Quantitative and qualitative comparison of three scoring systems for assessing recovery quality after general anaesthesia in horses

Objective To assess the reproducibility and repeatability of two commonly used recovery quality scoring systems and compare them with those of a novel system based on a greater number of objective criteria. Animals The video‐recorded recoveries of ten client‐owned horses selected from all recovery recordings taken between September 2005 and March 2006 at the Royal (Dick) School of Veterinary Studies. Materials and methods A digital versatile disc (DVD) was produced using edited video recordings of ten horses recovering from general anaesthesia. Twelve experienced equine anaesthetists (raters) studied the DVD on three occasions, and scored the recovery quality of each horse using one of three scoring systems (P, D or E) on each occasion. The process was repeated 6 months later (t = 6) to measure intra‐observer reliability (repeatability). At first use (t = 0) raters were asked to comment on the advantages and disadvantages of each system. Results Inter‐rater variability was limited for each system: at each observation period raters accounted for 0.3–4.4% variation. System P was insensitive to differences between recoveries. In system D, score variability increased as recovery quality deteriorated. Intra‐rater variability varied with system: using system P, raters provided consistent scores between the observation periods for some, but not all horses (‘horse’ and ‘rater’ accounted for 9.7% and 1.9% of variation respectively). Raters were less consistent between t = 0 and t = 6 using system D, but each horse was scored with similar consistency. System E produced little variation at the level of horse (1.0%) and rater (1.9%). Raters broadly agreed on the principle advantages and disadvantages of the three systems. Conclusions and clinical relevance The systems examined showed reliability and reproducibility but practicality and simplicity of use appeared to be inextricably linked with imprecision. Further work is required to produce a suitable recovery quality scoring system.     

Molecular Epidemiology of Contagious Bovine Pleuropneumonia in Tanzania Based on Amplified Fragment Length Polymorphism and Pulsed‐field Gel Electrophoresis Analysis

The genetic diversity of 60 field strains of Mycoplasma mycoides ssp. mycoides, small colony type (M. mycoides SC), comprising 56 isolates from cattle in Tanzania, one from Kenya, two from Botswana and one from Portugal, as well as the type (PG1^T) and vaccine (T₁‐SR49) strains, was investigated. The strains were analysed for variations in the EcoRI and Csp6I restriction sites in the genomic DNA using the amplified fragment length polymorphism (AFLP) technique, and variations in the BamHI restriction sites using pulsed‐field gel electrophoresis (PFGE). Six AFLP types were detected among the analysed strains. The AFLP profiles of the type and vaccine strains were indistinguishable from each other. Indistinguishable AFLP profiles were found for 55 Tanzanian field strains, one of them isolated in 1990 and the other 54 isolated in 1998/1999, although one strain isolated in 1999 showed a different profile. Strains from different countries revealed different AFLP profiles. Six PFGE types were detected among the analysed strains, with all the 56 Tanzanian field strains displaying indistinguishable PFGE profiles. Strains from different countries revealed different PFGE profiles, and so did the type and vaccine strains. The strong genomic homogeneity among M. mycoides SC strains associated with outbreaks of contagious bovine pleuropneumonia in different regions of Tanzania suggests that the outbreaks of the disease in the 1990–99 period might have been caused by a single epidemic clone. Moreover, this study has demonstrated that AFLP and PFGE are potential tools for molecular epidemiological studies of M. mycoides SC infections.     

Assessment of three methods for multilocus fragment typing of Cryptosporidium parvum from domestic ruminants in north west Spain

The performance of three different methods, capillary electrophoresis (CE), high resolution slab-gel electrophoresis and sequencing, for PCR fragment size analysis of two Cryptosporidium parvum microsatellite regions, ML1 and ML2, was investigated by analysing 27 isolates from calves and 14 from lambs. To assess genetic variability of this protozoan in domestic ruminants in north west Spain, results were combined with sequence analysis of the 60 kDa glycoprotein (GP60) gene creating a multilocus type and analysed by farm and host species. CE showed greater overall typability (T), discriminatory power and ease of use than slab-gel electrophoresis and sequencing which were both affected by PCR stutter, especially at ML2. CE fragment sizes were consistently 4 bp longer compared to sequencing which is considered the gold standard for allele sizing but which gave the lowest typability; CE sizes were therefore adjusted. Only three alleles were identified at the ML1 locus (ML1-238, ML1-229 and ML1-226). The ML2 locus was more polymorphic and eight alleles were found (ML2-235, ML2-233, ML2-231, ML2-229, ML2-227, ML2-225, ML2-201 and ML2-176). Adjusted ML1 and ML2 CE fragment sizes were combined with GP60 subtype for 37 of the 41 C. parvum isolates which were typable at all three loci (T=0.90): nine multilocus types (MLTs) were identified. The discriminatory power of the 3-locus typing method was 0.83. Greater genetic variability was observed in calf isolates (7 MLTs) than in those from lambs (4 MLTs) although more calf isolates were studied. The most common MLT in cattle was MLT1 (ML1-238, ML2-231, GP60 subtype IIaA15G2R1), while MLT3 (ML1-238, ML2-227, GP60 IIaA16G3R1) was predominant in lambs. Our findings demonstrate that high discrimination can be achieved by means of multilocus typing. CE appears to be an economic and rapid option for performing microsatellite fragment size analysis offering good typability, discrimination and ease of use but may require calibration to sequenced standards.     

Molecular evidence for the infection of zoo chimpanzees by pig Ascaris

We here describe the transmission of the pig roundworm, Ascaris suum to chimpanzees maintained in the Copenhagen Zoo, Denmark. Using a technique for whole genome fingerprinting, amplified fragment length polymorphism (AFLP) and the technique PCR restricted fragment length polymorphism (PCR-RFLP) of the internal transcribed spacer region (ITS) of the nuclear ribosomal DNA, the worms from the chimpanzees were compared with Ascaris spp obtained from humans and pigs in order to identify the source of the infection. By the use of different distance and clustering based methods on the AFLP data set the worms from the chimpanzees were assigned to the same cluster as that of the worms from pigs. The PCR-RFLP analysis supported the AFLP results. Therefore, the zoo chimpanzees have required Ascaris infections by cross-infection from pigs. Pigs as a potential source of Ascaris infections for both captive and wild chimpanzees and other animals, therefore needs to be considered and appropriate steps taken to prevent such infection.     

A Clinical Index for Disease Activity in Cats with Chronic Enteropathy

Background: There is a need for a clinically useful, quantitative index for measurement of disease activity in cats with chronic enteropathy (CE). Objective: To develop a numerical activity index that is of practical value to clinicians treating CE in cats. Animals: Eighty‐two cats with CE. Methods: Retrospective case review of 59 cats diagnosed with inflammatory bowel disease (IBD). Prospective validation study of 23 cats having either IBD or food‐responsive enteropathy (FRE). Multivariate regression analysis was used to identify which combination of clinical and laboratory variables were best associated with intestinal inflammation of IBD. This combination of variables was expressed in a score that was used as an activity index for the prospective assessment of disease activity and of the effect of treatment in cats with IBD or FRE. Results: The combination of gastrointestinal signs, endoscopic abnormalities, serum total protein, serum alanine transaminase/alkaline phosphatase activity, and serum phosphorous concentration had the best correlation with histopathologic inflammation and comprise the feline chronic enteropathy activity index (FCEAI). Positive treatment responses in cats with CE were accompanied by significant (P < .05) reductions in FCEAI scores after treatment. Conclusions and Clinical Importance: The FCEAI is a simple numerical measure of inflammatory activity in cats with CE. The scoring index can be reliably used in the initial assessment of disease severity for both IBD and FRE and as a measure of clinical response to treatment for these disorders.     

Endemic type of animal trypanosomiasis is not associated with lower genotype variability of Trypanosoma congolense isolates circulating in livestock

In order to verify whether the low impact on livestock production in endemic areas is related to a low number of trypanosome strains circulating in livestock, 37 Trypanosoma congolense isolates collected from cattle in 11 sites in an endemic trypanosomiasis area in Eastern Zambia were characterised for genotype variability using a modified amplified fragment length polymorphism technique (AFLP). Isolates were further cloned to evaluate the occurrence of mixed infections in individuals. The results obtained revealed a high genotype diversity (94.6%) among these isolates. Apart from one site, all isolates gave different AFLP profiles in each of the sites. When clones were compared, three (8%) of the 37 isolates had mixed infections. These results indicate the circulation of a high number of strains in this trypanosomiasis endemic area despite the low impact the disease has on livestock production.     

基于AFLP的燕麦遗传多样性研究

采用ＡＦＬＰ分子标记技术对来自中国、加拿大、澳大利亚和欧洲等国家的３４个有代表性的饲用燕麦品种及８个裸燕麦品种进行遗传多样性分析,从３０ 对ＥｃｏＲＩ／ＭｓｅＩ引物组合中筛选出５ 对多态性和清晰度较高的引物组合,共获得２６８条带,其中多态带１８５条,平均多态性检出率为６９．０％,Ｅ ＡＧＧ／Ｍ ＣＴＡ 的扩增效率最高达７８．６％。由Ｎｅｉ’ｓ指数（ｈ）估测,供试品种平均变异为０．１６６４,平均Ｓｈａｎｎｏｎ’ｓ信息指数估计值为０．２２０６;４２个燕麦品种间遗传相似系数为０．４８８１～０．９８８０,遗传距离的变化范围是０．０１２０～０．７１７２。通过ＡＦＬＰ 数据构建燕麦的ＵＰＧＭＡ 系统树,在遗传相似系数０．７４８水平,所有供试品种可聚为６类。皮燕麦与裸燕麦在遗传相似度０．５９处被明显分为２类,与形态学分类结果一致。由ＡＦＬＰ揭示的品种间遗传关系与品种实际来源基本一致,品种的遗传距离与其地理分布关系密切。     

Capillary electrophoresis for the detection of bilirubin in rat serum

Background: The rat is used often to assess the toxicity of new chemical entities in preclinical drug development. Bilirubin concentration in rat serum is routinely determined by colorimetric methods, but false positive results due to hemolyzed serum or direct interferences by test compounds may occur. Capillary electrophoresis (CE) is an automated method that requires small sample volume and facilitates the direct detection of bilirubin. Objective: The purpose of this study was to evaluate a CE method for detecting bilirubin and albumin‐bound bilirubin in rat serum and to measure potential interference by hemolysis and specific test compounds. Methods: Serum samples from male Sprague Dawley rats (n=20) were used in the study. Results obtained on a Beckman P/ACE MDQ CE instrument equipped with a UV‐detector were compared with those obtained using a colorimetric method on a Hitachi 912 analyzer. Bilirubin standards were used to evaluate the detection and stability of bilirubin in rat serum, and vials with ultrafiltration membranes were used to separate albumin‐bound bilirubin. Intraday and interday coefficients of variation (CV), linearity, and the effects of added hemoglobin and a test compound on CE results were determined. Results: The CE method was capable of detecting bilirubin and albumin‐bound bilirubin in rat serum samples with reproducible results and good accuracy. CVs were <3% and linearity of the CE assay was high (R²=0.9951). Abnormally high bilirubin peaks due to the presence of hemoglobin or the test compound were easily distinguished by means of CE. Conclusion: CE is a good alternative to the colorimetric methods currently used for the determination of bilirubin in rat serum.     

Development and validation of the Canine Atopic Dermatitis Lesion Index,a scale for the rapid scoring of lesion severity in canine atopic dermatitis

Background – The third iteration of the Canine Atopic Dermatitis Extent and Severity Index (CADESI‐03) is the only tool rigorously validated for canine atopic dermatitis (CAD) lesion scoring. The CADESI‐03 requires 248 evaluations, limiting its widespread use. Hypothesis/Objectives – The goal of the study was to develop and validate a practical method of grading CAD lesions that requires scoring only the frequently affected body regions. Animals – Fifty‐seven privately owned atopic dogs were used in the study. Methods – The Canine Atopic Dermatitis Lesion Index (CADLI) was evaluated in an open, multicentre reliability study. Validity was assessed with expert opinion (content validity) and comparison of CADLI with existing disease severity measures (construct and criterion validity). Reliability was evaluated by analysing repeated observations of each dog. Convenience was assessed in terms of the time required to complete the scale. Results – The CADLI scores correlated with overall assessment scores (r = 0.60, P < 0.001, linear mixed model) and pruritus severity scores (r = 0.53, P < 0.001, linear mixed model), establishing construct validity. The CADLI was strongly correlated with CADESI‐03 (r = 0.84, P < 0.001, linear mixed model), establishing criterion validity. The CADLI values obtained by two observers correlated very strongly (r = 0.91, P < 0.001), as did the repeat values for the same observer (r = 0.98, P < 0.001). The mean time to complete the CADLI was less than that required for CADESI‐03 (1.9 and 12.6 min, respectively), a highly significant difference (P < 0.001). Conclusion and clinical importance – The CADLI was found to be an effective measure of CAD lesion severity, strongly correlating with CADESI‐03. The convenience of CADLI makes it suitable for use in both clinical research and practice.     

Mapping of expressed sequence tag markers with a cDNA-amplified fragment length polymorphism method in Japanese quail (Coturnix japonica)

In our previous study, a Kobe‐NIBS Japanese quail (KNQ) linkage map was constructed mainly using amplified fragment length polymorphism (AFLP) markers. In order to compare chicken and quail chromosomes, we developed expressed sequence tag (EST) markers derived from cDNA‐AFLP fragments and localized these markers on the linkage map. Using a total of 128 AFLP primer combinations, 24 polymorphic bands were obtained between a neurofilament‐deficient mutant quail line male and a muscular disorder quail line female, which were the parents of the KNQ resource family. Nine of the 24 markers were mapped by linkage analysis. These markers were mapped to seven linkage groups, namely 1, 2, 3, 6, 8, 15 and 42. A subsequent homology search using chicken genome sequences strongly suggests that these linkage groups correspond with chicken chromosomes 1, 2, 3, 5, 15, 23 and 26.     

MAGNETIC RESONANCE IMAGING SCORING OF AN EXPERIMENTAL MODEL OF POST‐TRAUMATIC OSTEOARTHRITIS IN THE EQUINE CARPUS

Magnetic resonance imaging (MRI) is the most sensitive imaging modality to detect the early changes of osteoarthritis. Currently, there is no quantifiable method to tract these pathological changes over time in the horse. The objective of this experimental study was to characterize the progression of MRI changes in an equine model of post‐traumatic osteoarthritis using a semiquantitative scoring system for whole‐organ evaluation of the middle carpal joint. On day 0, an osteochondral fragment was created in one middle carpal joint (OCI) and the contralateral joint (CON) was sham‐operated in 10 horses. On day 14, study horses resumed exercise on a high‐speed treadmill until the completion of the study (day 98). High‐field MRI examinations were performed on days 0 (preosteochondral fragmentation), 14, and 98 and scored by three blinded observers using consensus agreement. Images were scored based on 15 independent articular features, and scores were compared between and within‐groups. On days 14 and 98, OCI joints had significantly (P ≤ 0.05) higher whole‐organ median scores (29.0 and 31.5, respectively), compared to CON joints (21.5 and 20.0, respectively). On day 14, OCI joints showed significant increases in high‐signal bone lesion scores, and osteochondral fragment number and size. On day 98, high‐signal bone lesion, low‐signal bone lesion, osteophyte formation, cartilage signal abnormality, subchondral bone irregularity, joint effusion, and synovial thickening scores were significantly increased in OCI joints. Study results suggest that the MRI whole‐organ scoring system reported here may be used to identify onset and progression of pathological changes following osteochondral injury.     

Silage fermentation and ruminal degradation of stylo prepared with lactic acid bacteria and cellulase

In order to improve the silage fermentation of stylo (Stylosanthes guianensis ) in tropical areas, stylo silages were prepared with commercial additives Lactobacillus plantarum Chikuso‐1 (CH 1), L. rhamnasus Snow Lact L (SN ), Acremonium cellulase (CE ) and their combination as SN +CE or CH 1 + CE , and the fermentation quality, chemical composition and ruminal degradation of these silages were studied. Stylo silages treated with lactic acid bacteria (LAB ) or cellulase, the pH value and NH ₃‐N ? total‐N were significantly (P  <  0.05) decreased while the ruminal degradability of dry matter (DM ), crude protein (CP ), neutral detergent fiber (aNDF om) and acid detergent fiber (ADF om) were significantly (P  <  0.05) increased compared to control. Compared to LAB or cellulase‐treated silages, the DM , CP contents and relative feed value (RFV ), and the ruminal degradability in LAB plus cellulase‐treated silages were significantly (P  <  0.05) higher, but the aNDF om content was significantly (P  <  0.05) lower. CH 1 + CE treatment was more effective in silage fermentation and ruminal degradation than SN +CE treatment. The results confirmed that LAB or LAB plus cellulase treatment could improve the fermentation quality, chemical composition and ruminal degradation of stylo silage. Moreover, the combined treatment with LAB and cellulase may have beneficial synergistic effects on ruminal degradation.     

Pharmacokinetic/pharmacodynamic modelling of robenacoxib in a feline tissue cage model of inflammation

Pelligand, L., King, J. N., Toutain, P. L., Elliott, J., Lees, P. Pharmacokinetic/pharmacodynamic modelling of robenacoxib in a feline tissue cage model of inflammation. J. vet. Pharmacol. Therap.  35 , 19–32. Robenacoxib is a novel nonsteroidal anti‐inflammatory drug developed for use in cats. It is a highly selective COX‐2 inhibitor. Results from previous feline studies showed that, despite a short half‐life in blood, the effect of robenacoxib persisted for 24 h in clinical studies. A tissue cage model of acute inflammation was used to determine robenacoxib’s pharmacokinetics and its ex vivo and in vivo selectivity for COX‐1 and COX‐2 using serum TxB₂ and exudate PGE₂ as surrogate markers for enzyme activity, respectively. After intravenous, subcutaneous and oral administration (2 mg/kg), the clearance of robenacoxib from blood was rapid (0.54–0.71 L·h/kg). The mean residence time (MRT) in blood was short (0.4, 1.9 and 3.3 h after intravenous, subcutaneous and oral administration, respectively), but in exudate MRT was approximately 24 h regardless of the route of administration. Robenacoxib inhibition of COX‐1 in blood was transient, occurring only at high concentrations, but inhibition of COX‐2 in exudate persisted to 24 h. The potency ratio (IC₅₀ COX‐1: IC₅₀ COX‐2) was 171:1, and slopes of the concentration–effect relationship were 1.36 and 1.12 for COX‐1 and COX‐2, respectively. These data highlight the enzymatic selectivity and inflamed tissue selectivity of robenacoxib and support the current recommendation of once‐daily administration.     

Identification of tamaraw (Bubalus mindorensis) from natural habitat‐derived fecal samples by PCR‐RFLP analysis of cytochrome b gene

Fecal DNA analysis is a useful tool for the investigation of endangered species. Tamaraw (Bubalus mindorensis) is endemic to the Philippine island of Mindoro but knowledge of its genetic and ecological information is limited. In this study, we developed a species identification method for tamaraw by fecal DNA analysis. Eighteen feces presumed to be from tamaraw were collected in Mount Iglit‐Baco National Park and species‐known feces from domestic buffaloes and cattle were obtained from a farm. Additionally, one species‐unknown fecal sample was obtained in Mount Aruyan Preserve, where the sighting of tamaraw has not been reported in recent years. Based on DNA sequence data previously reported, the genus Bubalus‐ and tamaraw‐specific primers for PCR of cytochrome b gene were newly designed. The Bubalus‐specific primer yielded a 976 bp fragment of cytochrome b for all fecal samples from tamaraw and domestic buffaloes, but not for cattle, whereas the tamaraw‐specific primer yielded a 582 bp fragment for all tamaraw fecal samples and for one of the four domestic buffalo samples. PCR‐RFLP (restriction fragment length polymorphism) analysis of the 976 bp PCR fragment with AvrII or BsaXI provided distinct differences between tamaraw and domestic buffalo. PCR‐RFLP analysis also showed that the species‐unknown sample obtained in Mount Aruyan Preserve, originates from tamaraw.     

Recreational sandboxes for children and dogs can be a source of epidemic ribotypes of Clostridium difficile

Different studies have suggested that the sand of public playgrounds could have a role in the transmission of infections, particularly in children. Furthermore, free access of pets and other animals to the playgrounds might increase such a risk. We studied the presence of Clostridium difficile in 20 pairs of sandboxes for children and dogs located in different playgrounds within the Madrid region (Spain). Clostridium difficile isolation was performed by enrichment and selective culture procedures. The genetic (ribotype and amplified fragment length polymorphism [AFLP]) diversity and antibiotic susceptibility of isolates was also studied. Overall, 52.5% (21/40) of samples were positive for the presence of C. difficile. Eight of the 20 available isolates belonged to the toxigenic ribotypes 014 (n = 5) and 106 (n = 2), both regarded as epidemic, and CD047 (n = 1). The other 12 isolates were non‐toxigenic, and belonged to ribotypes 009 (n = 5), 039 (n = 4), and 067, 151 and CD048 (one isolate each). Nevertheless, all isolates (even those of a same ribotype) were classified into different AFLP genotypes indicating non‐relatedness. In conclusion, our results revealed the presence of epidemic ribotypes of C. difficile in children's and dog's sandboxes located nearby, which constitutes a major health risk.     

A comparison of 12‐ and 19‐week CHOP protocols using non‐randomized,contemporaneous controls

This study is a concurrent comparison of two versions of CHOP protocols, a 19‐week CHOP and a comparatively overall dose‐intense 12‐week CHOP. The 12‐week protocol was designed to be 58% more dose intense than the 19‐week protocol for both doxorubicin and cyclophosphamide; however, it was 21% less dose intense for vincristine (VCR). Forty‐seven dogs were included for evaluation, and the characteristics of each population were similar. For dogs receiving the 19‐week CHOP protocol, 89.5% experienced a complete response, with a median progression‐free survival (PFS) of 245 days and median overall survival (OS) of 347 days. For dogs receiving the 12‐week CHOP protocol, 89.3% experienced a complete response, with a median PFS of 141 days and median OS of 229 days. When evaluated by Log‐rank analysis, the difference of PFS (P = 0.047) and OS (P = 0.013) between the groups were statistically significant. In summary, these data suggest that despite overall increased dose‐intensity, dogs receiving treatment with a 12‐week CHOP protocol experience less durable remission than our standard 19‐week protocol in this population. Additional prospective investigation will be required to explore the implication that VCR dose intensity and/or shorter overall temporal drug exposure in this protocol may result in diminished efficacy.     

Preliminary evaluation of subjective scoring systems for assessment of postoperative pain in horses

This study examined the performance of two subjective pain scoring systems for evaluating equine postoperative pain, and investigated differences in pain scoring tendencies of veterinarians and grooms. Fifteen horses were included in the study. Group 1 (n = 8) had chronic lameness and was admitted for elective arthroscopy under general anaesthesia, on one or two femoropatellar, femorotibial or tibiotarsal joint or digital flexor tendons. The anaesthetic protocol for each horse was similar but not standardized. Multi‐modal peri‐operative analgesia comprised: romifidine (100 µg kg^?1 IV); flunixin (1.1 mg kg^?1 IV); ketamine (2.2 mg kg^?1 IV); morphine (0.12 mg kg^?1 IV); phenylbutazone (4 mg kg^?1 IV/PO). Group 2 (n = 7) included pain free controls. At 6 hours post‐recovery from anaesthesia (PR) (group 1) or at 20.00 hours (group 2 with one limb bandaged), horses were filmed undisturbed in their stables for 90 seconds (dynamic behaviour, DB); thereafter, the surgery site and pharynx of each horse were palpated (and filmed) in a standardized manner (interactive behaviour, IB). Two observer groups, seven veterinarians and eight grooms, watched video footage of each horse and assigned pain scores using a visual analog scale (VAS) and a numerical rating scale (NRS). Observers assigned a pain score (VAS and NRS) for DB and IB separately and overall. Statistical analysis (Minitab 13.0, Wilcoxon signed rank and Mann–Whitney U‐tests) investigated differences in pain scores attributed to groups 1 and 2 horses, compared pain scores assigned by veterinarians and grooms, and examined differences in the performance of VAS and NRS techniques. There were significant differences in the pain scores assigned by veterinarians and grooms to groups 1 and 2 horses. When using DB or IB separately (but not combined) to score perceived pain, grooms assigned higher scores to group 1 than group 2 (U = 81.5, p < 0.05; U = 82.0, p < 0.05) using the VAS. There was no difference in NRS scores attributed by grooms to groups 1 and 2. Using DB and IB separately or combined, there was no difference in pain scores attributed to groups 1 and 2 by veterinarians using either VAS or NRS scoring systems. Using separate VAS scores for DB (W = 32.5, p < 0.05) and IB (W = 26.5, p < 0.05) and using combined (DB + IB) VAS scores, grooms awarded higher pain scores (W = 27.0, p < 0.05) than veterinarians to group 1. Using the NRS, vets and grooms did not score pain differently for group 1. For group 2, grooms scored pain significantly higher than vets when using the VAS to score IB separately (W = 21.0, p < 0.05); no other differences between grooms and veterinarians in pain scoring of group 2 (NRS or VAS, DB and IB separately or combined) were identified. The performance of subjective pain scoring systems for assessment of equine postoperative pain varies according to the scale used, the behaviour evaluated (dynamic or interactive) and the observer group. While data suggest that grooms distinguished post‐surgery horses from controls more successfully than vets and assigned higher pain scores to these horses, the specific behavioral criteria on which scores were assigned requires future investigation and identification.     

Application of single‐step genomic best linear unbiased prediction with a multiple‐lactation random regression test‐day model for Japanese Holsteins

This study aimed to evaluate a validation reliability of single‐step genomic best linear unbiased prediction (ssGBLUP) with a multiple‐lactation random regression test‐day model and investigate an effect of adding genotyped cows on the reliability. Two data sets for test‐day records from the first three lactations were used: full data from February 1975 to December 2015 (60 850 534 records from 2 853 810 cows) and reduced data cut off in 2011 (53 091 066 records from 2 502 307 cows). We used marker genotypes of 4480 bulls and 608 cows. Genomic enhanced breeding values (GEBV) of 305‐day milk yield in all the lactations were estimated for at least 535 young bulls using two marker data sets: bull genotypes only and both bulls and cows genotypes. The realized reliability (R²) from linear regression analysis was used as an indicator of validation reliability. Using only genotyped bulls, R² was ranged from 0.41 to 0.46 and it was always higher than parent averages. The very similar R² were observed when genotyped cows were added. An application of ssGBLUP to a multiple‐lactation random regression model is feasible and adding a limited number of genotyped cows has no significant effect on reliability of GEBV for genotyped bulls.     

FC‐48 Reliability of the Canine Atopic Dermatitis Extent and Severity Scoring Index

Objective assessment of canine atopic dermatitis severity is very difficult and only a few scoring systems have been developed. The most commonly used is the Canine Atopic Dermatitis Extent and Severity Scoring Index (CADESI), adapted from the human SCORAD. Despite wide use of this score in clinical trials, no validation has been performed to our knowledge. The aim of this study was to determine the reliability of the CADESI in clinical practice. First, a set of 28 pictures taken from dogs diagnosed with atopic dermatitis was scored by six different investigators for three items: erythema, lichenification and excoriation. Next, 23 dogs with clinical signs compatible with atopic dermatitis were graded by two investigators using the CADESI. Erythema, lichenification and excoriation were assessed on 39 areas. With the pictures, significant correlations (Spearman's r, P < 0.05) were found for each combination of investigators for erythema and lichenification, but only in 10 of 15 combinations for excoriation. Interobserver agreement ranged between poor and fair (0.221 < Cohen's κ <0.508, mean = 0.395). For the living animals, significant correlations (P < 0.0001), but poor interobserver agreement, were found for the three items (κ_erythema = 0.366, κ_{lichenification} = 0.385, and κ_excoration = 0.226). A significant correlation (P < 0.05) was found for each location, and interobserver agreement varied between very poor and good (0.16 < κ < 0.66). These results suggest that erythema and lichenification are reliably assessed, but that grading excoriation is more difficult. Also, the assessment of severity varied depending on the site studied. Funding: Self‐funded.     

Amplified fragment length polymorphism to detect clonal diversity and distribution of Mycobacterium avium subspecies paratuberculosis in selected Minnesota dairy cattle.

The molecular ecology of Mycobacterium avium subspecies paratuberculosis (MAP), the causative agent of Johne's disease, is not well understood in the United States. In this study, a DNA fingerprinting method, amplified fragment length polymorphism (AFLP), was used to subtype the pathogen and assess the clonal diversity of MAP in Minnesota dairy herds. Fifty-six fecal culture test-positive isolates from various Minnesota counties and culture dates were analyzed in this study. The AFLP identified 11 profiles with 50% of isolates representing 1 major profile. The major profile was distributed across the state. The genetic diversity of bovine MAP clones in Minnesota based on AFLP analysis of this data appears to be relatively low.