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The Efficacy of Clove Oil for Anaesthesia of Eight Species of Australian Tropical Freshwater Teleosts

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The Efficacy of Clove Oil for Anaesthesia of Eight Species of Australian Tropical Freshwater Teleosts LIMNOLOGY and Limnol. Oceanogr.: Methods 13, 2015, 463–475 VC 2015 The Authors Limnology and Oceanography: Methods published by Wiley Periodicals, Inc. on behalf of Association for the OCEANOGRAPHY: METHODS Sciences of Limnology and Oceanography doi: 10.1002/lom3.10040 The efficacy of clove oil for anaesthesia of eight species of Australian tropical freshwater teleosts Frederieke J. Kroon†* 1CSIRO Division of Ecosystem Sciences, Atherton, Queensland, Australia Abstract The efficacy of clove oil for anaesthesia was examined on eight species of Australian tropical freshwater fishes: the native Mulgrave goby (Glossogobius bellendenensis, Gobiidae), empire gudgeon (Hypseleotris compressa) and sleepy cod (Oxyeleotris lineolatus) (both Eleotrididae), Eastern rainbowfish (Melanotaenia splendida, Melano- taeniidae), Pacific blue-eye (Pseudomugil signifer, Pseudomugilidae), and eel-tailed catfish (Tandanus tandanus, Plotosidae), and the non-native species spotted tilapia (Pelmatolapia mariae, Cichlidae) and guppy (Poecilia retic- ulata, Poecilidae). Induction to anaesthesia, recovery from anaesthesia and survival of fish were determined for five concentrations (20 mg/L, 30 mg/L, 40 mg/L, 80 mg/L, and 160 mg/L). A 20 mg/L clove oil concentration was not sufficient to induce a loss of reflex reactivity within 10 min for most individuals (> 85%) in the eight species examined. Effective clove oil concentrations for anaesthesia induction and recovery (40–80 mg/L) for seven out of the eight species examined overlap with those reported for other tropical freshwater fish species. Differences in induction and recovery times among treatments were not always independent of fish size for G. bellendenensis, M. splendida, O. lineolatus, and P. mariae, emphasizing the importance of taking biological and environmental factors into account when determining and applying effective clove oil concentrations for anaesthesia in fish. In contrast, long recovery times and mortality for T. tandanus in even relatively low concen- trations suggests that clove oil may not be an appropriate anaesthetic for at least some fish species. In the last two decades, clove-oil-derived eugenol has been freshwater fish species (Keene et al. 1998; Waterstrat 1999; recognized as an effective anaesthetic for fish in aquaculture Sladky et al. 2001; Jayathilake et al. 2003; Small 2003; Cooke and aquatic research (Keene et al. 1998; Ross and Ross 2008). et al. 2004; Hoskonen and Pirhonen 2004; Roubach et al. Clove oil has a variety of biological effects, including anaes- 2005; Kaiser et al. 2006; Simoes~ et al. 2011; Morato-Fernandes thetic properties, with eugenol (4-allyl-2-methoxyphenol) et al. 2013), as well as for freshwater life history stages of eels being the primary component (Chaieb et al. 2007). Clove oil (Walsh and Pease 2002) and salmonids (Woody et al. 2002; is derived from the flower buds, i.e., cloves, of the clove tree Iversen et al. 2003; Hoskonen and Pirhonen 2004). Syzygium aromaticum (L.) Merrill and Perry (Myrtaceae) The freshwater fish fauna of Australia is unique in the (Chaieb et al. 2007). Effective concentrations of clove oil for world (Allen et al. 2003), with a high incidence of endemism anaesthetics have been determined for a variety of marine in the southern, central, and western provinces, as well as (Soto and Burhanuddin 1995; Munday and Wilson 1997; Grif- the North East Queensland region (Unmack 2001). This fiths 2000; King et al. 2005; Mylonas et al. 2005; Cunha and uniqueness attracts freshwater biologists to conduct both Rosa 2006; Park et al. 2008; Gullian and Villanueva 2009) and field and laboratory research on this fauna, with new fish species being discovered on a regular basis (Pusey et al. 2004; Thuesen et al. 2011; Welsh et al. 2014). However, effective † Present address: Australian Institute of Marine Science, Townsville, clove oil concentrations for anaesthetics have been pub- Queensland, Australia *Correspondence: [email protected] lished for only two Australian freshwater fish species, namely This is an open access article under the terms of the Creative Commons the tropical Western rainbowfish (Melanotaenia australis, Mel- Attribution-NonCommercial-NoDerivs License, which permits use and dis- anotaeniidae) (Castelnau 1875) (Young 2009) and the sub- tribution in any medium, provided the original work is properly cited, the tropical longfinned eel (Anguilla reinhardtii, Anguillidae) use is non-commercial and no modifications or adaptations are made. Steindachner 1867 (Walsh and Pease 2002). Interestingly, The copyright line for this article was changed on 23 September 2015 after the effective concentrations for these two species are higher original online publication. compared to non-Australian freshwater fish species from 463 Kroon Clove oil anaesthesia in Australian fish Table 1. Australian tropical freshwater fish species submitted to anaesthesia with clove oil. For each fish species, the mean length (total length, TL) 6 standard error (SE) and range (minimum–maximum), the number of individuals tested for each clove oil concen- tration, the total number of fish tested, and the mean water temperature of acclimation aquaria 6 SE and range (minimum–maxi- mum), are given. Number of fish tested for each clove TL (mm) oil concentration Temperature (8C) Range 20 30 40 80 160 Total # Range Fish species Mean 6 SE (min–max) mg/L mg/L mg/L mg/L mg/L of fish Mean 6 SE (min–max) Glossogobius 45.8 6 1.5 34.8–72.5 76977 3625.3 6 0.1 24.5–26.0 bellendenensis Hypseleotris compressa 44.4 6 1.4 23.0–73.5 15 19 19 20 16 89 25.4 6 0.2 25.0–27.5 Oxyeleotris lineolatus 69.5 6 21.5 34.0–583.0 55555 2525.1 6 0.1 25.0–26.0 Melanotaenia splendida 84.5 6 1.6 49.0–125.0 24 16 24 24 14 102 25.0 6 0.2 22.0–26.5 Pseudomugil signifer 36.4 6 0.8 25.8–49.6 10 10 11 10 12 53 24.5 6 0.1 23.5–25.0 Tandanus tandanus 93.5 6 3.8 67.9–127.1 30645 1825.1 6 0.1 25.0–26.0 Pelmatolapia mariae* 66.1 6 4.2 28.9–128.8 12 19 13 14 12 70 24.8 6 0.0 24.0–26.0 Poecilia reticulata* 24.5 6 0.7 16.7–40.7 4 10 10 10 12 46 24.7 6 0.1 24.0–26.0 *Denotes species non-native to Australia. temperate (Keene et al. 1998; Hoskonen and Pirhonen 2004), belonging to families Gobiidae (Mulgrave goby, Glossogobius subtropical (Waterstrat 1999; Small 2003; Cooke et al. 2004; bellendenensis Hoese and Allen 2009), Eleotridae (empire Mylonas et al. 2005; Morato-Fernandes et al. 2013), and gudgeon, Hypseleotris compressa (Krefft 1864), sleepy cod, tropical regions (Sladky et al. 2001; Jayathilake et al. 2003; Oxyeleotris lineolatus (Steindacher 1867)), Melanotaeniidae Roubach et al. 2005; Kaiser et al. 2006; Simoes~ et al. 2011). (Eastern rainbowfish, Melanotaenia splendida (Peters 1866)), This highlights that freshwater biologists studying Australian Pseudomugilidae (Pacific blue-eye, Pseudomugil signifer Kner fish cannot necessarily rely on clove oil concentrations 1865), Plotosidae (eel-tailed catfish, Tandanus tandanus determined for non-Australian fish, and that further work is Mitchell 1838), Cichlidae (spotted tilapia, Pelmatolapia needed to facilitate the collection, handling and transporta- mariae Boulenger 1899), and Poecilidae (guppy, Poecilia retic- tion of Australian freshwater fish. This is particularly perti- ulata Peters 1859) were examined (Table 1). All species exam- nent for Australia’s tropical north where research on the ined are native to Australia, except for P. mariae and P. freshwater fish fauna is urgently required to minimise the reticulata. potential impacts of proposed agricultural expansion (Slezak The following criteria were used to select the eight spe- 2014). cies: importance to recreational fisheries (e.g., T. tandanus, O. In this study, induction to anaesthesia, recovery from lineolatus), importance to aquarium trade and hobby (e.g., H. anaesthesia and survival rates were determined for eight Aus- compressa, M. splendida, P. signifer), and importance to con- tralian tropical freshwater fish species exposed to five differ- servation, such as endemic (e.g., G. bellendenensis) and non- ent concentrations of clove oil (20 mg/L, 30 mg/L, 40 mg/L, native (e.g., P. mariae, P. reticulata) species. In Queensland, 80 mg/L, and 160 mg/L). The induction and recovery times the six native species are not listed as threatened or endan- were compared for six native species (Pusey et al. 2004), and gered under the Nature Conservation (Wildlife) Regulation two non-native species with established populations in tropi- 2006, while Tilapia spp. are declared noxious fish under the cal Australia (Kroon et al. 2015). Moreover, the effect of fish Fisheries Regulation 2008. Both P. mariae and P. reticulata size on induction and recovery times was examined for all have established populations in Queensland (Department of eight species. The results will contribute to the use of effec- Primary Industries 2001), including in the Wet Tropics tive clove oil concentrations for anaesthesia in both field region (Kroon et al. 2015). and laboratory research on tropical freshwater fish. Fish were collected using seine nets, funnel traps, and backpack electrofishing, in the Barron and Mulgrave river basins in the Wet Tropics region in 2011 and 2012. Since Materials and procedures our collections, a new species of Tandanus has been identi- Description and collection of study species fied to co-occur with T. tandanus in the Mulgrave-Russell This study was conducted at the CSIRO laboratory in river drainage, namely Tandanus tropicanus (Plotosidae) Atherton (latitude 178150, longitude 1458290) in the Wet (Welsh et al. 2014). Our study occurred before the identifica- Tropics region of North Queensland, Australia. Eight species tion of T. tropicanus; hence we use the name T. tandanus. 464 Kroon Clove oil anaesthesia in Australian fish Fish were transported to the laboratory the same day, and 1985; Ross and Ross 2008).
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