Open Access

Use of propofol as an anesthetic and its efficacy on some hematological values of ornamental fish Carassius auratus

SpringerPlus20132:76

DOI: 10.1186/2193-1801-2-76

Received: 29 November 2012

Accepted: 25 February 2013

Published: 4 March 2013

Abstract

The aim of this study was to determine the level of anesthesia attained in Carassius auratus using a propofol bath administration and using values of haematological profile of blood and examinations, to assess the effects of the fish exposure to that anaesthetic. Acute toxicity values of propofol for gold fish were found 96 h LC50 6.353 mg/L, 96 h LC1 2.966 mg/L and 96 h LC99 13.609 mg/L. Time to induce anesthesia in propofol experiment was significantly higher than Clove oil (p < 0.05), but there was no significant difference in recovery time between the experiments. No significant decrease was found in Total RBC, WBC, HCT, MCH, MCV and leukogram indices (p > 0.05). MCHC (%) level of propofol experiment (13.93 ± 1.36) showed significant (p < 0.05) decrease than Clove oil anesthesia (94.95 ± 24.50) and control (62.46 ± 21.90). Hb(g/dl) content (5.20 ± 0.73) showed decrease in propofol exposure compared with control (15.41 ± 4.76) and clove oil experiment (25.39 ± 5.73) (p < 0.05).

Keywords

Anesthesia Gold fish Hematology Ornamental Propofol

Introduction

In recent years, different types of anesthetics are used to aid in the capture, handling, artificial reproduction, surgery procedures and transport of fish as an anti-stress in modern aquaculture (Roubach et al. 2005).

A few number of anesthetics have proved effective in anaesthetisation of fish with its own advantages and drawbacks (Vel&#x00ED;&#x0161;ek et al. 2006). Till now, only MS-222 (tricaine methanesulfonate) is registered for use on food fish in the U.S. and the United Kingdom. However, aquaculture industry needs more compounds to be evaluated experimentally (Coyle et al. 2004) and introduce on ornamental and food fish.

Some anaesthetics reduce or block the activation of the hypothalamic-pituitary-interrenal (HPI) axis associated with stressors and thus decrease or prevent the release of the stress hormone cortisol to the bloodstream of fish (Hoskonen and Pirhonen 2006).

Anesthetics act on the central nervous system in such a way with placing the fish into an anaesthetic solution that is absorbed through the gills and enters the arterial blood, then with the returning of the anaesthetised fish to the fresh water, the anaesthetics or their metabolites are excreted via the gills (Ross and Ross 1999).

Propofol (2,6 diisopropyl phenol) is an ultra-short–acting sedative agent with no analgesic properties, which provides sedative and anesthetic effects (Kay and Stephenson 1980). Propofol is being widely used as an anaesthetic drug in human patients (Andrews et al., 1997). It is reported to reduce both sympathetic and parasympathetic tone; however, it is not clear whether the changes in heart rate variability are associated with depth of anesthesia. It is generally considered safe for use in animals with renal or hepatic disease also most in instances of mild to moderate heart disease with appropriate monitoring and support.

Propofol is a short-acting, rapidly metabolized agent, which is characterized by a virtual lack of any cumulative effect and by rapid recovery after its administration in bolus doses or by continuous infusion. It provides a reliable, rapid and smooth induction of anaesthesia, adequate hypnosis and analgesia for surgical interventions and minimal suppression of vital organ functions. Moreover, recovery is observed to be rapid, uncomplicated and complete.

Propofol has been used for inducing anesthesia in reptiles such as green iguanas (Knotkova et al. 2005(Fleming et al.) and some fish species such as Acipencer oxyrinchus De soti 2003) and in spotted bamboo sharks () (Miller et al.Chiloscyllium plagiosum 2005), but the efficacy and safety of any anaesthetic agent vary among species, life stages and environmental conditions, and more studies are needed to take advantage of this anesthetic in ornamental fish.

The aim of this study was to compare the effectiveness of propofol to that of the commonly employed clove oil, as an anesthetic for Carassius auratus. Gold fish were exposed to varying doses of propofol to determine the 96-h LC50, as well as exposure to both propofol and eugenol to observe differences in anesthesia onset and recovery times, to determine the proper dosage and to evaluate selected blood parameters during anaesthesia with propofol in gold fish (Carassius auratus).

Method and material

In the study, propofol (pofol 1%) manufactured by the Dongkook Pharm Company (Choong cheong Book-Do, Korea) in 50 mL containers was used. Present research performed with the approval of an appropriate ethics committee INTL K3525.A35 B37 2000.

Experimental fish

Approximately 207 sexually immature, gold fish were used, with an average weight of 8 ± 2 g (mean ± SD) and a mean fork length of 100 ± 20 mm. The fish population was distributed equally among ten 50–L holding tanks, each maintained at 22°C with well aeration. The fish were maintained on a lighting regimen representative of the local natural environment (13 L: 11 D) and fed twice daily to satiation with commercially available flaked tropical fish food. Tanks were siphoned once every second day and approximately 2 L of water was exchanged during each cleaning.

Acute toxicity of propofol

Acute toxicity of propofol was ascertained by the OECD 203 “Fish, acute toxicity test” for the 96 h LC50 trials. At first Experimental fish (n = 72) were exposed to concentrations 0.5, 1, 2, 4, 8 and 16, mg/L dissolved in dechlorinated tap water and controls were placed in dechlorinated tap water with no tested substance added in five glass aquaria (50 cm × 26 cm × 30 cm) filled to a volume of 20 L. Twelve gold fish were randomly used for each concentration and for the control group in 2 replicate. The fish and its behavior, water temperature, pH and oxygen saturation were monitored throughout the tests at individual concentrations and in the control aquarium. The total mortalities, behaviors, temperature, and oxygen saturation were recorded every hour for the first 12 h of the experiment, every 3 h for the next 12 h, and every 6 h for the remaining 72 h. Fish were considered dead when there were no opercular beats observed for 15 continuously monitored min. This complete experimental protocol was replicated three times.

Mean lethal concentration at 96 h LC50 also 96 h LC1and 96 h LC99 was calculated from mortality rates over the period of 96 hours by the EPA probit analysis program version 1.5 software.

Onset and recovery from anesthesia

The observations of stages 5-anesthesia onset were made using propofol and clove oil under the same experimental conditions. A 20- L experimental aquarium was maintained at a temperature of 22°C with oxygen saturation greater than 85%. Gold fish (n = 135) were randomly distributed into the experimental tank at the treatment concentrations of either 30 ppm clove oil (Valisek et al. 2005) and 7 ppm propofol . Three replicates of 15 fish were used for each anesthetic concentration treatment of propofol, clove oil and control.

The times to achieve stage 5 of anesthesia were also recorded. Once an individual fish had reached the onset of stage 5 anesthesia, a dip net was used to immediately remove it from the tank. The fish was then transferred to a 20-L, well-oxygenated ‘recovery’ tank (i.e., no anesthesia present) maintained at 22°C and observed until it fully recovered. During this recovery period, the fish behavior was observed and times to recovery were recorded.

Once a fish had been used for a treatment, it was left in the recovery aquarium for approximately 1 day prior to being transferred back to a 50-L recovery holding tank for the remainder of a 14-day observational recovery period. Any abnormal behavior or mortalities were recorded during this 14-day recovery period. Anesthesia and recovery stages are presented in Table 1.
Table 1

Stages of anesthesia with clove oil in fish (Keene et al., 1998, modified from McFarland, 1959; Jolly et al. 1972) and Stages of recovery from anesthesia in fish (Keene et al., 1998modified from Hikasa et al., 1986) in fish

Stage

Behavior in anesthesia stages

Behavior in recovery stages

1

Normal Reacts to external stimuli; opercular rate and muscle tone normal

Decreased opercular movement

2

Light sedation Slight loss of reactivity to external visual and tactile stimuli; opercular rate slightly decreased; equilibrium normal

Partial recovery of equilibrium; partial recovery of swimming motion

3

Deep sedation Total loss of reactivity to external stimuli except very strong pressure;

Total recovery of equilibrium

4

slight decrease in opercular rate; equilibrium normal Partial loss Partial loss of muscle tone; increased opercular rate; reacts of equilibrium only to strong tactile and vibrational stimuli

Reappearance of avoidance swimming motion; reaction to external stimuli; behavioral response still stolid

5

Total loss Total loss of muscle tone and equilibrium; slow but regular of equilibrium opercular rate; loss of spinal reflexes

Swimming, rarely striking head firmly to sides or against bank of the tank

6

Loss of reflex Total loss of reactivity; opercular movements slow and reactivity irregular; heart rate very slow; loss of all reflexes

Total behavioral recovery; normal swimming

Table 2

Induction and recovery times for gold fish anesthetized with 7 ppm propofol and 30 ppm clove oil

Time (min)

propofol

Clove oil

Anesthesia (Stage 5)

7.40 ± .40*

4.26 ± .60

Recovery

8.52 ± .82

7.95 ± 1.21

Results are expressed as the mean ± SE; N = 36 for each anesthetic.

*p < 0.05.

Haematological blood profile

For the haematological blood profile tests, in experimental I twenty of gold fish anesthetized with 7 ppm propofol were examined immediately after 10 min anaesthesia. In experiment II twenty of gold fish anesthetized with 30 ppm clove oil were examined immediately after 10 min anaesthesia and in Control group twenty of fish without any anesthesia were hematologicaly tested. Heparinized injection needles were used to take samples of blood from coudal vein of fish. To stabilize blood samples, aqueous solution of heparin sodium salt at 0.01 mL per mL of blood was used (Svobodova et al. 1991).

The indices used to evaluate the haematological profile included the erythrocyte count (Er), haemoglobin concentration (Hb), haematocrit (PCV), mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), erythrocyte haemoglobin (MCH), leukocyte count (Leuko) and the differential leukocyte count (Svobodova et al. 1991).

Results of haematological examinations were tested by the variance analysis using the Statgraphics (ANOVA–Tukey Test) software.

Results

Acute toxicity of propofol

During the 96-hour LC50 tests, the mean water temperature was 22°C, pH was 7.5 and water oxygen levels were 75-85% saturation. Based on tests of acute toxicity to gold fish, the 96-hour lethal concentrations of propofol were determined (96 h LC50 6.353 mg/L, 96 h LC1 2.966 mg/L and 96 h LC99 13.609 mg/L). 96-h LC50 propofol of gold fish placed into the higher concentration (16 ppm) initially exhibited irritation, as evidenced by rapidly darting about the aquaria, the mortality of this concentration was 100% after 30 min. The first three concentrations (0.5. 1 and 2 ppm) survived the 96 h trial with 0% mortality. In concentration of 4 ppm deep sedation was observed after 10 min, with only one mortality after 24 h of exposure.

Anesthesia and recovery

The time required to induce anesthesia using propofol (7 ppm) and clove oil (30 ppm) is shown in Table 2.

The time required to induce anesthesia using propofol was significantly higher than Clove oil (p < 0.05), but there was no significant difference in recovery time between the experiments. No mortality in anesthesia group was observed.

Haematological parameters

Changes in the haematological parameters of gold fish in the control group and those exposed to propofol and clove oil are presented in Tables 3 and 4.
Table 3

Effects of propofol and clove oil anaesthesia on haematological indices in gold fish

Indices

Propofol

Clove oil

Control

Experiments

   

Erythrocyte (×106)

3.20 ± 0.63a

4.33 ± 0.30a

3.43 ± 0.43a

Hb (g/dL)

5.20 ± 0.73a

25.39 ± 5.73b

15.41 ± 4.76b

HCT (%)

31.80 ± 2.90a

27.40 ± 1.28a

26.60 ± 2.00a

MCV (fl)

137.4 ± 31.15a

64.98 ± 6.68a

83.42 ± 14.86a

MCH (pg)

19.25 ± 4.34a

59.60 ± 1.43a

47.84 ± 15.14a

MCHC (%)

13.93 ± 1.36a

94.95 ± 24.50b

62.46 ± 21.90b

Leuko ( ×104)

5.60 ± 0.97a

2.37 ± 0.95a

2.97 ± 0.63a

Groups with different alphabetic superscripts differ significantly at p < 0.05 (ANOVA).

Results are expressed as the mean ± SE; N = 20 for each concentration of anesthetic.

Table 4

Effects of propofol and clove oil anaesthesia on differential leukocyte counts in gold fish

Indices (%)

Propofol

Clove oil

Control

Experiments

   

Lymphocytes

92.80 ± 0.66a

92.00 ± 0.54a

93.80 ± 0.37a

Monocytes

0.20 ± 0.20a

0.00 ± 0.00a

0.00 ± 0.00a

Neutrophil

6.60 ± 0.60a

7.80 ± 0.37a

6.00 ± 0.44a

Eosinophil

0.40 ± 0.24a

0.20 ± 0.20a

0.20 ± 0.20a

Groups with different alphabetic superscripts differ significantly at p < 0.05 (ANOVA).

Results are expressed as the mean ± SE; N = 20 for each concentration of anesthetic.

No significant decrease was found in Total RBC, HCT, WBC, MCH, MCV and leukogram indices (p > 0.05). MCHC level of propofol experiment (13.93 ± 1.36) showed significant (p < 0.05) decrease than Clove oil experiment (94.95 ± 24.50) and control (62.46 ± 21.90). Decrease in Hb content (5.20 ± 0.73) was observed in propofol exposure compared with control (15.41 ± 4.76) and clove oil experiment (25.39 ± 5.73) (p < 0.05).

Discussion

Anaesthetics are necessary for many procedures in aquaculture. The analysis of blood parameters is one of the most valuable methods of anaesthetics evaluation, because it has been shown that the physiological effects of anaesthetics are species-specific and age-dependent (Anver Celik 2004). Because species may differ widely in their response to anaesthetics, screening of their use is necessary.

In this study acute toxicity of propofol to Carassius auratus is investigated from the point of view of propofol use as an anaesthetic, with anaesthetizing baths.

Hematological parameters can provide needed information on the physiological status of fishes, and help the aquaculture and research personnel to make proper decisions to increase the survival of fishes

Propofol (Diprivan®, Rapinovet®, Propoflo®), an alkyl phenol hypnotic has been investigated as a widely used intravenous anaesthetic in veterinary practice. Use of propofol as a sole anaesthetic produced effective general anaesthesia in different domestic animals (Duke et al., 1997; Lin et al., 1997; Carroll et al., 1998; Bayan et al., 2002; Zama et al., 2003; 2005) Propofol used as an anaesthetic agent in lizards showed a rapid onset of action. Following intravenous administration in green iguanas, the onset of anaesthesia maybe expected within several minutes (Bennett et al. 1998).

Guénette et al. (2008) determined the level of anesthesia attained in Xenopus laevis frogs using a propofol bath administration. An appropriate anesthetic dose was determined to be 88 mg/L for 15 min.

There are a few experimental papers reporting the effect of propofol on the fish species. According to FDA guidelines acute toxicity to rainbow trout (Onchorhynchus mykiss) and bluegill sunfish (Lepomis macrochirus) (FDA guideline 4.11, flow-through –no aeration) is 96 h LC50 = 0.37 mg/L and 96 h LC50 = 0.62 mg/L respectively. The bio concentration factor (BCF) has been determined for carp, Cyprinus carpio, and the results reported as: (BCF) 28 day = 27 (at 2 μg/L) and (BCF) 28 day = 26 (at 0.2 μg/L).

The bio degradability of propofol has been assessed according to the OECD guideline 301 F, the results showed >91% removal of propofol from the aqueous phase.

Fleming et al. (2003) evaluated propofol for short–term immobilization of Gulf mexico sturgeon (Acipencer Oxyrinchus de soti) and it was observed that the group receiving intra venous propofol (6.5 mg/kg body weight, i.v.) was in a light plane of anesthesia within 5 min after drug administration.

The effect of propofol on haematological parameters is reported in sheep (Brzeski et al., 1994), dogs (Gill et al. 1996), ewes (Handel et al. 1991), rabbits (Mazaheri-Khameneh et al. 2012) and horses (Mama et al., 1998), no other data on the blood profiles in the fish species anaesthetized with propofol are available in the literature.

Gold fish exposed to propofol showed lower Hb and MCHC. It is known that propofol induces moderate systemic hypotension, arterial vasodilatation and venodilatation (Branson and Gross, 1994).The lower concentration of Hb, could be explained by haemodynamic changes and re-distribution of blood cellular elements in the vascular bed.

In conclusion, the result of this study indicated that propofol (7 ppm) can induce safe and valid anaesthesia in gold fish. However, it seems that further studies on different dosage, also measuring more haematological and biochemical parameters in the gold fish and other (non-food) ornamental fish following anaesthesia with propofol are needed.

Declarations

Authors’ Affiliations

(1)
Department of fisheries and natural resource, agriculture faculty, Gonbad Kavous University

References

  1. Andrews DT, Leslie K, Sessler DI, Bjorksten AR: The arterial blood propofol concentration preventing movement in 50% of healthy women after skin incision. Anesth Analg 1997, 85: 414-419.Google Scholar
  2. Anver Celik E: Blood chemistry (electrolytes, lipoprotein and enzymes) values of black scorpion fish ( Scorpaena porcus 1758) in the Dardanelles. Turkey J Biol Sci 2004, 4: 716-719.View ArticleGoogle Scholar
  3. Bayan H, Sarma KK, Chakravarty P: Biochemical and haematological changes during propofol anaesthesia in canine. Indian J Vet Surg 2002, 23: 95-96.Google Scholar
  4. Bennett RA, Schumacher J, Hedjazi-Haring K, Newlel SM: Cardiopulmonary and anaesthetic effect of propofol administered intraosseously to green iguanas. J Am Vet Med Assoc 1998, 212: 93-98.Google Scholar
  5. Branson KR, Gross ME: Propofol in veterinary medicine. J Am Vet Med Assoc 1994, 204: 1888-1890.Google Scholar
  6. Brzeski W, Depta A, Jalynksi M, Chyczewski M: General anaesthesia in sheep with the use of diprivan-propofol. Med Weter 1994, 50: 215-217.Google Scholar
  7. Carroll GL, Hooper RN, Slater MR, Hartsfield SM, Matthews NS: Detomidine-butorphanol-propofol for carotid artery translocation and castration or ovariectomy in goats. Vet Surg 1998, 27: 75-82.View ArticleGoogle Scholar
  8. Coyle SD, Durborow RM, Tidwell JH: Anesthetics in Aquaculture. Texas: SRAC Publication No. 3900; 2004:6.Google Scholar
  9. Duke T, Egger CM, Ferguson JG, Frketic MM: Cardiopulmonary effects of propofol infusion in llamas. Am J Vet Res 1997, 58: 153-156.Google Scholar
  10. Fleming GJ, Heard DJ, Floyd RF, Riggs A: Evaluation of propofol and medetomidine-ketamine for short-term immobilization of Gulf of Mexico sturgeon ( Acipenser oxyrinchus de soti ). J Zoo Wild Med 2003, 34: 153-158.View ArticleGoogle Scholar
  11. Gill JR, Rodriguez JF, Ezquerra LJ, Vives MA, Jimenez J, Uson JM: Development of anaesthesia and changes in the blood parameters in dogs medicated with propofol. Medicina Veterinaria 1996, 13: 242-246.Google Scholar
  12. Guénette SA, Beaudry F, Vachon P: Anesthetic properties of propofol in African clawed frogs ( Xenopus laevis ). J Am Assoc Lab Anim Sci 2008, 47(5):35-38.Google Scholar
  13. Handel IG, Staddon GE, Weaver BMQ, Cruz JI: Changes in packed cell volume during anaesthesia. In Proceedings of the 4th International Congress of Veterinary Anaesthesia. Utrecht, Netherlands: J. Vet. Aanesth; 1991:25-31. 347–352Google Scholar
  14. Hikasa Y, Takase K, Ogasawara T, Ogasawara S: Anaesthesia and recovery with tricane methanesulphonate, eugenol and thiopental sodium in the carp (Cyprinus carpio). Japanese Journal of Veterinary Science 1986, 48: 341-351.View ArticleGoogle Scholar
  15. Hoskonen P, Pirhonen J: The effect of clove oil sedation on oxygen consumption of six temperate-zone fish species. Aquacult Res 2006, 35: 1002-1005.View ArticleGoogle Scholar
  16. Jolly DW, Mawdesley-Thomas LE, Bucke D: Anesthesia of fish Vet Rec. 1972, 91: 424-426.Google Scholar
  17. Kay B, Stephenson DK: ICI 35868 (Diprivan): A new intravenous anesthetic. A comparison with althesin Anaesthesia 1980, 35: 1182-1187.Google Scholar
  18. Keene JL, Noakes DLG, Moccia RD, Soto CG: The efficacy of clove oil as an anaesthetic for rainbow trout, Oncorhynchus mykiss (Walbaum). Aquacult Res 1998, 29: 89-101.View ArticleGoogle Scholar
  19. Knotkova Z, Pejrilova S, Trnkova S, Matouskova O, Knotek Z: Influence of reproductive season upon plasma biochemistry values in green iguanas. Acta Veterinaria Brno 2005, 74: 515-520.View ArticleGoogle Scholar
  20. Lin HC, Purohit RC, Powe TA: Anaesthesia in sheep with propofol or with xylazine-ketamine followed by halothane. Vet Surg 1997, 26: 247-252.View ArticleGoogle Scholar
  21. Mama KR, Steffey EP, Pascoe PJ, Kollias BC: Comparison of two techniques for total intravenous anesthesia in horses. Am J Vet Res 1998, 59: 1292-1298.Google Scholar
  22. Mazaheri-Khameneh R, Sarrafzadeh-Rezaei F, Asri-Rezaei S, Dalir-Naghadeh B: Evaluation of clinical and paraclinical effects of intraosseous vs intravenous administration of propofol on general anesthesia in rabbits. Vet Res Forum 2012, 3(2):103-109.Google Scholar
  23. McFarland WN: A study of the effects of anesthetics on the behavior and physiology of fishes. Publ Inst Mar Sci Univ Texas 1959, 6: 337-342.Google Scholar
  24. Miller SM, Mitchell MA, Heatley JJ, Wolf T, Lapuz F, Smith JA: Clinical and cardiorespiratory effects of propofol in the spotted bamboo shark ( Chiloscyllium plagiosum ). J Zoo Wild Anim Med 2005, 36: 673-676.View ArticleGoogle Scholar
  25. Ross LG, Ross B: Anaesthetic and Sedative Techniques for Aquatic Animals. 2nd edition. Oxford: Blackwell Science Ltd.; 1999:159.Google Scholar
  26. Roubach R, Gomes LC, Fonseca FAL, Val AL: Eugenol as an efficacious anaesthetic for tambaqui, Colossoma macropomum (Cuvier). Aquacult Res 2005, 36: 1056-1061.View ArticleGoogle Scholar
  27. Svobodova Z, Pravda D, Palackova J: Unified methods of haematological examination of fish. Methods No. 22 edition. Vodnany: Res Inst of Fish Cult and Hydrob; 1991:31.Google Scholar
  28. Valisek J, Svobodová Z, Piac4ková V, Groch L, Nepejchalová L: Effects of clove oil anaesthesia on common carp ( Cyprinus carpio L.). Vet Med 2005, 50(6):269-275.Google Scholar
  29. Velíšek J, Wlasow T, Gomulka P, Svobodová Z, Novotný L, Ziomek E: Effects of clove oil anaesthesia on european catfish ( Silurus glanis L.). Acta Vet Brno 2006, 75: 99-106.View ArticleGoogle Scholar
  30. Zama MMS, Singh NK, Gupta AK, Kumar S, Kalita A 27th Annual Congress of Indian Soc. Vet. Surg. In Propofol anaesthesia in adult sheep: Clinical, Haematological and Biochemical studies. : GBPUA & T. Pantnagar; 2003.Google Scholar
  31. Zama MMS, Harbans L, Gupta AK, Bhadwal MS 29th Annual Congress of Ind. Soc. Vet. Surg. In Blood gas and electrolyte changes during propofol anaesthesia in buffalo calves. U.P: IVRI, Izatnagar; 2005.Google Scholar

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© GholipourKanani and Ahadizadeh; licensee Springer. 2013

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