Haematological and biochemical parameters during the laying period in common pheasant hens housed in enhanced cages.

The development of selected haematological and biochemical parameters during the laying period was monitored in common pheasant hens housed in an enhanced cage system. The cages were enhanced by the addition of two perches and a shelter formed by strips of cloth hanging in the corner of the cage. The results showed significant changes in the haematological and biochemical parameters monitored during egg laying. At the time when laying capacity approached a maximum, a decrease was observed (P < 0.05) in haematocrit, erythrocytes, and haemoglobin values, whereas monocytes, eosinophils, the heterophil/lymphocyte ratio, phosphorus, and calcium exhibited an increase (P < 0.05). At the end of the laying period, an increase (P < 0.05) was recorded in the count of leukocytes, heterophils, lymphocytes and basophils, the heterophil to lymphocyte ratio, and the concentrations of aspartate aminotransferase, cholesterol, phosphorus, and calcium, whereas lower values (P < 0.05) were recorded for haematocrit and plasma total protein in comparison with the values of the indicators at the beginning of the laying period. The results provide new information about dynamic changes in selected haematological and biochemical parameters in clinically healthy common pheasant hens during the laying period.

The influence of crate height on selected biochemical indices in captive-reared mallards.

The aim of this study was to investigate the effects of crate type (particularly its height) on selected biochemical indices in captive-reared mallards (Anas platyrhynchos). The physiological changes in response to 2-h crating in crates of either 20 cm or 26 cm height were monitored in 6-week-old mallards. Plasma corticosterone concentrations showed an increase (P = 0.036) in mallards crated in crates of 26 cm height in comparison with control non-crated birds, whereas no difference in plasma corticosterone levels was found between mallards crated in crates of 20 cm height and control non-crated birds. Lower concentrations of plasma triglycerides (P < 0.05) and uric acid (P < 0.05) were found in crated mallards in comparison with control non-crated birds; the height of the crates had no effect. Mallards crated in crates of 26 cm height also exhibited a higher (P = 0.032) plasma lactate dehydrogenase concentration in comparison with control non-crated birds, whereas no difference (P > 0.05) was found in lactate dehydrogenase concentrations in mallards crated in crates of 20 cm height. Our results suggest that crating mallards in lower crates (20 cm) may be less stressful than keeping them in crates allowing vertical movements of the birds.

The effects of subchronic exposure to metribuzin on Danio rerio.

The aim of this study was to assess the impact of metribuzin in surface waters on fish under experimental conditions. The effects of subchronic exposure to metribuzin on fish growth and the development of histopathological changes in selected organs (gill, kidney, liver) and on activity of some biochemical markers (CYP450, EROD) in Danio rerio were investigated during a 28-day toxicity test. Juvenile growth tests were performed on D. rerio according to OECD method number 215. Fish at an initial age of 30 days were exposed to a range of metribuzin concentrations (1.5, 5, 16, 33, and 53 mg L⁻¹). Exposure to metribuzin at 53 mg L⁻¹ was associated with increased mortality. Negative effects with regard to total body weight, length, and the inhibition of specific growth rate were induced at concentrations of 33 and 53 mg L⁻¹. Histopathological examination revealed pathological lesions in the liver in pesticide-exposed fish only at the highest concentration of 53 mg L⁻¹ of metribuzin. Based on the results of growth rate, biochemical markers (CYP450, EROD), and histopathological examination, the lowest observed effect concentration (LOEC) value was 33 mg L⁻¹ and the no observed effect concentration (NOEC) value was 16 mg L⁻¹.

Comparison of acute toxicity of ketoprofen to juvenile and embryonic stages of Danio rerio.

OBJECTIVES: Ketoprofen is a common human medicine from a class of non-steroidal anti-inflammatory drugs (NSAIDs), which is provably detected in surface waters in concentrations ordinarily in µg.L-1. The aim of this study was to compare the acute toxicity of ketoprofen to embryonic and juvenile stages of aquarium fish - zebrafish (Danio rerio). METHODS: Tests were performed according to the methods of the Organisation for Economic Co-operation and Development (OECD) No. 203 (Fish, acute toxicity test) and OECD No. 212 (Fish, short-term toxicity test on embryo and sac-fry stages). RESULTS: The results showed (mean ± SD) LC50 value of ketoprofen to be 632.30 ± 10.10 mg.L-1 in juvenile zebrafish and 6.44 ± 2.22 mg.L-1 in embryonic stages of zebrafish. The results revealed statistically significantly higher sensitivity (p<0.01) of the embryonic stages of zebrafish to ketoprofen compared to its juveniles. The susceptibility of embryos depends on many factors, especially yet improperly developed enzymatic system in embryos, different ways of the absorption of the substance into the organism or differences in metabolism pathways. CONCLUSIONS: The acute toxicity of ketoprofen for juvenile stages of zebrafish is low, but the substance seems to be toxic for embryonic stages.

Effects of subchronic exposure to simazine on zebrafish (Danio rerio).

OBJECTIVES: Simazine is a triazine herbicide which has been used for a long period in agriculture and in the aquatic environment for control of weeds and algae. The aim of this study was to investigate the effects of subchronic exposure to simazine on growth and the development of histopathological changes in selected organs (gills, kidney, liver) in Danio rerio. METHODS: Juvenile growth tests were performed on D. rerio according to the OECD method No. 215. Fish at the age of 20 days were exposed to the environmental simazine concentration commonly detected in Czech rivers (0.06 µg.L-1) and a range of sublethal concentrations of simazine (0.6, 6.0 and 60.0 µg.L-1) for 28 days. RESULTS: There were no significant differences (p<0.05) between the specific growth rates (r) of the test groups and those of both control groups. Histopathological examination revealed pathological changes in fish exposed to a simazine concentration of 60.0 µg.L-1. The values of NOEC and LOEC of simazine were 6.0 µg.L-1 and 60.0 µg.L-1. CONCLUSIONS: The environmental concentration of simazine in Czech rivers did not have any effects on the growth and development of histopathological changes in D. rerio.

The effects of chronic intermittent noise exposure on broiler chicken performance.

Effects of different noise levels (70 or 80 dB) that broilers were exposed to during the entire fattening period and also the effect of the timing of the first exposure to intermittent noise in the course of fattening (day 1 vs. day 7) were monitored. After 7 days of exposure to intermittent noise, experimental chickens already showed a significant decrease in live body weight in comparison with the control group. The difference between the group exposed to intermittent noise at 70 dB and the group exposed to 80 dB levels was not statistically significant, although the mean live body weight of broilers in the latter was lower during the entire fattening period. The chickens exposed to intermittent noise from day 1 of age showed lower mean live weight throughout the fattening period compared to chickens exposed to the same level of intermittent noise only from day 7 of age, although at the end of fattening the difference was statistically significant only in chickens exposed to the higher level of intermittent noise (80 dB).

Responses of peripheral blood leukocytes to chronic intermittent noise exposure in broilers.

The purpose of the study was to specify full leukocytic responses in broilers to chronic intermittent noise exposure at two different levels (70 and 80 dB) and to assess the effect of the timing of the beginning of exposure to intermittent noise (from day 1 or day 7 of fattening). Noise treatment of 80 dB resulted in a significant (P < 0.05) elevation of heterophil to lymphocyte ratio (HLR) indicating a stress response of the broilers to the noise stimulus at the level of 80 dB in the course of fattening, in contrast to the 70 dB noise that did not cause any significant changes in HLR when compared to the control without noise treatment. Noise treatment of both 70 and 80 dB intensities also resulted in a significant elevation of basophil granulocytes. We found a different leukocytic response in broilers exposed to chronic intermittent noise from the first day of fattening in comparison to broilers exposed to this noise stimulus from the seventh day. There was a significantly smaller total number of leukocytes (P < 0.01) and significantly smaller differential counts of heterophils (P < 0.01), lymphocytes (P < 0.01) and basophils (P < 0.05) in broilers treated with noise (both 70 dB and 80 dB) from day 7 (group Day7) than in broilers treated with noise from day 1 (group Day 1). Differential counts of eosinophils and monocytes were not affected by the time of exposure to intermittent noise in this study.

Terbutryn toxicity to Danio rerio: effects of subchronic exposure on fish growth.

OBJECTIVES: The aim of this study was to investigate effects of subchronic exposure to sublethal levels of terbutryn on growth and on histopathological changes in Danio rerio. DESIGN: The acute toxicity tests were performed on the juvenile stage of Danio rerio according to OECD No. 203 Fish, Acute Toxicity Test. The juvenile growth tests were performed on D. rerio according to the OECD method No. 215. Fish at the age of 20 days were exposed to the terbutryn environmental concentration commonly detected in the Czech rivers (0.02 microg L-1) and the range of sublethal concentrations of terbutryn (0.06, 0.2, 0.6 and 2 mg L-1) for 28 days. RESULTS: The 96hLC50 terbutryn mean value for the juvenile stage of D. rerio was 5.71 +/- 0.46 mg L-1. A significant decrease (p < 0.01) in the fish growth caused by terbutryn was observed in the concentration of 0.6 mg L-1. The value of NOEC was 0.2 mg L-1 and LOEC was 0.6 mg L-1 of terbutryn. We found the damage to tubular system of kidneys in the concentration of 0.6 mg L-1 of terbutryn. CONCLUSION: The environmental concentration of terbutryn in the Czech rivers did not have any effects on growth and on histopathological changes in D. rerio and this concentration is lower than determined LOEC and NOEC values of terbutryn.

Comparison of the sensitivity of different fish species to medical substances.

OBJECTIVES: Formaldehyde, sodium chloride and potassium permanganate belong to the commonly used substances for fish treatment. The aim is to define and compare their acute toxicity and therapeutic index between two fish species - Danio rerio and Poecilia reticulata. DESIGN: To determine acute toxicity of these compounds, the semistatic method was implemented in compliance with the OECD No. 203 (Fish acute toxicity test). In each test series, 4 acute toxicity tests were performed. The results were subjected to the probit analysis to determine the 96hLC50 values. Therapeutic index (TI) was calculated with respect to short-term LC50 and effective therapeutic concentrations commonly used for fish treatment. RESULTS: The mean 96hLC50 values for sodium chloride were 21.69 +/- 0.92 g l-1 (TI = 1.4) for P. reticulata and 10.39 +/- 0.12 g l-1 (TI = 1.0) for D. rerio. The acute toxicity of formaldehyde for D. rerio expressed as 96LC50 was 0.12 +/- 0.003 ml l-1 (TI = 5.24) and for P. reticulata 0.1 +/- 0.003 ml l-1 (TI = 4.9). The acute toxicity of potassium permanganate for D. rerio expressed as 96LC50 was 1.25 +/- 0.15 mg l-1(TI = 1.5) and for P. reticulata 1.43 +/- 0.05 mg l-1 (TI = 2). CONCLUSION: P. reticulata showed significantly higher (p < 0.05) tolerance to sodium chloride than D. rerio, whereas D. rerio showed significantly higher (p < 0.05) tolerance to formaldehyde than P. reticulata. The acute toxicity of potassium permanganate was comparable for both fish species. Calculated therapeutic indexes of all tested substances were low; therefore it is important to conduct preliminary tolerance tests before application of treatment baths.

Comparison of acute toxicity of 2-phenoxyethanol and clove oil to juvenile and embryonic stages of Danio rerio.

OBJECTIVES: Anaesthetics are used in aquaculture to prevent stress and mechanical damage to fish during handling or the treatment of fish in breeding, blood sampling and other veterinary interventions. Clove oil and 2-phenoxyethanol are used in the Czech Republic in a water bath for the short-term immobilization of the fish. DESIGN: Acute toxicity tests were performed on aquarium fish Danio rerio, which is considered to be one of the model organisms most commonly used in toxicity testing. The semi-static method according to OECD No. 203 (Fish acute toxicity test) was used for testing juvenile fish. Embryo toxicity tests were performed in zebrafish embryos (D. rerio) in compliance with the OECD No. 212 methodology (Fish, short-term toxicity test on embryo and sac-fry stages). The results obtained (the number of dead individuals at particular test concentrations) were subjected to a probit analysis using the EKO-TOX 5.2 programme in order to determine LC50 clove oil and 2-phenoxyethanol values. The statistical significance of the difference between LC50 values in juvenile and embryonic stages of D. rerio was tested using the Mann-Whitney non-parametric test implemented in the Unistat 5.1 programme. RESULTS: The LC50 clove oil mean value was 18.8 +/- 5.52 mg.L-1 in juvenile D. rerio, and 15.64 +/- 3.30 mg.L-1 in embryonic stages of D. rerio. The LC50 2-phenoxyethanol mean value was 338.22 +/- 15.22 mg.L-1 in juvenile D. rerio, whereas in embryonic stages of D. rerio it was 486.35 +/- 25.53 mg.L-1. CONCLUSIONS: The study proved statistically significantly higher (p<0.01) sensitivity in juvenile fish to 2-phenoxyethanol compared to the embryonic stages. Acute toxicity values of clove oil for juvenile and embryonic stages were comparable.

Effects of different crating periods on selected biochemical indices in broiler chickens.

Effects of different crating periods on selected biochemical indices were monitored in a group of unsexed ROSS 308 broiler chickens aged 42 days. 120 randomly selected chickens were weighted and placed in 15 heavy duty plastic crates with reduction in floor space approx. 160 cm2/kg of body weight of broilers (average body weight was 2.75 kg). Five crates were kept for 4 hours (Group 4h), another five crates for 8 hours (Group 8 h) and another five crates for 12 hours (Group 12h). After tested periods of crating the broilers were released and blood samples were withdrawn from Vena bosilico in 24 randomly selected birds from each group (Groups 4 h, 8 h, 12 h respectively) and also in other 24 randomly selected non-crated control birds. The results of our experiment proved that duration of crating significantly influenced the stress response in broiler chickens. The duration of crating period was positively correlated with the plasma corticosterone level and negatively correlated with the lactate dehydrogenase, cholesterol, triglycerides, glucose and lactate levels in the blood of broiler chickens.

Comparison of the sensitivity of Danio rerio and Poecilia reticulata to silver nitrate in short-term tests.

The aim of this study is to assess the acute toxicity of silver nitrate in adult zebra fish and adult guppies and to compare the sensitivity of these species to this compound. Silver is a naturally occurring element in our environment and it combines with other elements such as sulfide, chloride, and nitrate. Silver, in the form of silver nitrate, is one of the most toxic metals affecting freshwater fish. Industry, particularly photographical and electrotechnical, is the major contributor of silver that is released into the environment. Tests of acute toxicity were performed on the most common species of aquarium fish, Danio rerio and Poecilia reticulata. Both zebra fish and guppies were exposed to progressive concentrations of silver nitrate; a semi-static method according to OECD 203 was used. In each test series, 6 tests of acute toxicity were conducted, with 10 fish used for each separate concentration and for the control group. The results (number of fish deaths in the individual test concentrations) were subjected to probit analysis (EKO-TOX 5.1 software) to determine the 96hLC(50) AgNO(3) values. The 96hLC(50) AgNO(3) value for the zebra fish was (mean±SEM) 15±0.52 µg/l and for the guppies was (mean±SEM) 17.14±5.43 µg/l. We didn't find any statistically significant difference between the sensitivity of zebra fish and guppies. The results reported in this study are in agreement with LC(50) values published in peer-reviewed literature, and conclude that AgNO(3) is one of the most toxic compounds known to fishery.

Variation in the mortality rate of turkeys during transport to the slaughterhouse with travel distance and month.

Failure to comply with animal welfare requirements during the transport of turkeys to the slaughterhouse increases stress in animals, which is manifested by increased mortality rate during transport. The numbers of turkeys that died during transport or soon after arrival may serve as an important parameter to indicate the level of animal welfare during transport of turkeys. The number of turkeys that died during transport to slaughterhouses in the Czech Republic in the period from 1997 to 2004 was investigated. The mortality rate found was 0.28% +/- 0.06% but varied with travel distance. The lowest mortality rate was found in case of travel distance below 50 km (0.18% +/- 0.08%) while long travel distances resulted in considerable increase in the mortality rates of turkeys (between 0.28% +/- 0.07 and 0.37% +/- 0.10%). The mortality rate of transported turkeys was also affected by the particular month of the year. Thus, the highest overall mortality rate occurred at long travel distances during winter months, i.e. in December (0.34% +/- 0.18%), January (0.32% +/- 0.06%), and February (0.36% +/- 0.07%). The comparison of individual years has shown a long-term trend towards a decrease in turkeys' mortality during transportation to slaughterhouses from 0.32% in 1998 to 0.20% in 2004. The decrease was statistically significant (Spearman's rank correlation coefficient r = -0.86, p < 0.01). This trend can be evaluated as positive.

Effects of reduction in floor space during crating on haematological indices in broilers.

In the perspective of animal protection and welfare, the most common and potentially the most frightening events encountered by the domestic fowl are procedures connected with transport. This study provides an evidence that reduction in floor space during crating, which is approximately 10% above requirements given to EEC Directives (accompanied with mixed social and heat stress due to high animal density), induces markedly elevated stress in broilers which is manifested by changes in haematological indices. The experiment was performed on ROSS 308 broilers (total number 70, mean body weight 3.05 kg). At 42 days of age, the broilers were crated for 2-hours either with reduction in floor space to 115 cm2/kg (S115) or 105 cm2/kg (S105) of body weight. The total erythrocyte and leukocyte count, haematocrit, haemoglobin, MCV (mean cell value), MCH (mean cell haemoglobin) and MCHC (mean cell haemoglobin concentration) values were monitored and differential leukocyte count was assessed 20 h following crating. When compared with uncrated control, S105 broilers exhibited highly significant increase in haemoglobin level, MCH and MCHC values, significant increase in MCV values and significant decrease in total erythrocyte count. S115 broilers did not manifest any significant changes in haematological parameters compared with control. When compared with S115 broilers, S105 broilers manifested significantly elevated haemoglobin level and MCV values, highly significantly elevated MCH and MCHC values and depressed erythrocyte count. It follows from results that the reduction in floor space to 115 cm2/kg for 2 hours did not induce any significant changes in both total and differential leukocyte counts in blood of broilers when compared with control chickens. However, reduction in floor space to 105 cm2/kg resulted in highly significant increase in heterophil counts and subsequently also HLR (heterophil/lymphocyte ratio) was significantly elevated. When compared S105 and S115 broilers, mean values of heterophil counts and HLR ratio were higher due to reduction in floor space to 105 cm2/kg. The differences, however, were not statistically significant.