E-MAIL

9:21 am, June 13, Sunday

Phase IV results: 2010

In this phase there were followed:

–          surveillance audit of farms in terms of fulfilling conditions for fish protection and welfare;

–          surveillance audit of transport of fish in terms of ensuring the protection and welfare conditions;

–          surveillance audit of slaughter units in terms of ensuring the protection and welfare conditions;

–          submission of proposals for welfare assessment systems in fish farms, during transport and slaughter applicable in our country.

I) surveillance audit of farms in terms of fulfilling conditions for fish protection and welfare

It was assessed fish welfare in two fish farms: a sturgeon farm in the center of the country (Black Sea sturgeon) and a private carp farm from  Comişani area, Dambovita county, basen on both water quality (the living environment of fish) and blood biochemical profile.

►In the sturgeons farm there were harvested water samples from three checkpoints: fishpond chute, center and outlet by usig PSB 4 sampler. From the samples there were analyzed: physical parameters (temperature, turbidity); chemical parameters (dissolved oxygen, chlorine, pH, ammonia, nitrites, nitrates, phosphates, phenols, detergents, iron, copper, sulfates) and microbiological ones (total count – TC and total coliform – CF). The physical chemical water parameters were determined by using Spectroquant Nova 60 photocolorimeter. The micriobiological parameters determination was done by cultivation on dehydrated (dry) media Compact Dry TC (total count – aerobic plate count) and CF (total coliforms). The method consisted in the aseptic spread of 1 ml water samples in the middle of the media, which diffuses automatically and evenly into all over the sheet and transform it into gel. The incubation period is one day at 25-35 °C temperature for total coliform and two days at 35-37 °C for total count. Results interpretation was done according to reference values from abroad literatures for sturgeons and to provisions of the 161/2006 Order of the Environment and Water Resources Management Ministry.

Concerning the physical water parameters, the turbidity was acceptable, the Sechi disk being visible at 80 cm depth and the temperature record a value of 22 °C, framing into the normal limits for sturgeons.

The results for chemical water parameters in the assessed farm are shown in table 1 and 2.

Table no. 1

Average values of water quality parameters in the sturgeons farm: oxygen, chlorine, nitrates, nitrites and phosphates

Assessed parameters Oxygenmg/l Chlorinemg/l pH Nitratesmg/l Nitritesmg/l Phosphatesmg/l
Fishpond chute 8 0.15 7.7 9 0.023 0.7
Center of the pond 7.9 0.09 7.7 15 0.036 1.3
Water outlet 7.2 0.13 7.4 14 0.056 0.8
Admitted limits 9 0.3 7-8 10 0.1 2

Table no. 2

Average values of water quality parameters in the sturgeons farm: iron, copper, ammonia, sulfates, phenols and detergents

Assessed parameters Ironmg/l Coppermg/l Ammonia Sulfatesmg/l Phenolsmg/l Detergentsmg/l
Fishpond chute 0.05 0.04 undetected 30 0.1 0.03
Center of the pond 0.05 0.03 undetected 32 0.21 0.03
Water outlet 0.05 0.03 undetected 30 0.2 0.02
Admitted limits 1 0.3 0.05 2-7 1-2 0.1

Disolved oxygen average values in the water sampled ranges within the normal limits of 7-8 mg/l according to Order 161/2006 and within the normal limits of less than 9 mg/l according to the reference values for sturgeon.

Chlorine in water samples presented values (0.15 mg/l, 0.09 mg/l and 0.13 mg/l) that frame within the corresponding values of class I of water quality in accordance with Order 161/2006, but also within the normal limits of less than 0.3 mg/l reference value for sturgeon.

The pH of the water for ponds supply must record the value of 7, i.e. to provide a neutral reaction. Acid reaction of water, especially in the presence of chemicals, affects in a negative way the processes of breathing and feeding in fish, which contributes to a poor nutritional base use and to a slowdown of fish development. Also in the acidic environment fish has a reduced mobility, it seems fearful, coming to the surface and piping air, although the oxygen level is normal. Alkaline environment is again unfavorable. It is unbearable, a pH higher than 9 can lead to death of the sturgeon. In this case, the pH values range into the Class I of water quality, according to Order 161/2006, but also within the reference values for sturgeon.

Nitrates, according to Order 161/2006, frame within the normal range (less than 50 mg/l) but exceed the normal limits, if there are taken into account the reference values for sturgeon (max 10 mg/l). Higher values than normal up to 5-6 times can lead to death of the population.

Nitrites in water samples recorded values (less than 0.1 mg/l) corresponding to class I of water quality in accordance with Order 161/2006, as well as reference values for sturgeon.

Phosphorus shows values (0.7, 1.3, 0.8 mg/l) which frame within the normal range according to Order 161/2006 (2 mg/l) and the normal limits for sturgeon (2 mg/l). For the fishery waters phosphorus to 1 mg is considered optimal, and in 2 mg/l it is related to the processes of water contamination.

Iron values recorded 0.5 mg/l, ranging within the limits provided by Order 161/2006, but also within normal limits for sturgeon. At values higher than 3 mg / l of iron salts in water fish die.

Copper in water samples presented the same value in two sampling points, namely the center and the outlet (0.3 mg/l), but recorded a value slightly increased in the chute (0.4 mg/l). It can be argued however that these values are appropriate to class II of water quality according to Order 161/2006 and a little beyond the normal values for sturgeon (less than 0.3 mg/l).

Ammonia recorded insignificant values, within the Class I of water quality according to Order 161/2006 and fitting the reference values for sturgeon (up to 0.05 mg/l).

Sulfate in water samples recorded important exceeding of the normal values (30, 32, 30 mg/l), both provided by the Order 161/2006 (2-7 mg / l) and for sturgeons farming (less than 8 mg/l). An increase in sulfate can damage the skin and resulting in neuronal disorders, inducing a poor welfare.

Phenols had values (0.1, 0.2 mg/l) in two sampling points (the chute and the outlet) that framing within both the normal values in terms of water quality classes specified in the Order 161/2006 and the provided standards for sturgeon (1-2 mg/l), but a slightly higher value than reference (0.21 mg/l) in the center of the pond.

Detergents in water samples showed values (0.03 mg/l, 0.03 mg/l, 0.02 mg/l) which framing within the corresponding values of class I of water quality according to the Order 161/2006, but also the standard for sturgeon (less than 0.1 mg/l).

The results of microbiological parameters analyze of water in the sturgeons’ farm are shown in table no. 3

Table no. 3

Microbiological parameters of water

Sampling point TC/ml water CF/ml (total coliforms)
Chute 720 428
Middle 1005 920
Outlet 900 700

From the data presented in table no. 3 it can be conclude that water has a fairly large microbiological load, which may be due to a lack of water sanitation programs under its regular cleaning. This can be harmful and may increase the frequency of skin and respiratory diseases, resulting in poor welfare of sturgeons.

In order to establish the biochemical profile of sturgeon there were taken blood samples from 10 sujects (male and female), after a manual restraint, by vein puncture with ventral point of election. From the blood samples the following parameters were determined: blood urea nitrogen (BUN), creatinine (CREA), uric acid (URIC), calcium (CA), total protein (TP), albumin (ALB), globulin (GLOB – by difference between TL and ALB), ALT, aspartat aminotransferase (AST), gamma glutamyl transferase (GGT), triglycerides (TRIG), glucose (GLU), lactate dehydrogenase (LDH), phosphorus (PHOS), magnesium (MG), alkalin phosphatase (ALP), total bilirubin (TBIL), cholesterol (CHOL), ammonia (NH3), amylase (AMYL), lipase (LIPA), creatine kinase (CK), by using the Vettest 8008 device. Results interpretation was done according to reference values provided by the Department of Biochemistry, School of Veterinary Medicine – University of Teheran.

The obtained values are presented in table no 4.

Table no. 4

Average values of biochemical parameters – farm 1 in sturgeon

Assessed parameters Obtained value (males) Obtained value (females) Reference value
males females
BUN 3.5 mg/dl 3.0 mg/dl 3.78 +/- 0.84 mg/dl 3.69 +/- 0.64 mg/dl
Creatinine 0.0 mg/dl 0.0 mg/dl 0.340 +/- 0.060 mg/dl 0.344 +/- 0.048 mg/dl
Uric acid 0.1 mg/dl 0.1 mg/dl 0.03 +/- 0.005 mg/dl 0.02 +/- 0.003 mg/dl
Calcium 8.4 mg/dl 9.4 mg/dl 8.52 +/- 2.76 mg/dl 8.52 +/- 2.76 mg/dl
Total protein 4 g/dl 4.3 g/dl 5.50 +/- 0.94 g/dl 4.51 +/- 1 g/dl
Albumin 1 g/dl 1.5 g/dl 1.26 +/- 0.29 g/dl 1.26 +/- 0.29 g/dl
Globulin 3.2 g/dl 2.9 g/dl 4.50 +/- 0.69 g/dl 3.63 +/- 0.84 g/dl
ALT 100 U/L 139 U/L 100.65 +/- 1.18 U/L 100.65 +/- 1.18 U/L
AST 200 U/L 654 U/L 265.6 +/- 56.55 U/L 265.6 +/- 56.55 U/L
GGT 0.0 U/L 0.0 U/L 0.02 +/- 0.035 U/L 0.02 +/- 0.035 U/L
Triglycerides 450 mg/dl 375 mg/dl 699.6 +/- 22.94 mg/dl 699.6 +/- 22.94 mg/dl
Glucose 100 mg/dl 56 mg/dl 120.54 +/- 26.74 mg/dl 61.62 +/- 15.13 mg/dl
LDH 2100 U/L 2800 U/L 2007.15 +/- 521.97 U/L 2007.15 +/- 521.97 U/L
Phosphorus 12.6 mg/dl 12.4 mg/dl 9.009 +/- 2.07 mg/dl 12.39 +/- 0.267 mg/dl
Magnesium 2 mg/dl 1.89 mg/dl 3.67 +/- 0.85 mg/dl 2.79 +/- 0.63 mg/dl
ALP 100 U/L 244 U/L 69.05 +/- 13.04 U/L 69.05 +/- 13.04 U/L
Total bilirubin 0.1 mg/dl 0.1 mg/dl 0.616 +/- 0.0234 mg/dl 0.616 +/- 0.0234 mg/dl
Cholesterol 100 mg/dl 92 mg/dl 90 +/- 40 mg/dl 90 +/- 40 mg/dl
NH3 250 milmol/l 267 milmol/l 300 +/- 170 mmol/l 300 +/- 170 mmol/l
Amylase 0.0 U/L 0.0 U/L 0.001 +/- 0.0003 U/L 0.001 +/- 0.0003 U/L
Lipase 0.0 U/L 0.0 U/L 0.004 +/- 0.001 U/L 0.004 +/- 0.001 U/L
CK 2000 U/L 2036 U/L 2700 +/- 1500 U/L 2700 +/- 1500 U/L

The data shows that males recorded higher values than females for the following parameters: total protein, BUN, uric acid, globulin, magnesium, phosphorus and glucose.

Biochemical parameters for the two sexes (BUN, creatinine, uric acid, calcium, total protein, albumin, globulins, ALT, AST, GGT, triglycerides, carbohydrates, LDH, phosphorus, magnesium, ALP, total bilirubin, cholesterol, NH3, amylase, lipase, CK) are within the standard limits for acipenserid.

For creatinine, lipase, amylase and GGT there were no results, because values were smaller than the detection limits of the kits.

ALT and AST recorded overvalues in females (ALT = 139 U / L, AST = 654 U / l), while in males the values were within the normal range (ALT = 100.65 +/- 1.18 U/L, AST = 265.60 +/- 56.55 U/L).

Triglycerides in serum sampled from both sexes were recorded lower values (375mg/dl, respectively 450 mg / dl) than the normal range (699.6 + / – 22, 94 mg / dL). These values may be due to the stress caused by the restraining and blood collection methods.

Glucose, both in males and females, recorded low values (56 mg / dl and 100 mg / dl) compared with normal values (61,62 +/- 15.13 mg/dl in females and 120.54 +/- 27.74 mg/dl in males) which also may be due to the stress and intense muscular activity that fish have suffered before harvesting.

Magnesium recorded slightly lower values (1.89 mg / dl; 2 mg / dl) than the normal limits (2.79 + / – 0.63 mg / dl females and 3.67 + / – 0.85 mg / dl males).

Alkaline phosphatase recorded much higher values for both sexes (244 U/L and respectively 100 U/l) than the normal limits (69.05 +/- 13.04 U/L). This could be caused by the stress they have been exposed, to nutrition or coping problems, considering the fact that some individuals were newly brought for populating the pool.

BUN presented values (3.5 mg / dl; 3 mg / dl) that framing within the reference values for sturgeon obtained from scientific research on the Black Sea sturgeon (3.69 + / – 0.64 mg / dl females and 3.78 + /- 0.84 mg/dl males).

Uric acid recorded values of 0.1 mg / dl which framing within the normal range for sturgeons (0.02 + / – 0.003 mg / dl females and 0.03 + / – 0.005 mg / dl males).

Calcium recorded values (9.4 mg / dl – 8.4 mg / dl) which framing within the normal range for sturgeon (8.52 + / – 2.76 mg / dl).

Total protein recorded values (4.3 g / dl; 4 g / dl) within the normal range for sturgeon (4.51 + / – 1g/dl females and 5.5 + / – 0.94 mg / dl males).

Globulins in blood samples recorded averages values of 2.9 g / dl, respectively 3.2 g / dl, slightly lower than the admitted limits for sturgeon (3.63 + / – 0.83 g / dl females and 4.5 + / – 0.69 g / dl males.

LDH, phosphorus, total bilirubin, cholesterol, ammonia and creatine kinase average values were within normal limits.

In conclusion, due to overvalues recorded for nitrate (by 1.5 times in the center of the basin and the outlet point), copper and phenols (water falling within class II of quality in terms of their concentration) and sulphates (the most significant overruns –  by 6 times compared with the admitted limit, the water falling in the fourth class of quality for sulfates), the welfare of farmed sturgeon is average.  Biochemical blood picture shows ALT overvalues in females by 1.4 times and alkaline phosphatase in both sexes of 3.5 times.  Welfare problems may arise if there are not taken immediate remedial action.

►In the private fish pond from Comisani area – Dambovita County water quality was monitored, by collecting water samples from 3 points of the pond: admission, center and draining. From the above samples, there were analyzed: pH, O2, ammonia (NH4+), nitrates (NO3-), nitrites (NO2-), total phosphorus (P), iron (Fe), residual chlorine, sulfates (SO42-), copper (Cu) and detergents. Results interpretation was made according to the reference values suggested by Schlotfeldt H. J., 1995.

The results of water quality parameters analysis are shown in table no. 5.

Table no. 5

The values of water quality parameters – Comisani fish pond

Sampling point No. of samples Assessed parameters
pH O2 mg/l NH4 mg/l NO2 mg/l NO3 mg/l P mg/l Iron mg/l Free chlorine mg/l SO4 mg/l Copper mg/l Cationic detergents mg/l
Water admission 10 7.9 15.3 < 0.02 0.027 1.0 0.8 0.22 0.26 77.0 0.13 0.01
Center of the fishpond 10 7.9 21.0 0.01 0.027 2.0 0.8 0.19 0.33 72.0 0.14 0.02
Draining 10 7.9 24.0 0.01 0.030 1.0 1.2 0.1 0.32 71.0 0.16 0.02
Admitted Limits(Schlotfeldt H. J. 1995) 5.5 – 8.0 Min 4 Max 1.0 Max 0.02 5.0 0.6 – 1 Max      2 0.01 – 0.03 80 I 150 II mg/dl Max 0.3 0.05 mg/l

Analyzing the data in the table it can be noticed that the water pH in all 3 collecting points recorded values close to the maximum admitted limits for carp.

Water reaction is important as it is influencing physical chemical and biological processes as well as the toxicity level of metals and nitrites in the water.

Dissolved oxygen in all samples has values in normal range (over 4 mg/l – the minimum admitted limit), proper for the carp rearing.

Of the nutrients category there were assessed ammonia (NH4+), nitrates (NO3-), nitrites (NO2-) and total phosphorus (P). The results in the table show that ammonia, nitrates and nitrites recorded values within the admitted limits for carp, while total phosphorus exceeded the admitted limit in the samples collected from the water draining point by approximately 2 times.

Phosphorus is the second most important chemical element from fisheries in producing organic substance, fish ponds productivity depending on it.

Iron, chlorine and sulfates belong to the general ions class of surface waters quality. The obtained values for iron and sulfates in all water samples range within the admitted limits for carps. Free chlorine recorded overvalues in all collecting points by approximately 10 times.

High chlorine concentrations added to a pH value beyond the optimal level for carp could lead to irritations and gills’ damage.

The high chlorine concentration value revealed in the collected samples does not have a negative influence on fish life since the pH values are within the normal range and water temperature is under 20°C.

Of the metals group as surface waters quality indicators, copper was also assessed, which has recorded values up to the maximum admitted limit (0.3 mg/l) for carp.

Concerning the detergents (toxic substances – indicators of surface waters quality), it can be noticed that all the obtained values following the assessment are proper for carp rearing.

Serum biochemical parameters were determined based on samples from three individuals, the results being shown in table no. 6.

Table no. 6

Values of biochemical parameters in carp from Comisani fishpond, Dambovita county

Assessed parameters Obtained values: carp 1 Obtained values: carp 2 Obtained values: carp 3 Reference values
Blood urea nitrogen(BUN) 3 mg/dl 2 mg/dl 3 mg/dl 1,9-3,6 mg/dl
Creatinine(CREA) 0,1 mg/dl 0 mg/dl 0,1 mg/dl 0,07-0,09 mg/dl
Uric acid(URIC) 1,6 mg/dl 1,2 mg/dl 3,6 mg/dl 1,3-2,5 mg/dl
Calcium(CA) 11 mg/dl 10,1 mg/dl 12,4 mg/dl 8-9 mg/dl
Magnesium(MG) 5,2 mg/dl 3,62 mg/dl 3,62 mg/dl 2,6-3,4 mg/dl
Total protein(TP) 4,3 g/dl 2,8 g/dl 3,6 g/dl 2,5-3,5 g/dl
Albumin(ALB) 1,4 g/dl 0,5 g/dl 0,9 g/dl 0,76-0,85 g/dl
Alanine aminotransferase(ALT) 13 U/l 9,8 U/l 9,9 U/l 8,9-9,9 U/l
Aspartate aminotransferase(AST) 347 U/l 115 U/l 127 U/l 121-124 U/l
Triglycerides(TRIG) 164 mg/dl 170 mg/dl 130 mg/dl N/A
Glucose(GLU) 174 mg/dl 178 mg/dl 158 mg/dl 30-47 (96) mg/dl
Lactate dehydrogenase(LDH) 2565 U/l 2499 U/l 4853 U/l 860-1200 U/l
Alkaline phosphatase(ALKP) 72 U/l 28 U/l 38 U/l 14-24 U/l
Cholesterol(CHOL) 200 mg/dl 99 mg/dl 90 mg/dl 127-184 mg/dl

Analyzing the data in the table, it can be noticed that blood urea nitrogen, creatinine and uric acid are in normal range, so the excretory function (renal and gills excretion) is normal. However, in the third collected carp was registered an exceeding of uric acid reference value with 44% (3,6 mg/dl), related most likely with dehydration and muscular lesions caused by angling.

Regarding serum ionogram, calcium concentrations (10,1 to 12,4 mg/dl) were with 12,2 – 37,7% higher than reference value in all carps while magnesium registered a significant increase only in first individual (with 52,94%).

For total protein, albumin, alanine aminotransferase and aspartate aminotransferase there were noticed increasing of normal range only in the first collected carp, with 22,8%, 64,7%, 31,3%, respectively 179,8%. In conclusions, the welfare level in this individual is poor than in the others collected, the transaminase high activity pointing to severe hepatopancreatic lesions.

Glucose, lactate dehydrogenase and alkaline phosphatase registered overvalues in all collected carps: with 64,58-85,4%; 108,24-304,4% and 16,6-200%. The pronounced change of LDH level shows a high level of stress associated with capture and restraining.

Cholesterol level recorded an insignificant increase in the first collected carp (with 8,69%), while in the other the level is lower, most likely as results of malnutrition.

If the most changes of serum biochemical parameters are caused by fish capture and restraining, in the first individual the increasing is pronounced and is certainly related with welfare issues.

Water quality in Comisani pond is adequate for carp rearing in most parameters, except for free chlorine, and reflects high welfare of fish in relation to their living environment.  However, as shown by analysis of blood chemistry panel, there are individuals with welfare problems.

II) surveillance audit of transport of fish in terms of ensuring the protection and welfare conditions

There was assessed the welfare of carp which were transportated by car for 4 hours in order to populate one of the ponds of Comisani farm.  Fish were transported in doubled plastic bags (to avoid loss of content): 19 l water and 20 l oxygen introduced under pressure (compressor) for each individual of average weight 800 g (transport method described by Orlov).

Regarding water quality, the results revealed small exceedings in ammonia and phosphorus (table no. 7).

Blood biochemical parameters were determined by sampling blood from five individuals and by testing it using Vettest 8008 device.  It has been highlighted the transport stress on the studied fish lot based on the significantly increase creatinine and uric acid (kidney and gill excretory deficiency), calcium and magnesium, total protein and albumin, slightly high ALT and extremely high AST, hyperglycemia and increased  LDH activity.  The remarcable overvalues of some parameters can be explained by summation of transportation stress to the one caused by fish handling and blood sampling.  Ionograme changing and plasma protein level are caused by hemoconcentration due to dehydration and intense muscular effort during the journey and blood collection maneuvers performed on fish.

There is a high variability of values among individuals who have been sampled.

Based on the studied parameters it can be concluded that the welfare of the transported fish is average.

Table no. 7

The values of water quality – crap transport

No. of samples Assessed parameters
pH O2 mg/l NH4 mg/l NO2 mg/l NO3 mg/l P mg/l Iron mg/l Residual  Cl mg/l Cu mg/l Cationic detergents mg/l
Water admission 7.9 6.4 1.9 0.01 3.0 1.2 1.7 0.01 0.09 0.01
Center of the fishpond 7.9 8.9 1.8 0.01 4.4 1.2 1.5 0 0.11 0.02
Admitted limits(Schlotfeldt H. J.,1995) 5.5 – 8.0 Min 4 Max 1.0 Max 0.02 5.0 0.6 – 1 Max      2 0.01 – 0.03 Max 0.3 0.05 mg/l

Table no. 8

Serum biochemical values – transport carp

Analyzed parameters Carp 1 Carp 2 Carp 3 Carp 4 Carp 5 Average values Reference values
Blood urea nitrogen(BUN) 3 mg/dl 3 mg/dl 3 mg/dl 3 mg/dl 3 mg/dl 3 mg/dl 1,9-3,6 mg/dl
Creatinine (CREA) 0,2 mg/dl 0,2 mg/dl 0,1 mg/dl 0,2 mg/dl 0,1 mg/dl 0,6 mg/dl 0,07-0,09 mg/dl
Uric acid (URIC) 4,6 mg/dl 3,6 mg/dl 1,5 mg/dl 3,7 mg/dl 1,7 mg/dl 3,02 mg/dl 1,3-2,5 mg/dl
Calcium(CA) 11 mg/dl 11,2 mg/dl 12 mg/dl 11,2 mg/dl 14,1 mg/dl 11,9 mg/dl 8-9 mg/dl
Magnezium(MG) 5,2 mg/dl 5,2 mg/dl 5,2 mg/dl 5,2 mg/dl 5,2 mg/dl 5,2 mg/dl 2,6-3,4 mg/dl
Total Protein(TP) 3 g/dl 2,7 g/dl 5,5 g/dl 2,7 g/dl 5,8 g/dl 3,94 g/dl 2,5-3,5 g/dl
Albumin(ALB) 0,8 g/dl 0,6 g/dl 2,5 g/dl 0,6 g/dl 2,2 g/dl 1,34 g/dl 0,76-0,85 g/dl
Alanine aminotransferase (ALT) 21 U/l 10 U/l 10 U/l 10 U/l 10 U/l 12,2 U/l 8,9-9,9 U/l
Aspartate aminotransferase (AST) 723 U/l 374 U/l 852 U/l 334 U/l 147 U/l 486 U/l 121-124 U/l
Glucose(GLU) 185 mg/dl 142 mg/dl 81 mg/dl 146 mg/dl 75 mg/dl 125,8 mg/dl 30-47 (96) mg/dl
Lactate dehydrogenase (LDH) 2800 U/l 2800 U/l 2800 U/l 2800 U/l 2800 U/l 2800 U/l 860-1200 U/l
Alkaline Phosphatase (ALKP) 15 U/l 10 U/l 22 U/l 10 U/l 28 U/l 17 U/l 14-24 U/l

III) surveillance audit of slaughter units in terms of ensuring the protection and welfare conditions

Fish welfare was assessed at one slaughter point in Bucharest. After receiving the fish, some of them were kept in large basins (5 m3) until slaughter.

There has been assessed the welfare of a Prussian carp group based on the following elements:

– Behavior (feeding, swimming and exploring the environment: direction, speed etc..), which showed no major changes;

– Storage density: which joined in the standards;

– General clinical examination of fish: the state of fins, gills and the surface of the skin (lesions, discoloration of the gills – which would show the hypoxia and  skin surface – hipermelanosis is associated to stress), the presence and quantity of mucus in the skin, the frequency of opercular movements  etc.  To the specimens followed, there were no signs of illness or major modifications;

– Analysis of blood serum parameters.  There were two sets of samples taken from two individuals each during slauthering: a group of two fish on which there wasn’t applied any method of stunning (mechanical, electrical, chemical) and a group of two fish  chemically stunned through benzocaine (concentration of 100 mg / l) in water.

The results are summarized in the following table.

Table no. 9

Serum biochemical values – slaughter center

Analyzed parameters Prussian carp 1 Prussian carp 2 Average values: C1, C2 Prussian carp 3 Prussian carp 4 Average values: C3, C4
Blood urea nitrogen (BUN) 2 mg/dl 2 mg/dl 2 mg/dl 1 mg/dl 1 mg/dl 1 mg/dl
Creatinine (CREA) 0,5 mg/dl 0,6 mg/dl 0,55 mg/dl 0,3 mg/dl 0,4 mg/dl 0,35 mg/dl
Calcium (CA) 10,9 mg/dl 11,2 mg/dl 11,05 mg/dl 13 mg/dl 12,8 mg/dl 12,9 mg/dl
Magnezium(MG) 5,2 mg/dl 4,95 mg/dl 5,1 mg/dl >5,2 mg/dl >5,2 mg/dl >5,2 mg/dl
Total Protein (TP) 3,8 g/dl 3 g/dl 3,4 g/dl 2,8 g/dl 2,6 g/dl 2,7 g/dl
Albumin(ALB) 1,2 g/dl 0,8 g/dl 1 g/dl 0,7 g/dl 0,6 g/dl 0,65 g/dl
Alanine aminotransferase (ALT) 521 U/l 264 U/l 392,5 U/l 83 U/l 19 U/l 51 U/l
Aspartate aminotransferase (AST) >800 U/l >800 U/l >800 U/l 643 U/l 430 U/l 536,5 U/l
Lactate dehydrogenase (LDH) 2800 U/l 2800 U/l 2800 U/l 2700 U/l 846 U/l 1773 U/l
Alkaline Phosphatase (ALKP) 235 U/l 47 U/l 141 U/l 10 U/l 24 U/l 17 U/l

As seen from the table, for individuals in the second group (Prussian carp 3 and 4) compared with those in group 1, there is a significantly reduced enzyme activity (average value of lactate dehydrogenase – related to intense muscular effort  and muscle lesions associated with handling stress – decreases by 1027 U / L, with a maximum of 1954 U / l; alkaline phosphatase and liver transaminases have substantially lower values).  Also, in the two fish which received chemical stunning (benzocaine) were observed much lower values for the indicators related to the excretory function (urea nitrogen, creatinine), which shows that the stimulus was within the eustress limits compared to their congeners, which exceeded the capacity of adaptation.

IV) submission of proposals for welfare assessment systems in fish farms, during transport and slaughter applicable in our country.

As useful indicators to assess fish welfare in farms, should be established:

– Water quality from chemical and microbiological point of view, the most important factor, due to the fact thet it represents the living environment of fish;

– Groups’ density, according to rearing standards;

– Fish behavior: exploring and feeding (fish dynamic at feeding time, presence of abnormal behavior – which may indicate an inappropriate environment or existence of disease: jumps to the water surface, the behavior of “piping”, isolation of individuals and the presence of individuals with adynamia);

– Clinical examination of the fish (skin, gills lesions, fins’ injuries etc);

– Gain;

– Incidence of disease based on history and health records on the farm;

– Blood chemistry panel, which best reflect the issues of welfare.

Among these factors, the most easily established are water quality and blood chemistry panel.

As useful indicators to assess fish welfare during transportation, should be established:

–               Transport conditions (speed, duration, vibration, vehicles and tanks’ quality etc.) – based on questionnaires, but this indicator is subjective;

–               Stocking density of fish during transportation;

–               Water quality;

–               Fish behavior at the time of receipt (movements intensity reflecting the presence of any welfare problems);

–               The transfer mode of fish from vehicles into the ponds having in view the system and management practices (personnel training level, time);

–               Blood biochemical panel (short-term welfare issues are reflected in increased enzyme level – LDH, ALT, AST, hyperglycemia and excretory troubles- BUN, URIC, CREA).

As useful indicators to assess fish welfare in slaughter units, should be established:

–               Behavior (feeding, swimming and exploring the environment: direction, speed, etc.);

–               Stocking density compared with the standards;

–               General clinical examination of fish: the state of fins, gills and the skin surface, the presence and quantity of mucus in the skin, the frequency of opercular movements;

–               Blood biochemical panel.

Running as many studies in this area (by as many joint research teams, including veterinarians, hygienists, animal scientists etc.), following the above-mentioned indicators and the creation of a widest possible database will allow the desgn of an integrative numerical assessment system, which will be implemented nationally, allowing comparisons between different situations that may arise in the field.