling. This management will continue at the RTA Ponds. Develop measures to reduce threats of Chytrid, Plague Minnow, noxious weeds and predation by feral cats and foxes. Ponds will be drained or flooded as required. Draining of the ponds is observed at the RTA ponds to be an effective method to manage Plague Minnow which predate tadpoles. Ponds would be periodically drained and flooded as part of the management of these ponds in response to observations of Plague Minnow. Fences are to be maintained to exclude vertebrate mammal predators Control threats such as foxes and cats. The frog exclusion fencing would also need to be maintained to ensure that the fence retains integrity. Presence of predatory fish should be monitored and if present, the ponds should be drained to kill these fish. RTA ponds and adjacent habitat Contractor - CDS Project Ecologist Construction Annual or bi-annual salt flushing would be required to minimise the prevalence of chytrid. Salt flushing has worked well at the RTA ponds. Caution should be applied when salt loading into ponds is proposed because the threshold was found to be close to the concentrations required to limit chytrid growth and infection. A study has found that exposure to salt concentrations of 5 parts per trillion (ppt) and chytrid infection had a cumulative effect on Litoria peronii (Stockwell et al. 2012). Salt flushing should therefore not exceed 5 ppt. Frog under pass near cycleway The proposed frog movement corridor from the new Frog Habitat Area at Marsh Street to the cycleway underpass, through the underpass and on to the golf course and RTA ponds involves traversing two "pinch points". The cycleway underpass is heavily used by cyclists and pedestrians and the frog crossing must be physically separated from the From RTA Ponds under M5 East adjacent to cycleway to west of the M5 East Contractor - CDS Project Ecologist Construction and post-marsh Street habitat creation ECO LOGICAL AUSTRALIA PTY LTD 26

33 Mitigation measure Description Where Who Timing cycleway/pedestrian footpath. The proposed frog corridor would parallel with the footpath (but be separated from the footpath by a frog fence) from Marsh Street to the underpass. At the underpass, the frog corridor will pass under the M5 on the western side of the oil trap pond (the footpath passes on the eastern side of this pond). Up to this point it is possible to completely isolate the frog corridor from the cycleway footpath (Figure 4). Once the frog corridor emerges on the northern side of the underpass, it needs to cross the cycleway (and the SWSOOS) before it can continue again away from the cycleway. A low level frog underpass beneath the cycle path would need to be constructed (Figure 5). This would consist of constructing an open box culvert 2 metres wide and 60 centimetres high. The culvert would span the cycle path and the top of the culvert would be covered with a reinforced mesh. As frogs are only likely to use this underpass at night the underpass will not need to be lit. Small open basins would be constructed on the northern side of the frog underpass and one would be constructed under the M5 East at the southern end of the underpass to encourage frog movement to the entrance and exit points of the underpass. Once frogs have made this short passage under the cycleway, they reenter a fence corridor that leads then to a ramp that gets them over the SWSOOS and then to the main east-west thoroughfare beside the golf course. This design separates the frogs from cyclists and pedestrians and requires only a short underground (although it will be open to the night sky) passage. Frog tunnels of this type work overseas but are untested in Australia (where frogs have a reluctance to use enclosed culverts or enclosed ECO LOGICAL AUSTRALIA PTY LTD 27

34 Mitigation measure Description Where Who Timing tunnels). By having an open roof and only a short tunnel, there is more likelihood that the frogs will make the crossing. ECO LOGICAL AUSTRALIA PTY LTD 28

35 Figure 4: Approximate location of frog underpass under M5 East Figure 5: Frog underpass design concept ECO LOGICAL AUSTRALIA PTY LTD 29

36 4.4.3 Habitat creation at Marsh St and the establishment of a captive breeding population New Green and Golden Bell Frog habitat comprising at least three new ponds will be created and managed in perpetuity for the species on Roads and Maritime land at Marsh Street. See Figure 6. A habitat creation and captive breeding plan was prepared which: Detailed the results of further population survey to be undertaken from October 2015 to February The purpose of this survey was to improve the understanding of the population structure in the RTA ponds and adjacent habitat including the number, age, breeding status and sex ratio of the population. Detailed how a captive population would be established including the objectives of the program, details of the suitable conservation facility to host the animals, husbandry techniques, welfare protocols, hygiene protocols, collection and transportation protocols, duration of the program and final release proposals. Detailed how the new habitat will be prepared, populated and managed in perpetuity at Marsh Street. This included arrangements to secure a suitable water supply to the facility, fencing and site security protocols, weed removal and site remediation. New breeding habitat would involve the construction of at least three ponds capable of being managed as frog habitat over the long term Detailed the long-term management framework to apply to the lands, which would include exploring the option of entering into a BioBanking agreement over the land. Detailed the long term monitoring protocols, which would be established to address biologically meaningful KPIs. The breeding program adopts a leading practice approach to captive husbandry, disease control and maintenance of the genetic diversity of the population. The objective of the captive breeding program is to provide a level of security, especially for such a small population. For example, outcrossing with other populations would be considered if this would improve genetic and survival for the frogs. The habitat creation and captive breeding plan was reviewed by two independent frog experts and their advice was considered in the finalisation of the plan. The plan was prepared prior to the commencement of construction activities within the vicinity of the RTA ponds. ECO LOGICAL AUSTRALIA PTY LTD 30

37 Figure 6: Area proposed for habitat at Marsh Street ECO LOGICAL AUSTRALIA PTY LTD 31

38 (blank page) ECO LOGICAL AUSTRALIA PTY LTD 32

39 5 Monitoring and Reporting 5.1 Monitoring implementation of mitigation strategies A report detailing the implementation of the actions set out in Table 2 and Table 3 of this plan will be prepared by the contractor on a quarterly basis during construction. This report must be provided to OEH and DP&E. The following is an outline of the adaptive monitoring program that will be undertaken to assess the effectiveness of the construction and operational mitigation measures for the Arncliffe population of the Green and Golden Bell Frog Monitoring effectiveness of management and mitigation measures during construction A number of mitigation measures would be implemented to minimise indirect impacts to the Green and Golden Bell Frog Population at Arncliffe. These measures will be monitored to determine their effectiveness in managing these impacts. Table 4 outlines the monitoring program to assess the effectiveness of the mitigation measures during the construction period. ECO LOGICAL AUSTRALIA PTY LTD 33

40 Table 4: Monitoring requirements to assess effectiveness of mitigation measures Mitigation measure to be assessed Element monitored Where monitored Frequency of monitoring Who monitors Threshold trigger Action response Frog exclusion fence Fence integrity Entire extent of frog exclusion zone at the RTA ponds and construction zone, Arncliffe Weekly Project ecologist Damage to fence Fence breach Fix fence within 24 hours of detection of damage Remedial action to be carried out by CDS contractor Fix holes and / or damage to fence Remove predators via appropriate Control non-aquatic predator threat Presence of vertebrate predators using sand pads Within RTA ponds and up to four locations outside the construction zone Weekly Project ecologist Detect predator footprints in sand pads method that limits harm to bell frogs If predators other than fox and cats are detected, other control measures would need to be considered. This must be done in consultation with OEH and Roads and Maritime. Sediment and erosion control Sediment and erosion fencing integrity RTA Ponds and construction zone and wherever control fences are located at the Arncliffe works area Weekly and following heavy rain events (i.e. over 20 mm within 24 hour period) Construction environmental manager Fence or control measures breached by runoff Fix management measures (i.e. fence or bales) within 24 hours of detection of breach Remedial action to be carried out by CDS contractor Light spill control onto RTA ponds Light reaching RTA ponds at night from construction zone lamps RTA Ponds At first lighting of lamps and then during population monitoring (i.e. September to March) Construction environmental manager Project ecologist Light reaching pond from construction zone lamps Adjust directional lighting away from pond ECO LOGICAL AUSTRALIA PTY LTD 34

41 Mitigation measure to be assessed Element monitored Where monitored Frequency of monitoring Who monitors Threshold trigger Action response Presence of Gambusia holbrooki via direct visual assessment RTA Ponds Monthly Project ecologist OR appointed frog ecologist Presence of Gambusia fish in RTA ponds Empty ponds and refill with suitable water supply Maintain RTA ponds Presence of chytrid RTA ponds During frog surveys (i.e. September to March) Project ecologist OR appointed frog ecologist Presence of lesions, evidence of declining frog health attributable to chytrid Salt water flushing of RTA ponds to a maximum of 5 ppt Remedial action to be undertaken by CDS Noticeable presence or increase in dust RTA Ponds and exclusion zone Weekly Project ecologist The opinion of the project ecologist is that the level of dust may be affecting GGBF Review dust suppression measures with Construction Environmental Manager Increase frequency of spraying with town water supply ECO LOGICAL AUSTRALIA PTY LTD 35

42 5.1.2 Population monitoring during operation Population monitoring will be put in place to monitor any impacts of the project on frogs within the RTA ponds and within adjacent habitat during the construction and operation of the project. The monitoring aims to assess the ongoing survival of the Arncliffe population at the Kogarah Golf Course and within the RTA ponds. The population monitoring must occur at the RTA Ponds and the Kogarah Golf Course for at least three generations from completion of site rehabilitation. It is anticipated that the site would be rehabilitated by late March That is, from the time that the temporary works area is rehabilitated for a period of six to nine years or longer. This will be dependent on the successful breeding and survival of the frogs as generation length would potentially vary depending on favourable conditions. The frog survey methodology will be based on Pollock s robust design (Pollock 1982). This design would allow the most rigorous population estimates to be made as well as provide data on emigration/immigration, population dynamics and turnover. This method relies on repeated sampling in sites in quick succession (to decrease the impact of immigration or emigration). It is also suitable for an enclosed population. The survey area would be divided into zones (RTA ponds, enhancement area and golf course east, golf course north areas, new frog underpass and cycleway near Eve Street wetland). Animals would be captured during nocturnal surveys, marked and released within each survey zone. This needs to be done until recapture rates are sufficiently high and a set threshold level of 20 percent recaptures within each secondary survey period and preferably closer to percent. All precincts would need to be sampled in this way. This may mean surveying at one wetland or precinct for longer than six nights to achieve the recapture rates required. All captured Green and Golden Bell Frogs would be measured, weighed, sexed and inspected for reproductive condition and signs of illness or injury. Frogs larger than 40 millimetres snout-vent length would be micro-chipped and tissue samples will be taken by web punching between the third and fourth hind toe. The frogs would then be released. At the end of each period, a population estimate using MARK would be calculated. A suitably qualified and experienced statistician would analyse this data. This is to ensure that the analysis is conducted by a person who understands the design requirements for the survey and also the species ecology. Population monitoring surveys will be conducted monthly between August and April each season; in addition opportunistic surveys will be conducted when weather conditions are optimal for frog population estimates and frog recaptures. Table 5 provides further details about monitoring procedures. Pollock s robust design and the use of MARK have been recommended by the independent scientific reviewers of this plan. Monitoring can commence at the new Marsh Street habitat once it has been created and frogs are introduced to this habitat area. Monitoring design would follow that of the RTA Pond; the Pollock s robust design to be adopted. Monitoring of other suitable frog habitat in the vicinity of the Marsh Street frog habitat area should be considered and discussed with the expert frog ecologist. ECO LOGICAL AUSTRALIA PTY LTD 36

43 Table 5: Population monitoring and corrective actions for Green and Golden Bell Frog at Arncliffe What monitored How monitored Where Who monitors Trigger threshold Remedial action Establish Robust Design Pollock s Mark-recapture frogs in Green and Golden Bell Frog population size RTA Ponds, habitat enhancement area and Golf Course east Analyse data using MARK Animals would be captured during nocturnal surveys, marked and released within each survey zone. This needs to be done until recapture rates are sufficiently high and a set threshold level of 20 percent recaptures within each secondary survey period and preferably closer to percent. All precincts would need to be sampled in this way. This may mean surveying at one wetland or precinct for longer than six nights to achieve the recapture rates required. RTA Ponds Habitat enhancement area Kogarah Golf Course east Cycleway underpass New Frog Habitat at Marsh Street (once established) Other suitable frog habitat areas as discussed with expert frog ecologist Project ecologist OR appointed frog ecologist If in the opinion of the expert the situation is declining significantly as population estimate is already low The Arncliffe population is declining despite good and favourable conditions at other bell frog sites in the Sydney Basin (and that those other bell frog populations are not in decline) Assess whether food supply is sufficient; if not, introduce or increase food supply via compost bins Assess whether predation by Gambusia or other vertebrate pests is occurring; remediate as per Table 4 Assess whether decline is due to chytrid via standard chytrid assessment methods; remediate as per Table 4 Consider further habitat enhancement Introduce stock from captive breeding program Initiate translocation to Marsh Street frog habitat area Monitoring should be ECO LOGICAL AUSTRALIA PTY LTD 37

44 What monitored How monitored Where Who monitors Trigger threshold Remedial action conducted each month between August and April. The RTA ponds and golf course ponds would be Tadpole presence to indicate successful breeding sampled each month between September and March each year using long-handled sampling nets for tadpoles. Any tadpoles captured will be measured, staged (using standard staging in Gosner (1960)), identified (using Anstis 2013) and released. RTA ponds Remaining Kogarah Golf Course Ponds Project ecologist OR appointed frog ecologist No tadpoles present Assess whether predation by Gambusia is occurring; remediate as per Table 4 Release tadpoles with hind limb buds from the captive colony into RTA ponds ECO LOGICAL AUSTRALIA PTY LTD 38

45 5.1.3 Population performance criteria during construction and operation Table 5 outlines the performance criteria against which the ongoing survival of the Arncliffe population at the Kogarah Golf Course would be measured, details of contingency measures and corrective actions that would be implemented in the event of reductions in population numbers. The performance criteria for the population at Arncliffe consist of two foci: That the population size estimate does not decline significantly (in the opinion of the frog expert) beyond the current population size That there is successful annual breeding during spring-summer for each year the works are in place Goals and performance indicators for mitigation measures Table 4 outlines the performance indicators for the mitigation measures. For each measure, the method of monitoring, the timing or frequency, the responsibility has also been presented. The table includes remedial actions and the trigger thresholds for each measure. The goals for the mitigation measures are: The frog exclusion fence is to be intact during the entire period of construction works Non-aquatic predators are not present in the frog habitat area or the RTA ponds Sediment and erosion has been controlled so that it does not impact on the RTA ponds, the enhanced habitat area or any frog habitat area outside the construction zone Light spill onto RTA ponds is from the construction zone is controlled so that the ponds are not impacted by construction zone light during the night The RTA ponds are maintained to manage the presence of Gambusia holbrooki to the extent they are not present The RTA ponds are maintained to manage the presence of chytrid in the Green and Golden Bell Frog population That dust from the construction zone does not impact the RTA ponds No frog mortality on the M5 East cycleway Contingency measures and corrective actions for declines in the frog population Table 5 outlines the corrective actions to be taken in the event that the population declines beyond the current size estimate. An insurance population would be established as a captive colony as outlined in the Habitat Creation and Captive Breeding Plan. The release of progeny from the captive population would be predicated on an assessment of the capacity of the RTA ponds and surrounding areas to accommodate these frogs. A number of factors would need to be monitored and managed prior to the release of any captive bred individuals. These factors include the abundance of food, the presence of non-aquatic vertebrate predators, the presence of aquatic predators, a decline in health of the frogs as an indicator of chytrid Monitor densities, distribution and habitat use The monitoring would be designed to estimate the population size, use of habitat and movement of frogs at the RTA ponds and the Kogarah Golf Course. The Pollock s Robust design must be used to determine a population estimate (size) and the data will be interrogated using MARK. The precincts for survey are the RTA ponds and habitat enhancement area, golf course east and golf course north. These precincts may need to be altered based on the advice of the expert frog ecologist. ECO LOGICAL AUSTRALIA PTY LTD 39

46 Distribution and therefore habitat areas in which the frogs are found would be recorded. For example if frogs are found in ponds these would be sheltering or breeding habitat. Habitat use would therefore be based on the number of Green and Golden Bell frogs that utilise each area, how often each habitat area contains Bell frogs and whether Bell frogs are present in particular habitat areas over successive monitoring periods. 5.2 Evidence of consultation with OEH This plan of management has been reviewed by OEH. Ongoing discussions with OEH will be required throughout the project. Evidence can be found at Appendix B. 5.3 Review of Plan of Management This plan of management must be reviewed annually. The review must be conducted after the population monitoring has been completed and after population size at the RTA ponds and Kogarah Golf Course has been estimated. The annual review must be carried out by the agencies (OEH, DP&E and Roads and Maritime) with input from an independent frog expert. Information included in the review would be all of the monitoring results, details of any corrective actions, details of any action or activity carried out to increase the security of the population at Arncliffe. Where information of monitoring of other artificial Green and Golden Bell Frog habitats is publicly available, this information should be included in the annual report and revisions of the Plan of Management. The plan must be amended if the review indicates that elements of the plan require updating. These should reflect the effectiveness of mitigation and the ongoing survival of the population at Arncliffe. 5.4 Annual reporting of monitoring Results of the monitoring, review and any amendments must be reported annually to the Secretary of the Department of Planning and Environment and provided to OEH. This should be done after the monitoring results have informed the next iteration of the plan; the plan has been reviewed and amended. The monitoring results should be included on the Roads and Maritime website. ECO LOGICAL AUSTRALIA PTY LTD 40

47 (blank page) ECO LOGICAL AUSTRALIA PTY LTD 41

48 References Anstis, M Tadpoles and frogs Australia. New Holland Publishers, Chatswood, NSW. Biosphere Consulting Plan of Management for the RTA Ponds and Frog Habitat Area, Arncliffe. Report to the NSW Roads and Traffic Authority. Cogger, H.G Reptiles and Amphibians of Australia - 6th edition. Sydney, NSW: Reed New Holland. Cumberland Ecology Green and Golden Bell Frog Management Plan for Cooks Cove. A report prepared for Boyd Cook Cove. Department of the Environment Species Profile and Threats Database, Department of Environment, Canberra. Available Department of the Environment, Water, Heritage and the Arts (DEWHA) Significant impact guidelines for the vulnerable Green and Golden Bell Frog (Litoria aurea). Department of Sustainability, Environment, Water, Population and Communities (SEWPaC) EPBC Act Policy Statement - Translocation of listed threatened species assessment under chapter 4 of the EPBC Act. Gosner, K.L A simplified table for staging anuran embryos and larvae with notes on identification. Herpetologica, 16, NSW Department of Environment and Conservation (DEC) Green and Golden Bell Frog Litoria aurea (Lesson 1829) Recovery Plan. Draft Recovery Plan. NSW Department of Environment and Climate Change (NSW) (DECC) 2008a. Management Plan for the Green and Golden Bell Frog Key Population of the Lower Cooks River. Department of Environment and Climate Change (NSW), Sydney. NSW Department of Environment and Climate Change (DECC) 2008b. Best practice guidelines Green and Golden Bell Frog habitat. NSW Department of Environment and Climate Change (DECC) 2008c. Protecting and restoring Green and Golden Bell Frog habitat. NSW Department of Environment and Climate Change (DECC) 2008d. Hygiene protocol for the control of disease in frogs. Information Circular Number 6. DECC (NSW), Sydney South. NSW National Parks and Wildlife Service (NPWS) Policy and Procedure Statement No 9 policy for the translocation of threatened fauna in NSW. NSW National Parks and Wildlife Service (NPWS) Environmental impact assessment guidelines: Green and Golden Bell Frog Litoria aurea (Lesson 1829). Available online Office of Environment and Heritage 2014d. Infection of frogs by amphibian chytrid causing the disease chytridiomycosis - key threatening process listing. Available: ECO LOGICAL AUSTRALIA PTY LTD 42

49 [Accessed January 2014]. Office of Environment and Heritage (OEH), Threatened Species Profile Database (online). Available: Pollock, K. H A capture recapture design robust to unequal probability of capture. Journal of Wildlife Management, 46, Pyke, G.H. & A.W. White, Habitat requirements for the Green and Golden Bell Frog Litoria aurea (Anura:Hylidae). Australian Zoologist. 30(2): Pyke, G.H. & A.W. White A review of the biology of the green and golden bell frog Litoria aurea. Australian Zoologist. 31(4): NSW Roads and Traffic Authority (RTA) Guidelines for the Management of Acid Sulfate Materials: Acid Sulfate Soils, Acid Sulfate Rock and Monosulfidic Black Ooze. Available online NSW Roads and Traffic Authority (RTA) Biodiversity Guidelines: Protecting and managing biodiversity on RTA projects. Available online Stockwell, M.P., Clulow, J. and Mahony, M.J Sodium Chloride Inhibits the Growth and Infective Capacity of the Amphibian Chytrid Fungus and Increases Host Survival Rates. PLoS ONE 7, e doi: /journal.pone Stone, Y., Ahern, C. R., and Blunden, B Acid Sulfate Soils Manual Acid Sulfate Soil Management Advisory Committee, Wollongbar, NSW. White, A Management Plan for the Green and Golden Bell Frogs (Litoria aurea) at Arncliffe. Report prepared for the NSW Roads and Traffic Authority. White, A Monitoring reports for Green and Golden Bell Frog at Arncliffe. Reports to NSW Roads and Maritime Services. White, A Arncliffe Green and Golden Bell Frogs. Advice prepared for NSW Roads and Maritime Services. ECO LOGICAL AUSTRALIA PTY LTD 43

50 Appendix A Unanticipated find procedure Step One: Frogs observed durting course of construction Step Two: STOP WORK IMMEDIATELY in the vicinity of the sighting and notify your supervisor. Try to photgraph the frog to assist in confirmaiton of species. DO NOT recommence work until directed by the approved Environmental Represenative. Step Three: Supervisor to immediately notify the Environmental Manager Step Four: Environmental Manager to inform: o Project Director o Construction Manager o Environmental Represenative (ER)(engaged CDS) o Roads and Maritime Environmental Representative o Project Ecologist (PE)(engaged by CDS, with direct expereince in Green and Golden Bell Frog) Step Five: CDS Project Ecologist or CDS Environmental Representative place frog in re-sealable plastic bag or clean holding container. Pour approximately 5 10 mm bottled spring water into bag or container. PE or ER to transport container / bg to site office and commence assessing the health of the frog. Details to be taken (e.g. lifecycle stage, sex, locaiton and date where found, tag number, weight, snout-vent-length). Frog to be microchipped if not already tagged. Step Six: If frogs are detected between winter and spring, they should be checked for over-winter activity. If frogs are in torpor, they must not be released. Over-wintering frogs should be retained in an approved over-winter facility or be included in the captive breeding colony (Symbio Zoo). If frogs are detected between spring and autumn, they should be released by the PE in the RTA Ponds or the Marsh Street habitat area (if this area is available and suitable). Step Seven: PE and ER declare works okay to re-commence. ECO LOGICAL AUSTRALIA PTY LTD 44

51 Appendix B Consultation with OEH The following is a tabulated account of consultation with OEH during the process of updating the Plan of Management. Consultation details Initial phone call with Ray Giddins from OEH to explain process and timeframes received from Ray Giddins outlining initial points for consideration at meeting planned for 20/4/16 Date and time 19/4/ pm 19/4/ pm Meeting held with Ray Giddins (OEH) and Meredith Henderson (ELA) to discuss updates to GGBF plan and expectations regarding amendments 20/4/ am Meredith Henderson provided first draft of updated Plan of Management to SMC and Ray Giddins for comment Ray Giddins provided comment on updated Plan of Management (see note below) Meredith Henderson issued final Plan of Management with OEH and SMC comments incorporated 20/4/ pm 20/4/ pm 22/4/16 Ray Giddins provided the following feedback on the updated Plan of Management in the form of an on 20/4/ pm: 1. Added a row to Table 3. Maintain RTA ponds Noticeable presence increase dust or in RTA ponds and exclusion zone Weekly Project ecologist The opinion of the project ecologist is that the level of dust may be affecting GGBF Review dust suppression measures with. 2. Corrected typographical errors in sections 5.1 and 5.2. RMS accepted all of the changes made by Ray Giddins. Upon reviewing his changes on the following draft, Ray Giddings stated in an on 22/4/16 at 11.38am that All of the additions we ve discussed this week are in the document to my satisfaction. ECO LOGICAL AUSTRALIA PTY LTD 45

52 Appendix C Hygiene protocol Murray et al 2011 ECO LOGICAL AUSTRALIA PTY LTD 46

53 A REPORT FOR THE AUSTRALIAN GOVERNMENT DEPARTMENT OF SUSTAINABILITY, ENVIRONMENT, WATER, POPULATION AND COMMUNITIES Hygiene protocols for the control of diseases in Australian frogs June 2011 Prepared by: 1 Kris Murray, 1 Lee Skerratt, 2 Gerry Marantelli, 1 Lee Berger, 3 David Hunter, 4 Michael Mahony and 5 Harry Hines Author Affiliations 1 School of Public Health, Tropical Medicine and Rehabilitation Sciences, James Cook University, Queensland 2 Amphibian Research Centre, PO Box 1365 Pearcedale, Victoria Environment Protection and Regulation, New South Wales Office of Environment and Heritage 4 School of Environmental and Life Sciences, The University of Newcastle, NSW 5 Ecological Assessment Unit, Conservation Management Branch, Queensland Parks and Wildlife Service Hygiene protocols for the control of diseases in Australian frogs June

54 The authors gratefully acknowledge: Chris Banks, Dr John Clulow, Dr Graeme Gillespie, Professor Rick Speare and Russel Traher,for their contributions to components of the original project resulting in the production of this final report. Cover photo: Taudactylus eungellensis Eungella day frog. K. Murray Commonwealth of Australia (2011). The views and opinions expressed in this publication are those of the authors and do not necessarily reflect those of the Australian Government or the Minister for Sustainability, Environment, Water, Population and Communities. While reasonable efforts have been made to ensure that the contents of this publication are factually correct, the Commonwealth does not accept responsibility for the accuracy or completeness of the contents, and shall not be liable for any loss or damage that may be occasioned directly or indirectly through the use of, or reliance on, the contents of this publication. This work is copyright. You may download, display, print and reproduce this material in unaltered form only (retaining this notice) for your personal, non-commercial use or use within your organisation. Apart from any use as permitted under the Copyright Act 1968, all other rights are reserved. Requests and enquiries concerning reproduction and rights should be addressed to Department of Sustainability, Environment, Water, Populations and Communities, Public Affairs, GPO Box 787 Canberra ACT 2601 or Funding for this project (Procurement Reference Number: ) was provided by the Australian Government Department for Sustainability, Environment, Water, Population and Communities. This project progresses the implementation of the Threat abatement plan for infection of amphibians with chytrid fungus resulting in chytridiomycosis (Commonwealth of Australia, 2006). This report should be cited as: Murray, K., Skerratt, L., Marantelli, G., Berger, L., Hunter, D., Mahony, M. and Hines, H Hygiene protocols for the control of diseases in Australian frogs. A report for the Australian Government Department of Sustainability, Environment, Water, Population and Communities. Hygiene protocols for the control of diseases in Australian frogs June

55 Table of Contents 1. Who should use this document? Objectives Introduction Key disease issues in amphibian populations Fungi Batrachochytrium dendrobatidis Mucor amphiborium Oomycetes Viruses Bacteria Myxozoa Mesomycetozoa Alveolates Zoonotic Diseases Salmonella Leptospira Spirometra erinacei National and border biosecurity World Organisation for Animal Health (OIE) AUSVETPLAN and AQUAVETPLAN Key Threatening Process and Threat Abatement Plan (TAP) Biosecurity Australia Hygiene management In-situ (site) hygiene management Defining a site Determining the order of visitation of multiple field sites On-site hygiene Principles of cleaning and disinfection Handling of frogs in the field Housing frogs and tadpoles Marking, invasive and surgical procedures Sealing wounds Equipment Return of captive animals to the wild Displaced frogs Cane toads Sick and dead animals Hygiene protocol checklist and field kit Important Australian contacts Sick and dead frogs References Hygiene protocols for the control of diseases in Australian frogs June

56 Hygiene protocols for the control of diseases in Australian frogs 1. Who should use this document? This protocol is intended for use nationally by conservation agencies, zoos, scientific research staff, industry organisations (e.g., the pet industry), wildlife consultants, fauna surveyors, students, frog keepers, wildlife rescue and carer groups, frog interest groups/societies and other key interest groups who regularly deal with or are likely to encounter frogs. This protocol outlines the expectations of the Department of Sustainability, Environment, Water, Population and Communities (DSEWPaC) regarding precautionary procedures to be employed when working with frogs in Australia. The protocols were developed in collaboration with recognised experts in the fields of wildlife health, husbandry, research and conservation. The intention is to promote implementation of hygiene procedures by all individuals working with Australian amphibians. DSEWPaC recognises that some variation from the protocol may be appropriate for particular research and frog handling activities. Such variation should accompany any licence applications or renewals submitted to the relevant regulatory bodies for independent consideration. Variations should follow a risk analysis process which broadly involves hazard identification, risk assessment, risk management and risk communication. Where ex-situ activities are proposed, these guidelines should be used in conjunction with the Guidelines for captive breeding, raising and restocking programs for Australian frogs, which can be found here: 2. Objectives The objectives of the hygiene protocols are to: Improve the control of diseases in Australian frogs Improve preparedness for an emergency response to new amphibian disease incursions in Australia Recommend best-practice procedures for personnel, researchers, consultants and other frog enthusiasts or individuals who handle frogs Suggest workable strategies for those regularly working or considering working in the field with frogs or where frogs may exist Provide background information and guidance to people who provide advice or supervise frog related activities Inform regulatory bodies and animal care and ethics committees for their consideration when granting permit approvals Hygiene protocols for the control of diseases in Australian frogs June

57 3. Introduction Amphibians have declined globally. In the first global amphibian assessment, at least 43% of amphibian species with sufficient data were found to have declined in recent decades, 34 species were extinct and a further 88 were possibly extinct (Stuart et al. 2004). In 2010, approximately 30% of amphibians were threatened globally ( Diseases are responsible for many amphibian declines and extinctions and their risk needs to be addressed. Laurance et al. (1996) first proposed the epidemic disease hypothesis to account for Australian amphibian declines. Shortly after, an unknown chytridiomycete fungus was seen infecting the skin of sick and dying frogs collected from montane rain-forests in Queensland and Panama during mass mortality events associated with significant population declines (Berger et al. 1998; Longcore et al. 1999). The fungus was subsequently found to be highly pathogenic to amphibians in laboratory trials by inducing development of skin pathology, morbidity and mortality similar to that seen in the wild frogs. The disease was called chytridiomycosis and the fungus described as a new species Batrachochytrium dendrobatidis (Bd), also known as the amphibian chytrid fungus. Bd has been found infecting over 350 species in two amphibian orders (Anura and Caudata) from all continents where amphibians occur ( Sixty-three (~28%) of Australia s 223 (as listed by IUCN 2008) amphibian species are now known to be wild hosts for Bd (Murray et al. 2010a; Murray et al. 2010b), and over half of Australia s species may be naturally susceptible to Bd in the wild (Murray et al. 2011; Murray and Skerratt in press). While the discovery of chytridiomycosis has sparked renewed appreciation for the role that diseases can play in threatening wildlife populations and species, it is not the only disease currently affecting amphibians, nor is it likely to be the last. Ranavirus, for example, has been observed to induce mass mortality events in frog and salamander populations in the UK and North America. In response to these global threats, the World Organisation for Animal Health (OIE) has listed both chytridiomycosis and ranavirus as notifiable diseases to help control their spread. Similarly, numerous conferences and reports have been assembled to produce standards in managing diseases in wild and captive amphibian populations. Together, these measures highlight the importance of developing agreed hygiene protocols for the control of diseases in Australian frogs. This document fulfils this role. 4. Key disease issues in amphibian populations Here we review the most significant diseases of amphibians, including some that have zoonotic potential and some that have not been detected in Australia. There are many described diseases of amphibians but only a few are known to be an important threat to wild amphibians or other taxa including humans. Some become an issue in captive amphibian populations where management is inadequate. As research on this topic is limited, there are also likely to be many unknown diseases of amphibians which may pose a risk. Disinfection methods have not been validated for all pathogens. Any risk management strategy to minimise the impact of diseases of amphibians should take into account this uncertainty. For detailed reviews see Hemingway et al. (2009) and Berger et al (2009) for diseases in wild populations and Wright and Whitaker (2001) that also includes diseases in captivity. Hygiene protocols for the control of diseases in Australian frogs June

58 4.1. Fungi Batrachochytrium dendrobatidis Batrachochytrium dendrobatidis (Bd) is a fungal pathogen capable of driving amphibian species to perilously low numbers or extinction. In Australia, the oldest record of Bd is from a museum frog specimen collected in south-east Queensland near Brisbane in 1978 (Department of the Environment and Heritage 2006a), which coincides with sudden frog declines in a number of species and two species extinctions in the region (Berger et al. 1998; Hines et al. 1999). Subsequent amphibian declines in central coastal Queensland ( ) and the Wet Tropics ( ) suggest that B. dendrobatidis spread north to its current northern limit at Big Tableland near Cooktown (Laurance et al. 1996; Berger et al. 1999; Skerratt et al. 2010). In southern Australia, the spread of B. dendrobatidis is poorly documented but its distribution extends down the entire east coast to Tasmania (first detected in 2004) (Obendorf and Dalton 2006; Pauza and Driessen 2008). Two separate foci occur in other states, one in southwest Western Australia, where the earliest record dates to 1985, and another around Adelaide in South Australia (earliest record 1995) (Murray et al. 2010a). The Northern Territory is currently considered amphibian chytrid free (Skerratt et al. 2008; Skerratt et al. 2010; Murray et al. 2011). In the majority of infected animals for most of the time, clinical signs of chytridiomycosis are absent. The period of showing signs is typically short and mostly limited to those amphibians that die. Central nervous system signs predominate, including behavioural change, slow and uncoordinated movement, abnormal sitting posture, tetanic spasms, loss of righting reflex and paralysis. Skin changes associated with chytridiomycosis are typically microscopic and not detectable at the clinical level with any degree of confidence, although abnormal skin shedding occurs (skin shed more frequently and in smaller amounts) and erythema (tissue reddening) of ventral surfaces and digits may be seen. For what to do if you encounter a sick or dead amphibian in Australia, see section 6.7. below. For a detailed factsheet about chytridiomycosis, see the Australian Wildlife Health Network website ( Mucor amphiborium This fungus is an important cause of morbidity and mortality in platypus in Tasmania and amphibians are a potential reservoir host (Gust et al. 2009). Amphibian mucormycosis is a systemic disease caused by the fungus, Mucor amphibiorum. Severely infected amphibians have fungi disseminated through their internal organs and skin. The fungi incite formation of granulomas that consist of inflammatory cells and fibrous tissue. At postmortem, the liver contains small pale nodules up to about 5 mm in diameter and usually in massive numbers. These nodules can also be seen in other organs such as the kidney, lung, mesentery, urinary bladder, subcutaneous sinuses and skin. The microscopic fungi are found inside these nodules. M. amphibiorum is a primary pathogen and can infect normal amphibians, but in the wild it appears to cause only sporadic infections. Possibly the usual inoculating dose in the wild is not high enough to cause epidemic disease. In captivity it can cause fatal outbreaks in collections. For more information on mucormycosis, see Hygiene protocols for the control of diseases in Australian frogs June

59 Oomycetes Water moulds (family Saprolegniaceae, phylum Oomycota) are ubiquitous in surface water. High levels of infection with Saprolegnia ferax caused mortality of Western toad (Bufo boreas) egg masses in northwestern United States and were sufficient to affect local populations (Kiesecker et al. 2001). Epidemics may be associated with fish stocking or environmental cofactors Viruses There are a number of viruses that are known to cause disease and mortality in amphibians, including ranaviruses, frog erythrocytic virus, Lucké tumor herpesvirus, herpes-like virus of skin, calicivirus and leucocyte viruses (Hemingway et al. 2009). In Europe and America the most important of these for their ability to cause mass mortalities and potentially population declines are the ranaviruses (Hyatt et al. 2000). Ranaviruses have been identified in a range of ectothermic vertebrates, including fish, amphibians (frogs, toads, salamanders) and reptiles (lizards, turtles, snakes). Some species can infect a broad host range across all these taxa. Ranaviral disease is an emerging infectious disease overseas as it is being detected over an increasing geographic range and in more species (Hemingway et al. 2009). While ranaviral disease in wild amphibians has not been frequently observed in Australia, antibodies to ranaviruses have been detected widely (NSW, Qld, NT) in cane toads (Bufo marinus) (Zupanovic et al. 1998). Bohle iridoviris (BIV) was first found causing death in wild caught metamorphs of Limnodynastes ornatus and has since been detected in wild, moribund adult Litoria caerulea from Townsville and captive juvenile Pseudophryne coriacea from Sydney (Speare et al. 2001; Cullen and Owens 2002). Laboratory studies in Australia have also shown that cane toads (Bufo marinus) and a range of native frogs are susceptible to BIV (Speare et al. 2001). Tadpoles appear the most susceptible, while juvenile frogs were more susceptible than adults. Data on the geographical origin and time of emergence or introduction of ranaviruses in Australia is not known. Ranaviruses not currently found in Australia can cause disease in native Australian amphibians in experimental challenges; for example, Venezuelan Guatopo virus was able to kill Litoria caerulea in experimental trials ( We need to prevent the introduction of pathogenic ranaviruses into Australia. Clinical signs of acute ranaviral disease may be seen in tadpoles, metamorphs, juveniles and adults. In general, amphibians infected with ranavirus may show decreased activity, ascites (accumulation of fluid in the peritoneal cavity), anasarca (accumulation of serous fluid in various tissues and cavities of the body), skin ulceration, focal and systemic haemorrhages and death. For what to do if you encounter a sick or dead amphibian in Australia, see section 6.7. below. For a detailed factsheet about ranaviral disease, see the Australian Wildlife Health Network website ( Bacteria The range of bacteria reported as causing disease in amphibians is small. Bacterial septicaemia can cause significant disease in captivity. Infection with Aeromonas spp., nonhaemolytic group B Streptococcus, Flavobacteria and chlamydia have caused outbreaks in captive amphibians and Mycobacteria can cause chronic problems. Another group of bacteria can be carried by amphibians with minimal effect and are potentially capable of causing Hygiene protocols for the control of diseases in Australian frogs June

60 infections in humans (zoonotic diseases). Salmonella and Leptospira are in this category and are a potential risk to humans, livestock and domestic pets, see below Myxozoa Myxosporean parasites (Myxidium spp.) in the brain and liver of declining Australian frogs, the Green and Golden Bell frog (Litoria aurea) and the Southern Bell frog (Litoria raniformis), have recently been reported to be associated with disease and may have a significant impact on wild frogs (Hartigan et al. 2011) Mesomycetozoa Ichthyophonus sp. occurs the USA where it is often an incidental finding in tadpoles, frogs and salamanders but may cause morbidity and mortality. It infects muscles and adult frogs with massive infections become lethargic and emaciated. Massive acute lethal infections with numerous mortalities occur infrequently in ranid larvae (D. Green, unpubl., Mikaelian et al. 2000) 4.6. Alveolates A Perkinsus-like organism is a major cause of mortality events in tadpoles in the US. Occurs predominantly in tadpoles of Rana spp. and may cause mortality rates of 80-99% in a pond over the course of 2-6 weeks (Davis et al. 2007). Weakly swimming, bloated and floating tadpoles are found Zoonotic Diseases Guidelines for preventing human exposure to amphibian disease are available at the Centre for Disease Control website Salmonella Amphibians may carry pathogenic Salmonella species, but rarely show signs of disease (Anver and Pond 1984). Prevalence of salmonellas isolated in clinically normal amphibians is generally greater than 10% and bacterial levels can be high (Sharma et al. 1974). In Australia, Salmonella were isolated from 12.7% (19/150) of B. marinus collected from the wild and 9 serotypes were identified. All nine had previously been isolated in Australia from humans and livestock (O'Shea et al. 1990). An outbreak of gastroenteritis in humans near Rockhampton possibly originated from green tree frogs (Litoria caerulea) contaminating drinking water in rainwater tanks (Taylor et al. 2000). Some strains of salmonellae are cosmopolitan while others are not found in Australia, but could be imported Leptospira Leptospira are spirochaetal bacteria that usually invade the kidney of vertebrates and are excreted in the urine. Humans and domestic animals are susceptible to various strains of Leptospira usually from the species Leptospira interrogans. Serious acute and chronic disease occasionally with death can result. Little is known about the occurrence of Leptospira in amphibians, and on their significance as reservoir hosts for leptospirosis in humans. No studies appear to have been done on leptospires in amphibians in Australia. However in Hygiene protocols for the control of diseases in Australian frogs June

61 Barbados, toads (Bufo marinus) and frogs (Eleutherodactylus johnstonei) were found to be reservoirs for serovars of Leptospira pathogenic to humans (Gravekamp 1991) Spirometra erinacei The adult stage of the tape worm Spirometra erinacei inhabits the small intestine of carnivores such as the cat, dog, fox and dingo. The first larval stage occurs in copepods and the second larval stage (spargana) are long, flat white worms that can infect amphibians and other vertebrates in muscles and under the skin. Sparganosis occurs in around 5% of Australian frogs and heavy burdens are associated with severe disease (Berger et al. 2009). Sparganosis is a public health problem in Asia, usually occurring as subcutaneous or intramuscular infections. Humans become infected by drinking water with infected copepods, eating undercooked frogs, and the worms can also migrate from frog flesh into skin wounds 5. National and border biosecurity Unregulated trade in animals, as well as unintentional shipment, is suspected to have been a major contributor to the spread of emerging infectious diseases such as chytridiomycosis (Skerratt et al. 2007). There are numerous bodies and regulatory levels that attempt to provide guidance about how to minimise the risk of pathogen spread and transmission in amphibians World Organisation for Animal Health (OIE) The World Organisation for Animal Health (OIE) lists key diseases as notifiable to promote the reporting and management of diseases among member countries. Preventing the spread of amphibian diseases across international borders is important, and both chytridiomycosis (Article 8.1.1) and ranavirus (Article 8.2.1:) are now listed as notifiable diseases in the OIE Aquatic Animal Health Code ( To access these codes, follow these links: Chytridiomycosis: Ranavirus: The codes outline recommendations for the Importation or transit of aquatic animals and aquatic animal products for any purpose from a country, zone or compartment : Provided commodities are treated in a manner that inactivates the disease agent (Bd or ranaviruses), Competent Authorities should not require any disease conditions when authorising the above activities, regardless of the disease status of the exporting country However, in cases where it could otherwise reasonably be expected that commodities pose a risk of Bd or ranavirus transmission, a risk assessment should be carried out in accordance with the recommendations in the Aquatic Code. The exporting country would then be notified of the outcome of the risk assessment before trade commences.. Where commodities do not meet this condition and/or a reasonable risk remains, there are additional requirements that depend on the disease status of the country, zone or compartment. Hygiene protocols for the control of diseases in Australian frogs June

62 Freedom from disease: Importation of live aquatic animals from a country, zone or compartment declared free from disease (Bd or ranavirus) requires an international aquatic animal health certificate issued by the Competent Authority confirming disease free status. A country may make a self declaration of freedom from disease (Bd or ranaviruses) if one of the following conditions is met: 1. It has no amphibians or other susceptible species AND basic biosecurity conditions have been continuously met for a period of 2 years 2. There has been no observed occurrence of the disease for at least the past 10 years despite conditions that are conducive to its clinical expression AND basic biosecurity conditions have been continuously met for a period of 10 years 3. Targeted surveillance has been in place for at least the past 2 years without detection of disease (Bd or ranaviruses) AND basic biosecurity conditions have been continuously met for a period of 2 years 4. For a country that previously made a self declaration of freedom from disease, it may regain that status after detection of the disease if the affected area was declared an infected zone and a protection zone was established AND infected populations have been destroyed or removed from the infected zone by means that minimise the risk of further spread of the disease AND the appropriate disinfection procedures have been completed AND if the conditions of 3.) above are met. A zone or compartment may also be declared free from disease by the Competent Authority if it meets similar conditions to the above. Where a zone or compartment extends over more than one country, declarations must be made by all the Competent Authorities involved. A disease free status can be maintained if basic biosecurity conditions are continuously maintained. Targeted surveillance may be discontinued provided conditions that are conducive to clinical expression of disease exist. However, in infected countries and in all other cases where conditions are not conducive to clinical expression of disease, zones or compartments can only maintain a disease free status if targeted surveillance is maintained. Unknown or known infected country, zone or compartment: For the importation of live aquatic animals and aquatic animal products for any purpose (e.g., aquaculture, processing for human consumption, use in animal feed, agricultural, laboratory, zoo, pet trade, industrial or pharmaceutical use): In general, the Competent Authority of the importing country should require an international aquatic animal health certificate stating the commodities have been appropriately treated to inactivate disease agents OR undertake a risk assessment and apply appropriate risk mitigation measures Hygiene protocols for the control of diseases in Australian frogs June

63 The risk assessment and risk mitigation measures will vary with purpose of the importation or transit of commodities. Please see the Aquatic Code at the links provided above for more details AUSVETPLAN and AQUAVETPLAN In Australia, management of animal disease emergencies normally defaults to protocols outlined in the Australian Veterinary Emergency Plan (AUSVETPLAN - or the Australian Aquatic Veterinary Emergency Plan (AQUAVETPLAN - However, few of the diseases for which specific plans have been developed concern diseases of free-ranging wildlife. No amphibian diseases are currently included in AUSVETPLAN or AQUAVETPLAN Key Threatening Process and Threat Abatement Plan (TAP) Chytridiomycosis was listed as a Key Threatening Process in Australia in A Threat Abatement Plan (TAP) for infection of amphibians with chytrid fungus resulting in chytridiomycosis was subsequently prepared by representatives of the Commonwealth Government. These documents can be accessed here: Key Threatening Process: TAP: TAP Background document: Recommendation of the TAP proposes that a risk-based approach be used for chytridiomycosis using AUSVETPLAN as a model (Department of the Environment and Heritage 2006b). However, this has not progressed. Nation-wide mapping protocols and disease risk models have been developed as suggested in the TAP and should serve as the basis for cost-sharing arrangements between states and for setting research and management priorities (Skerratt et al. 2008; Murray et al. 2010a; Murray et al. 2010b; Skerratt et al. 2010; Murray et al. 2011). Implementing this step remains a priority Biosecurity Australia Risk analysis performed by Biosecurity Australia in Quarantine requirements for the importation of amphibians or their eggs into zoological facilities and Quarantine requirements for the importation of amphibians or their eggs for laboratory purposes (Animal Biosecurity Policy Memorandum 2003/26) does not list chytridiomycosis as a risk since it is endemic in Australia. However, this disregards the risk of importation into chytrid free areas or the introduction of novel strains. Although chytridiomycosis is not specifically mentioned, the general hygiene strategies recommended should still prevent the release of imported strains of B. dendrobatidis during the initial two years. After two years the amphibians can be released without testing for B. dendrobatidis. However, if being released into a chytrid free area, the same requirements imposed on Australian bred amphibians under the Threat Abatement Plan would apply. Hygiene protocols for the control of diseases in Australian frogs June

64 Risk analysis performed by Biosecurity Australia in Quarantine requirements for the importation of amphibians or their eggs into zoological facilities and Quarantine requirements for the importation of amphibians or their eggs for laboratory purposes (Animal Biosecurity Policy Memorandum 2003/26) mentions ranaviruses: The veterinary certificate must certify that for both live amphibians or amphibian eggs, as far as can be determined, no case of ranavirus infection (including frog virus 3, Redwood Park virus, Regina ranavirus), or ranid herpesviruses has been diagnosed at the premises of origin during the 12 months prior to certification. Importation of amphibians must meet the requirements of two Commonwealth departments, 1) Department of Agriculture, Fisheries and Forestry (DAFF) and 2) the DSEWPaC. The relevant documents can be accessed here: DAFF: Zoological facilities - Laboratory purposes - DSEWPaC: This site also has the requirements for export of amphibians from Australia. 6. Hygiene management Hygiene management issues can be broadly classed into in-situ (field based) and ex-situ (facility based) categories. While general isolation and disinfection hygiene management principles apply to both, greater detail on Guidelines for captive breeding, raising and restocking programs for Australian frogs can be found here: In-situ (site) hygiene management Individuals studying frogs often travel and collect samples of frogs from multiple sites. Numerous hygiene guidelines for handling wild frogs exist, including Daszak et al. (2001), NSW NPWS (2008), NWHC (2001), Speare et al. (2004) and CCADC (2008). Most recently, Phillott et al. (2010) provide a detailed review and synthesis of hygiene considerations that aim to minimise the risk of exposure of amphibians to pathogens in field studies. It is important to recognise that humans may aid in the: transmission (passing of disease from an infected to an uninfected individual), and spread (movement of disease geographically) of diseases, within and among amphibian populations For researchers working with amphibians or within areas where amphibians may occur, the risk of disease transmission within these habitats and the spread of disease among populations may be increased due to: Hygiene protocols for the control of diseases in Australian frogs June

65 movement of frogs or personnel between isolated areas of habitat or between captive husbandry and laboratory facilities and the field handling of amphibians It is therefore essential that personnel working with amphibians or within amphibian habitats take care to minimise disease transmission and spread. In order to do this, it is important that frog workers recognise the boundaries between sites/populations. This is especially important where rare, geographically restricted or threatened amphibian species are concerned and when the spread of diseases can have serious consequences for species survival. Phillott et al. (2010) recommend that field researchers evaluate their activities to determine the relative risk of pathogen transmission and spread compared with background levels (i.e., the risk posed by other mechanisms of disease transmission or pathogen dispersal) and implement appropriate strategies to minimise this risk during field studies. For a hygiene protocol checklist and suggested field kit see section 7. The risk of transmission and spread should also be evaluated by researchers, animal ethics committees and government agencies issuing permits Defining a site Defining the boundary of a site may not be straightforward. In some places, the boundary between sites will be obvious but in others it may not. Undertaking work at a number of sites or conducting routine monitoring at a series of sites within walking distance creates obvious difficulties with boundary definitions. It is likely that defining the boundary between sites will differ among localities. In general: watershed and geographical barriers should be used to designate separate sites river/stream tributaries should be considered separate sites wetlands, ponds, lakes etc. separated by dry land should be considered separate sites upstream locations separated by considerable distance (e.g., 500 m) should be considered separate sites any obvious break, barrier or change in habitats should be treated as separate sites, particularly if there is no known interchange of frogs between sites Determining the order of visitation of multiple field sites When a field trip encompasses several field sites, or a number of locations are being visited in succession, the order of visitation should be determined according to the presence of known pathogens and diseases. Areas known to be absent of disease should be visited first, followed by areas of unknown status, followed by known infected areas Hygiene protocols for the control of diseases in Australian frogs June

66 On-site hygiene When travelling from site to site it is recommended that the following hygiene precautions be taken to minimise the possibility of transfer of disease from personnel, footwear, equipment and/or vehicles. A list of suitable disinfectants, their required concentrations and exposure times for various purposes is summarised by Phillott et al. (2010) and is reproduced in Table 1 below. Personnel Hands, arms, knees etc. should be cleaned to remove debris and washed or wiped with a suitable disinfectant. It is preferable to do this before entering the vehicle or moving to another site. Footwear and clothing Footwear must be thoroughly cleaned and disinfected at the commencement of fieldwork and between each sampling site. This can be achieved by initially scraping boots clear of mud and standing the soles in a disinfecting solution. The remainder of the boot should be rinsed or sprayed with a disinfecting solution. Clothing that has significant contact with frogs and the environment should also be subjected to changing or cleaning Disinfecting solutions should be prevented from entering any water bodies. Several changes of footwear/clothing bagged between sites might be a practical alternative to on-site cleaning. In high value sites, dedicated equipment and clothing stored at the entry to the site may be desirable. (e.g., in a lockbox) Equipment Equipment such as nets, balances, callipers, bags, scalpels, headlamps, torches, wetsuits and waders etc. that are used at one site must be cleaned and disinfected before re-use at another site Disposable items should be used where practical/possible Non-disposable equipment should be used only once during a particular field exercise and disinfected later or disinfected at the site between uses using procedures outlined below in Table 1. Vehicles Transmission of disease from vehicles is generally unlikely to be a problem. However, if a vehicle is used to traverse a known frog site and could result in mud and water being transferred to other bodies of water or frog sites, then wheels and tyres should be cleaned and disinfected. This is particularly important where vehicles are used in areas not normally frequented by other vehicles. Disinfection should be carried out at a safe distance from water bodies to minimise the risk of chemical contamination. Hygiene protocols for the control of diseases in Australian frogs June

67 Principles of cleaning and disinfection Designing an effective disinfection protocol requires understanding of the properties of disinfectants and target pathogens, and practical consideration of the equipment or processes requiring disinfection. As well as understanding the efficacy of various disinfecting processes, it is important to consider the safety of any disinfection protocol to the environment and the animals on which they will be used. Key distinctions include: Cleaning: The physical removal of all visible organic and inorganic debris from items Disinfection: A physical (e.g., UV light) or chemical (e.g., bleach) process to reduce the numbers and/or viability of microorganisms (e.g., bacteria, fungi or viruses) on an object, surface or material Sterilization: A physical or chemical process that removes all microorganisms from an object, surface or material Thorough cleaning and disinfection reduces most of the risk of transferring amphibian pathogens. Sterilization of objects is labour intensive and less practical for most routine applications. Cleaning alone does not render an object free of pathogens. However, it is important to thoroughly clean objects prior to disinfection or sterilization. Thorough cleaning physically removes many or most pathogens that are trapped in organic debris Thorough cleaning makes successful disinfection more likely Cleaning allows disinfectants to directly contact the surfaces of an object Warm or hot water improves the ability to remove organic materials from objects Regular cleaning of all items used should be performed Use of detergents aid cleaning by loosening organic material from the surface of objects and help to break apart biofilms of microorganisms that can resist disinfection Thorough rinsing of detergents from objects is essential after cleaning Disinfection of an item by application of an appropriate chemical agent after cleaning reduces pathogen numbers and viability and minimises potential for disease transmission. Things to consider include: Efficacy of the disinfectant and the type of pathogens that must be eliminated. For example, some microorganisms such as Mycobacterium spp. or Cryptosporidium spp. are very resistant to most common disinfectants The potential for toxicity to amphibians that are exposed to the disinfectant. Amphibians are very sensitive to some disinfectant residues and thorough rinsing of all disinfectants is required after use Concerns about human exposure to disinfectants and about discharge of disinfectants into the environment Safety for use on different materials. Some disinfectants may be corrosive to materials or tools used in amphibian facilities Ease of use and disposal Cost Hygiene protocols for the control of diseases in Australian frogs June

68 Table 1. Disinfection strategies suitable for killing Batrachochytrium dendrobatidis, Mucor amphibiorum and ranaviruses in field studies. From Phillott et al. (2010) and Webb et al. (submitted). Application Disinfectant Strength Time Target pathogen Surgical equipment Benzalkonium 1 mg ml 1 1 min B. dendrobatidis and other instruments (e.g. scales, callipers) chloride Ethanol 70% 1 min B. dendrobatidis Ranaviruses Collection equipment and containers Footwear Cloth (e.g. carry bags, clothes) Sodium hypochlorite (bleach contains 4% sodium hypochlorite) Path X or quaternary ammonium compound 128 1% 1 min B. dendrobatidis 3% 1 min Ranaviruses 1 in 500 dilution 0.5 min B. dendrobatidis 1 in 100 dilution 10 min M. amphibiorum Trigene 1 in 5000 dilution 1 min B. dendrobatidis F10 1 in 1500 dilution 1 min B. dendrobatidis Virkon 2 mg ml 1 1 min B. dendrobatidis 1% 1 min Ranaviruses Nolvasan 0.75% 1 min Ranaviruses Potassium 1% 10 min B. dendrobatidis permanganate Complete drying >3 h B. dendrobatidis Heat 60 C 30 min B. dendrobatidis Ranaviruses Heat 37 C 8 h B. dendrobatidis Sterilising UV light 1 min Ranaviruses only Sodium 1% 1 min B. dendrobatidis hypochlorite (bleach contains 4% sodium hypochlorite) 3% 1 min Ranaviruses Path X or 1 in 500 dilution 0.5 min B. dendrobatidis quaternary ammonium 1 in 100 dilution 10 min M. amphibiorum compound 128 Trigene 1 in 5000 dilution 1 min B. dendrobatidis F10 1 in 1500 dilution 1 min B. dendrobatidis Phytoclean (30% 0.075% 1 min B. dendrobatidis benzalkonium chloride) 5% 1 min M. amphibiorum Complete drying >3 h B. dendrobatidis Hot wash 60 C or 30 min B. dendrobatidis greater Ranaviruses Hygiene protocols for the control of diseases in Australian frogs June

69 6.2. Handling of frogs in the field The spread of pathogens may occur as a result of handling frogs. In addition to spreading disease among captured frogs, handling may stress animals making them more susceptible to infection from other sources or more likely to succumb to infection. Capture, handling and housing of wild amphibians should be minimised or avoided where possible Where handling is necessary, care must be taken to ensure individuals do not have their exposure to pathogens elevated over their background exposure levels. Direct transfer of pathogens during capture and handling of successive adult amphibians can be reduced by using: single-use gloves (latex, nitrile or vinyl), and/or single-use lightweight plastic bags adequate cleaning of hands and handling equipment Many researchers use disposable plastic bags to catch and/or restrain frogs followed by handling/processing with disposable gloves. As some tadpoles may suffer lethal effects when exposed to latex, nitrile or vinyl gloves (Cashins et al. 2008), researchers should only use gloves that have been proven or rendered safe (e.g., by rinsing with water) for the study species. In situations where gloves are not available or suitable: hand washing with 70% ethanol (allowing hands to dry) between handling individual frogs is acceptable (note, repeated use on human skin is not recommended). Alcohol is toxic to frogs so hands must be washed thoroughly in water after treatment with alcohol If 70% ethanol is not available or suitable, the minimum treatment is handwashing in the water to which the amphibian is normally exposed. In situations where amphibians must be held temporarily: Individuals should be housed in single-use containers (e.g. plastic bags) or in containers disinfected between each animal Adults should not be held in groups Tadpoles from the same water body may be housed for short periods in a common container, although overcrowding should be avoided Longer holding times (>60 min) will require changes to water and the provision of appropriate food (>24 h). Tadpoles should always be treated with care to prevent damage on capture and with movement of water within holding containers. If animals must be removed from the field for greater periods and later returned, it should always be to the same site. Hygiene protocols for the control of diseases in Australian frogs June

70 6.3. Housing frogs and tadpoles Frogs and tadpoles should only be removed from a site when absolutely necessary. Detailed Guidelines for captive breeding, raising and restocking programs for Australian frogs can be found at: See also A Manual for Control of Infectious Diseases in Amphibian Survival Assurance Colonies and Reintroduction Programs (Pessier and Mendelson 2010) at: mphibian%22 When frogs or tadpoles are to be collected and held for a period of time, the following measures are recommended: Isolate animals obtained at different sites Aquaria set up to hold frogs should not share water, equipment or any filtration system. Splashes of water from adjacent enclosures or drops of water on nets may transfer pathogens between enclosures Ensure that tanks, aquaria and any associated equipment are disinfected prior to housing frogs or tadpoles Tanks and equipment should be cleaned, disinfected and dried after frogs/tadpoles are removed 6.4. Marking, invasive and surgical procedures Strict hygiene standards must be maintained during amphibian marking procedures including implanting internal radio transmitters, passive integrated transponder (PIT) tags, visible implant alphanumeric (VIA) tags, visible implant elastomer (VIE) tags and toe tipping or clipping. Due to the high permeability of amphibian skin, special disinfectants are required. The only suitable, commercially available preparation for disinfecting wounds is: Bactine spray (active ingredient 0.14% w/w benzalkonium chloride and 2.6% w/w lidocaine hydrochloride in a non-alcohol base) Chlorhexidine (0.75% diluted from 2% Nolvasan ) is also suitable for surgical disinfection Alcohol, phenol and iodine based disinfectants should not be used because they are potentially toxic and can destroy mucus and wax that prevent dehydration and microbial infection of amphibian skin. Contrary to the recommendations of previous hygiene protocols, Betadine or other povidone-iodine products are not recommended for use as disinfectants for amphibians until species-specific toxicity has been determined (Phillott et al. 2010). Toe tipping (removal of most distal phalange) or toe clipping (amputation of a greater proportion of the digit): should occur through the interphalangeal joints Hygiene protocols for the control of diseases in Australian frogs June

71 Scissors should be sterilised in 70% ethanol and dried before use on frogs in the field For studies in which diagnostic testing of disease is important, the diagnostic test step (e.g., swabbing for Bd) should be undertaken before any other processing step to minimise the potential for false-positives due to cross contamination PIT, VIE and VIA tags should be inserted with a sterile, single-use applicator Sealing wounds A cryanoacrylate compound such as Vetbond (active ingredient n-butyl cryanoacrylate) as a tissue adhesive after toe tipping or clipping is recommended. Vetbond can also be used to seal incisions made during subdermal injection of VIA, VIE and PIT tags A disinfectant such as Bactine should be applied before the adhesive to avoid trapping microbes Less expensive industrial adhesives ( superglues ) should not be used as a replacement for surgical tissue glues However, this procedure may only be possible in larger amphibians. In smaller animals, it can be difficult to isolate toes for application and internal marking devices such as PIT tags may be unsuitable. Moisture can interfere with setting times and adhesion so care must be taken to ensure setting has occurred before release. Problems may be experienced in their application to stream- or pond-dwelling amphibians, but can be avoided by using a small piece of sterile absorbent dressing to draw surplus water from the wound before application of the adhesive (Phillott et al. 2010) Equipment Equipment used in marking or surgery should be appropriately disinfected Disposable sterile instruments should be used where practical/possible Instruments should be disinfected or changed in between each frog All used disinfecting solutions, gloves and other disposable items should be stored in a sharps or other waste container and disposed of or sterilised appropriately at the completion of fieldwork Disinfecting solutions must not come into contact with frogs or be permitted to contaminate any water bodies 6.5. Return of captive animals to the wild In general, if wild frogs or tadpoles are housed for any period of time in a captive situation (e.g. laboratory, zoo or captive breeding facility), they should not be returned to the wild Exceptions to this can occur if they have been kept in isolation, their captive history is free of undiagnosed morbidity or mortality and they have had rigorous pathogen screening before release. This is usually beyond the means of most studies. Detailed Guidelines for captive breeding, raising and restocking programs for Australian frogs can be found at: Hygiene protocols for the control of diseases in Australian frogs June

72 See also A Manual for Control of Infectious Diseases in Amphibian Survival Assurance Colonies and Reintroduction Programs (Pessier and Mendelson 2010) at: mphibian% Displaced frogs Displaced frogs should be treated as if they are infected and should not be transported anywhere for release to the wild Displaced frogs are native frog species and introduced cane toads (Bufo marinus) that have been unintentionally transported from one place to another. This may typically occur with the transport of fresh produce and landscaping supplies. Banana Box frog is the term used to describe several native frog species (usually Litoria gracilenta, L. fallax, L. caerulea, L. rubella, L. infrafrenata and L. bicolor) commonly transported in fruit and vegetable shipments and landscaping supplies. There is risk of spread of disease if these frogs are transferred from place to place. When encountering a displaced frog: Contact a licensed wildlife carer organisation to collect the animal. The frog may then undergo a quarantine period along with an approved disinfection treatment Post-quarantine, and dependant on local state legislation and policies, the frog may be transferred to a licensed frog keeper once permission from the relevant regulatory body has been received. Licensed carer groups are to record and receipt frogs obtained and disposed of in this way. Frogs held by licensed frog keepers are not to be released to the wild except with relevant regulatory body approval Displaced frogs may also be made available to recognised institutions for research projects, display purposes or offered to a museum as scientific specimens once approval has been provided by the relevant regulatory body. Frogs encountered on roads, around dwellings and gardens or in swimming pools should not be considered as displaced frogs unless they are of a species not local to the area Local frogs encountered in these situations should be assisted off roads, away from dwellings, or out of swimming pools preferably to the nearest area of vegetation or suitable habitat Cane toads Cane toads are known amphibian disease carriers and should not be knowingly transported or released to the wild. If a cane toad is discovered it should be humanely euthanized in accordance with the recommended Animal Welfare procedures. Care should be taken to avoid euthanasia of native species due to mistaken identity. Hygiene protocols for the control of diseases in Australian frogs June

73 6.7. Sick and dead animals Dead amphibians or live animals showing clinical signs of disease must be regarded as having a high infection risk to healthy animals and rigorous hygiene measures are required. Sick and dead frogs should be collected and sent for disease diagnosis No effective and practical field treatment for chytridiomycosis has been demonstrated. Similarly, no treatment regimes for ranaviral infection of frogs have been described. The collection of sick and dead frogs for expert diagnosis may improve disease surveillance activities, which can help detect disease introduction and enable emergency responses. It is also useful to assess the risk of pathogen transmission to other individuals or spread to other populations. A procedure for the preparation and transport of a sick or dead frog is given below. Adherence to this procedure will ensure the animal is maintained in a suitable condition for pathological examination and assist determining the extent of the disease and the number of species affected. For more information about sick and dead amphibians, see Collection: Do not use bare hands to handle sick or dead frogs Disposable gloves should be worn when handling sick or dead frogs New gloves and a clean plastic bag should be used for each frog specimen to prevent cross-contamination If the frog is dead, keep the specimen cool and preserve as soon as possible to avoid decomposition Preserving specimens: Specimens can be preserved/fixed in 70% ethanol or 10% buffered formalin Cut open the belly and place the frog in about 10 times its own volume of preservative Where no preservative is available, specimens can also be frozen. If numerous frogs are collected, some should be preserved and some should be frozen. Portions of a dead frog can also be sent for analysis (e.g., a preserved foot, leg or a portion of abdominal skin) Transportation: If the frog is alive and likely to survive transportation, place the frog into either a moistened cloth bag with some damp leaf litter or into a plastic bag with damp leaf litter and partially inflated before sealing Remember to keep all frogs separated during transportation If the frog is alive but unlikely to survive transportation (death appears imminent), euthanize the frog and place the specimen in a freezer or preservative. Once frozen/preserved the specimen is ready for shipment All containers should be labelled showing at least the species (if known), date and collection location Preserved samples can be sent in jars or wrapped in wet cloth, sealed in bags and placed inside a padded box Send frozen samples in an esky with dry ice Hygiene protocols for the control of diseases in Australian frogs June

74 Place live or frozen specimens into a small Styrofoam esky. Seal esky with packaging tape before sending Send the package by courier and declare any hazardous or flammable contents (e.g., 70% ethanol) 7. Hygiene protocol checklist and field kit The following checklist and field kit are designed to assist with minimising the risk of transferring pathogens between frogs and sites in field studies (follows NSW 2008) Have you considered the following questions before handling frogs in the field: Has your proposed field trip been sufficiently well planned to consider hygiene issues? Have you considered the boundaries between sites (particularly where endangered species or populations at risk are known to occur)? Have footwear disinfection procedures been considered and a strategy adopted? Have you planned the equipment you will be using and developed a disinfection strategy? Are you are planning to visit sites where vehicle disinfection will be needed? If so, do you have a plan to deal with vehicle disinfection? Have handling procedures been planned to minimise the risk of frog to frog pathogen transmission? Do you have a planned disinfection procedure to deal with equipment, apparel and direct contact with frogs? If you answered NO to any of these questions please re-read the relevant section of the Hygiene Protocols for the Control of Disease in Australian Frogs and apply a suitable strategy. Field hygiene kit When planning to survey frogs in the field a portable field hygiene kit should be assembled to assist with implementing the hygiene protocols. Recommended contents of a field hygiene kit would include: Plastic box to store field equipment Small Styrofoam esky Disposable gloves Disinfectant spray bottle (atomiser spray) and/or wash bottle for disinfectants Disinfecting solutions Scraper or scrubbing brush for cleaning mud off footwear, vehicles etc. Bucket for mixing disinfecting solutions and soaking Plastic bags, large and small for hygienic temporary animal handling/holding Sharps or other container for safe waste disposal Materials for dealing with sick and dead frogs (see section 6.7.) Hygiene protocols for the control of diseases in Australian frogs June

75 Detailed Guidelines for captive breeding, raising and restocking programs for Australian frogs can be found at: See also A Manual for Control of Infectious Diseases in Amphibian Survival Assurance Colonies and Reintroduction Programs (Pessier and Mendelson 2010) at: mphibian%22 8. Important Australian contacts 8.1. Sick and dead frogs To arrange receipt and analyse sick and dead frogs, make contact with experts at any of the organisations below prior to dispatching package: Australian Registry of Wildlife Health Taronga Conservation Society, Australia PO Box 20 MOSMAN NSW 2088 Phone: School of Public Health, Tropical Medicine and Rehabilitation Sciences James Cook University Douglas Campus TOWNSVILLE QLD 4811 Phone: School of Biological Sciences University of Newcastle CALLAGHAN NSW 2308 Phone: Hygiene protocols for the control of diseases in Australian frogs June

76 9. References Anver, M. R. and C. L. Pond (1984). Biology and diseases of amphibians. Laboratory Animal Medicine. B. J. C. J. G. Fox, F. M. Loew. New York, Academic Press: Berger, L., J. E. Longcore, R. Speare, A. Hyatt and L. F. Skerratt (2009). Fungal Diseases in Amphibians. Amphibian Biology, Volume 8 Amphibian Decline: Disease, Parasites, Maladies, and Pollution. H. Heatwole and J. W. Wilkinson, Surrey Beatty & Sons. NSW. Berger, L., R. Speare, P. Daszak, D. E. Green, A. A. Cunningham, C. L. Goggin, R. Slocombe, M. A. Ragan, A. D. Hyatt, K. R. McDonald, H. B. Hines, K. R. Lips, G. Marantelli and H. Parkes (1998). "Chytridiomycosis causes amphibian mortality associated with population declines in the rain forests of Australia and Central America." Proceedings of the National Academy of Sciences of the United States of America 95(15): Berger, L., R. Speare and A. Hyatt (1999). Chytrid fungi and amphibian declines: Overview, implications and future directions. Declines and Disappearances of Australian Frogs. A. Campbell. Canberra, Environment Australia: Cashins, S., R. Alford and L. F. Skerratt (2008). "Lethal effect of latex, nitrile, and vinyl gloves on tadpoles." Herpetological Review 39: CCADC (2008). "Decontamination protocol to reduce the risk of spreading infectious amphibian diseases in freshwater systems. (California Centre for Amphibian Disease Control). Cullen, B. R. and L. Owens (2002). "Experimental challenge and clinical cases of Bohle iridovirus (BIV) in native Australian anurans." Diseases of Aquatic Organisms 49(2): Daszak, P., A. A. Cunningham and H. A.D. (2001). Draft guidelines for international translocation of amphibians with respect to infectious diseases. Attachment 6. In: Speare R and Steering Committee of Getting the Jump on Amphibian Disease. Developing management strategies to control amphibian diseases: Decreasing the risks due to communicable diseases. School of Public Health and Tropical Medicine, James Cook University: Townsville. 2001: Davis, A. K., M. J. Yabsley, M. K. Keel and J. C. Maerz (2007). "Discovery of a novel alveolate pathogen affecting southern leopard frogs in Georgia: Description of the disease and host effects." Ecohealth 4(3): Department of the Environment and Heritage. (2006a). "Background document for the Threat Abatement Plan: Infection of amphibians with chytrid fungus resulting in chytridiomycosis." from chytrid-background.pdf. Department of the Environment and Heritage. (2006b). "Threat Abatement Plan: Infection of amphibians with chytrid fungus resulting in chytridiomycosis." 2007, from Gravekamp, C., Korver, H., Montgomery, J., Everard, C.O., Carrington, D., Ellis, W.A., Terpstra, W.J. (1991). "Leptospires isolated from toads and frogs on the Island of Barbados." Zentralblatt fur Bakteriologie 275: Gust, N., J. Griffiths, M. Driessen, A. Philips, N. Stewart and D. Geraghty (2009). "Distribution, prevalence and persistence of mucormycosis in Tasmanian platypuses (Ornithorhynchus anatinus)." Australian Journal of Zoology 57(4): Hartigan, A., I. Fiala, D. I, M. Jirků, B. Okimoto, K. Rose, D. N. Phalen and Š. J. (2011). "A suspected parasite spill-back of two novel Myxidium spp. (Myxosporea) causing Hygiene protocols for the control of diseases in Australian frogs June

77 disease in Australian endemic frogs found in the invasive cane toad." PLoS One 6: e Hemingway, V., J. Brunner, R. Speare and L. Berger (2009). Viral and bacterial diseases of amphibians. Amphibian Biology, Volume 8 Amphibian Decline: Disease, Parasites, Maladies, and Pollution. H. Heatwole and J. W. Wilkinson, Surrey Beatty & Sons. NSW. Hines, H., M. Mahony and K. McDonald (1999). An assessment of frog declines in wet subtropical Australia Declines and Disappearances of Australian Frogs. A. Campbell. Canberra, Environment Australia: Hyatt, A. D., A. R. Gould, Z. Zupanovic, A. A. Cunningham, S. Hengstberger, R. J. Whittington, J. Kattenbelt and B. E. H. Coupar (2000). "Comparative studies of piscine and amphibian iridoviruses." Archives of Virology 145(2): Kiesecker, J., A. R. Blaustein and C. L. Miller (2001). "Transfer of a pathogen from fish to amphibians." Conservation Biology 15: Laurance, W., K. McDonald and R. Speare (1996). "Catastrophic declines of Australian rain forest frogs: support for the epidemic disease hypothesis." Conservation Biology 10: Longcore, J. E., A. P. Pessier and D. K. Nichols (1999). "Batrachochytrium dendrobatidis gen et sp nov, a chytrid pathogenic to amphibians." Mycologia 91(2): Mikaelian, I., M. Ouellet, B. Pauli, J. Rodrigue, J. C. Harshbarger and D. M. Green (2000). "Ichthyophonus like infection in wild amphibians from Québec, Canada." Diseases of Aquatic Organisms 40: Murray, K. A., R. Retallick, K. McDonald, D. Mendez, K. Aplin, P. Kirkpatrick, L. Berger, D. Hunter, H. B. Hines, R. Campbell, M. Pauza, M. Driessen, R. Speare, S. J. Richards, M. Mahony, A. Freeman, A. D. Phillott, J.-M. Hero, K. Kriger, D. Driscoll, A. Felton, R. Puschendorf and L. F. Skerratt (2010a). "The distribution and host range of the pandemic disease chytridiomycosis in Australia spanning surveys from 1956 to 2007." Ecology 91(5): Murray, K. A., R. W. R. Retallick, R. Puschendorf, L. F. Skerratt, D. Rosauer, H. McCallum, L. Berger, R. Speare and J. VanDerWal (2011). "Assessing spatial patterns of disease risk to biodiversity: implications for the management of the amphibian pathogen, Batrachochytrium dendrobatidis." Journal of Applied Ecology 48(1): Murray, K. A., D. Rosauer, H. McCallum and L. F. Skerratt (2010b). "Integrating species traits with extrinsic threats: closing the gap between predicting and preventing species declines." Proceedings of the Royal Society B-Biological Sciences Published online October (doi: /rspb ). Murray, K. A. and L. F. Skerratt (in press). "Predicting wild hosts for amphibian chytridiomycosis: integrating host life-history traits with pathogen environmental requirements." Human and Ecological Risk Assessment. NSW, N. P. a. W. S. (2008). Hygiene protocol for the control of disease in frogs. Threatened Species Management: Information Circular No : p 218. NWHC (2001). "Toe-clipping of frogs and toads standard operating procedure no NWHC (National Wildlife Health Center), Madison, WI O'Shea, P., R. Speare and A. D. Thomas (1990). "Salmonellas from the cane toad, Bufo marinus." Australian Veterinary Journal 67: 310. Obendorf, D. and A. Dalton (2006). "A survey for the presence of the amphibian chytrid fungus (Batrachochytrium dendrobatidis) in Tasmania." Papers and Proceedings of the Royal Society of Tasmania 140: Pauza, M. and M. Driessen (2008). Distribution and potential spread of amphibian chytrid fungus Batrachochytrium dendrobatidis in the Tasmanian Wilderness World Heritage Hygiene protocols for the control of diseases in Australian frogs June

78 Area, Biodiversity Conservation Branch, Department of Primary Industries and Water, Tasmania. Pessier, A. and J. R. Mendelson (2010). A manual for control of infectious diseases in amphibian survival assurance colonies and reintroduction programs. Apple Valley, MN, IUCN/SSC Conservation Breeding Specialist Group:. Phillott, A. D., R. Speare, H. B. Hines, E. Meyer, L. F. Skerratt, K. R. McDonald, S. D. Cashins, D. Mendez and L. Berger (2010). "Minimising exposure of amphibians to pathogens during field studies." Diseases of Aquatic Organisms. Sharma, V. K., Y. K. Kaura and I. P. Singh (1974). "Frogs as carriers of Salmonella and Edwardsiella." Antonie von Leeuwenhoek 40: Skerratt, L. F., L. Berger, H. B. Hines, K. R. McDonald, D. Mendez and R. Speare (2008). "Survey protocol for detecting chytridiomycosis in all Australian frog populations." Diseases of Aquatic Organisms 80(2): Skerratt, L. F., L. Berger, R. Speare, S. Cashins, K. R. McDonald, A. D. Phillott, H. B. Hines and N. Kenyon (2007). "Spread of chytridiomycosis has caused the rapid global decline and extinction of frogs." Ecohealth 4(2): Skerratt, L. F., K. R. McDonald, H. B. Hines, L. Berger, D. Mendez, A. Phillott, S. D. Cashins, K. A. Murray and R. Speare (2010). "Validation of the mapping protocol for Batrachochytrium dendrobatidis in Queensland, Australia " Diseases of Aquatic Organisms 92: Speare, R., L. Berger, L. F. Skerratt, R. Alford, D. Mendez, S. Cashins, N. Kenyon, K. Hauselberger and J. Rowley (2004). Hygiene Protocol for handling amphibians in field studies. A. D. Group. Townsville, James Cook University: 4. Speare, R., o. G. t. J. o. A. D. Core Working Group and o. G. t. J. o. A. Steering Committee (2001). Developing Management Strategies to Control Amphibian Diseases: Decreasing the Risks Due to communicable Diseases. S. C. o. G. t. J. o. A. D. (eds Speare R. Townsville, School of Public Health and Tropical Medicine, James Cook University. Stuart, S. N., J. S. Chanson, N. A. Cox, B. E. Young, A. S. L. Rodrigues, D. L. Fischman and R. W. Waller (2004). "Status and trends of amphibian declines and extinctions worldwide." Science 306(5702): Taylor, R., D. Sloan, T. Cooper, B. Morton and I. Hunter (2000). "A waterborne outbreak of Salmonella Saintpaul." Communicable Diseases Intelligence 24: Webb, R., A. Philips, L. Berger, J. Connolly and R. Speare (submitted). "Controlling the spread of wildlife diseases: In vitro efficacy of disinfectants against the pathogenic fungi Batrachochytrium dendrobatidis and Mucor amphibiorum." Diseases of Aquatic Organisms. Wright, K. M. and B. R. Whitaker (2001). Amphibian Medicine and Captive Husbandry. Malabar, Florida, Krieger Publishing. Zupanovic, Z., G. Lopez, A. D. Hyatt, B. Green, G. Bartran, H. Parkes, R. J. Whittington and R. Speare (1998). "Giant toads Bufo marinus in Australia and Venezuela have antibodies against 'ranaviruses'." Diseases of Aquatic Organisms 32(1): 1-8. Hygiene protocols for the control of diseases in Australian frogs June

79 Appendix D Consultation with DP&E Report name Agency Date 29/04//2016 Comment number Ecological Australia Green and Golden Bell Frog Plan of Management Arncliffe (GGBFPOM) April 2016 Department of Planning and Environment Document section/ref CoA DP&E Comment 1. Section 2.1 This section states that under the TSC Act, public authorities must not make decisions that are inconsistent with the provisions of the draft species Recovery Plan. The GGBFPOM must provide details of how the POM addresses the provisions of the GGBF Recovery Plan. 2. Section 3 This section must provide details of the Arncliffe GGBF population (RTA ponds) and its environment, to follow section Section 4.1 Add the following potential indirect impact to the RTA ponds introduction of frog pathogen by construction personnel and construction equipment and machinery. 4. Section 4.2 The discussion of GGBF habitat includes dispersal habitat. A frog passage was provided beneath the M5 East Motorway to facilitate movement between the golf course and habitat to the west and south (Eve and Marsh Street wetlands). Details of the frog passage (size, length, etc) should be provided and the location of the passage shown in Figure Table 1 The proposed mitigation measures include the provision of fencing to exclude frogs from construction areas, noise wall fence adjacent to the RTA ponds and frog exclusion fencing around the construction zone. The POM must provide details of the fencing and how it would be installed. 6. Table 1 The mitigation measures include pre-clearance survey and salvage. Details are provided of measures to handle frogs should they be encountered. The POM states that water bodies will need to be decommissioned prior to work commencing. Frogs encountered during decommissioning would be collected in accordance with the process described in Table 1. However, no details are provided on how tadpoles encountered would be handled. Details of the handling of tadpoles encountered during water body decommissioning must be provided. Reference in PoM Section 2.1 New section Section 4.1 Section 4.2; figure 1 New Table 2 and Figure 3 New Table 2 ECO LOGICAL AUSTRALIA PTY LTD 47

80 Report name Agency Date 29/04//2016 Comment number Ecological Australia Green and Golden Bell Frog Plan of Management Arncliffe (GGBFPOM) April 2016 Department of Planning and Environment Document section/ref CoA DP&E Comment 7. Table 2 The Hygiene Protocol must be included in the POM. The introduction of frog pathogens, in particular Chytrid fungus is identified as a key threatening process leading to the decline of the GGBF. The measures to prevent/address the introduction of the pathogen to the GGBF must be included in the POM and not developed separate to and outside the POM. 8. Table 2 Details of the management measures to maintain the RTA ponds must be provided in the POM. This includes details of the water supply, water level management and salt water supply. 9. Table 2 Details of measures to reduce predatory fish, noxious weeds and predation by cats, rats and foxes must be provided in the POM. 10. Section 5.1 The quarterly report on the implementation of the actions in Table 1 and 2, during construction, must be provided to DPE and OEH. 11. Table 3 The monitoring requirements do not include monitoring of the water quality of the existing RTA ponds. The POM must provide details of water quality monitoring. 12. Section Population monitoring must be undertaken at the new habitat at Marsh Street following its occupation by frogs. 13. Table 4 Monitoring would be established in accordance with Pollocks Robust Design and monitoring data analysed using MARK. The POM must provide details of this methodology and the suitability of these for determining the population status of the GGBF. Reference in PoM Table 3 and Appendix C Table 3 Table 3 Section 5.1 Table 3 Table 4 Section Population monitoring must include the existing Marsh Street and Eve Street wetlands. 14. Table 4 The Department considers that monitoring of the frog passageway beneath the M5 East Motorway must be included in population monitoring. This would inform the current usage of the underpass (it is understood the passage was last monitored in 2007). Once the new habitat was created at Marsh Section ECO LOGICAL AUSTRALIA PTY LTD 48

81 Report name Agency Date 29/04//2016 Comment number Ecological Australia Green and Golden Bell Frog Plan of Management Arncliffe (GGBFPOM) April 2016 Department of Planning and Environment Document section/ref CoA DP&E Comment Street and the ponds populated by frogs, the passage would become more important in east west movements between the new habitat and the Kogarah Golf Course. Should monitoring show the passage is not being used, then actions to improve accessibility to the underpass would need to be considered, including the provision of new connectivity. The monitoring must include the Eve Street cycleway. 15. Section Include the following as a goal for mitigation no frog mortality on the Eve Street cycleway underpass of the M5 East Motorway. Once the new March Street habitat is created and occupied, movement between the new ponds and the Kogarah Golf Course would be expected to occur, given there is very limited foraging habitat on the western side of the M5 East Motorway. Anecdotal evidence has been cited that frogs use the cycleway. Given that the cycleway is a hostile environment for frogs, measures to provide suitable frog movement must be considered in the event of frog on the cycleway. Reference in PoM New Table 3 Section Section 5.4 The annual monitoring reports must be provided to OEH. Section Appendix A Unanticipated finds procedure the procedure makes a number of references to the approved Environmental Representative (ER), the Roads and Maritime Environmental Representative and the Environmental Representative. Is the RMS ER and the approved ER separate positions? Is the approved ER and the ER the ER under condition D1 of the approval? 18. Appendix A Step six states frogs detected should be released by the Project Ecologist the Taronga Zoo this position should be clarified. Is the project ecologist an employee of the Taronga Zoo. Is the project ecologist the position created by the Contractor or a RMS ecologist? Does the project ecologist have expertise in the handling of the GGBF? 19. Appendix B The consultation with OEH must include comments made by OEH on the POM and how RMS has responded to the issues raised. Appendix A Appendix A Appendix B ECO LOGICAL AUSTRALIA PTY LTD 49

82 13 Tasman Place Flora & Fauna Assessment HEAD OFFICE Suite 2, Level Old Princes Highway Sutherland NSW 2232 T F SYDNEY Level Sussex Street Sydney NSW 2000 T F HUSKISSON Unit 1, 51 Owen Street Huskisson NSW 2540 T F CANBERRA Level 2 11 London Circuit Canberra ACT 2601 T F NEWCASTLE Suites 28 & 29, Level 7 19 Bolton Street Newcastle NSW 2300 T F NAROOMA 5/20 Canty Street Narooma NSW 2546 T F COFFS HARBOUR 35 Orlando Street Coffs Harbour Jetty NSW 2450 T F PERTH Suite 1 & 2 49 Ord Street West Perth WA 6005 T F ARMIDALE 92 Taylor Street Armidale NSW 2350 T F WOLLONGONG Suite 204, Level 2 62 Moore Street Austinmer NSW 2515 T F MUDGEE Unit 1, Level 1 79 Market Street Mudgee NSW 2850 T F GOSFORD Suite 5, Baker One 1-5 Baker Street Gosford NSW 2250 T F DARWIN BRISBANE 16/56 Marina Boulevard Suite 1 Level 3 Cullen Bay NT Adelaide Street Brisbane QLD ECO LOGICAL AUSTRALIA PTY LTD 50 T F T F