Binding of soluble immune complexes to fractionated Atlantic salmon (Salmo salar L.) leucocytes.

Binding of a fluorescent-labelled soluble immune complex to different types of Atlantic salmon leucocytes was investigated using flow cytometry. Peripheral blood leucocytes (PBL) were separated into sIg+ and sIg enriched populations by magnetic activated cell sorting, blood neutrophils were identified by electronic gating, and kidney macrophages were selected by plastic-adherence. About 60% of both sIg+ and sIg- enriched PBL, 44% of neutrophils and 34% of macrophages bound the soluble immune complexes.     

Graft-versus-host reaction (GVHR) in clonal amago salmon,Oncorhynchus rhodurus

The graft-versus-host reaction (GVHR) was demonstrated in a salmonid model system of clonal diploid and triploid amago salmon. Triploid operculum grafts on clonal diploid evoked an acute rejection within 12 days. Grafts exchanged among triploid amago salmon exhibited prolonged survival for 18 days. In contrast, diploid grafts on triploid, and allografts among clonal diploid amago salmon were accepted. A typical GVHR was induced in triploid recipients by intraperitonal injection of head kidney cells from sensitised diploid donors. The clinical signs of graft-versus-host disease (GVHD) were observed in the recipients after 1 week of cell injection as a loss of appetite and appearance of solid faeces, followed by haemorrhage, local swelling of ventral skin and an enlarged spleen. Three of six fish died within 1 month. Water temperature and frequency of sensitisation are critical to induce GVHR. Diploid donors had to be sensitised three times at 20 degrees C to induce the typical GVHR. GVHR was most effectively induced by head kidney cells, followed by peripheral blood leucocytes (PBL) and spleen cells. Ploidy analysis by flow cytometry revealed that the donor head kidney cells greatly increased in the recipient liver, head kidney and spleen, and reached the peak after 9 days of donor cell injection. The results in the present study are quite similar to the findings in ginbuna and ginbuna-gold fish hybrid system, suggesting the presence of T cells in salmonid as well as cyprinid fish.     

Assessing the Suitability of a Partial Water Reuse System for Rearing Juvenile Chinook Salmon for Stocking in Washington State

Abstract

To assess the suitability of water reuse technology for raising Pacific salmon Oncorhynchus spp. for stocking purposes, fish health and welfare were compared between two groups of juvenile Chinook salmon O. tshawytscha from the same spawn: one group was reared in a pilot partial water reuse system (circular tanks), and the other group was reared in a flow-through raceway. This observational study was carried out over a 21-week period in Washington State. Reuse and raceway fish were sampled repeatedly for pathogen screening and histopathology; fin erosion and whole-blood characteristics were also evaluated. By the study's end, no listed pathogens were isolated from either cohort, and survival was 99.3% and 99.0% in the reuse and raceway groups, respectively. Condition factor was 1.28 in raceway fish and 1.14 in reuse fish; this difference may have been attributable to occasional differences in feeding rates between the cohorts. Fin indices (i.e., length of the longest dorsal or caudal fin ray, standardized by fork length) were lower in reuse fish than in raceway fish, but fin erosion was not grossly apparent in either cohort. The most consistent histological lesion was gill epithelial hypertrophy in reuse fish; however, blood analyses did not suggest any corresponding physiological imbalances. Overall, results suggest that water reuse technology can be employed in rearing juvenile anadromous salmonids for stocking purposes.

Received December 3, 2009; accepted February 14, 2011     

Fin Condition of Atlantic Salmon Reared at High Densities in Heated Water

Abstract

We assessed the condition of dorsal and pectoral fins of parr of Atlantic salmon Salmo salar that were reared for 70 d at 17.5°C to final densities of 14–55 kg/M² (rearing-unit bottom area) or 80–310 kg/m³ (rearing-unit volume). Pectoral fins were not significantly affected by rearing density, but dorsal fins were more eroded in fish reared at the highest density than in fish checked before the rearing experiments or in those reared at lower densities. Densities as great as 41 kg/m² (229 kg/m³) did not affect fin quality of Atlantic salmon reared at 17.5°C.     

Effect of two thermal regimes on the muscle growth dynamics of sea bass larvae,Dicentrarchus labrax L

Muscle growth was studied in larvae of sea bass, Dicentrarchus labrax L., reared at two temperatures: real ambient temperature ( congruent with 15 degrees C during vitelline phase and increased gradually) and 19 degrees C from fertilization until the end of larval development. Muscle cellularity, body length and body weight were measured. Early temperature influenced larval development and so, pre-larval phase finished earlier at 19 degrees C than at ambient temperature (4 and 6 days, respectively). Temperature also affected muscle growth such that at hatching and at mouth opening hypertrophy of muscle fibres was greater at 19 degrees C (P < 0.05), whereas hyperplasia was similar in both groups. After 25 days, the cross-sectional area of the white muscle was greater at 19 degrees C (P < 0.05), which was mainly associated with a higher proliferation of new white muscle fibres. At this stage the body length was also higher at 19 degrees C. Metamorphosis finished earlier in fish reared at 19 degrees C (52 days) than at natural temperature (82 days). At this developmental stage body length and cross-sectional area of the myotome were similar in both groups. However, muscle cellularity differed between groups. Thus, hypertrophy of muscle fibres was higher in fish reared at ambient temperature (P < 0.05), whereas proliferation of new muscle fibres was higher at 19 degrees C (P > 0.05).     

Effect of slaughter by removal from water on visual evoked activity in the brain and reflex movement of rainbow trout (Oncorhynchus mykiss)

The effects of removal from water and death by anoxia on brain function and reflex movement were examined in conscious trout. Visual evoked responses (VERS) were measured in fish with a body temperature of 20 degrees C, 14 degrees C and 2 degrees C. On average the fish took 2.6 minutes at 20 degrees C, 3.0 minutes at 14 degrees C and 9.6 minutes at 2 degrees C to lose VERS, and 11.5, 27.7 and 197.6 minutes, respectively, to lose all movement. VERS were absent from fish anaesthetised with MS222. From a humanitarian standpoint these results indicate that fish killed by anoxia could be exposed to periods of distress or suffering, and that cooling them in ice prolongs this period.     

Assessing the suitability of a partial water reuse system for rearing juvenile Chinook salmon for stocking in Washington State

To assess the suitability of water reuse technology for raising Pacific salmon Oncorhynchus spp. for stocking purposes, fish health and welfare were compared between two groups of juvenile Chinook salmon O. tshawytscha from the same spawn: one group was reared in a pilot partial water reuse system (circular tanks), and the other group was reared in a flow-through raceway. This observational study was carried out over a 21-week period in Washington State. Reuse and raceway fish were sampled repeatedly for pathogen screening and histopathology; fin erosion and whole-blood characteristics were also evaluated. By the study's end, no listed pathogens were isolated from either cohort, and survival was 99.3% and 99.0% in the reuse and raceway groups, respectively. Condition factor was 1.28 in raceway fish and 1.14 in reuse fish; this difference may have been attributable to occasional differences in feeding rates between the cohorts. Fin indices (i.e., length of the longest dorsal or caudal fin ray, standardized by fork length) were lower in reuse fish than in raceway fish, but fin erosion was not grossly apparent in either cohort. The most consistent histological lesion was gill epithelial hypertrophy in reuse fish; however, blood analyses did not suggest any corresponding physiological imbalances. Overall, results suggest that water reuse technology can be employed in rearing juvenile anadromous salmonids for stocking purposes.     

Metabolism and residue depletion of albendazole and its metabolites in rainbow trout, tilapia and Atlantic salmon after oral administration

Metabolic and residue depletion profiles of albendazole (ABZ) and its major metabolites in three fish species, rainbow trout, tilapia and Atlantic salmon are reported. Based on these profiles, similarities (or dissimilarities) between species will determine the potential to group fish species. ABZ at 10 mg/kg body weight was incorporated into fish food formulated in a gelatin base or in gel capsule and fed as a single dose to six fish from each species. Rainbow trout were held three each in a partitioned 600-L tank. Tilapia and Atlantic salmon were housed in separate 20-L tanks. Samples of muscle with adhering skin were collected at 8, 12, 18, 24, 48, 72, and 96 h postdose from trout kept at 12 degrees C, at 4, 8, 12, 24, 48, 72, 96, 120, and 144 h postdose from tilapia kept at 25 degrees C and at 8, 14, 24, 48, 72, and 96 h postdose from Atlantic salmon kept at 15 degrees C. The samples were homogenized in dry ice and subjected to extraction and cleanup procedures. The final extracts were analyzed for parent drug ABZ and its major metabolites, albendazole sulfoxide (ABZ-SO), albendazole sulfone (ABZ-SO2) and albendazole aminosulfone using high-performance liquid chromatography with fluorescence detection. ABZ was depleted by 24 h in trout and tilapia and by 48 h in salmon; ABZ-SO, a pharmacologically active metabolite, was depleted by 48 h in tilapia, by 72 h in rainbow trout and was present until 96 h in salmon; and low levels of ABZ-SO2 and albendazole aminosulfone, both inactive metabolites, were detectable at least till 96 h in all three fish species.     

Pathology of ocular lesions associated with gas supersaturation in white seabass

Cultured juvenile white seabass Atractoscion nobilis (WSB) can suffer from intraocular emphysemas and exophthalmia in the hatchery environment. To identify the cause, two size-groups of WSB were exposed to five gas saturation levels, ranging from 98% to 122% total gas pressure (TGP), over a 96-h exposure period in 18 degrees C and 23 degrees C seawater. Histological examination revealed that the gross and subgross lesions associated with gas supersaturation included corneal and orbital emphysema, along with subretinal, optic nerve, and iridial hemorrhage. Corneal emphysema was the most prominent gross lesion, with the severity and prevalence increasing between size-groups and water temperatures as TGP increased. Following the same pattern was orbital emphysema, which affected more than 93% of the fish examined and caused hemorrhage in the subretinal space, around the optic nerve, in the iris, or a combination thereof. Iridial hemorrhage occurred in 91% of the fish examined and decreased significantly with fish size. The prevalence and severity of hemorrhage in the subretinal space increased significantly with TGP and fish size but not with temperature. Optic nerve hemorrhage was absent in small fish exposed at 18 degrees C but increased significantly with temperature and fish size. The reverse was true for the large fish.     

Early-age cold conditioning of broilers: effects of timing and temperature

1. The aim of the present study was to determine the effects of early-age cold conditioning (CC) on performance, ascites mortality, thyroid hormones status and immune response (leucocytes count) of broiler chickens.

2. A total of 336 chicks at 2 and 3?d of age were exposed to 20 or 25°C (for 3 or 6?h) in a 2?×?2?×?2 factorial experiment, while a control group was kept under normal temperature conditions (30°C). Thereafter, both control and cold conditioned birds were reared under standard conditions until 42?d of age.

3. The results showed that performance (weight gain and feed efficiency) was improved by CC at the end of the rearing period. Carcase traits (breast, thigh and abdominal fat percentage) were not affected by different treatments. Heart weight was lower in cold conditioned birds accompanied with lower ascites mortality. Total leucocyte count was higher in CC birds. Higher concentrations of thyroxin (T₄) were found in plasma of treated groups, while triiodothyronine (T₃) to T₄ ratio was decreased.

4. In conclusion, it seems that early age CC improves performance and reduces ascites mortality of broilers through altered thyroid hormones metabolism and leucocyte function. According to the results, the best timing for CC of broilers was 20°C for 6?h at the age of 2?d, and no significant benefit was observed by repeated CC.     

Ethically chosen studies with early-weaned piglets during their raising in pens with different applications of straw. 1. The effects of different applications of straw and different floor conditions in areas of uniform size

In extension of earlier experiments in housing systems with perforated floors, utilization rates of solid floor with litter, straw in a rack and deep litter, as well as the behaviour and physical condition, especially of claws, were investigated. Each pen had an area of 0.45 m2/piglet. In further experiments the influences of race (DL instead of DL x Piétrain), kind of rearing (without straw instead of rearing with straw), lowering temperature and the effect of a preferred perforated floor in comparison with solid floor were investigated. Deep litter was preferred for activity and lying behaviour only in the case of low temperature (14-18 degrees C) whereas solid floor with litter was preferred in the case of higher temperature (19-25 degrees C). The perforated floor was equivalent to solid floor on the condition that the area dimension was sufficient, temperature 3 degrees C higher than in the other experiments and straw in a rack was offered. However, the utilization rates of solid floor increased in the case of lowering temperature by 4 degrees C. There was no significant influences of the different race or kind of rearing. In the experiments there were significant signs of a more harmonious condition of the animals and a shorter, respectively no period where the animals had to adapt to the housing conditions. These findings were in contrast to those of earlier investigations. The possibility of acting with straw led to an undisturbed circadian rhythm and to a higher rate of standing within the total activity which was as high as in earlier studies. Further social companions and environmental objects lost attractiveness.(ABSTRACT TRUNCATED AT 250 WORDS)     

Studies on the modulation of leucocyte subpopulations and immunoglobulins following intramammary infusion of beta 1,3-glucan into the bovine udder during the dry period

Inflammatory and immunological reactions after intramammary infusion of beta 1,3-glucan were studied during the steady dry period and involution phase of the bovine udder. The effects of a single intramammary infusion of two different doses (100 and 200 mg) of beta 1,3-glucan were evaluated during the steady dry period. In a second study, the effects of beta 1,3-glucan at drying off were studied by using two treatment regimens; a single infusion at drying off, compared with two infusions of the compound, at drying off and again 2 weeks later. Total and differential leucocyte counts were measured in both blood and udder secretions. Additionally, the expression of receptors for CD14 and MHC class II on leucocytes, and the expression of receptors for CD4, CD8, WC1, IL2R and B-cells on lymphocytes was measured in mammary secretions by flow cytometric analyses. The concentrations of immunoglobulins in udder secretions were measured by radial immunodiffusion. The results showed that a single intramammary infusion of beta 1,3-glucan during the steady dry period causes transient enhancement of some aspects of the inflammatory and immune responses. The increases in somatic cell counts, numbers of monocytes/macrophages, and in proportions of CD14+ and MHC class II+ leucocytes in udder secretions were dose-dependent. Infusion of beta 1,3-glucan also slightly increased the proportion of CD4+ lymphocytes and the concentrations of IgG1 and IgG2 in dry secretions. Infusion of beta 1,3-glucan at drying off seemed to accelerate the involution process through an increase in somatic cells, particularly in the numbers of macrophages, in mammary secretions. The numbers of lymphocytes and polymorphonuclear leucocytes, the proportions of IL2R+ lymphocytes, the proportions of CD14+ or MHC class II+ leucocytes and the concentrations of IgG1 and IgG2 also increased in comparison with untreated controls. Moreover, a second infusion of beta 1,3-glucan tended to prolong this response, indicating that this might be an effective means of enhancing the mammary defence against udder infections closer to calving. In conclusion, the results indicate the intramammary infusion of beta 1,3-glucan could be used to enhance the defence mechanisms of the bovine udder against infections, especially during early involution.     

Decreased mononuclear cell response to mitogens in artificially reared neonatal pigs.

The influence of two rearing methods for neonatal pigs on mononuclear cell responses to mitogens was studied. Littermate pigs were reared artificially or on the sow. Artificially reared pigs displayed a leukocytosis (P less than 0.05) characterized by an increase (P less than 0.05) in mature and immature neutrophils. Skin-test responses to intradermal injections of phytohemagglutinin were less (P less than 0.05) in artificially reared pigs than in sow-reared controls. Lymphocyte proliferative responses to mitogenic stimulation were approximately 50% lower (P less than 0.05) in artificially reared pigs than in sow-reared pigs. These data suggest that artificial rearing lowers in vivo and in vitro lymphocyte responses in neonatal pigs.     

Leucocyte counts in the healthy English Thoroughbred in training

Total and differential leucocyte counts have been determined by electronic counting techniques in 474, two-to four-year-old healthy Thoroughbreds in training. The ranges of values observed (particularly those for neutrophils and lymphocytes) were narrower than previously described. Absolute and percentage lymphocyte values were significantly decreased with advancing age causing a relative increase in percentage neutrophils. The fall in absolute lymphocyte numbers was the main cause for a significant decrease in total leucocytes with age. Frequency plots for each variable showed that, with the exception of the eosinophil count, data from other leucocyte cell types following either a normal or lognormal distribution.     

Effects of ambient temperature and restricted feeding on the growth of feathers in growing turkeys

1. Male turkeys were reared to 6 weeks of age at 15 degrees C and 26 degrees C and fed ad libitum or restricted to 0.5 of the body weight of birds fed ad libitum. Basal metabolic rate was determined by indirect calorimetry at an ambient temperature of 20 degrees C. 2. Turkeys at 15 degrees C were lighter than those kept at 26 degrees C. Feather lengths and weight were similar in both groups. Fasting heat production corrected for both metabolic body size and activity was greater in turkeys reared at 15 degrees C than those at 26 degrees C. 3. Cranial breast feathers were significantly longer in restricted birds than in those fed ad libitum in contrast to a proportional decrease in the lengths of other feathers of 0.1 to 0.3. Feather weight as a proportion of body weight was 0.072 in restricted turkeys compared with 0.046 in birds fed ad libitum. There was no difference in basal metabolic rate between ad libitum and restricted turkeys. 4. It was concluded that feather growth was maintained in preference to body and muscle growth and that rearing birds at 15 degrees C did not improve breast feather cover. It is suggested that the growth of breast feathers in turkeys fed ad libitum is impaired.     

The persistence of infectious pancreatic necrosis virus in Atlantic salmon

Atlantic salmon leucocytes from Infectious Pancreatic Necrosis Virus (IPNV) carriers showed a suppressed response to phytohemagglutinin stimulation compared with uninfected controls. A significant degree of inhibition of DNA synthesis was observed using 3H-thymidine incorporation. IPNV was isolated from 41% of the stimulated leucocyte cultures supernatants, while only 6% of the unstimulated cultures were found to be positive.     

Some observations on the effects of age of calves on the phagocytosis and killing of Staphylococcus aureus by polymorphonuclear leucocytes

The effects of age on bovine polymorphonuclear (PMN) cell function were investigated by comparing the efficiencies of phagocytosis and killing of Staphylococcus aureus by peripheral blood leucocytes sequentially obtained from 15 calves between the ages of less than 1 and 84 days. One group of seven calves was kept in a controlled environmental chamber with air temperature of 5 degrees C and 58% relative humidity (RH) and another group of eight calves was kept at 16 degrees C and 58% RH. The calves were given a diet a liquid milk substitute and dry food, and were weaned abruptly from the liquid diet at 35 days of age. The in-vitro efficiencies of phagocytosis, and of killing, Staphylococcus aureus by peripheral blood leucocytes were similar for calves in air temperatures of 5 degrees C and 16 degrees C (P greater than 0.05). Peripheral blood leucocytes obtained from calves of less than 1 day of age were more efficient in phagocytosing S. aureus than those obtained when the same calves were 14-84 days of age (P less than 0.001). Peripheral blood leucocytes obtained when the calves were 42 and 56 days of age were significantly less efficient in phagocytosing and killing S. aureus than those obtained when the same calves were less than 1, 14, 28, 70 and 84 days of age (P less than 0.001).     

Studies on the Modulation of Leucocyte Subpopulations and Immunoglobulins following Intramammary Infusion of β1,3‐glucan into the Bovine Udder during the Dry Period

Inflammatory and immunological reactions after intramammary infusion of β1,3‐glucan were studied during the steady dry period and involution phase of the bovine udder. The effects of a single intramammary infusion of two different doses (100 and 200 mg) of β1,3‐glucan were evaluated during the steady dry period. In a second study, the effects of β1,3‐glucan at drying off were studied by using two treatment regimens; a single infusion at drying off, compared with two infusions of the compound, at drying off and again 2 weeks later. Total and differential leucocyte counts were measured in both blood and udder secretions. Additionally, the expression of receptors for CD14 and MHC class II on leucocytes, and the expression of receptors for CD4, CD8, WC1, IL2R and B‐cells on lymphocytes was measured in mammary secretions by flow cytometric analyses. The concentrations of immunoglobulins in udder secretions were measured by radial immunodiffusion. The results showed that a single intramammary infusion of β1,3‐glucan during the steady dry period causes transient enhancement of some aspects of the inflammatory and immune responses. The increases in somatic cell counts, numbers of monocytes/macrophages, and in proportions of CD14 + and MHC class II + leucocytes in udder secretions were dose‐dependent. Infusion of β1,3‐glucan also slightly increased the proportion of CD4 + lymphocytes and the concentrations of IgG₁ and IgG₂ in dry secretions. Infusion of β1,3‐glucan at drying off seemed to accelerate the involution process through an increase in somatic cells, particularly in the numbers of macrophages, in mammary secretions. The numbers of lymphocytes and polymorphonuclear leucocytes, the proportions of IL2R + lymphocytes, the proportions of CD14 + or MHC class II + leucocytes and the concentrations of IgG₁ and IgG₂ also increased in comparison with untreated controls. Moreover, a second infusion of β1,3‐glucan tended to prolong this response, indicating that this might be an effective means of enhancing the mammary defence against udder infections closer to calving. In conclusion, the results indicate that intramammary infusion of β1,3‐glucan could be used to enhance the defence mechanisms of the bovine udder against infections, especially during early involution.     

Transmission of Loma salmonae (Microsporea) to chinook salmon in sea water.

Transmission studies were conducted to determine if Loma salmonae was transmissible in sea water. Transmission of L. salmonae to chinook salmon (Oncorhynchus tshawytscha) held in sea water was achieved by exposing fish to macerated, infected gill tissue. Fish were exposed in seawater in a flow-through aquarium, and the infection was detected as soon as 5 wk after exposure. Heavily infected fish exhibited numerous xenomas in the branchial arteries, central venous sinusoids, and within the blood channels of the lamellae. The pathological changes were similar to those seen in pen-reared salmon with L. salmonae infections. The infection was not observed in Atlantic salmon (Salmo salar), Pacific herring (Clupea pallasi, family Clupeidae), or shiner perch (Cymatogaster aggregata, family Embiotocidae), experimentally exposed using identical methods. This study suggests that L. salmonae is transmissible to chinook salmon in seawater netpens. Fish farmers and fish health specialists should consider this possibility when developing and implementing strategies to control the infection.     

Isolation and cryopreservation of functionally competent equine leucocytes

Sufficient numbers of functionally competent polymorphonuclear neutrophil granulocytes (PMN) seem to be of major importance during the course of equine endometritis. In this study, we wanted to establish a method for cryopreservation of functionally competent neutrophils for an intended local endometritis therapy in mares. The separation of leucocytes by hypotonic lysis of whole blood from clinically healthy mares was superior to the separation by dextrose sedimentation. After suspension of the cells in the cryoprotective solution [equine plasma with 5% (v/v) dimethyl sulphoxide (DMSO)], the leucocytes were frozen in liquid nitrogen. A temperature gradient with low cooling velocity (1 degree C/min between 4 and -70 degrees C) resulted in highest numbers of viable cells after thawing. Thawed PMN had a high phagocytic capacity for opsonized streptococci. Their ability to generate reactive oxygen species (ROS) after stimulation with a phorbol ester was even higher than that of freshly isolated PMN and was preserved up to 6 h after thawing. The results of this study indicate that cryopreservation of PMN may provide viable and functionally competent neutrophils for therapeutic use in mares susceptible to endometritis.