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Researchers who use animals in science must balance statistical power with the need to satisfy the three Rs, whereby researchers are required to reduce numbers of animals, refine what they experience, and use alternatives to (replace) higher animals where possible. In repeated sampling over time-series studies, there is potential loss of power as well as ethics implications posed by repeated sampling of individual animals, where this cannot be managed or avoided. Here, we consider the mathematics of repeated sampling from three perspectives: that of the population at large, from the experience of the individual, and the conditional probability of sampled individuals being sampled again. The calculations are illustrated using four theoretical case studies across veterinary epidemiology with different practical implications and a provided R Shiny tool for researchers. Despite the availability of exact calculations, it is necessary to also consider the biological factors which may affect capture and recapture rates in sampling studies such as animal personality and response to capture. Researchers must also choose their question carefully to avoid inappropriate framing of ethical concerns around repeated sampling., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

We analyse the network structure of the British salmonid aquaculture industry from the perspective of infectious disease control. We combine for the first time live fish transport (or movement) data covering England and Wales with data covering Scotland and include network layers representing potential transmission by rivers, sea water and local transmission via human or animal vectors in the immediate vicinity of each farm or fishery site. We find that 7.2% of all live fish transports cross the England-Scotland border and network analysis shows that 87% of English and Welsh nodes and 72% of Scottish nodes are reachable from cross-border connections via live fish transports alone. Consequently, from a disease-control perspective, the contact structures of England and Wales and of Scotland should not be considered in isolation. We also show that large epidemics require the live fish movement network and so control strategies targeting movements can be very effective. While there is relatively low risk of widespread epidemics on the live fish transport network alone, the potential risk is substantially amplified by the combined interaction of multiple network layers., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

Bovine tuberculosis (bTB) is a chronic zoonosis with major health and economic impact on the cattle industry. Despite extensive control measures in cattle and culling trials in wildlife, the reasons behind the expansion of areas with high incidence of bTB breakdowns in Great Britain remain unexplained. By balancing the importance of cattle movements and local transmission on the observed pattern of cattle outbreaks, we identify areas at elevated risk of infection from specific Mycobacterium bovis genotypes. We show that elevated-risk areas (ERAs) were historically more extensive than previously understood, and that cattle movements alone are insufficient for ERA spread, suggesting the involvement of other factors. For all genotypes, we find that, while the absolute risk of infection is higher in ERAs compared to areas with intermittent risk, the statistically significant risk factors are remarkably similar in both, suggesting that these risk factors can be used to identify incipient ERAs before this is indicated by elevated incidence alone. Our findings identify research priorities for understanding bTB dynamics, improving surveillance and guiding management to prevent further ERA expansion., (Crown Copyright © 2018. Published by Elsevier B.V. All rights reserved.)

The performance of a visual image analysis (VIA) system was tested with regards to its potential to determine in vivo carcass composition and conformation, either alone, or in conjunction with other in vivo measures such as live weight and backfat depth. Pigs of both sexes of a commercial type were reared and slaughtered at weights ranging from 50 to 120 kg. Feeding was ad libitum on diets ranging from 0.14 to 0.19 kgkg(-1) crude protein content to produce animals of a range of body condition. Two analyses were carried out: the first analysis addressed the relationship between dimensionless carcass and VIA indices; the second analysis assessed the relationship between carcass composition and VIA body shape using detrended carcass and VIA data, which were produced by removal of allometric growth trends. A statistically significant relationship (P<0.05) between in vivo VIA body size and shape and carcass muscle dimensions and composition was found for most body regions. Adjusted R(2) statistics ranged between 0.13 and 0.54 for relative fat weights and between 0.14 and 0.51 for relative lean weights. The predictive power of the regression models, indicated by R(2)-like statistics for prediction, was approximately 70% of the adjusted R(2) values. The descriptive and predictive powers of the corresponding models generally strengthened if VIA indices were combined with other in vivo measurements. The relationships between in vivo and carcass measures remained statistically significant (P<0.05) after removal of the growth trends, although adjusted R(2) statistics generally decreased. The predictive power of models corresponding to the detrended measures was, however, weak. The results show in vivo VIA measurements to be useful in the estimation of muscle size, carcass conformation and composition, all of which are of significant importance to the pig production, marketing and processing industries.

In vitro measurements of skin absorption are an increasingly important aspect of regulatory studies, product support claims, and formulation screening. However, such measurements are significantly affected by skin variability. The purpose of this study was to determine inter- and intralaboratory variation in diffusion cell measurements caused by factors other than skin. This was attained through the use of an artificial (silicone rubber) rate-limiting membrane and the provision of materials including a standard penetrant, methyl paraben (MP), and a minimally prescriptive protocol to each of the 18 participating laboratories. "Standardized" calculations of MP flux were determined from the data submitted by each laboratory by applying a predefined mathematical model. This was deemed necessary to eliminate any interlaboratory variation caused by different methods of flux calculations. Average fluxes of MP calculated and reported by each laboratory (60 +/- 27 microg cm(-2) h(-1), n = 25, range 27-101) were in agreement with the standardized calculations of MP flux (60 +/- 21 microg cm(-2) h(-1), range 19-120). The coefficient of variation between laboratories was approximately 35% and was manifest as a fourfold difference between the lowest and highest average flux values and a sixfold difference between the lowest and highest individual flux values. Intralaboratory variation was lower, averaging 10% for five individuals using the same equipment within a single laboratory. Further studies should be performed to clarify the exact components responsible for nonskin-related variability in diffusion cell measurements. It is clear that further developments of in vitro methodologies for measuring skin absorption are required., (Copyright 2005 Wiley-Liss, Inc. and the American Pharmacists Association.)

The growth of carcass tissues and developmental changes in tissue distribution were studied in three pig types which were selected to have morphologies that could be described as attenuated, blocky and flabby. These were achieved by incorporating, respectively, Landrace, Pietrain and Meishan genes to give commercial 'types' (designated L, P and M,) exhibiting some of the phenotypic qualities of these breeds. Twenty-five female pigs of each type with an average start weight of 27.2 kg were fed ad libitum and slaughtered over a (nominal) live weight range of 35-115 kg. Relations were quantified using the logarithmic transformation of data in the allometric model. Significant type differences in relative growth rates (the b coefficient or slope in the logarithmic plot) were not common, occurring in 14% of the relationships examined, whereas 61% of differences in the constant term (a or intercept in the logarithmic plot) were significant. Increase in carcass weight with age was not different between the types but in relation to slaughter live weight (dressing percentage) P had the highest value, M the lowest. Carcass dimensions showed that, relative to body length, P had the widest ham and shoulder over the whole size range whereas M had a deep (ventral-dorsal) shoulder and wide belly, attributes of shape that would be regarded as undesirable by the meat trade. Relative to carcass weight, L was only slightly longer (3 mm) in the body than M at the heavy end of the weight range but markedly longer (29 mm) than P. These differences in carcass conformation were also evident in the shape of pelvic limb muscles which, at a given length, were lightest and narrowest in M and, in some cases, heavier and wider in P than in L. Pelvic limb volume relative to limb length was greatest in P and least in M. Carcass composition (at a given prepared side weight) of P was characterized by low fat and high lean weights, and a high lean to bone ratio. Subcutaneous fat b was lowest in P, indicating that P was early maturing, but M carcasses had the most subcutaneous and intermuscular fat, also indicative of early maturing. There was, thus, no simple relation between maturity characteristics and carcass composition. M carcasses also had the greatest weight of skin. P had a light hindloin and heavy pelvic limb, M a heavy shoulder, hindloin and flank but a light pelvic limb; L had a light shoulder but a heavy foreloin. The distribution of individual tissues mirrored these differences in joint weights to a varying extent; in most cases the match (relatively heavy/light) was with a single tissue. These data, thus, indicated some pig type differences in tissue distribution and of particular significance was the relatively light lean mass in the pelvic limb of M. The results showed important differences in carcass quality between commercially available pig types differing in conformation.

The inclusion of 1% casein or bovine serum albumin in buffer used to reactivate enzymes subjected to sodium dodecyl sulfate (SDS)-polyacrylamide electrophoresis resulted in accelerated removal of SDS and restoration of nuclease and beta-galactosidase enzyme activities. Nuclease and beta-galactosidase activities which are absent from gels after longer wash procedures are detectable with this technique. Enzyme activity in gels prepared with SDS which contained inhibitory contaminants was partially restored by the casein wash procedure. The threshold of detection of two-dimensionally separated deoxyribonuclease I using the casein wash procedure was 1 picogram.

The irritant properties of 3 methylated derivatives of dibenz[b,f]1,4-oxazepine (CR) have been investigated, using the guinea-pig blepharospasm test and isolated cat eye preparations, and compared with the irritancy of the unsubstituted CR molecule. In both tests, the 11-methyl derivative was non-irritant whereas 3 Me-CR and 9 Me-CR retained some irritancy although both were less potent than CR itself. The possible significance of these results are discussed in relation to earlier biochemical studies.

A retrospective comparison was made of three methods of central-nervous-system prophylaxis in childhood acute lymphoblastic leukaemia; (1) intrathecal methotrexate only; (2) intermediate-dose methotrexate infusion and intrathecal methotrexate and, (3) 2400 rads cranial irradiation and intrathecal methotrexate. The incidence of primary meningeal relapse was statistically significantly lower in both standard-risk patients (age grear than 24 months and less than or equal to 120 months white-cell count less than 20,000) and increased-risk patients (age less than or equal to 24 months or greater than 120 months and/or white-cell count greater than 20,000) whose central-nervous-system prophylaxis included cranial irradiation. The disease-free and overall survival of irradiated increased-risk patients was significantly better than that of unirradiated increased-risk patients. The disease-free survival of standard-risk patients who received intermediate-dose methotrexate was statistically superior to that of the remaining standard-risk patients. There were no significant differences in overall survival between the three groups of standard-risk patients.

Wilms' tumor is the model of the treatment of a pediatric solid tumor. Initially it appeared that multi-modality therapy, consisting of transabodominal nephrectomy, post-operative radiation therapy to the tumor bed and adjuvant, single agent chemotherapy provided the highest likelihood of disease-free survival. The identification of important prognostic factors, such as histology, tumor weight, lymph node involvement and age at diagnosis has led to a re-examination of the treatment of Wilms' tumor. Future therapeutic developments will include the administration of less therapy to some well defined groups of patients, and the exploration of new programs for patients who have been demonstrated to have a poor prognosis using currently accepted treatment techniques.

A two-dimensional zymogram procedure for the analysis of nucleases is described. Isoelectric focusing (IEF) and nonequilibrium pH gradient electrophoresis (NEPHGE) were compared as first dimensions in combination with sodium dodecyl sulfate (SDS) electrophoresis as the second dimension in analyzing nucleases in lysates of Bacillus subtilis. All renaturable nucleases detected following SDS electrophoresis alone were resolved in NEPHGE-SDS electrophoresis gels whereas, in IEF gels, most either were at the basic end or were not present in the second-dimension gels. This method of analysis has revealed a complexity in nuclease species in B. subtilis not previously recognized. Eighty-three discreet nuclease activities have been detected in B. subtilis lysates. Using purified deoxyribonuclease I (bovine pancreas), as little as 10 pg of nuclease can be detected.