Abstract: In the study in which the effects of clove oil and phenoxyethanol in fish farming are examined, the most cultured rainbow trout (Oncorhynchus mykiss) and brown trout (Salmo trutta fario) are used as material. As a result of the treatment, blood biochemical hematology and enxyme activity values are studied. The interaction of treatment, species and treatment species are found insignificant among hematological parameters; hemoglobin, hematocrite, erythrocyte, average hemoglobin amount per erythrocyte (MCH), average volume of erythrocyte (MCV) and average hemoglobin concentration per erythrocyte (MCHC) (p>0.05). Although, leukocyte values are significantly (p<0.01) effected by treatment and treatment species interaction the effect of species is insignificant. While, the effect of treatment and species are insignificant in erythrocyte sedimentation ratio compared with control group, the interaction between treatment and species is significant (p<0.05). While, the effect of treatment and species on the number of thrombocyte is significant, the effect of treatment x type interaction is detected to be insignificant.

INTRODUCTION

Anesthetics are physical or chemical agents that act on an animal by initially inducing a calming effect and subsequently inducing loss of equilibrium, mobility, consciousness and reflex action (Summer and Smith, 1990). The welfare of fish has been the focus of less research than that of higher vertebrates such as birds and mammals (Braithwaite and Huntingford, 2004).

An ideal anesthetic should induce anesthesia rapidly with minimum hyperactivity or stress. It should be easy to administer and should maintain the animal in the chosen state. When the animal is removed from the anesthetic, recovery should be rapid. The anesthetic should be effective at low doses and the toxic dose should greatly exceed the effective dose so that there is a wide margin of safety (Coyle et al., 2004).

Clove oil is a dark-brown liquid, a distillate of flowers, stalks and leaves of the clove tree Eugenia aromatica (Sato and Burhanuddin, 1995). According to Isaacs (1983); Briozzo et al. (1989) and Keene et al. (1998), clove oil is distilled from stems, leaves and flower buds of Eugenia caryophyllata and its active ingredient, i.e., eugenol (4-allyl-2-methoxyphenol), makes up 70-90% by weight of clove oil. Clove oil also contains eugenol acetate (>17%) and kariofilen 5 (>12%).

Clove oil is highly effective even at low doses. Keene et al. (1998) report that it induced anaesthesia faster and at lower concentrations than MS-222 while Munday and Wilson (1997) found clove oil only marginally less effective than quinaldine and more effective than 3 other chemicals, except at high doses. Clove oil provides a much calmer induction to anaesthesia than the other chemicals. Recovery time after clove oil anaesthesia is substantially longer than recovery time from other anaesthetics.

Clove oil is much less expensive than other chemicals. For example, Keene et al. (1998) showed clove oil to be <1.15 the price of MS-222 when preparing solutions of each capable of inducing stage 5 anaesthesia in <3 min in rainbow trout.

At present, 2-phenoxyethanol is used in the Czech Republic for short-term immobilization of fish before artificial spawning and whenever fish is handled outside water. The generally recommended concentration is 0.20 mL of water bath. For big breeding fish, the recommended concentration is 0.30 mL^-1 of water bath. At the recommended concentrations, anaesthesia is induced within 5-10 min. When transferred to clean water, fish will recover within 10 min (Svoboda and Kolarova, 1999; Hamackova et al. 2001). 2-Phenoxyethanol is an opaque, oily liquid. This drug is moderately soluble in water but freely soluble in ethanol.

Table 1:	Stages of anesthesia and recovery

The solution is bactericidal and fungicidal and is therefore, useful during surgery. It is relatively inexpensive and remains active in the diluted state for at least 3 days. About 2-phenoxyethanol has a relatively large margrin of safety and has been reported to produce a range of effects from light sedation to surgical anesthesia at concentrations of 100-600 mg L^-1. Concentrations of 300-400 mg L^-1 are useful for short procedures and lower concentrations of 100-200 mg L^-1 are considered safe for prolonged sedation such as during transport. About 2-phenoxyethanol is not approved by FDA for use on food fish in the U. S (Coyle et al., 2004).

Because the efficacy of most anesthetics are affected by species, body size, the density of fish in the bath as well as water quality (e.g., hardness, temperature or salinity), it is imperative that preliminary tests be performed with small numbers of the fish to determine the optimal dosage and exposure time. Due care should be taken to control the level of anesthesia desired, through the application of the appropriate concentration and to maintain constant observation of the fish as they go through the various stages of anesthesia (Table 1).

Since hematological parameters reflect the poor condition of fish more quickly than other commonly measured parameters and since they respond quickly to changes in environmental conditions (Atkinson and Judd, 1978; Atamanalp and Yanik, 2003), they have been widely used for the description of healthy fish (Blaxhall, 1972) for monitoring stress responses (Soivio and Oikari, 1976) and for predicting systematic relationships and the physiological adaptations of animals. The aim of the study was to investigate acute toxicity of clove oil and 2-phenoxyethanol for brown trout and rainbow trout using values of haematology profiles of blood to assess the effects of the fish exposure to the anaesthetics.

MATERIALS AND METHODS

The active substance of 2-phenoxyethanol is ethylene glycol monophenyl ether. Its summary formula is C₈H₁₀O₂, the molar weight 138.17 g•l-1, density 1.107-1.108 g•dm-3, peroxide content <0.005% and the boiling temperature is 245°C. The anaesthetic is slightly soluble in water (26.7 g•l-1) at 25°C but readily soluble in ethanol. The anaesthetic affects fish through the skin and gills. A group of 48 fish with an average weight of 180±25 g, reared in well water with a constant temperature of 8.5°C at the farm located at the Research and Extension Center in Ataturk University was transferred to the Central Laboratory in the Aquarium Fish Rearing Facility and was exposed to a anesthesia dose of 0.5 ppm of clove oil and 0.2 ppm of 2-phenoxyethanol 4 weeks in circular fiberglass tanks 780l in volume (100 cm diameter and 100 cm depth) under natural light conditions with a constant flow (1.5 l min^-1) of aerated dechlorinated tap water at 9-11°C and with no recirculation. The dissolved oxygen and pH levels and total water hardness were 8-9 ppm, 7.8 and 102 mg in CaCO₃, respectively. The tanks were aerated with an air pump. Eight fish were placed into each of 6 tanks, one for testing the clove oil, 2-phenoxyethanol and the other as the control. At the end of each day exposure, 8 fish from the control tank, 8 fish from the clove oil treatment tank and 8 fish from the 2-phenoxyethanol tank (Ahmad et al., 1995; Shakoori et al., 1996) were taken out and their blood was subjected to hematological analysis (Aziz et al., 1993; Shakoori et al., 1996; Santhakumar et al., 1999; Atamanalp et al., 2002). Approximately 2 cc venous blood was drawn from each subject for the determination of hematological parameters using heparin as an anticoagulant and for the estimation of the Red Blood Cell (RBC) count (Blaxhall and Daisley, 1973; Atamanalp et al., 2002), the total White Blood Cell (WBC) count (Blaxhall and Daisley, 1973; Atamanalp et al., 2002), the Hemoglobin (Hb) concentration (Kocabatmaz and Ekingen, 1984) and the Packed Cell Volume (PCV) (Schalm et al., 1975) whereas the Mean Corpuscular Volume (MCV), the Mean Corpuscular Hemoglobin (MCH) and the Mean Corpuscular Hemoglobin Concentration (MCHC) were calculated according to Reddy and Bashamohideen (1989).

Statistical analysis: The results are presented as means ±SD. Differences between parameters were analyzed by one-way Analysis Of Variance (ANOVA) and significant means were subjected to a multiple comparison test (Duncan) at the α = 0.05 level (26).

RESULTS AND DISCUSSION

Hematological parameters: Effects of clove oil and 2-phenoxyethanol anaesthesia on haematological indices on rainbow trout and brown trout as shown in Table 2.

Table 2:	Effects of clove oil and 2-phenoxyethanol anaesthesia on hematological indices on rainbow trout and brown trout
*p<0.05; **p<0.01

The number of erythrocyte (RBC): Average number of erythrocyte in the blood samples of brown trout and rainbow trout in the control group are detected to be 0.649x10⁶ mm^-3, 0.703x10⁶ mm^-3. This value shows compatibility with the value (min 0.538x10⁶ mm^-3, max 1.185x10⁶ mm^-3 ort 0.782x10⁶ mm^-3) Kocabatmaz and Ekingen (1984) and Atamanalp et al. (2002) have expressed for a healthy rainbow trout. Ezzat et al. (1974) reported this value to be 1.8x10⁶ mm^-3 in Tilapia zilli, Satake et al. (1986) to be between 0.66-2.01x10⁶ mm^-3 in armoured cat fish (Hypostomus paulinus), Favaretto et al. (1978) to be 0.69x10⁶ mm^-3 in cultured Hypostomus regani, Satake et al. (1986) to be 1.04x10⁶ mm^-3 wild Hypostomus regain, Torres et al. (1986) to be 1.00x10⁶ mm^-3 in Hypostomus punctatus, Santhakumar et al. (1999) to be 4.09x10⁶ mm^-3 in Anabas testudineus.

Average number of erythrocyte in rainbow trout and brown trout which are 2-phenoxyethanol anesthetics are found to be 0.608±0.04 x 10⁶, 0.641±0.03x10⁶ and 0.716± 0.05x10⁶ mm^-3 and 0.657± 0.07x10⁶ mm^-3 in rainbow trout and brown trout which are applied with clove oil. According to the results, 2-fenoksietanol decreased the number of erythrocyte in rainbow trout and brown trout but clove oil increased.

Velisek et al. (2005) detected the erythrocyte values of rainbow trout which is applied with clove oil as 1.28±0.26x10⁶ mm^-3 right after the anesthetics and as 1.16±0.25x10⁶ mm^-3 24 h later. The value detected right after the anesthetics is lower compared with control group; values taken 24 h later are higher than control group 2-fenoksietanol application decreasing the number of erythrocyte in carp shows parallelism with the current study.

No significant increases are observed in RBC number, Hb value and Hct ratio of scald fish (Scophthalmus aquosus) from Scophthalmidae family that are exposed to chemicals when compared with control group. It is reported that in the study carried out with bull head (Siluris glanis) there is an increase in the number of erythrocyte right after and 24 h later the anesthetics with clove oil (Velisek et al., 2006). This situation calls complication with both the results and literature results as well. This complexity can be explained with the species of fish being different in the studies.

The number of leukocyte (WBC): Average number of leukocyte in the blood samples of brown trout and rainbow trout in the control group are detected to be 4.033±0.30x10⁴ mm^-3, 3.867±0.25x10⁴ mm^-3. This value shows suitability with the value (min 3.0x10⁴ mm^-3, max 6.5x10⁴ mm^-3 and avr. 4.6x10⁴ mm^-3) Kocabatmaz and Ekingen (1984) and Atamanalp et al. (2002) have expressed for a healthy rainbow trout (Oncorhynchus mykiss).

WBC change according to anesthetic materials applied is 4.350±0.38x10⁴ mm^-3 in 2-phenoxyethanol applied brown trout, 3.829±0.35x10⁴ mm^-3 in rainbow trout, 4.300±1.01x10⁴ mm^-3 in clove oil applied brown trout. According to these results it is detected that the number of leukocyte increased in brown trout as a result of phenoxyethanol anesthetics, decreased in rainbow trout treated with only phenoxyethanol.

From the average results gathered from study, it is detected that the difference between control group and clove oil and phenoxyethanol groups and the difference among groups do not have significance, however the effect of clove oil on rainbow trout is found to be significant.

Velisek et al. (2005) detected the erythrocyte values of rainbow trout as 23.70±4.69x10⁴ mm^-3 right after the anesthetics and as 19.60±9.96x10⁴ mm^-3 24 h later. No significant change is observed in the hematological profile of carp and rainbow trout which is anesthetized with clove oil. Significant amount of leukocyte decrease is detected right after and 24 h later the treatment with clove oil in bullhead (Siluris glanis) (Velisek et al., 2006).

In their studies Noyan (1980); Roberts (1989); Yilmaz and Otlu (1989) and Isoda and Fujimaki (1990) expressed that there is a decrease in the number of leukocytes in sick fish which are exposed to any kind of infection and increase related with defend in leukocyte cells. While there is no change in rainbow trout right after the anesthesia, an increase is detected 24 h later (Velisek et al., 2005). In the analyses carried out right after the anesthesia as a result of the treatment with 2 phenoxyethanol in carp and rainbow trout, significant increase is reported in the amount of leukocyte (Velisek et al., 2007).

The number of WBC detected in the study is lower than the value of Atamanalp and Yanik (2003) detected for the rainbow trout (Oncorhynchus mykiss), for Israeli bream (Oreochromis niloticus), Kocabatmaz and Ekingen (1984) for abant trout (Salmo trutta abanticus), Quentel and Obach (1992) for turbot (Scophthalmus maximus), higher than the value Blaxhall and Daisley (1973) found for brown trout (Salmo trutta), Das and Mukherjee (2003) for Indian carp (Labeo rahita), Hasiloglu and Atamanalp (2002) for chub (Leuciscus cephalus), Van Vuren and Hattingh (1978) for carp (Cyprinus carpio). The value is found similar to the value Kocabatmaz and Ekingen (1984) found for rainbow trout.

The number of trombocytes (Plt): Erythrocyte values of 0.967±0.20x10⁴ mm^-3 and 1.067±0.15x10⁴ mm^-3 detected for brown trout and rainbow trout in control group show compatibility with the average values (min 0.4x10⁴ mm^-3. max 2.41x10⁴ mm^-3 and avr. 0.94x10⁴ mm^-3) Kocabatmaz and Ekingen (1984) and Atamanalp et al. (2002) expressed. It is also expressed by the researchers that trombocyte values are effected significantly by the stress and the value of 2.1x10⁴ mm^-3 before stress increased to 4.3x10⁴ mm^-3 in rainbow trout after stress. Satake et al. (1986) expressed that this value is1.657±0.341x10⁴ mm^-3 for armored catfish (Hypostomus pulinus).

For the fish in the group of 2-phenoxyethanol anesthetics applied; trombocyte values are found to be 0.700±0.08x10⁴ mm^-3 in brown trout. While, 0.957±0.20x10⁴ mm^-3 in rainbow trout. Here, 0.700±0.10x10⁴ mm^-3 in brown trout treated with clove oil and 0.833±0.05x10⁴ mm^-3 in rainbow trout.

As a result of the study it is detected that the number of trombocyte decreased in both species of fish at the end of the treatment. As a result of the statistical analysis carried out; it is detected that the difference between treatment and species are significant and treatment x type interaction is insignificant. In the study about the number of trombocyte, Lester and Budd (1983) detected the number of trombocyte as 26,103 mm³ in coho salmons, Casillas and Smith (1977) detected it as 21.103 mm³ before the stress and as 43.103 mm^-3 after the stress in rainbow trout. Velisek et al. (2006) reported that the change of trombocyte numbers in bullhead (Siluris glanis) is not significant after being exposed to 30 mg L^-1. clove oil concentration.

MCV: Average MCV value is detected to be 377.965±42.23 μm³ in control group brown trout and 261.010±3.54 μm³ in rainbow trout. As a result of the statistical analysis treatment, species and treatment x type interaction is found to be insignificant. MCV value is found to be 313.398±67.58 μm³, 316.66±45.56 μm³ in 2-phenoxyethanol anesthetics applied brown trout and rainbow trout and 326.517±90.12 μm³, 295.100±46.34 μm³ in clove oil applied brown trout and rainbow trout.

Compared with the control group while there is a decrease in MCV values in brown trout applied with 2-phenoxyethanol and clove oil there is an increase in rainbow trout. In their study with clove oil; Velisek et al. (2006) detected MCV values to be 344.05±57.67 μm³ right after the anesthetics and 339.79±56.96 μm³ 24 h later in bullhead (Siluris glanis). Compared with the control; there is an increase in the measurements right after the anesthetics and 24 h later the anesthetics. After the anesthetics with 2-phenoxyethanol, there is an increase in MCV values in carp and rainbow trout compared with the control group (Velisek et al., 2007). This situation can be explained with the species of fish and anesthetic materials being different in the studies.

Velisek et al. (2005) reported that there is a decrease in MCV values of rainbow trout after treatment with clove oil. This result shows incompatibility with current findings.

MCV value obtained in this study is higher than the values found for (Pagrus auratus) by Canfield et al. (1994), chub (Leuciscus cephalus) found by Hasiloglu and Atamanalp (2002); Martinez et al. (1994) for rainbow trout (Oncorhynchus mykiss), Mughal et al. (1993) and Shakoori et al. (1996) for grass carp (Ctenopharyngodon idella), Sandnes et al. (1988) for salmon (Salmo salar), Pages et al. (1995) for bream (Sparus aurata), Schutt et al. (1997) for swordtail fish (Xiphophorus helleri), Yamawaki et al. (1986) and Van Vuren and Hattingh (1978) for carp (Cyprinus carpio), Quentel and Obach (1992) for turbot (Scophthalmus maximus), Yone et al. (1986) for common seabream (Chrysophrys major); lower than the value Atamanalp et al. (2002) found for rainbow trout (Oncorhynchus mykiss).

MCH: Average MCH value is detected to be 111.730±8.089 pg in control group brown trout and 89.930±5.572 pg in rainbow trout. No significant differences are detected either between treatment and species or among the groups. Compared with the control group while there is a decrease in MCH values in brown trout applied with 2-phenoxyethanol and clove oil there is an increase in rainbow trout. Velisek et al. (2006) detected MCH values to be 49.15±11.55 pg right after the anesthetics and 51.06±9 pg 24 h later in rainbow trout after clove oil treatment. Compared with the control while there is an increase in the values right after the anesthetics, there is decrease 24 h later. In the studies carried out with carp there is an increase in the values right after the anesthetics and 24 h later the anesthetics. 45.60±4.24 and 5.25±6.28 pg values are detected in the treatments with 2-phenoxyethanol in rainbow trout and carps. It is reported that there is an increase of values compared with control group in both situations (Velisek et al., 2007).

As a result of the treatment with clove oil in bullhead (Siluris glanis), there is a decrease compared with control group and 24 h later there is an increase compared with the control group (Velisek et al., 2006). Average MCH value obtained in this study is higher than the values of Atamanalp et al. (2002) found for rainbow trout (Oncorhynchus mykiss), Hasiloglu and Atamanalp (2002) for chub (Leuciscus cephalus), Mughal et al. (1993) for grass carp (Ctenopharyngodon idella), Pages et al. (1995) for bream (Sparus aurata), Quentel and Obach (1992) for turbot (Scophthalmus maximus), Sandnes et al. (1988) for salmon (Salmo salar) and Yamawaki et al. (1986) for carp (Cyprinus carpio).

MCHC: Average MCH value is calculated as 29.625±1.66 g/100 mL in control group brown trout and 34.470±2.602 g/100 mL in rainbow trout. Although, the difference among species is significant in the sense of average MCHC values, treatment and treatment species interaction is found to be insignificant. This value is higher than the value (20.5-28.4 g/100 mL) Kocabatmaz and Ekingen (1984) and Atamanalp et al. (2002) found for healthy trout.

When MCHC values are observed it is detected that there is decrease in both the treatment groups and fish species. Velisek et al. (2006) calculated MCHC values to be 141.71±10.79 g/100 mL right after the anesthetics and 150.46±15.87 g/100 mL 24 h later in rainbow trout. There is a significant decrease in MCHC value after anesthetics with clove oil in fishcat. It is reported that the value right after the anesthetics is 236.99±6.96 g/100 mL in rainbow trout treated with 2-phenoxyethanol and 269.14±16.02 g/100 24 h later the treatment; and 139.56±15.09 g/100 mL and 141.66±13.74 g/100 mL, respectively in carp. These results are different from the results of the study.

Average MCHC value obtained in this study is higher than the values of found for rainbow trout (Oncorhynchus mykiss), Hasiloglu and Atamanalp (2002) for chub (Leuciscus cephalus), Mughal et al. (1993) for grass carp (Ctenopharyngodon idella), Pages et al. (1995) for bream (Sparus aurata), Quentel and Obach (1992) for turbot (Scophthalmus maximus), Sandnes et al. (1988) for salmon (Salmo salar) and Yamawaki et al. (1986) for carp (Cyprinus carpio), lower than the value found for turbot (Scophthalmus maximus) by Quentell and Obach (1992).

It reported that the change of hematological parameters among fish are influenced by the fish species as well as season and temperature, reproduction period and nutrition condition, size and weight of the fish, age, moreover salinity of the water, disease, toxic materials, heavy metal and industrial wastes and stress factors.

Hemoglobin: Average hemoglobin value is calculated as 6.950±0.35 g/100 mL in control group brown trout and 6.550±0.49 g/100 mL in rainbow trout. The difference among groups is not significant in the sense of average hemoglobin value. This value is between the value Kocabatmaz and Ekingen (1984) and Atamanalp et al. (2002) have reported (min. 4.3 max. 10.9 and avr. 6.5) for a healthy truot.

In this study the highest level is detected in clove oil treated brown trout (5.033±0.23 g/100 mL) and the lowest value is detected on clove oil treated rainbow trout (7.033±2.33 g/100 mL). These findings show that rainbow trout and brown trout give different reactions to clove oil.

While the hemoglobin value increase for clove oil treated rainbow trout there is no change in other treatments.

In order to detect the effects of infection on blood parameters of fish which is thought to be a stress factor; carp (Cyprinus carpio) is anesthetized (25% phenoxyethanol) and analyzed by taking blood samples. According to the values obtained, it is seen that hemoglobin values have increased (7.47+0.35 g dL^-1) compared with the control group (5.16+0.20 g dL^-1).

Goel et al. (1981) have carried out comparative hematological studies on some of the India fresh-water fish and observed the changes of hematological parameters of fish related to environmental effects. According to their findings with the decrease of oxygen content of water, erythrocyte and hemoglobin amount increase in blood.

Average Hb value obtained in this study is lower than the values of Canfield et al. (1994) for sea bass (Pagrus auratus), Martinez et al. (1994) for rainbow trout (Oncorhynchus mykiss), Hasiloglu and Atamanalp (2002) for chub (Leuciscus cephalus), Sandnes et al. (1988) for salmon (Salmo salar) and Yamawaki et al. (1986) for carp (Cyprinus carpio), similar to the values of (Kocabatmaz and Ekingen, 1984) for Abant trout (Salmo trutta abanticus), Pages et al. (1995) for bream (Sparus aurata), Van Vuren and Hattingh (1978) for swordtail fish (Xiphophorus helleri), carp (Cyprinus carpio). The value is higher than the value of Atamanalp et al. (2002) found for rainbow trout (Oncorhynchus mykiss), Azizoglu and Cengizler for Israeli bream (Oreochromis niloticus), Blaxhall and Daisley (1973) for brown trout (Salmo trutta), Dawson for scaldfish (Scophthalmus aquasus), Lie et al. (1989) for gadoid fish (Gadus morhua), Martinez et al. (1994) for grass carp (Ctenopharyngodon idella) and turbot (Scophthalmus maximus) by Quentel and Obach (1992).

Haematocrit: Average haematocrit percentages are calculated as 23.50±2.121 in control group brown trout and 19.00±2.04 in rainbow trout. These values show compatibility with the value (Min. 19.0. Max. 41.3 and avr. 28.0) Kocabatmaz and Ekingen (1984) and Atamanalp et al. (2002) found for healthy trout. This value is calculated as 20.00±3.916 for phenoxyethanol treated brown trout and 19.14±2.34 for rainbow trout. Haematocrit levels are found to be 21.00±3.606 for clove oil treated salmo fontinalic and 21.00±2.00 for rainbow trout. Haematocrit value is not influenced from species, treatment and treatment x type interaction.

Although, both the anesthetics treatments increase haematocrit level in rainbow trout compared with control group, this ratio is too low for brown trout. Although, it is known that HT ratio changes according to the species of fish; average Ht ratio obtained in this study is lower than the values of Kocabatmaz and Ekingen (1984) and Atamanalp et al. (2002) for rainbow trout (Oncorhynchus mykiss), Chen et al. (2003) for Israeli bream (Oreochromis niloticus) for salmon (salmo salar), Blaxhall and Daisley (1973) for brown trout (Salmo trutta), Canfield et al. (1994) for sea bass (Pagrus auratus), Kocabatmaz and Ekingen (1984) for Abant trout (Salmo trutta abanticus), Pages et al. (1995) for bream (Sparus aurata), Van Vuren and Hattingh (1978) for swordtail fish (Xiphophorus helleri), Yone et al. (1986) for common sea bream (Chrysophrys major). These values are similar to the values of Dawson for turbot (Scophthalmus aquasus) and Nelson et al. (1996) for gadoid fish (Gadus morhua).

In the studies carried out with brown trout (Salmo trutta), garyling (Thymallus thymallus), carp (Condrostoma nasus), gray mullet (Leusiscus cephalus), barbus barbus (Barbus barbus) in natural environment, it is detected that hematological parameters such as haematocrit, hemoglobin values and number erythrocyte change due to biotic and abiotic factors, individual sexual differences and reproduction cycle is effective in this (Luskova, 1997). It is asserted that sex, healthiness and acclimation have effects on blood parameters and must be taken into regard while evaluating the hematological parameters of the species (Gabriel et al. 2004). It is asserted that haematocrit, hemoglobin and total plasma protein levels of C. gariepinus show significant differences in 14 and 41°C acclimation temperatures compared with 29°C control group (Adeyemo et al., 2003).

Erythrocyte sedimentation ratio: Sedimentation values are calculated as 2.475±0.3304 mm h^-1 in control group brown trout and 2.275±0.2630 mm h^-1 in rainbow trout. These values show compatibility with the value (0.0-8.0 and 1.8-6.0 mm h^-1) Kocabatmaz and Ekingen (1984) and Atamanalp et al. (2002) found for healthy trout. In the treatments carried out with phenoxyethanol and clove oil; the highest value is detected in clove oil treated rainbow trout with 2.750±0.3728 mm h^-1, the lowest value is detected in clove oil treated brown trout with 1.900± 0.2191 mm h^-1. As a result of statistical analysis the difference between treatment groups and species are found to be insignificant, the result of treatment x type interaction is significant.

While the sedimentation value decreases for brown trout as a result of clove oil and 2-phenoxyethanol treatment, it increases for rainbow trout. Kocabatmaz and Ekingen (1984) reported the sedimentation value as 0.8-2.3 mm h^-1 for common carp, 1.8-2.5 mm h^-1 for bullhead, 2.0-3.3 mm h^-1 for chub; reported it as 6.0±2.0 mm h^-1 for fresh water catfish (Heteropneustes fossilis).

Erythrocyte sedimentation ratio increase in the events such as acute infections, heavy metal toxicities and kidney deformations (Blaxhall and Daisley, 1973). Infection existence and sedimentation speed increase in fish (Kocabatmaz and Ekingen, 1984). It is thought that these values showing difference with the values given in literature is based on the fish species taken into experiment, gender and age of the fish used and the study conditions (Kocabatmaz and Ekingen, 1984). Changes in physical and chemical features of water cause change in the hematological parameters of fish as well.

CONCLUSION

It is expressed that Ph of water moving towards acidic side increase haematocrit and hemoglobin levels of fish and the number of erythrocyte and this may be based on the swelling of blood cells as a result of deformation in the regulation of plasma electrolyte with hemoconcentration (Heath, 1990).