Reversible defects in natural killer and memory CD8 T cell lineages in interleukin 15-deficient mice.

C57BL/6 mice genetically deficient in interleukin 15 (IL-15(-/-) mice) were generated by gene targeting. IL-15(-/-) mice displayed marked reductions in numbers of thymic and peripheral natural killer (NK) T cells, memory phenotype CD8(+) T cells, and distinct subpopulations of intestinal intraepithelial lymphocytes (IELs). The reduction but not absence of these populations in IL-15(-/-) mice likely reflects an important role for IL-15 for expansion and/or survival of these cells. IL-15(-/-) mice lacked NK cells, as assessed by both immunophenotyping and functional criteria, indicating an obligate role for IL-15 in the development and functional maturation of NK cells. Specific defects associated with IL-15 deficiency were reversed by in vivo administration of exogenous IL-15. Despite their immunological defects, IL-15(-/-) mice remained healthy when maintained under specific pathogen-free conditions. However, IL-15(-/-) mice are likely to have compromised host defense responses to various pathogens, as they were unable to mount a protective response to challenge with vaccinia virus. These data reveal critical roles for IL-15 in the development of specific lymphoid lineages. Moreover, the ability to rescue lymphoid defects in IL-15(-/-) mice by IL-15 administration represents a powerful means by which to further elucidate the biological roles of this cytokine.

Noise levels in lairages for cattle, sheep and pigs in abattoirs in England and Wales.

Levels of sound intensity were measured over periods of 24 hours in 34 abattoir lairages in England and Wales. The mean integrated range in 12 cattle lairages was 52 to 79 dB(A), in 11 sheep lairages, 45 to 76 dB(A) and in 11 pig lairages, 46 to 87 dB(A). In general, the pig lairages were the noisiest, with spot peak recordings of up to 110 dB(A). Typically, the sound intensities in all the lairages were 10 to 20 dB(A) higher during the working day than at night. In many sheep lairages, high intensities of sound were recorded frequently throughout the night, but others were very quiet, below 40 dB(A). Vocalisations were the major sources of noise in the pig and cattle lairages, but there were variations between them. There were high intensities of sound from handling systems (80 to 90 dB[A]), and ventilating fans (70 to 80 dB[A]).

Factors affecting the mortality of pigs being transported to slaughter.

A multilevel logistic regression model was used to identify and quantify the effects of factors affecting the mortality of pigs being transported to slaughter under commercial conditions. A survey of 739 journeys to 37 slaughterhouses in five eu countries was carried out, and information potentially related to the welfare and mortality of the pigs and the number of injuries were recorded. The average temperature during the journey, its duration, the average loading time per pig, the recorded injuries, fasting before transport, and the interaction between fasting and journey duration were used in the final model. The average mortality was 0.11 per cent and the average proportion of injured pigs was 0.36 per cent, and these figures were significantly correlated (P<0.001). In 29 per cent of the journeys the pigs were not fasted before being loaded, which doubled the risk of mortality irrespective of whether the pigs were injured or not. The risk of mortality increased with average temperature. In journeys with fasted pigs that did not have any recorded injury, average temperature was more important than the duration of the journey. The risk of mortality increased as the average time taken to load them decreased, and the risk was highest when the pigs were not fasted and when injuries were recorded. Other factors such as the country, loading density, availability of drinking water and type of ventilation did not affect the risk of mortality.

Aspects of meat and eating quality of broiler chickens reared under standard, maize-fed, free-range or organic systems.

1. The effects of rearing chickens using standard, maize-fed, free-range or organic production systems on meat quality and sensory characteristics were evaluated. The standard system used either Ross or Cobb birds with a slaughter age of approximately 40 d, the other systems in the trial used Hubbard birds with considerably older slaughter ages, up to 72 d in the case of organic systems. 2. Paired breast fillets from 120 birds, 30 from each rearing system, were used. The meat quality variables, ultimate pH, colour coordinates L*, a*, b* and water-holding capacity were measured and taste panel assessments were made using 8-point category scales of texture, juiciness, abnormal flavour, flavour liking and overall flavour. 3. There were significant differences between rearing systems, with fillet muscles from birds grown under the standard system having a higher ultimate pH. Differences were also seen in colour with fillets from birds reared under a standard system having a smaller hue angle than those grown using the maize-fed system which had the highest. 4. Fillets from birds reared in the standard system were rated by the taste panel as more tender and juicy. There were no significant differences in chicken flavour. Based on hedonic assessments of flavour liking and overall liking, by a small panel of assessors, meat from birds produced in the standard system was most preferred and that from organic systems the least preferred. Meat from free-range and maize-fed systems was intermediate in preference. This result reveals a trend, but does not infer consumer acceptance.

Prevalence and factors associated with it, of birds dead on arrival at the slaughterhouse and other rejection conditions in broiler chickens.

1. Standardised data on husbandry were recorded for a flock of birds in one house on each of 150 broiler farms in the UK during the 4 d prior to slaughter. 2. For each flock, the incidence of birds found dead on arrival (DoAs) and the Meat Hygiene Service carcase rejection records were recorded at the slaughterhouse. 3. The mean percentage of birds in each flock found DoA was 0.12% (range 0-0.64%) and the mean percentage of Total Carcase Rejects (TCRs) for each flock was 1.23% (range 0.07-5.51%). 4. A general linear model was developed to examine factors associated with flock percentage DoAs. Assuming a linear relationship, all other factors remaining the same, a one percentage point (PP) increase in small/emaciated birds will result in a 0.155 PP increase in DoAs and a 1 PP increase in wheat in diet 4 will result in a 0.003 PP decrease. An increase by one in the total number of vaccines administered will cause a 0.029 PP decrease in DoAs, a 1 g increase in live weight at slaughter will be associated with a 0.000043 PP increase and a 1 PP increase in mortality on farm would be associated with a 0.000044 PP increase. A 1 PP increase in Ross birds decreases DoAs by 0.0004 PPS: there is also a seasonal effect. 5. The model developed for flock percentage TCRs found that a 1 PP increase in wheat in diet 3 will result in a 0.052 PP decrease in TCRs and a 1 PP increase in Ross birds will cause a 0.009 PP decrease. A 1 PP increase in birds culled on farm will be associated with a 0.03 PP increase in TCRs and the diagnosis of disease during the flock cycle increases TCRs by 0.397 PPs. A one day increase in age at slaughter will result in a 0.046 PP increase in TCRs.

Microinjections of melanin concentrating hormone into the nucleus tractus solitarius of the rat elicit depressor and bradycardic responses.

The presence of melanin-concentrating hormone (MCH) containing processes, projecting from the lateral hypothalamus to the medial nucleus tractus solitarius (mNTS), has been reported in the rat. It was hypothesized that MCH acting within the mNTS may modulate the central regulation of cardiovascular function. This hypothesis was tested in urethane-anesthetized, artificially ventilated, adult male Wistar rats. Microinjections (100 nl) of MCH (0.25, 0.5, 0.75, and 1 mM) into the mNTS of anesthetized rats elicited decreases in mean arterial pressure (20.4+/-1.6, 50.7+/-3.3, 35.7+/-2.8 and 30.0+/-2.6 mm Hg, respectively). The decreases in heart rate in response to these concentrations of MCH were 40.0+/-8.7, 90.0+/-13.0, 48.0+/-7.3 and 48.0+/-8.0 beats/min, respectively. Maximum cardiovascular responses were elicited by a 0.5 mM concentration of MCH. Cardiovascular responses to MCH were similar in unanesthetized mid-collicular decerebrate rats. Control microinjections of normal saline (100 nl) did not elicit any cardiovascular response. Ipsilateral or bilateral vagotomy significantly attenuated MCH-induced bradycardia. Prior microinjections of PMC-3881-PI (2 mM; MCH-1 receptor antagonist) into the mNTS blocked the cardiovascular responses to microinjections of MCH. Microinjection of MCH (0.5 mM) into the mNTS decreased efferent greater splanchnic nerve activity. Direct application of MCH (0.5 mM; 4 nl) to barosensitive nucleus tractus solitarius (NTS) neurons increased their firing rate. These results indicate that: 1) MCH microinjections into the mNTS activate MCH-1 receptors and excite barosensitive NTS neurons, causing a decrease in efferent sympathetic activity and blood pressure, and 2) MCH-induced bradycardia is mediated via the activation of the vagus nerves.

The colour of the adductor muscle as a predictor of pork quality in the loin.

The relation between measurements of colour made in the m. adductor (AD) at 45min or 20 h post mortem and the quality, assessed subjectively in terms of colour and waterholding capacity, of the m. longissimus (LD) in the loin was examined. The study used data from 100 pig carcasses exhibiting a wide range of meat quality from extreme PSE (pale, soft and exudative) to extreme DFD (dark, firm and dry). The subjective assessments were confirmed by objective measures of paleness (reflectance) and waterholding capacity (drip loss in storage) in the LD. Lightness (L(∗)) measured at 20h post mortem in the AD was the best potential predictor of loin muscle quality, explaining 59% of the variation in subjective and objective quality measures. Comparable measurements at 45min post mortem explained between 21% and 44% of the variation. The equation that described the relation between AD Lightness (L(∗)) and subjectively assessed LD quality was derived. This could be used to transpose the AD L(∗) values from a population of slaughtered pigs into nominal subjective scores for the LD, allowing the frequency of the five subjective quality groups (extremely DFD, slightly DFD, normal, slightly PSE, and extremely PSE) in the population of carcasses to be defined.

Estimating the body temperature of groups of pigs by thermal imaging.

Measurements on 28 pens of pigs containing 384 animals to be slaughtered at a commercial abattoir showed that the mean ear temperatures of the pigs in each pen, measured with a thermal imaging camera, were significantly correlated (r=0.71, P<0.001) with the mean temperature of the blood the pigs lost at exsanguination. In measurements on 220 pigs in 16 of the 28 pens, the mean activity of serum creatine kinase was positively correlated with the mean ear temperature (r=0.55, P<0.05) and the mean concentration of serum cortisol was positively correlated with the mean blood temperature (r=0.50, P<0.05), suggesting that the hotter pigs were suffering from a higher level of stress.

Comparison of the effects of fan-assisted and natural ventilation of vehicles on the welfare of pigs being transported to slaughter.

A total of 417 pigs with an estimated average liveweight of 91 kg were transported to slaughter in three journeys, carried out in late June, each consisting of a three-hour drive followed by a one-hour holding period at the slaughter plant before they were unloaded. The pigs were accommodated on the lower two decks of a three-deck articulated lorry, each deck having five equal-sized pens; half were kept in naturally ventilated pens and half in pens with fan-assisted ventilation. The pigs' body temperature was recorded as the temperature of the blood lost at exsanguination, and as the temperature of the inner surface of the ear measured by thermal imaging. The pigs carried on the upper deck were hotter than those carried on the lower deck, and the pigs carried at the front of the vehicle were hotter than those at the back. There were small differences between the temperatures of the pigs in the different pens, but overall the temperatures of the pigs kept in pens with fan-assisted ventilation were no lower than those of the pigs kept in pens with natural ventilation. The higher temperature of the pigs transported in the front of the vehicle was associated with significantly higher serum concentrations of cortisol and activities of creatine kinase, and a tendency to higher albumin concentrations and osmolality, suggesting that they may have been exposed to more physical and psychological stress, and were possibly slightly more dehydrated, than the pigs in the rearmost pens. The type of ventilation had no effect on the blood composition of the pigs.

The response of pigs to being loaded or unloaded onto commercial animal transporters using three systems.

Groups of pigs were subjected to three different systems of loading and unloading to and from commercial animal transporters. The systems under trial were: the use of a hydraulic tail-lift, a tail board ramp at an angle of 18 degrees and a modular system. The module was a container 2 m x 2.4 m x 1 m high into which the pigs were loaded. The module could then be lifted on and off the lorry with a fork lift truck. Non-invasive monitoring techniques were used to study the responses of the pigs. A subjective handling score, time taken to load and unload, skin temperature, heart rate and salivary cortisol were recorded. Loading and unloading were subjectively assessed as being easiest and quickest using the modular system, which also appeared to be less physically demanding for the animals, as evidenced by lower heart rate and a reduced maximum heart rate. However, with the modular system, elevated cortisol was found during the loading, unloading and resting periods. This could be interpreted as indicating that the pigs were unable to settle in the module and suffered some small degree of stress, which might become more of a problem during prolonged transport. Of the other two systems, there appeared to be little difference between the hydraulic tail-lift and the ramp.

The genomic organization of Isopeptidase T-3 (ISOT-3), a new member of the ubiquitin specific protease family (UBP).

A novel Isopeptidase T gene (ISOT-3) has been identified on human mosome 3q26.2--q26.3. gene shows 67.3% nucleotide identity and 54.8% amino acid identity to n Isopeptidase (ISOT-1). Northern blot analysis has shown that ISOT-3 is highly essed in ovary and testes, low-level expression in six other tissues tested. In contrast, ISOT-1 is essed at high levels in brain, and there is no detectable expression in ovary. The exonic nization of these two genes highly conserved with only one variant intron position. Intron 15 in -3 is absent in ISOT-1, there is an alternate splice site at the same location. Although the --intron structure has been erved between the two genes, ISOT-3 has significantly larger intronic ons, and the overall of this gene is at least 90 kb compared to 15 kb for ISOT-1. These data suggest that both ISOT-1 and ISOT-3 have descended from a common ancestor. In addition, the low overall sequence identity and different expression patterns may reflect differences in substrate specificity.

Methods for detection of haematogenous dissemination of brain tissue after stunning of cattle with captive bolt guns.

Because of concern that the stunning of cattle with captive bolt guns (CBGs) could, if used on an animal with bovine spongiform encephalopathy (BSE), cause embolism of infective brain tissue and carcass contamination, the Ministry of Agriculture, Food and Fisheries commissioned research to assess the risk of haematogenous dissemination of CNS material after stunning. We have devised two methods to investigate this risk. The first involves the concentration of embolic tissue in buffy coat Cytoblocks that can be embedded for sectioning, microscopy and immunocytochemistry. The second method is an ELISA for the presynaptic protein, syntaxin 1B. The methods were validated by analysis of several bovine tissues, including blood samples deliberately contaminated with brain. We then studied jugular venous blood obtained before and after the stunning of 60 cattle with CBGs. Samples obtained, after stunning, from five of the cattle contained CNS tissue within the Cytoblocks and yielded positive syntaxin assays. Syntaxin was also detected in samples from one other animal that had been stunned with a pneumatically operated CBG. The described methods should allow an assessment of the risk of neuroembolism associated with different types of CBG and may also be useful in other contexts.

Variable effect of toxic shock toxins from different sources on neutrophil function in vitro.

Toxic shock syndrome toxins (TSST) are 23-30 kD proteins that have been isolated from incubation media of strains of Staphylococcus aureus cultured from patients with toxic shock syndrome (TSS). Injection of TSST into animals produces many of the symptoms that characterize TSS including shock, fever, and multiple organ failure. We found that addition of increasing concentrations of TSST-1-VP1035-16A, but not TSST-PEC, TSST-SEC, staphylococcal enterotoxin A or B, progressively decreased human neutrophil bactericidal activity against S. aureus, 502A in vitro. TSST-1-VP1035-16A, but not the other toxins, also decreased superoxide anion and hydrogen peroxide concentrations in mixtures containing neutrophils and phorbol myristate acetate (PMA) in vitro. The results indicate that various preparations of TSST have different effects on neutrophil function in vitro and, accordingly, may have different effects in other in vitro and in vivo models of TSS.

Potential effect of vibration during transport on glycogen reserves in broiler chickens.

Subjection of broiler chickens to random, narrow band vibration (2, 5, 10 Hz) for 1 h, simulating that experienced during normal road transport, did not significantly influence liver or muscle glycogen concentrations or muscle ultimate pH (pHu). Vibration for 3 h increased body temperature (P<0.05) and decreased (P<0.01) pHu in both 'white' pectoralis superficalis (PS) and 'red' biceps femoris (BF) muscles overall, but did not affect liver or muscle glycogen concentrations. However, higher vibration frequencies resulted in reduced (P<0.05) glycogen concentrations in liver and BF muscle. The conclusion was that vibration was unlikely to be the major cause of muscle glycogen depletion seen in transported broilers, but the reduction in pHu in the PS muscle after vibration may have been related to the similar effect seen in previous studies after normal transport.

The effects of the beta-adrenergic agonist salbutamol on meat quality in pigs.

The beta-adrenergic agonist Salbutamol was administered to pigs at 3 ppm in the feed between weaning and slaughter at 85 kg. Growth rate was not affected by Salbutamol. Treated pigs had a higher dressing percentage (2.6%) and produced carcasses that were less fat (17%) and had longissimus (LD) muscles of larger (11%) cross-sectional area. They also had smaller livers that contained less glycogen. The thinner backfat in treated animals was less firm and tended to separate from the underlying lean. However, these changes were attributable solely to the reduced fatness and there was no direct effect of Salbutamol. There were no differences in pH 45 min postmortem, percentage drip loss during storage or reflectance value of the LD between the two groups, indicating no greater propensity for Salbutamol-treated pigs to develop pale, soft, exudative muscle. However, treated pigs had higher final pH values in the muscles; this was reflected in slightly reduced hue and saturation values. These results suggest that the propensity of the pigs to develop dark, firm, dry meat was slightly increased. Salbutamol-treated pigs produced LD muscles that were slightly tougher (22%), had reduced concentration of heme pigments in the muscle, reduced plasma glucose and increased plasma creatine phosphokinase activity. Salbutamol improved lean meat yield but slightly increased the potential to produce dark, firm, dry meat and reduced tenderness.

Interactions between the beta-adrenergic agonist salbutamol and genotype on meat quality in pigs.

Salbutamol (2.7 ppm) fed to pigs between weaning and slaughter increased ADG (5%), dressing percentage (2%) and cross-sectional area of the longissimus (LD) muscle (14%). In fatter, White-line-sired pigs, but not in leaner, Meat-line-sired animals, it reduced backfat thickness (25%). Liver weight and glycogen concentration also were decreased. In neither genotype were there effects of Salbutamol on pH45, drip loss or reflectance value of the LD, but ultimate pH was elevated in semimembranosus (SM), adductor (AD) and supraspinatus (SS) muscles, resulting in reduced color saturation values. Total muscle heme pigment concentrations were reduced by 10 (SS) to 19% (LD and SM) and the intramuscular fat concentration of the AD, but not the LD, was reduced by 21%. Treated pigs had LD and SM muscles that when measured instrumentally were 15 and 8% tougher, respectively, after cooking, but the texture of the SS was not significantly altered by treatment. Salbutamol increased plasma lactate and creatine phosphokinase (CPK) concentrations. Compared with White-line-sired animals, Meat-line-sired pigs had a higher dressing percentage (2%) and LD muscles with larger cross-sectional area (13%). They also had higher circulating CPK levels.

A note on the use of subjective methods for assessing pig meat quality on the slaughterline.

The relationships between assessment of exudation from the intrascope hole, seen as a discolouration on the surface of the skin at 20h post mortem, subjective assessment of meat quality in the longissimus dorsi (LD), gluteus medius (GM) and rectus femoris (RF) muscles and objective measures of raw meat quality in the LD were determined on 100 pig carcasses representing a wide range of quality. The relationships between subjective exudation assessment and objective measures of quality were good with the best correlations being found with those measures indicating pale, soft, exudative (PSE), those with objective indicators of dark, firm, dry (DFD) being poorer. The relationships between subjective assessment of quality in the LD, GM and RF and all objective measures of meat quality in the LD were high. The appearance of any of the muscles could be used to predict overall meat quality in the carcasses after cutting. The subjective assessment of exudation was therefore a good, non-invasive predictor of severe PSE, but not DFD meat, which could be applied on the line after overnight chilling. After butchery, subjective assessment of LD, GM or RF muscles could be used to identify both PSE and DFD carcasses.

The relationships between initial pH, reflectance and exudation in pig muscle.

Using data collected from several hundred pigs, the relationships of drip loss during storage, and reflectance, to pH(45) in the M. longissimus dorsi (LD) were both found to be biphasic. Muscles with a lower pH(45) lost more drip and were paler. Below pH 6·1, however, decreasing pH(45) had only a small effect on the weight of drip lost but a large influence on reflectance. In contrast, above pH 6·1, increasing pH(45) had a large influence on drip loss but only a small effect on reflectance. There was also a biphasic relationship between drip loss and reflectance with the discontinuity at 9·9% drip. Differences in pH(u) explained less than 15% of the variation in exudation and reflectance.

The use of muscle protein solubility measurements to assess pig lean meat quality.

The relationships between sarcoplasmic, myofibrillar and total soluble protein (sarcoplasmic + myofibrillar) concentrations, and subjective (colour-structure score) and objective (drip loss and reflectance) measures of lean meat quality were determined using 100 samples of M. Longissimus dorsi that showed a wide range of quality. There was good agreement between the subjective and objective assessments of quality. Overall, the concentration of soluble sarcoplasmic proteins showed the highest correlations with quality assessments. The relationships with total soluble protein were poorer and were least good with myofibrillar protein concentration. Sarcoplasmic protein concentration was also the best potential predictor of meat quality when pale, soft, exudative (PSE) muscles were excluded from the sample. However, if dark, firm, dry (DFD) muscles were excluded instead, the concentration of total soluble protein was as good as, or better than, sarcoplasmic protein as a potential predictor of quality, particularly muscle reflectance.

An estimate of the incidence of dark cutting beef in the United Kingdom.

Eight slaughterplants with throughputs ranging from 20 to 300 animals per day were examined to estimate the incidence of dark cutting beef in the United Kingdom. Four thousand, eight hundred and sixteen animals were surveyed and information concerning animal category, source, season and preslaughter handling conditions recorded. Muscle samples were removed to estimate glycogen concentration and after incubation, ultimate pH. The overall incidence of dark cutting (pHu ≥ 6·0) was 4·1%. Increased incidence was associated with short (≤ 20 miles) and long (≥ 150 miles) transport distances. Slaughter on the day of arrival rather than overnight lairage also increased the incidence. Plants were classified into small (killing ≤ 50 animals per day) or large (killing ≥ 100 per day). Eighty per cent of the animals slaughtered passed through the large plants, and a higher incidence was also associated with these plants. Bulls had the highest incidence and heifers the lowest. A seasonal effect was recorded with the highest incidence found between July and October. The results, however, indicate that factors in addition to those examined are also important.