Anthelmintic Efficacies of Three Common Disinfectants and Extracts

View metadata, citation and similar papers at core.ac.uk brought to you by CORE

provided by Institute of Hydrobiology, Chinese Academy Of Sciences

Aquaculture 466 (2017) 72–77

Contents lists available at ScienceDirect

Aquaculture

journal homepage: www.elsevier.com/locate/aquaculture

Anthelmintic efﬁcacies of three common disinfectants and extracts of four traditional Chinese medicinal plants against Gyrodactylus kobayashii (Monogenea) in goldﬁsh (Carassius auratus)

Shun Zhou a,b,WenX.Lia, Ya Q. Wang a,b,HongZoua, Shan G. Wu a, Gui T. Wang a,b,⁎

a Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, Hubei Province, PR China b University of Chinese Academy of Sciences, Beijing 100049, PR China

article info abstract

Article history: Gyrodactylids are commonly occurring fish ectoparasites that cause high host mortality and considerable eco- Received 15 August 2016 nomic losses in aquaculture. With the goal of finding safe and effective alternative agents to control monogenean Received in revised form 27 September 2016 infections in aquaculture, we investigated the anthelmintic efficacy of three common disinfectants Accepted 28 September 2016 (trichloroisocyanuric acid, glutaraldehyde and potassium permanganate) and crude extracts of four herbal med- Available online 29 September 2016 icines (Cortex meliae, Radix zanthoxyli, Semen aesculi and Macleaya cordata) against Gyrodactylus kobayashii in goldfish. Also, we compared the anthelmintic efficacy of the above agents with formaldehyde, which is one of Keywords: Monogenean the most frequently-used agents to treat gyrodactyliasis. After 48-h exposure, trichloroisocyanuric acid had the fi Gyrodactylus kobayashii EC50 value of 0.12 mg/L and EC90 value of 0.65 mg/L, and exhibited a 100% ef cacy against G. kobayashii at Common disinfectants 1.0 mg/L. Glutaraldehyde and potassium permanganate were less effective, with EC50 values of 4.0 and 2.9 mg/

Trichloroisocyanuric acid L, and EC90 of 5.0 and 3.6 mg/L, respectively. For the extracts of herbal medicines, EC50 and EC90 values of meth- Herbal medicines anol extract of M. cordata were 8.6 and 25.5 mg/L, respectively, after 48-h exposure, and EC100 of ethyl acetate and Macleaya cordata petroleum ether extracts of M. cordata were 80 and 100 mg/L, respectively. The remaining extracts were either fi Gold sh highly toxic to fish, or exhibited very weak anthelmintic efficacy, even at higher concentrations. Acute toxicity

assay indicated that the LC50 of the promising agents (trichloroisocyanuric acid, glutaraldehyde, potassium permanganate and methanol extract of M. cordata) against goldﬁsh after 48-h exposure were 4.1, 8.8, 5.9 and

81.4 mg/L, which were 6.3, 1.8, 1.6 and 3.2 times higher than the corresponding EC90. The LC50/EC90 ratio for formaldehyde (2.1-fold) was higher than for glutaraldehyde and potassium permanganate, but lower than for trichloroisocyanuric acid and methanol extract of M. cordata. These results suggested that trichloroisocyanuric acid and methanol extract of M. cordata are both viable alternatives for formaldehyde in aquaculture. Statement of relevance: The objective of the present study was motivated by the urgent need to ﬁnd safe and effective alternative agents to control monogenean infections in aquaculture. © 2016 Elsevier B.V. All rights reserved.

1. Introduction symptoms to host fish, including extensive fin damage and thickening of the epidermis (Cone and Odense, 1984). The attachment organ deep- Gyrodactylids are commonly occurring monogenean ectoparasites ly buried into the host epidermis would also destroy the osmotic integ- on the skin, fins and gills of a great variety of marine, freshwater and rity of the epidermis. Such wounds facilitate potential mixed infection brackish fishes (Bakke et al., 2002). These viviparous epidermal para- with other parasites, as well as secondary bacterial and fungal infec- sites can directly transmit from one host to another during the contact tions, which can cause high host mortality and considerable economic between hosts, and have the capability of continuous transmission dur- losses in aquaculture (Bakke et al., 2007; Cone and Odense, 1984; ing the entire lifetime, which allows them to rapidly colonize new hosts Heggberget and Johnsen, 1982). (Boeger et al., 2005). Their short generation time and viviparous repro- To date, many chemical compounds and treatment protocols, in- duction in situ on the host can lead to exponential population growth in cluding formaldehyde, rotenone and praziquantel, were used to control intensive aquaculture (Cable and van Oosterhout, 2007; Jorgensen et al., Gyrodactylus infections with varying levels of success (Harris et al., 2009). Heavy infection with Gyrodactylus may lead to noticeable 2008; Rowland et al., 2006; Santamarina et al., 1991). However, these compounds are often accompanied by serious drawbacks, such as ad- verse environmental impacts, toxicity to the host, and even contamina- ⁎ Corresponding author at: Institute of Hydrobiology, Chinese Academy of Sciences, fi Wuhan 430072, Hubei Province, PR China. tion of sh products with drug residues (Srivastava et al., 2004). In E-mail address: [email protected] (G.T. Wang). addition, prolonged and frequent use of these chemicals can result in

Table 1 at 48-h intervals. Ectoparasite-free fish were kept in aquaria for Characterization of herbal medicines used in anthelmintic efficacy tests against 30 days after the cleaning procedure, then ten goldfish were randomly Gyrodactylus kobayashii. taken, anesthetized with 0.02% MS222 (tricaine methane sulfonate), Herbal medicine Plant name Test part Origin and examined to confirm the ectoparasite-free status under a stereomi- fi Cortex meliae Melia azedarach Bark Sichuan, China croscope. Gold sh-G. kobayashii model was established according to the Radix zanthoxyli Zanthorμlum nitidum Root Yunnan, China previously described technique (Harris, 1988). In brief, an uninfected Semen aesculi Aesculus chinensis Seed Zhejiang, China fish, anesthetized with 0.02% MS222, was placed near the caudal finof Macleaya cordata Macleaya cordata Whole plant Yunnan, China an infected fish, allowing an individual parasite to transfer. After the success of transmission and proliferation, two parasites were collected from the infected fish for morphological and molecular identification the evolution of drug resistance (Buchmann et al., 1992). Therefore, the to confirm they are G. kobayashii (Cable et al., 1999; Li et al., 2014). To development of effective and safe alternative agents to control the dis- obtain more infected goldfish, ectoparasite-free goldfish were intro- eases caused by Gyrodactylus in the intensive aquaculture is urgently duced periodically into the aquarium with goldfish infected with G. needed. kobayashii. Only those fish with medium intensity (40–60) and ap- Plant-based medicines are attracting increasing attention as an alter- peared to be healthy (normal swimming behavior and undamaged native strategy to control diseases in aquaculture, due to their demon- gill, fin and scale) were selected for the drug experiment. strable efficacy, environmental friendliness and biodegradability (Rahuman et al., 2008). For example, Levy et al. (2015) explored the 2.2. Preparation of plant materials and drugs possibility of using ginger (Zingiber officinale)totreatGyrodactylus turnbulli infection in guppy Poecilia reticulata. Several studies also re- The herbal medicines (C. meliae, R. zanthoxyli, S. aesculi and M. ported anthelmintic activity of garlic extracts/compounds against cordata)(seeTable 1) were purchased from a local drug store, washed Gyrodactylus spp. in guppies (Fridman et al., 2014; Schelkle et al., thoroughly, air-dried and finally oven-dried at 45 °C for 48 h. The dry 2013). Likewise, common disinfectants, such as hydrogen peroxide plant materials were then powdered, strained with filter net of 30– and sodium percarbonate, showed high anthelmintic efficacy to treat 40 mesh (450–600 μm), and freeze-dried at −54 °C to ensure the com- monogenean infections in aquaculture (Bowker et al., 2012; plete removal of water. Batches of 50 g of each prepared plant material Buchmann and Kristensson, 2003). were then extracted with four different solvents: methanol, ethyl ace- Therefore, with the goal of finding safe and effective alternative tate, petroleum ether or distilled water for 48 h (each 0.5 L), and the agents to control monogenean infections in aquaculture, in this study process was repeated three times. To obtain solidified crude extracts, we investigated the anthelmintic efficacy of three common disinfec- the extract solutions were filtered and concentrated under reduced tants (trichloroisocyanuric acid, glutaraldehyde and potassium perman- pressure in a vacuum rotary evaporator (RE-52aa, Shanghai Yarong) ganate) and crude extracts of four herbal medicines (Cortex meliae, until all solvents had evaporated. All solid extracts were dissolved in Radix zanthoxyli, Semen aesculi and Macleaya cordata) against G. dimethyl sulfoxide (DMSO) to obtain 0.5 mg/L (sample/solution) kobayashii in goldfish (Carassius auratus). Additionally, to assess their stocking solution for anthelmintic efficacy assay. Formalin (37% formal- applicability in practice, we would compare the anthelmintic efficacies dehyde reagent), 25% glutaraldehyde and potassium permanganate and toxicities of the above agents with one of the most frequently- were obtained from Sinopharm Chemical Reagent Co. Ltd., and used agents to treat gyrodactyliasis–formaldehyde. trichloroisocyanuric acid (98.5% purity, J&K®) was purchased from Bei- jing Bailingwei Chemical Technology Co. Ltd. The anthelmintic efficacy 2. Materials and methods of the above four compounds was also assessed.

2.1. Fish and parasites 2.3. In vivo anthelmintic activity against G. kobayashii

Goldfish with mean body weight of 4.32 ± 0.87 g were purchased Two goldfish infected with G. kobayashii were randomly taken from from a local fish farm in Wuhan city, China and kept in several 100-L the same batch of infected goldfish and placed in 20 × 12 × 10 cm tanks aquariums equipped with aerators (water temperature 19.0–21.0 °C, containing 0.5 L of test solution at 20 ± 1 °C for 48 h exposure. Two con- pH 6.9–7.3). After an acclimation for seven days, fish were treated trol groups were set up under the same conditions: one with no agents with three consecutive baths in 1:10,000 formalin solutions for 12 h, added, while the other one contained 0.2% DMSO. The latter was

120

100

Mean anthelmintic efficacy (%) 0

Concentration (mg/L)

Fig. 1. Mean anthelmintic efﬁcacy of four chemical compounds against Gyrodactylus kobayashii after 48 h of exposure. F, formaldehyde; T, trichloroisocyanuric acid; G, glutaraldehyde; P, potassium permanganate. Star indicates that the agent causes ﬁsh mortality. 74 S. Zhou et al. / Aquaculture 466 (2017) 72–77

120

100

Mean anthelminthic efficacy (%) 0

Concentration (mg/L)

Fig. 2. Mean anthelmintic efficacy of different Cortex meliae extracts against Gyrodactylus kobayashii after 48 h of exposure. MEE, methanol extract; EAE, ethyl acetate extract; PEE, petroleum ether extract; WAE, water extract. Star indicates that the extract causes fish mortality. included in order to eliminate the possible influence of DMSO on which fish mortalities begin to occur, to the concentration where mor- parasites. All treatments with different concentrations of the drugs tality is 100%. Control groups were conducted without the tested agents (Figs. 1–5) and the control groups were performed in five replicates. Ap- under the same conditions, and three replicate tanks were used for all propriate concentrations were estimated based on the preliminary ex- treatment groups. Fish mortalities in each tank were recorded after periments. Before treatment and after the 48-h treatment, the goldfish 48-h of exposure. To avoid deterioration of the water quality, fish was were anesthetized with 0.02% MS222 and the number of gyrodactylids not fed during the experiment and dead fish was removed periodically. in the caudal fin was counted under a stereomicroscope. Anthelmintic efficacies of each drug were calculated according to the 2.5. Statistical analyses following formula: E = (L − Lt)/L×100%forLN Lt,E=0forL≤ Lt.Eis anthelmintic efficacy, L is the number of G. kobayashii on the caudal fin The homogeneity of the replicates of the samples was assessed using before treatment, and Lt is the number of surviving parasites after the the Mann-Whitney U test. Median effective concentration (EC50,EC90) treatment. Mean anthelmintic efficacies of each drug with different con- and median lethal concentration (LC50,LC90) with the 95% confidence centrations were also recorded according to the five replicates. levels were calculated by Probit analysis. All analyses were performed using SPSS 19.0. 2.4. Acute toxicity tests 3. Results Based on the results of above experiment, the experimental agents or extracts with the best anthelmintic efficacy against G. kobayashii 3.1. The anthelmintic efficacy of experimental agents against G. kobayashii were selected to further evaluate the safety to goldfish: formaldehyde, in vivo trichloroisocyanuric acid, glutaraldehyde, potassium permanganate and three extracts of M. cordata (methanol, ethyl acetate, petroleum Trichloroisocyanuric acid was the most effective among the tested ether). Acute toxicity tests to goldfish were conducted in 26.5 × drugs, with the EC50 value of 0.12 mg/L and EC90 value of 0.65 mg/L 16.5 × 12.5 cm tanks at 20 ± 1 °C, containing ten uninfected goldfish after 48 h of exposure, and it exhibited 100% efficacy against G. and 2 L of aerated tap water, to which a variety of drugs were added kobayashii at 1.0 mg/L. Glutaraldehyde and potassium permanganate to achieve the desired experimental concentrations. Preliminary exper- were partly effective in eliminating G. kobayashii, with EC50 values of iments were performed to establish the range of concentrations at 4.0 and 2.9 mg/L, and EC90 values of 5.0 and 3.6 mg/L, respectively.

120

100

Mean anthelmintic efficacy (%) 0

Concentration (mg/L)

Fig. 3. Mean anthelmintic efﬁcacy of different Radix zanthoxyli extracts against Gyrodactylus kobayashii after 48 h of exposure. MEE, methanol extract; EAE, ethyl acetate extract; PEE, petroleum ether extract; WAE, water extract. Star indicates that the extract causes ﬁsh mortality. S. Zhou et al. / Aquaculture 466 (2017) 72–77 75

120

100

Mean anthelmintic efficacy (%) 0

Concentration (mg/L)

Fig. 4. Mean anthelmintic efﬁcacy of different Semen aesculi extracts against Gyrodactylus kobayashii after 48 h of exposure. MEE, methanol extract; EAE, ethyl acetate extract; PEE, petroleum ether extract; WAE, water extract. Star indicates that the extract causes ﬁsh mortality.

Formaldehyde had EC50 and EC90 values of 10.5 and 23.4 mg/L, respec- permanganate) were 48.1, 4.1, 8.8 and 5.9 mg/L, which were 2.1, 6.3, tively, and showed 100% efficacy against G. kobayashii at 35 mg/L 1.8 and 1.6 times higher than the corresponding EC90, respectively (Table 2; Fig. 1). (Table 3). For the methanol extract of M. cordata,theLC50 (81.4 mg/L) Among the herbal extracts, methanol extract of M. cordata was the was 3.2-fold larger than EC90. The ethyl acetate and petroleum ether ex- most effective, with EC50 and EC90 values of 8.6 and 25.5 mg/L, respec- tracts exhibited a very weak toxicity to goldfish, with only one and two tively, after 48-h exposure. Ethyl acetate and petroleum ether extracts fish died at the concentrations of 1000 and 2000 mg/L, respectively of M. cordata exhibited 100% anthelmintic efficacy at 80 and 100 mg/L, (Table 3). and had EC50 and EC90 values of 30.4 and 57.7 mg/L (ethyl acetate), and 43.2 and 87.2 mg/L (petroleum ether), respectively. The water ex- 4. Discussion tract exhibited a weak activity with anthelmintic efficacy of 70.5% at the high concentration of 300 mg/L (Table 2; Fig. 5). The results suggested that, among the tested drugs, For C. meliae, R. zanthoxyli and S. aesculi, only the methanol extract of trichloroisocyanuric acid is the most effective against G. kobayashii,

C. meliae displayed good anthelmintic efficacy after the exposure for with EC90 value of 0.65 mg/L after 48-h exposure. Trichloroisocyanuric 48 h, albeit at high concentrations: EC50 =237.8andEC90 = acid is widely used as a disinfectant, because its primary decomposition 381.8 mg/L. The remaining extracts were either highly toxic to fish, or product, hypochlorous acid, is an effective biocidal agent against bacte- exhibited very weak anthelmintic efficacy, even at higher concentra- ria, viruses and fungi (Azanza et al., 2001; Frerichs et al., 2000; tions (Table 2; Figs. 2–4). In the two control groups, there were both ob- Khomvilai et al., 2006). However, little is known about its biocidal activ- vious increases in mean intensity on the caudal of goldfish after two day ity against monogenean parasites, so the present study is the first at- (from 51.7 ± 7.5 to 81.5 ± 17.3 for no drug group, from 53.4 ± 8.8 to tempt to examine the anthelmintic efficacy of trichloroisocyanuric

71.5 ± 17.4 for DMSO group), which suggested no anthelmintic efﬁcacy acid against G. kobayashii. The toxic dose (LC50) is about six times higher against G. kobayashii. than the effective one (EC90), which is a much higher ratio than that of formaldehyde (2.1-fold). The active ingredient of trichloroisocyanuric 3.2. Acute toxicity of the experimental agents to the goldﬁsh acid, hypochlorous acid, penetrates the cell relatively easily, where it re- acts with amino groups of amino acids and proteins resulting in death of

The LC50 values after 48 h of exposure of the four chemicals (formal- parasites (Noguchi et al., 2002). In addition, the biocidal activity of dehyde, trichloroisocyanuric acid, glutaraldehyde and potassium trichloroisocyanuric acid can reduce the potential secondary bacterial,

120

100

Mean anthelmintic efficacy (%) 0

Concentration (mg/L)

Fig. 5. Mean anthelmintic efﬁcacy of different Macleaya cordata extracts against Gyrodactylus kobayashii after 48 h of exposure. MEE, methanol extract; EAE, ethyl acetate extract; PEE, petroleum ether extract; WAE, water extract. 76 S. Zhou et al. / Aquaculture 466 (2017) 72–77

Table 2

EC50 and EC90 of four chemicals and different extracts of four Chinese medicinal plants against Gyrodactylus kobayashii after 48 h of exposure. EC50, effective concentration with 50% an- 2 thelmintic efficacy; EC90, effective concentration with 90% anthelmintic efficacy; 95% CI, 95% confidence interval; χ ,chi-square;–, no solvent.

2 Drug or plant material Solvent EC50 (mg/L) 95% CI EC90 (mg/L) 95% CI χ Formaldehyde – 10.5 6.1–13.1 23.4 21.8–25.6 2.06 Trichloroisocyanuric acid – 0.12 0.01–0.21 0.65 0.58–0.74 3.69 Glutaraldehyde – 4.0 3.8–4.1 5.0 4.9–5.3 2.08 Potassium permanganate – 2.9 2.7–3.2 3.6 3.3–4.3 13.13 Cortex meliae Methanol 237.8 221.2–253.8 381.8 359.1–410.6 5.15 Petroleum ether 155.6 129.1–196.8 221.9 186.7–419.8 14.99 Radix zanthoxyli Petroleum ether 214.5 182.0–249.1 296.0 258.6–391.2 13.09 Semen aesculi Methanol 68.0 58.8–96.7 103.7 83.0–191.2 7.05 Ethyl acetate 369.5 0–738.3 777.8 542.6–4272.4 8.01 Petroleum ether 332.2 261.7–412.5 515.7 429.6–754.9 13.37 Water 165.1 144.7–242.0 232.6 191.5–517.5 11.97 Macleaya cordata Methanol 8.6 4.7–11.3 25.5 23.0–28.8 2.16 Ethyl acetate 30.4 22.7–40.6 57.7 48.6–74.4 6.21 Petroleum ether 43.2 7.9–61.7 87.2 67.0–175.2 17.97 Water 219.2 197.8–241.2 427.8 376.6–518.4 1.31

viral and fungal infections caused by G. kobayashii. These results indicat- resulted in 100% mortality at 10 mg/L. The discrepancy among these re- ed that trichloroisocyanuric acid could be an effective alternative agent sults indicated that insecticide activity of potassium permanganate is for controlling monogenean infections in aquaculture. highly dependent on the concentration and parasite species. Similarly, glutaraldehyde is widely used as a sterilizing agent against Macleaya cordata, widely distributed in the northwest and south- bacteria, fungi and viruses (Mainous et al., 2011; Muller et al., 2011; west of China, has been used as a traditional Chinese medicine for its Simoes et al., 2011). To date, little is available about the anthelmintic ef- analgesic and anti-inflammatory properties in humans. The extracts of ficacy of glutaraldehyde in aquaculture. In this study, we found it had a M. cordata exhibit an extensive range of bioactivities, including antimi- high anthelmintic efficacy against G. kobayashii of almost 100% at the crobial (Kosina et al., 2010), antiviral (Kerry et al., 1998), and insecticid- dose of 6.0 mg/L. However, exposure to 6.0 mg/L glutaraldehyde for al properties (Wang et al., 2012; Yao et al., 2010). The extract of M.

48 h caused fish mortality and the value (LC50/EC90) of glutaraldehyde cordata-sanguinarine was 100% effective against Ichthyophthirius was lower than that of formaldehyde, which is one of the most fre- multifiliis at the concentration of 0.7 mg/L after 4 h of exposure in in quently-used agents to treat the monogenean infections. This indicated vitro antiparasitic efficacy tests, and exposure to 0.9 mg/L sanguinarine that the anthelmintic efficacy of glutaraldehyde under safe concentra- for 48 h in in vivo antiparasitic efficacy tests could reduce 96.8% of I. tions was probably not better than that of formaldehyde. Even though multifiliis burden in grass carp (Ctenopharyngodon idella)(Yao et al., Wilson and Margolin (1999) reported that a 2.5% glutaraldehyde prod- 2010). Wang et al. (2012) also reported that three kinds of alkaloids uct was effective in halting the excystation of Cryptosporidium parvum from M. cordata, sanguinarine, chelerythrine and allocryptopine, sporozoites and subsequent infection after 10-h exposure, due to the were effective in killing the nematodes Bursaphelenchus xylophilus, observed toxicity to fish, glutaraldehyde should be used very cautiously Caenorhabditis elegans and Meloidogyne incognita at low concentrations in aquaculture. after a 24-h treatment. In our trial, methanol extract of M. cordata was In our trials, potassium permanganate was partly effective in found to be the most effective against G. kobayashii with the lowest controlling G. kobayashii. Even though it had a very high anthelmintic EC90 at 25.5 mg/L after a 48-h exposure and the toxic dose (LC50)of efficacy (100%) at 4.0 mg/L, which was also capable of causing fish methanol extract of M. cordata is about three times the EC90.Thesere- mortality. In treatment of protozoan infections, potassium permanga- sults indicated that methanol extract of M. cordata extracts is promising nate only slightly reduced the loads of Ichthyobodo necator on sunshine insecticidal agents but with some toxic to fish. In addition, of the three bass (female white bass Morone chrysops × male striped bass extracts of M. cordata,theEC90 and LC50 of methanol extract were M. saxatilis) after 2 days of exposure, and the parasite loads both the lowest. This means that more active ingredients in M. cordata raised again over the next three days (Mitchell et al., 2008). However, are easily extracted by methanol, and the active ingredients against Straus and Griffin(2001)demonstrated that the exposure to 0.9 mg/L G. kobayashii are also toxic to fish. potassium permanganate for 4 h resulted in over 95% mortality of Semen aesculi is also a traditional Chinese medicine used for the Ichthyophthirius multifiliis theronts, and the dose of 1.0 mg/L could pre- treatment of distention and pain in chest and abdomen. Liu et al. vent the initial infestation of juvenile channel catfish and blue tilapia by (2010) have reported that both methanol and aqueous extracts of S. this parasite. Hakalahti-Siren et al. (2008) also reported that both juve- aesculi were effective against Dactylogyrus intermedius (Monogenea), nile and adult fish lice Argulus coregoni were highly sensitive to potassi- with EC50 values of 5.23 and 12.29 mg/L (respectively), after 48-h expo- um permanganate treatments, exposure to which for over 30 min sure in in vivo anthelmintic efficacy assays. However, in our trial, we

Table 3 Acute toxicity for goldﬁsh exposed to four disinfectants (formaldehyde, trichloroisocyanuric acid, glutaraldehyde and potassium permanganate) and three extracts (methanol, ethyl ac- 2 etate and petroleum ether) of Macleaya cordata after 48 h of exposure. LC50, 50% lethal concentration; LC90, 90% lethal concentration; 95% CI, 95% conﬁdence interval; χ ,chi-square;–,not calculated.

2 Drugs LC50 (mg/L) 95% CI LC90 (mg/L) 95% CI χ Formaldehyde 48.1 44.5–51.6 61.5 57.1–68.9 3.01 Trichloroisocyanuric acid 4.1 3.7–4.5 5.6 5.1–6.4 5.27 Glutaraldehyde 8.8 8.1–9.5 11.6 10.7–13.3 4.61 Potassium permanganate 5.9 5.5–6.3 7.4 6.9–8.2 2.06 Methanol extract 81.4 78.0–84.9 94.2 89.9–101.7 2.46 Ethyl acetate extract N1000 –– – – Petroleum ether extract N1000 –– – – S. Zhou et al. / Aquaculture 466 (2017) 72–77 77 found a very weak anthelmintic activity of the extracts of S. aesculi Harris, P.D., 1988. Changes in the site specificity of Gyrodactylus turnbulli Harris, 1986 (Monogenea) during infections of individual guppies (Poecilia reticulata Peters, against G. kobayashii. The contrasting results might indicate that the an- 1859). Can. J. Zool. 66, 2854–2857. thelmintic efficacy of S. aesculi was highly parasitic species-specific. Harris, P.D., Shinn, A.P., Cable, J., Bakke, T.A., Bron, J., 2008. GyroDb: gyrodactylid monoge- In conclusion, trichloroisocyanuric acid and methanol extract of M. neans on the web. Trends Parasitol. 24, 109–111. fi Heggberget, T.G., Johnsen, B.O., 1982. Infestations by Gyrodactylus sp of Atlantic Salmon, cordata both exhibited high anthelmintic ef cacy against G. kobayashii Salmo salar L., in Norwegian rivers. J. Fish Biol. 21, 15–26. in goldfish. However, more researches such as further bioassay-guided Jorgensen, T.R., Raida, M.K., Kania, P.W., Buchmann, K., 2009. Response of rainbow trout isolation and purification of compounds responsible for the observed (Oncorhynchus mykiss)inskinandfin tissue during infection with a variant of – anthelmintic efficacy, the detailed mechanism of anthelmintic activity Gyrodactylus salaris (Monogenea: Gyrodactylidae). Folia Parasitol. 56, 251 258. Kerry, M.A., Duval, O., Waigh, R.D., Mackay, S.P., 1998. The role of the iminium bond in the and field experiments should be conducted before the application in inhibition of reverse transcriptase by quaternary benzophenanthridines. J. Pharm. aquaculture. Pharmacol. 50, 1307–1315. Khomvilai, C., Kashiwagi, M., Sangrungruang, C., Yoshioka, M., 2006. Preventive efficacy of sodium hypochlorite against water mold infection on eggs of chum salmon Onco- Compliance with ethical standards rhynchus keta. Fisheries Sci. 72, 28–32. Kosina, P., Gregorova, J., Gruz, J., Vacek, J., Kolar, M., Vogel, M., Roos, W., Naumann, K., All applicable international, national, and/or institutional guidelines Simanek, V., Ulrichova, J., 2010. Phytochemical and antimicrobial characterization of Macleaya cordata herb. Fitoterapia 81, 1006–1012. for the care and use of animals were followed. Levy, G., Zilberg, D., Paladini, G., Fridman, S., 2015. Efficacy of ginger-based treatments against infection with Gyrodactylus turnbulli in the guppy (Poecilia reticulata (Pe- ters)).Vet.Parasitol.209,235–241. Acknowledgements Li, R.R., Li, W.X., Wu, X.D., Wang, G.T., 2014. Identification of Gyrodactylus species in goldfish (Carassius auratus) through morphological study and the analysis of rDNA ITS se- This work was supported by the Earmarked Fund for China quence. Acta Hydrobiol Sin. 38, 903–909 (Chinese). Agriculture Research System (CARS-46-08), the Major Scientificand Liu, Y.T., Wang, F., Wang, G.X., Han, J., Wang, Y., Wang, Y.H., 2010. In vivo anthelmintic activity of crude extracts of Radix angelicae pubescentis, Fructus bruceae, Caulis Technological Innovation Project of Hubei Province (2013ABA004, spatholobi, Semen aesculi,andSemen pharbitidis against Dactylogyrus intermedius 2015ABA045), and the National Natural Science Foundation of China (Monogenea) in goldfish (Carassius auratus). Parasitol. Res. 106, 1233–1239. fi (31172409, 31272695). The authors also thank Dr. Jakovlić I. for his Mainous, M.E., Kuhn, D.D., Smith, S.A., 2011. Ef cacy of common aquaculture compounds for disinfection of Aeromonas hydrophila, A. salmonicida subsp. salmonicida, and A. help in English language editing. salmonicida subsp achromogenes at various temperatures. N. Am. J. Aquac. 73, 456–461. References Mitchell, A.J., Darwish, A., Fuller, A., 2008. Comparison of tank treatments with copper sul- fate and potassium permanganate for sunshine bass with Ichthyobodosis. J. Aquat. Anim. Health 20, 202–206. Azanza, M.P.V., Azanza, R.V., Gedaria, A.I., Sententa, H.G., Idjao, M.V., 2001. Decimal reduc- Muller, M., Ilardi, P., Avendano-Herrera, R., 2011. Efficacy of a commercial disinfectant tion times of Pyrodinium bahamense var. compressum and Escherichia coli in chlorine- against Vibrio ordalii, Vibrio anguillarum, Francisella sp. and Infectious Pancreatic Ne- and ultraviolet-treated seawater. Lett. Appl. Microbiol. 33, 371–376. crosisVirus(IPNV)pathogensofAtlanticsalmon(Salmo salar) farmed in Chile. Bakke, T.A., Harris, P.D., Cable, J., 2002. Host specificity dynamics: observations on Arch Med Vet. 43, 73–78. gyrodactylid monogeneans. Int. J. Parasitol. 32, 281–308. Noguchi, N., Nakada, A., Itoh, Y., Watanabe, A., Niki, E., 2002. Formation of active oxygen Bakke, T.A., Cable, J., Harris, P.D., 2007. The biology of gyrodactylid monogeneans: the species and lipid peroxidation induced by hypochlorite. Arch. Biochem. Biophys. “Russian-doll killers”. Adv. Parasitol. 64, 161–376. 397, 440–447. Boeger, W.A., Kritsky, D.C., Pie, M.R., Engers, K.B., 2005. Mode of transmission, host Rahuman, A.A., Gopalakrishnan, G., Venkatesan, P., Geetha, K., 2008. Larvicidal activity of switching, and escape from the red queen by viviparous gyrodactylids some Euphorbiaceae plant extracts against Aedes aegypti and Culex quinquefasciatus (monogenoidea). J. Parasitol. 91, 1000–1007. (Diptera: Culicidae). Parasitol. Res. 102, 867–873. Bowker, J.D., Carty, D., Dotson, M.M., 2012. Efficacy of 35% PEROX-AID (hydrogen perox- Rowland, S.J., Nixon, M., Landos, M., Mifsud, C., Read, P., Boyd, P., 2006. Effects of formalin ide) in reducing an infestation of Gyrodactylus salmonis in freshwater-reared rainbow on water quality and parasitic monogeneans on silver perch (Bidyanus bidyanus trout. N. Am. J. Aquac. 74, 154–159. Mitchell) in earthen ponds. Aquacult. Res. 37, 869–876. Buchmann, K., Kristensson, R.T., 2003. Efficacy of sodium percarbonate and formaldehyde Santamarina, M.T., Tojo, J., Ubeira, F.M., Quinteiro, P., Sanmartin, M.L., 1991. Anthelmintic bath treatments against Gyrodactylus derjavini infestations of rainbow trout. N. Am. treatment against Gyrodactylus sp. infecting rainbow trout Oncorhynchus mykiss.Dis. J. Aquac. 65, 25–27. Aquat. Organ. 10, 39–43. Buchmann, K., Roepstorff, A., Waller, P.J., 1992. Experimental selection of mebendazole- Schelkle, B., Snellgrove, D., Cable, J., 2013. In vitro and in vivo efficacy of garlic compounds resistant gill monogeneans from the European eel, Anguilla anguilla L. J. Fish Dis. 15, against Gyrodactylus turnbulli infecting the guppy (Poecilia reticulata). Vet. Parasitol. 393–400. 198, 96–101. Cable, J., van Oosterhout, C., 2007. The impact of parasites on the life history evolution of Simoes, L.C., Lemos, M., Araujo, P., Pereira, A.M., Simoes, M., 2011. The effects of glutaral- guppies (Poecilia reticulata): the effects of host size on parasite virulence. Int. dehyde on the control of single and dual biofilms of Bacillus cereus and Pseudomonas J. Parasitol. 37, 1449–1458. fluorescens. Biofouling 27, 337–346. Cable, J., Harris, P.D., Tinsley, R.C., Lazarus, C.M., 1999. Phylogenetic analysis of Srivastava, S., Sinha, R., Roy, D., 2004. Toxicological effects of malachite green. Aquat. Gyrodactylus spp. (Platyhelminthes: Monogenea) using ribosomal DNA sequences. Toxicol. 66, 319–329. Can. J. Zool. 77, 1439–1449. Straus, D.L., Griffin, B.R., 2001. Prevention of an initial infestation of Ichthyophthirius Cone, D.K., Odense, P.H., 1984. Pathology of 5 species of Gyrodactylus Nordmann, 1832 multifiliis in channel catfish and blue tilapia by potassium permanganate treatment. (Monogenea). Can. J. Zool. 62, 1084–1088. N. Am. J. Aquac. 63, 11–16. Frerichs, G.N., Tweedie, A., Starkey, W.G., Richards, R.H., 2000. Temperature, pH and elec- Wang, K., Luo, C., Liu, H., Xu, J., Sun, W., Zhou, L., 2012. Nematicidal activity of the alkaloids trolyte sensitivity, and heat, UV and disinfectant inactivation of sea bass from Macleaya cordata against certain nematodes. Afr. J. Agric. Res. 7, 5925–5929. (Dicentrarchus labrax) neuropathy nodavirus. Aquaculture 185, 13–24. Wilson, J.A., Margolin, A.B., 1999. The efficacy of three common hospital liquid germicides Fridman, S., Sinai, T., Zilberg, D., 2014. Efficacy of garlic based treatments against monoge- to inactivate Cryptosporidium parvum oocysts. J. Hosp. Infect. 42, 231–237. nean parasites infecting the guppy (Poecilia reticulata (Peters)). Vet. Parasitol. 203, Yao, J.Y., Shen, J.Y., Li, X.L., Xu, Y., Hao, G.J., Pan, X.Y., Wang, G.X., Yin, W.L., 2010. Effect of 51–58. sanguinarine from the leaves of Macleaya cordata against Ichthyophthirius multifiliis in Hakalahti-Siren, T., Mikheev, V.N., Valtonen, E.T., 2008. Control of freshwater fish louse grass carp (Ctenopharyngodon idella). Parasitol. Res. 107, 1035–1042. Argulus coregoni: a step towards an integrated management strategy. Dis. Aquat. Organ. 82, 67–77.