Review: Environmental enrichment builds functional capacity and improves resilience as an aspect of positive welfare in production animals.

The success of the animal in coping with challenges, and in harnessing opportunities to thrive, is central to its welfare. Functional capacity describes the capacity of molecules, cells, organs, body systems, the whole animal, and its community to buffer against the impacts of environmental perturbations. This buffering capacity determines the ability of the animal to maintain or regain functions in the face of environmental perturbations, which is recognised as resilience. The accuracy of physiological regulation and the maintenance of homeostatic balance underwrite the dynamic stability of outcomes such as biorhythms, feed intake, growth, milk yield, and egg production justifying their assessment as indicators of resilience. This narrative review examines the influence of environmental enrichments, especially during developmental stages in young animals, in building functional capacity and in its subsequent expression as resilience. Experience of enriched environments can build skills and competencies across multiple functional domains including but not limited to behaviour, immunity, and metabolism thereby increasing functional capacity and facilitating resilience within the context of challenges such as husbandry practices, social change, and infection. A quantitative method for measuring the distributed property of functional capacity may improve its assessment. Methods for analysing embedded energy (emergy) in ecosystems may have utility for this goal. We suggest functional capacity provides the common thread that links environmental enrichments with an ability to express resilience and may provide a novel and useful framework for measuring and reporting resilience. We conclude that the development of functional capacity and its subsequent expression as resilience is an aspect of positive animal welfare. The emergence of resilience from system dynamics highlights a need to shift from the study of physical and mental states to the study of physical and mental dynamics to describe the positive dimension of animal welfare.

Targeting improved resilience in Merino sheep - Correlations between immune competence and health and fitness traits.

Resilience can be defined as the ability of an animal to remain productive in the face of diverse environmental challenges. Several factors contribute to an animal's resilience including its ability to resist disease, cope with climatic extremes and respond to stressors. Immune competence, a proxy trait for general disease resistance, is expected to contribute to an animal's resilience. This research aimed to develop a practical method to assess immune competence in Merino sheep which would not restrict the future sale of tested animals, and to estimate genetic parameters associated with the novel trait. We also aimed to explore associations between immune competence and other industry-relevant disease resistance and fitness-related traits and to assess the ability of immune competence phenotypes to predict health outcomes. Here, the ability of Merino wethers (n = 1 339) to mount both an antibody-mediated and cell-mediated immune response was used to define their immune competence phenotype. For that purpose, antigens in a commercial vaccine were administered at the commencement of weaning and their responses were assessed. Univariate sire models were used to estimate variance components and heritabilities for immune competence and its component traits. Bivariate sire models were used to estimate genetic correlations between immune competence and a range of disease resistance and fitness-related traits. The heritability of immune competence and its component traits, antibody-mediated immune response and cell-mediated immune response were estimated at 0.49 ±â€¯0.14, 0.52 ±â€¯0.14 and 0.36 ±â€¯0.11, respectively. Immune competence was favourably genetically correlated with breech flystrike incidence (-0.44 ±â€¯0.39), worm egg count (-0.19 ±â€¯0.23), dag score (-0.26 ±â€¯0.31) and fitness compromise (-0.35 ±â€¯0.24) but not fleece rot (0.17 ±â€¯0.23). Results suggest that selection for immune competence has the potential to improve the resilience of Merino sheep; however, due to the large standard errors associated with correlation estimates reported here, further studies will be required in larger populations to validate associations between immune competence and disease resistance and fitness traits in Australian Merino sheep.

Effects of a topical anaesthetic formulation and systemic carprofen, given singly or in combination, on the cortisol and behavioural responses of Merino lambs to castration.

OBJECTIVE: To determine the effectiveness of a topical anaesthetic formulation (Tri-Solfen) with or without the administration of a non-steroidal anti-inflammatory drug (carprofen) on the pain and distress response associated with ring or surgical castration of ram lambs. PROCEDURES: Merino ram lambs (n = 78) were allocated to 10 treatment groups: 4 groups of knife-castrated lambs and 4 groups of ring-castrated lambs received carprofen (4 mg/kg SC) and Tri-Solfen; 2 control groups (sham) received carprofen at 0 or 4 mg/kg SC. Measurements included plasma cortisol and haptoglobin concentrations, haematology, and behaviour, including posture. RESULTS: Knife-castrated lambs had higher peak cortisol and integrated cortisol responses for the first 6 h after treatment and greater concentration s of circulating acute phase proteins than ring-castrated lambs, both of which were significantly different from the sham controls. Tri-Solfen applied to the knife castration wound significantly reduced both the peak plasma cortisol concentration and the integrated cortisol response for the first 6 h and improved lying behaviour in the first 12 h. Carprofen reduced the cortisol response to knife castration at 30 min, but elevated the cortisol responses at 24 and 48 h. Carprofen nearly halved the number of acute pain behaviours associated with ring castration. There were no significant additive or synergistic effects from combining the analgesic treatments. Tri-Solfen applied to the tail wound provided no detectible benefits during ring castration + tail docking. CONCLUSIONS: The physiological and behavioural responses suggest that ring castration has less impact on the lamb than knife castration. The specific analgesic treatments can provide modest amelioration of the pain and discomfort associated with castration. Alternative doses or application methods may enhance their efficacy.

Assessment of welfare of suckling lambs following intradermal injection of cetrimide as a non-surgical alternative to conventional mulesing.

OBJECTIVE: To assess in suckling lambs the impact of intradermal injection of cetrimide, a quaternary ammonium compound formulated to induce non-surgical mulesing, on some physiological and behavioural indicators of welfare. PROCEDURES: We allocated 32 suckling lambs (9-11 weeks old) to three groups: (1) control (n = 10), (2) conventional surgical mules (n = 11) and (3) non-surgical mules (n = 11). Non-surgical mulesing was induced by intradermal injection of 4% (w/w) cetrimide + 3% (w/w) polyvinylpyrrolidone in water. Lambs were run in pens of four together with their dams. Haematology, cortisol, beta-endorphin and haptoglobin levels, and rectal temperature were monitored at least daily for the first 7 days after treatment, then weekly until day 28. Body weight was measured weekly and behaviour was measured every 15 min for 12 h on the day of treatment, then on days 1, 2, 4, 6, 12, 21 and 28 following treatment. RESULTS: The intradermal treatment induced local tissue swelling, systemic signs of severe inflammation, including high fever (> 41.0 degrees C) and elevated blood cortisol levels, by 12 h. Rectal temperatures were significantly elevated until 6 days after treatment, cortisol levels were elevated until 4 days after treatment, haptoglobin levels for at least 7 days after treatment and the neutrophil to lymphocyte ratio until 5 days after treatment. Peak cortisol values were comparable in mulesed lambs and lambs receiving the intradermal treatment, whereas the areas under the curves for cortisol and temperature were greater in lambs receiving the intradermal treatment than in mulesed lambs. Beta-endorphin levels were significantly elevated in mulesed sheep at 12 h. There was no effect of intradermal treatment on average daily gain, fibre diameter or beta-endorphin concentration. Mulesed lambs spent 44% of the time in abnormal behaviours (hunched standing, stiff walking, pawing, lateral lying and lying intention) on the day of treatment. On the day after treatment, lambs receiving the intradermal treatment spent 11% of the time (comparable to mulesed lambs) in abnormal behaviours. In comparison, control lambs spent 0.4% of their time in abnormal behaviours on the same day. CONCLUSIONS: The welfare of suckling lambs that were non-surgically mulesed by intradermal injection of cetrimide was measurably poorer than control lambs.

Effect of the non-steroidal anti-inflammatory drug, carprofen, on weaned sheep following non-surgical mulesing by intradermal injection of cetrimide.

OBJECTIVE: To assess in weaned lambs the palliative effects of the non-steroidal anti-inflammatory drug, carprofen, following intradermal injection of cetrimide to induce non-surgical mulesing. PROCEDURES: We allocated 40 weaned lambs (20-22 weeks old) to four groups of 10 animals: (1) control, 2) conventional surgical mules, (3) intradermal treatment and (4) intradermal treatment + carprofen. Non-surgical mulesing was induced by intradermal injection of 4% (w/w) cetrimide + 3% (w/w) polyvinylpyrrolidone in water. In group 4, carprofen (4 mg/kg, SC) was administered 1 h before intradermal treatment. Five weaners, including an animal from each treatment, were run in each pen. Neutrophil to lymphocyte ratio, cortisol, beta-endorphin and haptoglobin levels and rectal temperature were monitored at least daily for the first 7 days after treatment, then weekly until day 28. Body weight was measured weekly and behaviour was measured every 15 min for 12 h on the day of treatment, then on days 1, 2, 4, 6, 12, 21 and 28 following treatment. RESULTS: The intradermal treatment resulted in high fever and elevated blood cortisol by 12 h. Rectal temperatures were significantly elevated until 5 days after treatment, cortisol was elevated until 3 days after treatment, haptoglobin for at least 7 days after treatment and the neutrophil to lymphocyte ratio until 4 days after treatment. Average daily gain was depressed in the week following treatment. Abnormal behaviours (hunched standing, stiff walking, pawing, lateral lying and lying intention) were increased on the day of treatment and for 6 days post treatment. Carprofen reduced the time spent in abnormal behaviours by approximately two-thirds but did not ameliorate the physiological responses to the intradermal treatment. CONCLUSIONS: In weaner sheep, carprofen ameliorated the behavioural responses, but was unable to provide relief from the intense and sustained physiological responses to non-surgical mulesing by intradermal injection of cetrimide. Systemic side-effects may be unavoidable with formulations based on quaternary ammonium compounds that are designed to reduce the risk of fly strike in sheep by remodelling breech tissue through induction of tissue necrosis.

Efficacy of meloxicam in a pain model in sheep.

OBJECTIVE: To evaluate the efficacy of the non-steroidal anti-inflammatory drug, meloxicam, in alleviating pain and inflammation and on production-related variables in a model of sterile acute inflammation in sheep. METHODS: Groups of 12 mature Merino ewes received 0, 0.5, 1.0 or 1.5 mg/kg meloxicam subcutaneously 90 min before injection of 0.1 mL turpentine subcutaneously on the anterior aspect of the proximal phalanx of a forelimb. Pain- and inflammation-related variables were assessed at -18, 3, 6, 9, 12, 24, 48 and 72 h relative to meloxicam administration. Daily feed intake and body weight change 7 days later were also assessed. Pain-related variables measured were weight borne on each forelimb, lameness score, time each forelimb was raised in a 20-s interval and tolerance to a noxious mechanical stimulus. Inflammation-related variables measured were skin temperature, limb circumference, body temperature, plasma haptoglobin concentration and peripheral blood leucocyte parameters. RESULTS: Meloxicam was effective in improving all pain-related variables. A dose-dependent response was seen between 0 and 1.0 mg/kg, with no additional benefit provided by 1.5 mg/kg. At a dose rate of 1.0 mg/kg, meloxicam improved weight borne on the turpentine-treated limb by 14%, reduced the time the treated limb was held in a non-weight-bearing posture by 46%, reduced the lameness score by 58% and improved tolerance to pressure by 52%. No significant effects of meloxicam on inflammatory variables or appetite were observed. CONCLUSIONS: Using a validated pain model, the data suggested that 1.0 mg/kg meloxicam provided significant analgesic benefits to sheep.

Fleece rot and dermatophilosis in sheep.

Fleece rot and dermatophilosis reduce health and production of sheep and predispose them to blow fly strike. This paper reviews aetiology, prevalence, pathogenesis, resistance, attempts to develop vaccines and prospects for new control strategies to these important skin diseases. Although the severity of fleece rot is associated with the abundance of Pseudomonas aeruginosa on skin, microbial ecology studies are providing new insights into the contribution of other bacteria to the disease. Wool traits and body conformation traits that predispose sheep to fleece rot and dermatophilosis are heritable and have been used as indirect selection criteria for resistance for many years. Selection against BoLA-DRB3-DQB class II haplotype in cattle can substantially reduce the prevalence of dermatophilosis and holds promise for identification of gene markers for resistance to these bacterial diseases in sheep. Immune responses in skin and systemic antibody responses to bacterial antigens are acquired through natural infection and contribute to resistance; however, prototype antibacterial vaccines have to date failed to provide protection against the diversity of isolates of Dermatophilus congolensis and Pseudomonas species present in the field. Opportunities for future control through breeding for resistance, vaccines and non-vaccine strategies for controlling the microbial ecology of fleece are discussed. In combination, control strategies need to reduce the risk of transmission, minimise exposure of animals to stressors that enhance the risk of infection, and enhance resistance though genetics or vaccines.

Adjusting worm egg counts for faecal moisture in sheep.

The number of eggs from gastrointestinal nematodes per gram of faeces (worm egg count WEC) is commonly used to determine the need for anti-parasite treatments and the breeding value of animals when selecting for worm resistance. Diarrhoea increases faecal moisture and may dilute the number of worm eggs observed. To quantify this effect, egg counts in sheep at pasture were simulated by dosing 15 animals with chromic oxide particles. The simulated WEC diminished as faecal moisture increased. When faeces were dried, simulated WEC per unit dry matter was not influenced by the amount of faecal moisture present prior to drying. The results suggest that adjustment for faecal moisture may provide an improved estimate of FEC. Drying faeces to calculate the WEC per unit dry matter would provide such an adjustment but may not be practical for industry application. In the past, the CSIRO McMaster Laboratory has used an adjustment factor developed by Gordon based on the classification of faecal consistency derived from the morphology of faeces. To examine the utility of an adjustment factor based on faecal consistency score (FCS), the relationships between FCS and simulated WEC and dry matter were examined. Dry matter and simulated WEC exhibited an exponential decline as FCS increased. The relationship between FCS and dry matter was further examined in 368 samples collected over 12 months from sheep at pasture, where it was observed that dry matter showed a linear decline as FCS increased. Adjustment factors based on dry matter were similar to those proposed by Gordon however adjustment factors predicted from simulated WEC diverged from the remainder for FCS>4. As no samples scored FCS 5 in the study of simulated FEC, the adjustment factors based on the larger study that included samples with FCS 5 was therefore considered more robust. Adjustment factors were given by the equation: WEC(estimated)=(WEC(observed)/(34.21-5.15 FCS))x29.06. This equation estimates for samples with FCS>1 the WEC that would be expected if the samples were FCS 1, the faecal consistency score for normal faeces. The impact of adjustment of observed WEC for faecal moisture predicted by FCS on decision points for treatment and on estimated breeding values requires further examination.

The effect of a topical anaesthetic formulation, systemic flunixin and carprofen, singly or in combination, on cortisol and behavioural responses of Merino lambs to mulesing.

OBJECTIVE: To determine the pain responses of lambs to mulesing, and the effectiveness of potential analgesic treatments. PROCEDURES: Merino lambs (n=64) were allocated at 5 weeks of age to eight treatment groups: 1) sham mules; 2) conventional mules; 3) topical anaesthetic, incorporating lignocaine, bupivicaine, adrenaline and cetrimide, applied immediately after mulesing; 4) flunixin + topical anaesthetic, with flunixin administered 2.5 mg/kg s.c. 90 min before mulesing; 5) carprofen + topical anaesthetic, with carprofen administered 4 mg/kg s.c. 90 min before mulesing; 6) carprofen, administered as above; 7) flunixin, administered as above; and 8) carprofen + flunixin, administered as above. Plasma cortisol was measured at 0, 0.5, 6, 12 and 24 h relative to mulesing. Animal behaviour, including posture, was recorded for 12 h after mulesing. RESULTS: The conventional mules lambs exhibited large increases in plasma cortisol, reduced lying and increased standing with a hunched back compared with sham mules animals. Topical anaesthetic reduced the cortisol peak to mulesing and hunched standing, and increased lying compared with the conventional mules treatment, but generally did not result in values equivalent to sham mules animals. Carprofen, flunixin, and carprofen + flunixin treatments did not reduce the cortisol response to mulesing but substantially ameliorated some changes in behavioural postures. Flunixin + topical anaesthetic reduced the cortisol peak following mulesing and substantially ameliorated most changes in behavioural postures. Carprofen + topical anaesthetic abolished the cortisol peak following mulesing and substantially ameliorated most changes in behavioural postures. All mulesed animals lost weight in the week after mulesing regardless of analgesic administration, but there were no significant differences in growth rate between any of the eight treatments over the 3 weeks after mulesing. CONCLUSIONS: Analgesics can moderate the pain response of lambs to mulesing. The welfare outcome for lambs of mulesing could be improved by use of a combination of local anaesthetic and long acting non-steroidal anti-inflammatory drug.

Migratory responses of ovine neutrophils to inflammatory mediators in vitro and in vivo.

The migration of 111In-labeled ovine neutrophils towards a range of inflammatory mediators was examined in vitro using a 48-well chemotaxis chamber. Typical curves were obtained for the chemotactic response to zymosan-activated plasma (ZAP, a source of C5a) and interleukin-8 (IL-8). In contrast, leukotriene B4 (LTB4), platelet-activating factor (PAF), interleukin-1 alpha (IL-1 alpha), tumor necrosis factor alpha (TNF-alpha), N-formyl-methionine-leucyl-phenylalanine (fMLP), and endotoxins from Escherichia coli and Pseudomonas aeruginosa failed to induce neutrophil migration in vitro. Of these mediators LTB4, ZAP, IL-1 alpha, TNF-alpha, IFN-gamma, and IL-8 have been reported to induce neutrophil accumulation in skin of sheep, and in the current study E. coli endotoxin was a potent inducer of 111In-labeled neutrophil accumulation and plasma leakage in skin. In contrast, PAF induced intense plasma leakage but failed to induce accumulation of 111In-labeled neutrophils in skin. Histologic examination of skin sites receiving PAF confirmed the failure of PAF to stimulate neutrophil extravasation. FMLP lacked inflammatory activity in skin. Coinjection of actinomycin D did not abrogate recruitment of neutrophils to skin sites receiving LTB4; thus neither induction of endothelial adhesion molecules nor synthesis of IL-8 was necessary for LTB4 to exhibit inflammatory activity in vivo.

Neutrophil accumulation and plasma leakage induced in vivo by neutrophil-activating peptide-1.

Neutrophil accumulation and plasma leakage induced in rabbit skin by neutrophil-activating peptide-1 (NAP-1, a 72 amino acid peptide produced by monocytes and a variety of tissue cells), E. coli endotoxin, and interleukin-1 (IL-1) were compared. Neutrophil accumulation at sites injected with NAP-1 was intense, rapid, and long-lasting; it reached a maximum rate during the first 30 min, continued at constant rate for 4-6 h, and remained detectable up to at least 8 h. In contrast, the neutrophil-attracting effect of endotoxin and IL-1 was slower in onset and more transient; it peaked in the first 2 h and declined to a very low level after 4 h. Plasma leakage induced by NAP-1 had a shorter time course than neutrophil accumulation and ceased after 6 h. Depletion of blood neutrophils by treatment with hydroxyurea prevented the plasma leakage induced by NAP-1 or endotoxin but not by histamine. Desensitization to NAP-1 was studied by restimulation of lesions. Following restimulation with NAP-1 after intervals from 6-10 h, there was diminished infiltration of neutrophils, while nearly normal responses were obtained after an interval of 24 h. Desensitization was dose dependent and affected both plasma leakage and neutrophil accumulation. In lesions initiated with NAP-1 there were normal responses following restimulation with endotoxin but marked desensitization to IL-1, suggesting that NAP-1 may contribute to inflammation induced by IL-1 but not by endotoxin. This study indicates that NAP-1 is a potent mediator of neutrophil accumulation in vivo, with characteristics similar to those reported for C5 fragments, but with a more protracted action.

Trial of human laser epilation technology for permanent wool removal in Merino sheep.

OBJECTIVE: To assess whether human laser epilation technology can permanently prevent wool growth in sheep. DESIGN: An observational study. METHODS: Two commercial human epilation lasers (Sharplan alexandrite 755 nm laser, and Lumenis LightSheer 800 nm diode laser) were tested at energies between 10 and 100 J/cm2 and pulse widths from 2 to 400 ms. Wool was clipped from flank, breech, pizzle and around the eyes of superfine Merino sheep with Oster clippers. After initial laser removal of residual wool to reveal bare skin, individual skin sites were treated with up to 15 cycles of laser irradiation. Behavioural responses during treatment, skin temperature immediately after treatment and skin and wool responses for 3 months after treatment were monitored. RESULTS: A clear transudate was evident on the skin surface within minutes. A dry superficial scab developed by 24 h and remained adherent for at least 6 weeks. When scabs were shed, there was evidence of scarring at sites receiving multiple treatment cycles and normal wool growth in unscarred skin. There was no evidence of laser energy level or pulse width affecting the response of skin and wool to treatment and no evidence of permanent inhibition of wool growth by laser treatment. Laser treatment was well tolerated by the sheep. CONCLUSIONS: Treatment of woolled skin with laser parameters that induce epilation by selective photothermolysis in humans failed to induce permanent inhibition of wool growth in sheep. Absence of melanin in wool may have contributed to the result.

Randomised trial of the bioavailability and efficacy of orally administered flunixin, carprofen and ketoprofen in a pain model in sheep.

OBJECTIVE: To determine the efficacy and bioavailability of non-steroidal anti-inflammatory drugs (NSAIDs) when administered orally to sheep. DESIGN: Randomised experimental design with four treatment groups: three NSAID groups and one control group (n = 10/group). The study animals were 40 18-month-old Merino ewes with an average weight of 31.4 ± 0.5 kg. METHODS: Treatment was given orally at 24 h intervals for 6 days at dose rates expected to achieve therapeutic levels in sheep: carprofen (8.0 mg/kg), ketoprofen (8.0 mg/kg) and flunixin (4.0 mg/kg). Oil of turpentine (0.1 mL) was injected into a forelimb of each sheep to induce inflammation and pain; responses (force plate pressure, skin temperature, limb circumference, haematology and plasma cortisol) were measured at 0, 3, 6, 9, 12, 24, 36, 48, 72 and 96 h post-injection. NSAID concentrations were determined by ultra-high-pressure liquid chromatography. RESULTS: The NSAIDs were detectable in ovine plasma 2 h after oral administration, with average concentrations of 4.5-8.4 µg/mL for ketoprofen, 2.6-4.1 µg/mL for flunixin and 30-80 µg/mL for carprofen. NSAID concentrations dropped 24 h after administration. Pain response to an oil of turpentine injection was assessed using the measures applied but no effect of the NSAIDs was observed. Although this pain model has been previously validated, the responses observed in this study differed from those in the previous study. CONCLUSIONS AND CLINICAL RELEVANCE: The three NSAIDs reached inferred therapeutic concentrations in blood at 2 h after oral administration. The oil of turpentine lameness model may need further validation.

Effect of local infusion of NSAID analgesics administered alone or in combination on the pain associated with band castration in calves.

OBJECTIVE: To evaluate the analgesic efficacy of flunixin alone or in combination with diclofenac administered locally to the scrotum at the time of band castration of calves. METHODS: Angus bull calves (n = 40; ≈7-9 weeks old) were allocated to four treatment groups (n = 10 per group) to examine the effects of two non-steroidal anti-inflammatory analgesics (NSAIDs) administered locally at the time of band castration: sham control; castration + flunixin; castration + flunixin + diclofenac; castration + saline. The NSAIDs and saline were administered subcutaneously into the scrotum under the band. Blood was sampled at -0.5, 0.5, 1.5, 3, 12, 24, 48 and 72 h relative to castration. Haematology parameters and plasma cortisol levels were determined in samples at all time points and plasma haptoglobin levels determined in samples collected at -0.5, 24, 48 and 72 h. Pain avoidance and postural behaviours were measured for 2 and 12 h, respectively, after castration. RESULTS: Band-castrated calves exhibited significantly higher peak cortisol and higher integrated cortisol responses during the first 6 h post-castration relative to sham controls. Individual active pain avoidance behaviours observed for 1 h post-castration were not significantly different between treatment groups; however; the sum of the total behaviours was significantly increased by castration (P = 0.023). Postural changes included increased abnormal ventral lying for all castrated groups and decreased normal standing and increased combined abnormal postures for the flunixin- and saline-treated groups. Growth rates of calves were not affected by treatments during weeks 1 and 2 post-castration; however, growth rates of castrated calves were significantly lower than those of sham-treated calves in week 3 post-castration (1.41 vs 0.84, 0.75 and 0.56±0.19 kg/day for sham, flunixin-, flunixin + diclofenac- and saline-treated groups, respectively). CONCLUSIONS: The administration of flunixin or flunixin + diclofenac intrascrotally at several sites at the time of banding did not improve the welfare of young Angus bull calves undergoing this method of castration when assessed by both physiological and behavioural parameters. In this study, band castration at 7-9 weeks of age had less effect compared with results for older calves reported previously. Further research is required to develop effective analgesic treatments that can be administered at the time of castration to large numbers of animals.

Chemotactic factor-specific desensitization of skin to infiltration by polymorphonuclear leukocytes.

The accumulation of polymorphonuclear leukocytes (PMNs) was measured in rabbits following single or repeated injections into skin sites of the inflammatory agents, zymosan-activated plasma and formylmethionyl-leucyl-phenylalanine. Fewer cells entered lesions restimulated with the same agent during the subsequent inflammatory response than simultaneously entered skin sites stimulated for the first time. Decreased reactivity of a restimulated site was specific to the initiating stimulus, developed within 2 to 4 h of initial stimulation and persisted for at least 8 h. The results indicate that PMN accumulation in acute inflammatory lesions is regulated locally by a chemotactic factor-specific mechanism which differs from PMN deactivation and concentration gradient mechanisms currently thought to regulate PMN migration in vivo.

The effect of immune and inflammatory mediators on growth of Lucilia cuprina larvae in vitro.

Immune and inflammatory responses occurring during dermal infestation by larvae of Lucilia cuprina can retard larval growth and development. This study examined the effect of 4 classes of humoral inflammatory mediators on larval growth in an in vitro assay. Mediators of plasma leakage (histamine, bradykinin, platelet-activating factor and serotonin), leucocyte chemotactic agonists (activated complement, leukotriene B4 and interleukin-8), effector molecules of immune responses (interleukin-1 beta, tumour necrosis factor-alpha and interferon-gamma) and endotoxin all failed to inhibit larval growth. In contrast, immunoglobulins isolated from immune serum caused marked retardation of larval growth. The results suggest that humoral mediators of inflammatory and immune responses do not play a role in immune defence against Lucilia cuprina.

Immunity to Haemonchus contortus and the cellular response to helminth antigens in the mammary gland of non-lactating sheep.

Cellular exudates induced by infusion with helminth antigens were examined in non-lactating mammary glands of ewes immune to infection with the abomasal nematode, Haemonchus contortus. Secondary immunological responsiveness was expressed in two ways. Firstly, antigens from adult H. contortus elicited larger eosinophil-rich cellular exudates in immune compared to non-immune ewes. In this situation, secondary responsiveness in the mammary gland must have been generated through abomasal infection with the parasite. Secondly, repeated infusion with the antigens from adult H. contortus increased the size of cellular exudates in both immune and non-immune ewes. Eosinophils predominated but numbers of macrophages and lymphocytes were also increased. In this second situation, secondary responsiveness must have been either supplemented in immune ewes or derived completely in non-immune ewes by contact with helminth antigens through the mammary gland. The helminth antigens which induce eosinophil exudates in the mammary gland may not be potently protective against H. contortus. Furthermore, eosinophil exudation may not be an in vivo correlate of immunity which is directly useful for discriminating protective antigens and applicable to vaccine development. Infusion with antigens from adult forms of either H. contortus or Trichostrongylus colubriformis elicited cellular exudates equally well in immune ewes primed by infusion with H. contortus adult antigens 7 days beforehand. In addition, antigens from infective larvae of H. contortus elicited cellular exudates more potently than antigens from adult worms. However, vaccination with irradiated larvae has shown that species-specific protective immunity for H. contortus is stronger than cross-protective immunity conferred by T. colubriformis.(ABSTRACT TRUNCATED AT 250 WORDS)

Some relationships between age, immune responsiveness and resistance to parasites in ruminants.

In the Australian livestock industries, susceptibility to infectious diseases is generally greater in young than in mature ruminants. The increased susceptibility is manifest as respiratory and intestinal infections (viral and bacterial) of calves, as well as fleece rot, flystrike and, especially, gastrointestinal parasitic infestations of young sheep. Lower resistance to infectious disease in young ruminants appears to be due largely to immunological hyporesponsiveness, and is not simply a consequence of their not having been exposed sufficiently to pathogens to develop active immunity. Young sheep have significantly lower proportions of CD4+ and CD8+ lymphocytes, but similar proportions of T19+ and B lymphocytes in blood, lymph and skin compared with mature sheep. Blood lymphocytes from young sheep produce less interferon-gamma in culture and young sheep invariably mount smaller antibody responses than do mature animals. Taken together, these findings begin to explain why young ruminants are more susceptible to infectious diseases in general, and to gastrointestinal parasites in particular, when compared to mature animals. Haematological markers of disease resistance, the prevalence of non-selected diseases and immune responses to vaccination were examined in the internal parasite-resistance flocks in Armidale NSW and the fleece rot/flystrike selection flocks at Trangie NSW. Any programme that seeks to improve resistance to parasitic or any other disease should have the capacity to make contemporary measurements of resistance to other diseases which are important in, or threaten, the production system.

Growth of Lucilia cuprina larvae following treatment of sheep divergently selected for fleece rot and fly strike with monoclonal antibodies to T lymphocyte subsets and interferon gamma.

Intensive lymphocytic infiltration of the underlying dermis occurs during cutaneous myiasis caused by larvae of the blow fly, Lucilia cuprina. To determine the effect of this infiltrate on larval growth, monoclonal antibodies (mAb) to CD4, CD8 or WC1 lymphocyte subset determinants were injected intravenously before and during experimental infection of sheep with larvae. The effect of intravenous injection of mAb to ovine interferon (IFN) gamma was also examined. The experiments were performed in 18-month-old maiden ewes with genetic resistance or susceptibility to the disease complex, bacterial dermatitis/cutaneous myiasis. mAbs induced profound depletion of CD8+ and WC1+ subpopulations from blood and skin at sites of larval growth. mAb to CD4+ gave only a moderate reduction in lymphocytes from blood or skin. mAb treatments did not modify larval growth or survival at 20 or 50 h after infection. Larval growth rates did not differ between resistant and susceptible genotypes. No evidence was found for a role of T lymphocyte subpopulations or the cytokine IFN, in modifying larval growth during the first 50 h of infection. It seems unlikely that T lymphocyte-dependent immunological effector mechanisms contribute to the lower prevalence of fly strike seen in the resistant genotype in the field.

Immunological responses of the lactating ovine udder following experimental challenge with Staphylococcus epidermidis.

The responses of five lactating ewes to experimental mammary infection with Staphylococcus epidermidis were examined. Infection caused an intense but transient influx of neutrophils into milk, which peaked at 8 h and was accompanied by mild fever and mild leukopaenia in blood. No other signs of systemic infection were observed. Number of staphylococci in milk decreased logarithmically until 24 h, were absent from three ewes at 48 h and then increased in number or re-emerged in four of the five ewes at 72 or 144 h. At all times milk appeared grossly normal. Expression of the adhesion molecules CD11b and CD18 increased on neutrophils in milk at 24 h then tended to decline over subsequent days. The proportion of lymphocytes positive for CD4, CD8, WC1 and MHCII tended to decrease from 24 to 72 h then increased at 144 h. Cytokines in milk were measured by ELISA. IL-8 was elevated in infected glands at 2 h, peaked at 24 h and remained elevated until the final sampling at 144 h. IL-6 was transiently elevated at 4 and 8 h while IL-1beta remained elevated from 8 until 144 h. The results suggest that the intense early neutrophil infiltrate eliminated most but not all bacteria and a state of subclinical infection ensued. After 24 h , leukocyte numbers in milk declined while cytokines, especially IL-8 remained elevated, suggesting that sensitivity or responsiveness of gland to inflammatory signals decreased as infection progressed. This attenuation of the host defence response may have contributed to the failure of the gland to eliminate bacteria and may be an important feature of the development of chronic and subclinical mastitis.