Neopterin and biopterin as biomarkers of immune system activity associated with crating in broiler chickens.

Neopterin and biopterin belong to a group of unconjugated pterin derivates. These biomolecules are present in many animal species and perform several functions. Pterin concentrations may provide additional information on the effect of stress on immune system activity. This study focused on an investigation of the effect of crating on plasma concentrations of neopterin and biopterin in broilers. The effects of 2 crating periods (2 and 4 h) were monitored in Hubbard broilers (n = 90) aged 42 d. After a given crating period, randomly selected chickens from each group were sampled immediately and the remaining chickens were sampled after 24 h. Plasma corticosterone increased (P < 0.001) immediately after 2 and 4 h crating, but no difference between the crated and the control noncrated broilers was found 24 h later. Immediately after crating, neopterin in 2- and 4-h broilers did not differ from the control, but 24 h later a decrease (P = 0.011) in plasma neopterin was found in 4-h broilers compared with the control. Simultaneously, 24 h after crating, neopterin levels in 2- and 4-h broilers decreased (P < 0.001) in comparison with the levels immediately after crating. Plasma biopterin was higher (P < 0.001) in 4-h broilers than in the control immediately after the crating. A time of sampling effect (P = 0.016) was found for the heterophil-to-lymphocyte ratio, with heterophil-to-lymphocyte ratio higher 24 h after crating in comparison with its level immediately after the crating. This study shows that crating may significantly affect the immune system of broiler chickens. This is corroborated by the increase in plasma biopterin concentrations in broilers immediately after crating and the decrease in plasma neopterin concentrations in broilers 24 h after crating. The correlations were found for widely used indicators of acute and chronic stress in birds [i.e., plasma corticosterone concentrations (biopterin) and the heterophil-to-lymphocyte ratio (neopterin), respectively].

Biochemical and haematological profile of pheasant hens during the laying period.

The present paper provides new experimental data on the biochemical and haematological profile of blood in pheasant hens, and points out the changes in both biochemical and haematological parameters that occur during the laying period. Significant effects of egg laying on both the biochemical and the haematological blood parameters of pheasant hens were found. Biochemical analyses revealed a significant increase in the metabolites cholesterol, uric acid, lactate, the enzyme aspartate aminotransferase (AST) and the minerals calcium and phosphorous, as well as a significant decrease in total protein, albumin and glucose in the course of the laying period. Haematological analyses revealed a significant increase in the count of leukocytes, lymphocytes, eosinophils, basophils and monocytes due to egg laying. In addition, the erythrocyte count and haemoglobin content significantly decreased in the middle of the laying period and then rebounded at the end of the laying period. The haematocrit content gradually decreased till the end of the laying period. All together, the results of this study underline the impact of the reproduction status of pheasant hens on basic blood parameters. The biochemical and haematological values presented in this study may be of help in assessing disease conditions in laying pheasant hens.

Changes in selected biochemical indices, leukocyte profile, and pterins as biomarkers of immune system activity due to antipecking measures in pheasants.

The physiological changes in response to beak trimming and spectacle usage as antipecking measures were monitored in 10-mo-old common pheasants (Phasianus colchicus). Short-term analysis conducted before the beginning of the laying period showed immediate increases of plasma corticosterone (P < 0.05) and lactate dehydrogenase (P < 0.001) concentrations and decrease of plasma triglycerides (P < 0.01) levels in response to both beak trimming and the application of spectacles. Beak-trimmed pheasants exhibited higher plasma corticosterone concentrations than pheasants fitted with spectacles (P < 0.001). To assess long-term changes, blood samples for biochemical (neopterin and biopterin determination) and hematological (leukocyte profile determination) examinations were taken from beak-trimmed, spectacles-fitted, and control pheasant hens housed in cages during their laying period. At the end of the laying period, hens fitted with spectacles exhibited lower concentrations of plasma neopterin (P = 0.005) and biopterin (P = 0.005) than beak-trimmed pheasant hens. Our findings suggest that the immune system was suppressed in spectacles-fitted pheasant hens as a result of chronic stress, as also indicated by the higher heterophil-to-lymphocyte ratio (P = 0.001) compared with beak-trimmed hens. Our study found a negative correlation (r = -0.31, P = 0.019) between the heterophil-to-lymphocyte ratio and plasma neopterin concentration. This study demonstrated that both beak trimming and use of spectacles are not only stressful procedures for pheasants, but long-term effects may also include a negative impact on the immune system.

Recovery ability of common carp (Cyprinus carpio) after a short-term exposure to terbuthylazine.

Effects of a high terbuthylazine concentration (3.3 mg/1) on Cyprinus carpio were studied using a commercial herbicide formulation Click 500 SC (terbuthylazine 500 g/l). The fish were exposed to the pesticide for 24 h and allowed to recover for 6 days. Biometric parameters, plasma biochemical parameters and biomarkers of oxidative stress as well as histopathological changes in selected tissues were assessed on day 1 and 7. After a 24-h exposure, there were significant alterations found in the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) as well as in the plasma concentrations of glucose, natrium, chlorides, calcium and phosphorus. Hepatosomatic index, plasma albumin and lactate reflected the treatment with a delay. Ion levels and ALT were found to be restored after a 6-day recovery period, which was too short for AST activity and glucose to diminish to the control levels. The histopathological examination revealed disorders in the gills of the exposed fish, however, the changes were not detected after a 6-day recovery period. The study shows high regeneration potential of the fish.

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

The aim of this study was to investigate the effects of subchronic exposure to atrazine on fish growth and the development of histopathological changes in selected organs (gill, kidney, liver) in Danio rerio. Juvenile growth tests were performed on D. rerio according to OECD method No. 215. For 28 days, fish at an initial age of 30 days were exposed to the environmental atrazine concentration commonly detected in Czech rivers (0.3 microg/L) and a range of sublethal concentrations of atrazine (3.0, 30.0 and 90.0 microg/L). The results showed decreasing growth rates and morphological changes in the liver (dystrophic lesions of hepatocytes) at 90.0 microg/L of atrazine. The environmental concentration of atrazine in Czech rivers did not have any effect on fish growth and development of histopathological changes in D. rerio. The value of NOEC was 30.0 microg/L and the value of LOEC was 90.0 microg/L.

Time course changes in selected biochemical indices of broilers in response to pretransport handling.

Two experiments were conducted to assess the stress response of broilers to catching and pretransport handling followed by different periods of crating. The short-term changes in selected biochemical indices were monitored at 1-min intervals within 10 min of crating after the catching and handling of broilers (experiment 1). These indices were further monitored at 15-min intervals for 2 h of crating after the catching and handling of broilers (experiment 2). Increased (P < 0.001) corticosterone concentrations were observed immediately after the broilers were caught, handled, and placed in crates. They continued to increase until 7 min after crating and then slowly decreased, but even at 120 min after handling, corticosterone concentrations were higher (P < 0.001) in crated broilers than in control broilers. In addition, lactate concentrations increased (P < 0.001) immediately in comparison with those of broilers with no additional handling except for catching and blood sampling, but 15 min later, the lactate concentrations had decreased to the precrating level. Lactate dehydrogenase concentrations increased (P = 0.042) 30 min after crating and continued to increase for the rest of the monitored period. A decreased level of cholesterol (P = 0.017) and increased concentration of uric acid (P = 0.041) were found 1 min after crating. The decrease in cholesterol was visible up to 9 min after crating; it then returned to its original value except for the period from 75 to 90 min after crating, when a decrease (P < 0.05) was again detected. Higher (P < 0.01) concentrations of uric acid were found continuously from 4 to 75 min after crating. Glucose concentrations were increased (P = 0.017) 2 min after crating, but the stress-induced increase was not consistent over the course of the next few minutes after crating. Glucose concentrations were not different from those of control broilers from 10 to 120 min after crating, although they showed a decreasing pattern. A decrease (P = 0.031) in triglyceride concentrations was detected 75 min after crating.

Changes in selected biochemical indices related to transport of broilers to slaughterhouse under different ambient temperatures.

The effect of transport distance on selected biochemical parameters (corticosterone, uric acid, triglycerides, total protein, glucose, and lactate) under various ambient temperatures was monitored in a group of unsexed Ross 308 broilers aged 42 d. Broilers were transported to the slaughterhouse over 3 different travel distances (10, 70, and 130 km). They were sampled before and after each transportation in 3 various periods with different ambient temperatures (-5 to +5°C, 10 to 20°C, and 25 to 35°C), which approximately correspond to temperature conditions during transport in individual seasons of the year (winter, fall, summer). The changes in biochemical parameters were specific in their dependence on the travel distance and the ambient temperature under which the broilers were transported. The highest corticosterone concentration was found in broilers before transport (i.e., immediately after catching, crating, and loading) at all ambient temperatures. The concentration of corticosterone was higher at winter temperatures than at summer and fall temperatures. Triglycerides decreased with travel distance, although this effect was detected under summer temperatures only. The concentration of total protein was higher only after 10 km of transport and then it decreased with travel distance at all monitored ambient temperatures. A highly significant decrease (P < 0.01) in the glucose level of broilers was observed after 130 km of transport when compared with broilers before transport at fall and winter temperatures. The effect of travel distance on lactate concentrations was the same at all monitored ambient temperatures, with the lactate level decreasing with travel distance. The results obtained indicate that pretransport handling procedures (catching, crating, and loading) may be more stressful for broilers than the transport itself. To improve broiler meat quality, it is necessary to meet the need for broilers to recover before being slaughtered. With regard to different seasons of the year, we can assume that transport under conditions of low ambient temperatures in winter represents a more stressful event than transport during fall and summer.

Stress in broilers resulting from shackling.

The aim of this study was to assess stress response of broilers to different periods of shackling. Stress effects of shackling were monitored in a group of male Ross 308 broilers (total number: 400) aged 42 d. Three shackling treatments were used in our experiment: shackling of broilers for 30 s (group T30), 60 s (group T60), and 120 s (group T120). Corticosterone plasma concentration was elevated in T60 broilers (P<0.05) and in T120 birds (P<0.01); glucose plasma concentration was increased (P<0.05) in both T60 and T120 broilers when compared with nonshackled control. Lactate concentrations increased in T30 birds (P<0.05) and in both T60 and T120 birds (P<0.01). Furthermore, T120 broilers exhibited an increase (P<0.01) in heterophil counts and heterophil:lymphocyte ratio. Duration of tonic immobility was increased (P<0.05) in T60 and T120 broilers. Number of attempts to induce tonic immobility decreased (P<0.01) in all test groups (T30, T60, T120). Duration of shackling period was positively correlated (P<0.001) with corticosterone, glucose and lactate level, tonic immobility duration, and heterophil:lymphocyte ratio. The number of inductions was negatively correlated (P<0.001) with duration of the shackling period. According to the results of our study, the act of shackling is a considerable traumatic procedure for broilers, and its stress effect is markedly dependent on duration of shackling period that the broiler chickens experience. It follows from our study that the optimal shackling period should be less than 60 s.

Changes in haematological profile of common pheasant (Phasianus colchicus) induced by transit to pheasantry.

The aim of this study was to assess haematological changes in hand-reared pheasants (Phasianus colchicus) transported from intensive housing facilities to a pheasantry. Selected haematological parameters were monitored in a group of 100 pheasants (50 males and 50 females) aged of 9 weeks that were transported for 4 hours by a covered lorry in crates, with a total body weight of 12 +/- 0.5 kg per crate (Group C12 - floor space: 290 cm2/kg) and with a total body weight of 18 +/- 0.5 kg per crate (Group C18 - floor space: 195 cm2/kg). Blood samples were taken from 10 randomly selected males and 10 females before transport (CON group) and 20 hours after transport (C12 and C18 groups). Examinations consisted in determining the total erythrocyte and leukocyte counts, haematocrit values, haemoglobin levels and differential leukocyte counts, whereby the proportions of heterophil, basophil and eosinophil granulocytes, lymphocytes and monocytes of the total leukocytes were computed. The changes in the parameters of red blood cell count were manifested by an increase (P < 0.01) in the haemoglobin level, MCH (mean cell haemoglobin) and MCHC (mean cell haemoglobin concentration) values and a decrease (P < 0.01) in the total erythrocyte count and haematocrit level in both C12 and C18 pheasants, when compared with the control group of non-transported pheasants. C18 pheasants exhibited also a significant increase (P < 0.05) in MCV (mean cell value) value. When analyzing differential leukocyte counts, C18 pheasants showed a decrease (P < 0.01) in heterophil counts and H/L ratio, whereas values in C12 pheasants did not differ from the non-transported control group. Individual counts of lymphocytes were decreased (P < 0.05) in C12 pheasants, whereas basophil counts were increased (P < 0.01) in both C12 and C18 pheasants. Total leukocyte count was decreased (P < 0.01) in C12 and C18 pheasants. In conclusion, the specific requirements of pheasants, as primarily wild animals, for the density in crates should be respected during transportation and they should be transported at lower densities than other poultry species, at least 290 cm2/kg live weight should be provided.