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1.  Effect of pre-weaning concentrate supplementation on peripheral distribution of leukocytes, functional activity of neutrophils, acute phase protein and behavioural responses of abruptly weaned and housed beef calves 
The effect of pre-weaning concentrate supplementation on peripheral distribution of leukocytes, functional activity of neutrophils, acute phase protein response, metabolic and behavioural response, and performance of abruptly weaned and housed beef calves was investigated. Calves were grazed with their dams until the end of the grazing season when they were weaned and housed (day (d) 0) in a concrete slatted floor shed, and offered grass silage ad libitum plus supplementary concentrates. Twenty-six days prior to weaning and housing, 20 singled suckled, pure-bred Simmental male (non-castrated), (n = 10, m) and female (n = 10, f) calves were assigned to one of two treatments (i) concentrate supplement (CS: n = 10 (5 m and 5 f), mean age (s.d.) 201 (12.8) d, mean weight (s.d.) 258 (20.2) kg) or (ii) no concentrate supplement (controls) (NCS: n = 10, (5 m and 5 f), mean age (s.d.) 201 (13.4) d, mean weight (s.d.) 257 (19.6) kg) pre-weaning.
There was a treatment × sampling time interaction (P < 0.05) for percentage CD4+ and WC1+ (γδ T cells) lymphocytes and concentration of plasma globulin. On d 2, percentage CD4+ lymphocytes decreased (P < 0.001) in both treatments. Subsequently on d 7, percentage of CD4+ lymphocytes increased (P < 0.01) in CS compared with d 0, whereas percentage of CD4+ lymphocytes in NCS did not differ (P > 0.05) from d 0. On d 2, WC1+ lymphocytes decreased (P < 0.05) in both treatments but the decrease was greater (P <0.05) in NCS than CS. Subsequently, percentages did not differ (P > 0.05) from pre-weaning baseline. On d 2, the increase in concentration of globulin was greater (P < 0.05) in CS compared with NCS, and subsequently there was no difference between treatments. Pre-weaning ADG was 1.07 (s.e.m.) (0.26) kg and 0.99 (s.e.m.) (0.26) kg for CS and NCS, respectively. Post-weaning, CS calves spent more time lying compared with NCS calves.
Calves supplemented with concentrate prior to weaning had a lesser reduction in WC1+ lymphocytes, increased percentage CD4+ lymphocytes and concentration of total protein, and spent more time lying post-weaning, compared with non-supplemented calves.
PMCID: PMC3286413  PMID: 22217360
2.  Examination of the bovine leukocyte environment using immunogenetic biomarkers to assess immunocompetence following exposure to weaning stress 
The molecular mechanisms by which stress induces the development of pathologies remains unclear, although it is recognised that one of the major factors affecting health as a consequence of stress is the involvement of the neuroendocrine system. In cattle, a number of necessary husbandry practices have been shown to activate the stress response, yet very little is known about the impact these have at the molecular level. The objectives of the study were to characterise, in male and female beef calves, the immune response to weaning stress in bovine leukocytes at the physiological and molecular levels and to assess the difference between calves weaned in the presence of the dam and those weaned and penned away from the dam.
Following exposure to weaning stress, total neutrophil number and neutrophil:lymphocyte (N:L) ratio increased (P < 0.01) in calves. Additionally, expression of pro-inflammatory cytokine genes, including IL-1β, IL-8, IFN-γ and TNFα, were up-regulated (P < 0.01). Furthermore, there was increased (P < 0.001) expression of the glucocorticoid receptor, GRα, the pro-apoptotic gene, Fas and the Gram-negative pattern recognition receptor, TLR4. Calves penned away from the dam post-weaning had increased (P < 0.01) neutrophil number and N:L ratio compared with calves penned next to the dam, and female calves had higher (P < 0.05) expression levels of IL-2, IL-8, IFN-γ and TNFα than male calves.
Weaning elicits an immediate and somewhat short-lived acute stress response in the calf. The effects serve to enhance, rather than suppress, the immune response by means of a heightened inflammatory response and cellular mobilization. The earlier and more profound increase in neutrophil number and N:L ratio together with reduced lymphocyte number in calves penned away compared with calves penned near their dams post-weaning suggests that the former may be more sensitive to weaning stress. The data also show a clear effect of gender in differential gene expression in response to stress with IFN-γ having increased expression in female calves compared with male calves over the course of the study. Additionally, this study has helped to characterise the inflammatory response to stress in calves and identify a number of novel candidate biomarkers suitable for investigation in future studies of stress.
PMCID: PMC3177877  PMID: 21834971
3.  An on-farm investigation of beef suckler herds using an animal welfare index (AWI) 
Beef suckler farms (194 farms throughout 13 counties) were assessed once with housed cattle and once with cattle at grass using an animal welfare index (AWI). Twenty-three of the 194 farms were revisited a year later and re-evaluated using the AWI and the Tier-Gerechtheits-Index 35L/2000 (TGI35L/2000). Thirty-three indicators were collected in five categories: locomotion (5 indicators); social interactions (between animals) (7), flooring (5), environment (7) and Stockpersonship (9). Three indicators relating to the size of the farm were also collected.
Improving animal welfare is an increasingly important aspect of livestock production systems predominantly due to increased consumer concern about the source of animal products. The objectives were (i) to evaluate animal welfare of Irish beef suckler herds using an animal welfare index (AWI), (ii) to examine correlations between parameters, how they influence the AWI and investigate the applicability of the parameters used, (iii) to investigate the impact of the activity of the farmer (full-time or part-time), the interest of the farmer and the number of animals on the AWI.
The mean AWI was 65% and ranged from 54% to 83%. The grazing period represented 16.5% of the total points of the AWI. Seventy percent of the farms were rated as "Very Good" or "Excellent". There was no difference (P > 0.05) in AWI between full-time and part-time farmers. Part-time farmers had greater (P = 0.01) "social interactions": calving (P = 0.03) and weaning (P < 0.001) scores. Full-time farmers had cleaner animals (P = 0.03) and their animals had less lameness (P = 0.01). The number of animals on-farm and the interest of the Stockperson were negatively and positively correlated (P = 0.001), respectively, with the AWI. A hierarchical classification was performed to examine how the indicators influenced the AWI.
The AWI was easily applicable for an on-farm evaluation of welfare. The Stockpersonship was an important factor in determining the AWI (11% of the total variation) more specifically, the interest of the farmer. Part and full-time farming did not differ (P > 0.05) in AWI scores. This method could, with further development, be used in countries with both intensive and/or extensive production systems and would require substantially less resources than animal-based methods.
PMCID: PMC3022571  PMID: 21144023
4.  The effect of road and sea transport on inflammatory, adrenocortical, metabolic and behavioural responses of weanling heifers 
The objective was to investigate the effect of transport on inflammatory, adrenocortical, metabolic and behavioural responses of weanling heifers transported from Ireland to Spain.
At the end of the grazing season, 60 Charolais × beef heifers (mean live weight 245, s.e. 4.3 kg and mean weaning age 219, s.e. 4.9 days) were abruptly weaned from their dams on day 0. The animals were assigned by live weight to two treatments, transport (T) (n = 40) (mean 246, s.e. 5.4 kg) and control (C) (n = 20) (mean 247, s.e. 7.2 kg) on day 0. The T animals were transported from Ireland to France on a roll-on roll-off ferry at a stocking density of 0.93 m2/animal and then by road for 9 h to a French lairage while C animals remained in Ireland and were not transported. On arrival at the French lairage (d 2), 20 T animals were unloaded (ULT) and rested for 12 h in the French lairage and 20 animals rested (RT) on the transporter. All animals had access to hay and water. After the rest period, the ULT animals were re-loaded. The subsequent journey by road from France to Spain was 9 h travel, 7 h rest (on the transporter) and a further 7 h travel by road. All T animals were blood sampled prior to transport (day (d) 0; baseline), on arrival in the French lairage (d 2), after 12 h rest in the French lairage (d 2), on arrival at the feedlot in Spain (d 4) and on d 6, d 8, d 10 and d 34. Control animals were blood sampled at the same times as T animals.
ULT and RT animals had lower (P < 0.05) live weight than C animals on d 6. WBC number was lower (P < 0.05) in control animals on d 10 and greater on d 34 compared with baseline values. RT and ULT animals had greater (P < 0.05) WBC number than baseline on d 2 (arrival in France) to d 34. Neutrophil % was unchanged in RT, ULT and control animals compared with baseline. Control, RT and ULT animals had lower haematocrit % (P < 0.05) throughout the study compared with d 0. There was no difference (P > 0.05) in plasma protein concentration between RT and ULT animals from day 2 to d 34. Plasma concentrations of urea were higher (P < 0.05) in RT animals from d 2 to d 34 compared with C animals. RT and ULT animals had lower (P < 0.05) non-esterified fatty acid (NEFA) and βeta-hydroxy-butyrate (ßHB) concentrations on d 10 and d 34 compared with d 0.
It is concluded that, within the conditions of the present study, the performance of the animals that remained on the transporter during the 12 h lairage period in France was not different post-transport from the transported animals that were unloaded and lairaged in France.
PMCID: PMC2917422  PMID: 20646267
5.  Characterisation of physiological and immunological responses in beef cows to abrupt weaning and subsequent housing 
Weaning involves the permanent separation of the calf from the dam and has been shown to be stressful for both. The objectives of this study were to characterise the effect of i) abrupt weaning and ii) subsequent housing on the extended physiological and immunological responses of beef cows. At weaning (day (d) 0, mean age of calf (s.d.) 212 (24.5) d), cows were abruptly separated from their calves and returned to the grazing area. After 35 d at pasture, cows were housed in a slatted floor shed and offered grass silage ad libitum plus a mineral-vitamin supplement daily. Rectal body temperature was recorded and blood samples were obtained on i) d 0 (weaning), 2, 7, 14, 21, 28, 35 and subsequently on ii) d 0 (housing), 2, 7, 14 and 21 for physiological, haematological and immunological measurements.
Post-weaning, concentration of cortisol and dehydroepiandrosterone were unchanged (P > 0.05). Rectal body temperature, neutrophil number and neutrophil: lymphocyte ratio increased (P < 0.01) on d 2 compared with pre-weaning baseline. Lymphocyte and neutrophil number decreased (P < 0.05) on d 2 to 7 and d 7 to 21, respectively, compared with pre-weaning baseline. Interferon-γ production decreased (P < 0.05) on d 2 compared with pre-weaning baseline. An increase (P < 0.05) in acute phase proteins, fibrinogen and haptoglobin was evident on d 2 to 35 compared with pre-weaning baseline. Concentration of glucose increased on d 2 to 28, whereas non-esterified fatty acid decreased on d 2 to 35 compared with pre-weaning baseline. Post-housing, concentrations of cortisol, rectal body temperature, total leukocyte number, and glucose were unchanged (P > 0.05). On d 2 post-housing, neutrophil number and neutrophil: lymphocyte ratio increased (P < 0.05), whereas lymphocyte number and concentrations of dehydroepiandrosterone, fibrinogen and non-esterified fatty acid decreased (P < 0.05) compared with pre-housing baseline. Concentration of haptoglobin increased (P < 0.05) on d 14 to 21 post-housing.
A transitory increase in neutrophil number and decrease in lymphocyte number, increased neutrophil:lymphocyte ratio coupled with decreased interferon-γ production, and increased concentration of acute phase proteins indicate a stress response in cows post-weaning, whereas post-housing, changes were less marked.
PMCID: PMC2917423  PMID: 20646268
6.  Effect of road transport for up to 24 hours followed by twenty-four hour recovery on live weight and physiological responses of bulls 
The transport of livestock can have major implications for their welfare, and there is strong public interest and scientific endeavour aimed at ensuring that the welfare of transported animals is optimal. The objective of the study was to investigate the effect of transport on live weight, physiological and haematological responses of bulls after road transport of 0, 6, 9, 12, 18 and 24 hours (h). Seventy-two Charolais bulls (mean weight (s.d.) 367 (35) kg), naïve to transport, were randomly assigned to one of six journey (J) times of 0 h, 6 h, 9 h, 12 h, 18 h and 24 h transport (n = 12 animals/treatment) at a stocking density of 1.02 m2/bull. Blood samples were collected by jugular venipuncture before transport (-0.25 h), immediately after (0 h) and at 1 h, 2 h, 4 h, 6 h, 8 h, 12 h and 24 h relative to time 0 h. The bulls were weighed before transport (- 24 h and - 0.25 h), immediately after (0 h), and at 4 h, 12 h and 24 h relative to time 0 h. Control animals were blood sampled before assignment (-0.25 h) to novel pens, after (24 h), and at 1 h, 2 h, 4 h, 6 h, 8 h, 12 h and 24 h relative to the 24 h sampling time point.
Bulls travelling for 6 h (280 km), 9 h (435 km), 12 h (582 km), 18 h (902 km) and 24 h (1192 km) lost 4.7, 4.5, 5.7 (P < 0.05), 6.6 (P < 0.05) and 7.5 (P < 0.05) percentage (%) live weight compared with baseline. Live weight re-gained to pre-transport levels during the 24 h recovery period. Lymphocyte percentages were lower (P < 0.05) and neutrophil percentages were greater (P < 0.05) in all animals. Blood protein, glucose and NEFA concentrations and creatine kinase activity were greater (P < 0.05) in the bulls following transport and returned to baseline within 24 h.
Under the conditions of the present study, transport of bulls on journeys by road, ranging from 6 h (280 km) to 24 h (1192 km) duration, affected live weight, haematological and physiological measurements of metabolism and inflammation. Our findings showed that live weight and some physiological and haematological responses of bulls returned to pre-transport levels within 24 h with animals having had access to feed and water.
PMCID: PMC2918586  PMID: 20646269
7.  Peripheral and gastrointestinal immune systems of healthy cattle raised outdoors at pasture or indoors on a concentrate-based ration 
Despite an increasing preference of consumers for beef produced from more extensive pasture-based production systems and potential human health benefits from the consumption of such beef, data regarding the health status of animals raised on pasture are limited. The objective of this study was to characterise specific aspects of the bovine peripheral and the gastrointestinal muscosal immune systems of cattle raised on an outdoor pasture system in comparison to animals raised on a conventional intensive indoor concentrate-based system.
A number of in vitro functional tests of immune cells suggested subtle differences between the animals on the outdoor versus indoor production systems. There was a decrease in the number of neutrophils and monocytes engaged in phagocytosis in outdoor cattle (P < 0.01 and P < 0.05, respectively) in comparison to those indoors. Following mitogen stimulation, a lower level of interferon-γ was produced in leukocytes from the outdoor animals (P < 0.05). There was evidence of a gastrointestinal nematode infection in the outdoor animals with elevated levels of serum pepsinogen (P < 0.001), a higher number of eosinophils (P < 0.05) and a higher level of interleukin-4 and stem cell factor mRNA expression (P < 0.05) in the outdoor animals in comparison to the indoor animals. Lower levels of copper and iodine were measured in the outdoor animals in comparison to indoor animals (P < 0.001).
Despite distinctly contrasting production systems, only subtle differences were identified in the peripheral immune parameters measured between cattle raised at pasture in comparison to animals raised on a conventional intensive indoor concentrate-based production system.
PMCID: PMC2864234  PMID: 20356390
8.  Temporal patterns of inflammatory gene expression in local tissues after banding or burdizzo castration in cattle 
Castration of male cattle has been shown to elicit inflammatory reactions and acute inflammation is initiated and sustained by the participation of cytokines.
Sixty continental × beef bulls (Mean age 12 ± (s.e.) 0.2 months; Mean weight 341 ± (s.e.) 3.0 kg) were blocked by weight and randomly assigned to one of three treatments (n = 20 animals per treatment): 1) untreated control (Con); 2) banding castration at 0 min (Band); 3) Burdizzo castration at 0 min (Burd). Samples of the testis, epididymis and scrotal skin were collected surgically from 5 animals from each group at 12 h, 24 h, 7 d, and 14 d post-treatment, and analysed using real-time PCR. A repeated measurement analysis (Proc GLM) was performed using SAS. If there was no treatment and time interaction, main effects of treatment by time were tested by ANOVA.
Electrophoresis data showed that by 7 d post-castration RNA isolated from all the testicle samples of the Burd castrated animals, the epididymis and middle scrotum samples from Band castrates were degraded. Transitory effects were observed in the gene expression of IFN-γ, IL-6, IL-8 and TNF-α at 12 h and 24 h post treatment. Burd castrates had greater (P < 0.05) testicular IFN-γ mRNA levels compared with Band and Con animals, but lower (P < 0.05) testicular TNF-α mRNA levels compared with Con animals. Band castrates had greater (P < 0.05) testicular IL-6 mRNA levels than Burd castrates at 12 h post-castration. Burd castrates had greater (P < 0.05) testicular IL-8 mRNA levels than Band and Con animals at 24 h post-castration. In the epididymis, Burd castrates had greater (P < 0.05) IL-6 mRNA (both at 12 h and 24 h post treatment) and IL-8 mRNA (12 h post treatment) levels compared with Band and Con animals; Burd castrates had greater (P = 0.049) IL-10 mRNA levels than Band castrates at 12 h post-castration.
Banding castration caused more inflammatory associated gene expression changes to the epididymis and scrotum than burdizzo. Burdizzo caused more severe acute inflammatory responses, in terms of pro-inflammatory cytokine gene expression, in the testis and epididymis than banding.
PMCID: PMC2758859  PMID: 19775432

Results 1-8 (8)