Accuracy of three diagnostic tests to detect tooth resorption in unowned unsocialised cats in Denmark.

OBJECTIVES: To estimate the relative diagnostic sensitivity and specificity of oral clinical examination, full-mouth dental radiography, and cone-beam CT for the detection of tooth resorption in cats, and to estimate the prevalence of tooth resorption in unowned, unsocialised cats in Denmark. MATERIALS AND METHODS: Cadavers of 144 adult cats underwent an oral examination, full-mouth dental radiography, and cone-beam CT. Sensitivity and specificity of the three tests, along with the true prevalence, overall and stratified by sex and tooth location, were estimated using latent class methods. RESULTS: We found cone-beam CT to be the superior image modality, with a sensitivity of 99.5% and a specificity of 99.8%. Dental radiography had a sensitivity of 78.9% and a specificity of 100%, and oral clinical examination had a sensitivity of only 36.0% and specificity of 99.9%. We estimated the prevalence of tooth resorption among unowned unsocialised cats in Denmark to be 40% of adult individuals, and 6.1% of teeth. CLINICAL SIGNIFICANCE: When dealing with tooth resorption, cone-beam CT can help the operator to find and treat affected teeth that could otherwise go undiagnosed. The prevalence of tooth resorption among unowned, unsocialised cats in Denmark does not appear to differ from other populations of cats.

Co-designing a Canadian adaptation of a lifestyle-oriented intervention aimed to improve daily functioning of individuals living with chronic pain: a multi-method study protocol of REVEAL(OT) Canada.

Background: Living with chronic pain (CP) often implies major lifestyle changes, including modifications of daily routines and work. Surprisingly, few validated and effective interventions specifically target functional outcomes in this population. Redesign your Everyday Activities and Lifestyle with Occupational Therapy [REVEAL(OT)] is a lifestyle-oriented intervention led by occupational therapists that directly targets the daily functional challenges of living with CP. The intervention was initially developed and studied as an add-on to standard treatment delivered by Danish multidisciplinary specialized pain clinics. Adapting, implementing, and evaluating REVEAL(OT) within the Canadian healthcare system will contribute to broadening the scope of treatments offered in specialized pain clinics that do not yet include occupational therapy. Objective: The proposed study aims to define and refine REVEAL(OT)/CA with partners (authors of original intervention, people with lived experience, clinicians, managers). Methods: This participatory action research will use a multi-method design and follow the ORBIT model for developing behavioral treatments for chronic diseases. A process of co-construction with partners and an advisory committee will take place in two Montreal specialized pain clinics. It consists of two related work packages (WPs). In WP1, a first series of focus groups with partners (n = 86) and workshops with the advisory committee will be conducted to co-develop the hypothetical pathway describing intervention components and their potential mechanisms of action on targeted outcomes, as well as the first version of the adapted intervention manual. WP2 will co-refine REVEAL(OT)/CA by exploring its acceptability, feasibility and mechanisms of action through intervention deliveries (at least twice in each of two specialized pain clinics; n ≥ 60 patients) and focus groups and/or individual interviews with participating patients and partners. At the end of this study, the intervention manual will be generated both in French and English. Discussion: This study will set the stage for subsequent implementation and effectiveness assessment projects and be an important step towards the deployment of interventions aiming to improve engagement in meaningful daily activities among adults living with CP. Registration: OSF Registries, osf.io/8gksa. Registered 3 August 2023, https://osf.io/8gksa.

Dairy cattle welfare - the relative effect of legislation, industry standards and labelled niche production in five European countries.

The only common European Union (EU) legislation set up specifically to ensure the welfare of dairy cattle is for calves. As a consequence, there is wide diversity in how dairy cattle welfare is ensured in EU countries. A few countries have legal requirements for dairy cattle welfare, while in others, it is left to industry standards or niche production requirements, typically linked to various premium labels. In this paper, we compared animal welfare provisions in dairy cattle production across five countries with different combinations of legislative and other approaches: Denmark, Germany, the Netherlands, Sweden, and the United Kingdom. Firstly, we aimed to map the diversity of animal welfare initiatives. Secondly, we used the Benchmark method of expert valuations and weightings of the relative importance of individual welfare provisions. We found that Denmark and Sweden have the highest level of dairy cattle welfare provisions as measured by the Benchmark method, partly due to high legislative welfare requirements, followed by the United Kingdom, which has an extensive industry standard with very high uptake. Germany and the Netherlands, on the other hand, have lower levels of documented welfare provisions, and correspondingly a Benchmark score closer to a baseline defined by legal requirements at EU level. We also found differences in what elements of animal welfare were focussed on. Some initiatives emphasised fulfilling the social needs of cattle, while others focused more on space and freedom to move. Also, the countries with the highest Benchmark score had a relatively high level of production of organic and other specialty dairy products. We found the effect of national legislation or ambitious industry standards on dairy cattle welfare to be much larger than previous studies have found in either pigs or poultry. At a time when the EU is considering stepping up its efforts to improve animal welfare in terms of common minimum standards, the results of this study could have important policy implications. The diversity in the level of dairy cattle welfare standards found across countries may speak in favour of having shared minimum standards, both at EU level and globally. However, even among countries with a similar Benchmark score, we found a difference in the kinds of welfare provisions at work, which may make full harmonisation of standards more challenging.

Examples of proper reporting for evaluation (Stage 2 validation) of diagnostic tests for diseases listed by the World Organisation for Animal Health.

Reporting and design standards are key indicators of the quality of diagnostic accuracy (validation) studies but, with the exception of aquatic animal diseases and paratuberculosis in ruminants, there is limited guidance for designing these studies in animals. There is, therefore, a need for generic guidelines that are based on disease characteristics, such as mode of transmission, latent period and pathogenesis. Comprehensive, clear and transparent reporting of primary test accuracy studies for diseases listed by the World Organisation for Animal Health (OIE) has value for the end users of diagnostic tests and, ultimately, for decision-makers, who require systematic reviews and meta-analysis of multiple tests for specified diseases and testing purposes. The recent publication of reporting standards for Bayesian latent class models, to analyse test-accuracy data from naturally occurring disease events, fills an important gap as these methods are being increasingly used for OIE-listed diseases. Adherence to design and reporting standards, as well as to guidelines, helps to ensure that research funding for test validation studies is used appropriately and that the strengths and limitations of single tests or test combinations are made clear to test users. The authors provide a review of key points that are often overlooked or misinterpreted in test validation studies, as well as two concrete examples of good practice for use as a reference point for future studies.

Les normes de notification et de conception sont des indicateurs essentiels de la qualité des études de validation des tests destinées à déterminer leur exactitude diagnostique ; or, en dehors des maladies des animaux aquatiques et de la paratuberculose chez les ruminants, il n'existe guère de lignes directrices pour concevoir ce type d'études pour les tests utilisés en santé animale. À la connaissance des auteurs, il n'existe pas non plus de normes de conception applicables aux études de validation en santé humaine. Par conséquent, il conviendrait de disposer de lignes directrices génériques fondées sur les caractéristiques des maladies telles que leurs modalités de transmission, leur période de latence et leur pathogénie. Une notification complète, claire et transparente des études d'exactitude des tests primaires pour les maladies listées par l'Organisation mondiale de la santé animale (OIE) serait une aide précieuse pour les utilisateurs finaux des tests de diagnostic, mais aussi pour les responsables de l'élaboration des politiques, dont les décisions reposent sur des examens et des méta-analyses systématiques couvrant un grand nombre de tests pour certaines maladies ou pour certains usages d'un test. La publication récente des normes de notification applicables aux modèles bayésiens à classe latente pour analyser les données de performance d'un test à partir de foyers naturels de maladie comble une lacune importante dans la mesure où ces méthodes sont de plus en plus utilisées pour les maladies listées par l'OIE. L'adhésion à des normes de conception et de notification ainsi qu'à des lignes directrices en la matière permettra de garantir que les fonds alloués aux études de validation des tests sont bien utilisés et que les atouts et les limitations de certains tests individuels ou associations de tests sont clairement perçus par les utilisateurs. Les auteurs passent en revue certains points essentiels qui sont souvent ignorés ou mal interprétés lors des études de validation des tests et proposent deux exemples concrets de bonnes pratiques qui pourront servir de références pour les études à venir.

Las normas de comunicación y diseño son indicadores básicos de la calidad de los estudios encaminados a determinar la exactitud de diagnóstico (validación) pero, con la excepción de las enfermedades de los animales acuáticos y la paratuberculosis en rumiantes, hay escasas directrices que se apliquen al diseño de esos estudios en animales. Además, hasta donde saben los autores, en el ámbito de la salud humana no hay normas de diseño. De ahí la necesidad de directrices genéricas que estén basadas en las características de las enfermedades, como modo de transmisión, período de latencia o patogénesis. La comunicación exhaustiva, clara y transparente de estudios primarios sobre la exactitud de pruebas de diagnóstico de enfermedades incluidas en las listas de la Organización Mundial de Sanidad Animal (OIE) reviste utilidad no solo para los usuarios finales de la prueba, sino también, en última instancia, para los órganos decisorios, que necesitan metaanálisis y estudios sistemáticos de múltiples pruebas que se apliquen a una u otra enfermedad y sirvan para una u otra finalidad. La reciente publicación de normas de comunicación de modelos bayesianos de clases latentes para analizar los datos de exactitud de pruebas a partir de episodios de enfermedad de origen natural viene a colmar una importante laguna, en la medida en que estos métodos se aplican cada vez más al diagnóstico de enfermedades incluidas en las listas de la OIE. El cumplimiento de las normas de diseño y comunicación, y también de las directrices, ayuda a garantizar que los fondos de investigación destinados a estudios de validación de pruebas sean utilizados debidamente y que el usuario final de una prueba reciba información clara sobre los puntos fuertes y las limitaciones de una prueba o combinación de pruebas. Los autores pasan revista a los principales aspectos que se suelen pasar por alto o malinterpretar en los estudios de validación de pruebas y ofrecen dos ejemplos concretos de buenas prácticas que se pueden utilizar como referencia en futuros estudios.

Changes in the soft-tissue thickness of the claw sole in Holstein heifers around calving.

Claw horn disruption lesions are a common cause of lameness in dairy cattle. It is commonly agreed that they develop due to excessive pressure from the distal phalanx on the horn-producing tissue in the sole. The distal phalanx is supported by the suspensory apparatus, a large proportion of which is made up of connective tissue. It has been hypothesized that increased laxity of the connective tissue due to hormonal changes around calving may lead to decreased support of the distal phalanx, resulting in a sinking of the bone and increased pressure on the horn-producing tissue. In this longitudinal study, our first objective was to determine whether the thickness of the soft tissue related to the claw sole (the combined depth of the digital cushion and corium; soft-tissue thickness, STT) in the weight-bearing hind claws of live heifers changed around calving (the period from approximately 3 wk before calving to 2 wk after calving). Our second objective was to evaluate the relationship between the body condition score (BCS) of recruited heifers and the STT estimates we obtained. We measured STT (defined as the distance between the distal phalanx and the sole horn) in the apex and the posterior part of the sole of the hind claws of 34 Holstein heifers 4 times over 3 to 4 wk around calving. We determined STT by ultrasonographic measurements through the sole horn on weight-bearing claws. The overall mean STT at site 1 was 3.3 mm [minimum-maximum (min-max): 1.8 to 5.2 mm], and the mean at site 2 was 3.7 mm (min-max: 2.2 to 5.8). The overall mean STT was thickest in the first examination before calving [site 1: 3.4 mm (min-max: 1.9 to 4.7); site 2: 4.1 mm (min-max: 2.7 to 5.8)] and thinnest at site 1 in the second examination after calving (3.2 mm, min-max: 1.8 to 5.2) and at site 2 at the first examination after calving (3.5 mm, min-max: 2.2 to 5.0). We observed an average reduction in STT of -0.2 mm (min-max: 0.8 to -0.9 mm) at site 1 during the study period, but we did not observe this reduction in all animals. At site 2, we found an average reduction in STT of -0.6 mm (min-max: 0.8 to -1.3 mm) from 10 d before calving to 5 d after calving, corresponding to a decrease of approximately 15% (min-max: 0 to 31%). We suggest that this reduction was caused by a rotation of the distal phalanx toward the posterior part of the sole horn in the weeks around calving. During the same period, heifer BCS decreased between 0.25 and 0.5 increments in the BCS scale, but we found no correlation between BCS and STT. However, the heifers experienced many changes related to management, feeding, and time budget during the study period, and the rotation of the distal phalanx and reduction of the STT might be explained not only by calving but also by other factors that affect heifers and their claws during the transition period.

Differential somatic cell count as an additional indicator for intramammary infections in dairy cows.

Mastitis, often caused by intramammary infection (IMI), is a significant problem in dairy farming globally. Somatic cell count (SCC) is widely used as a parameter for screening IMI in cows that are then treated or culled. We investigated the potential of a new parameter, differential SCC (DSCC), to detect IMI at cow level when SCC is already known. We achieved this using bacterial culture (BC) and PCR to detect 4 categories of pathogens (major, minor, other, and any) in 2 Danish dairy herds. Quarter milk samples were collected from monthly dairy herd improvement samplings over 1 yr and analyzed with BC, whereas cow-level dairy herd improvement samples were analyzed using PCR. Days in milk, parity, and IMI status had a significant effect on DSCC. Using DSCC in addition to SCC significantly improved the indication of IMI compared with using only SCC in the any pathogen category in both herds as well as the minor pathogens category in herd 2 when BC was used for detection. When PCR was used to detect IMI, the use of DSCC in addition to SCC was significant for the other pathogens category in herd 1 and the minor pathogens category in herd 2. Thus, our data revealed that DSCC can add significant information describing IMI status even when SCC is already known; however, this depends on the causative pathogen. Future studies may address how to use DSCC in practice as well as consider the availability of temporal data to potentially gain insight into the course of infection.

Composite or aseptic quarter milk samples: Sensitivity and specificity of PCR and bacterial culture of Staphylococcus aureus based on Bayesian latent class evaluation.

Bacterial culture (BC) and quantitative polymerase chain reaction (qPCR) are widely used as diagnostic tests for pathogens causing intra-mammary infections (IMI) and it is therefore important to evaluate their performance to optimize pathogen detection. However, with no reference standard, their sensitivity (Se) and specificity (Sp) are unknown. Furthermore, the Se and Sp can differ between quarter samples and composite samples, both of which can be used to test for IMI. Latent class analysis (LCA) offers a method for estimating the Se and Sp of two tests on the same population, but to our knowledge this has not yet been applied to qPCR and BC for the detection of Staph. aureus in both composite and aseptically collected quarter milk samples. This allows for a performance evaluation of not only the two diagnostic methods, but also the two different samples -both cow and quarter level. In this study, we used Bayesian LCA on a dataset from one sampling day at a Danish dairy herd to estimate the Se and Sp for detecting Staph. aureus at cow and quarter level. We used Ct cut-offs of 32 and 37 cycles for the qPCR. When using a cut-off of 37 cycles, the estimated Se and Sp for BC were 62.2% and 90.5% at cow level, and 90.7% and 93.5% at quarter level, respectively. Similarly, the estimated Se and Sp for qPCR at cow level were 82.7% and 84.1%, respectively, and 80.0% and 96.8% at quarter level. Sp was therefore higher for both BC and qPCR at quarter level. Se was also highest for BC at quarter level, but the opposite was true for qPCR. The same pattern was found using a Ct cut-off at 32 cycles, but with different estimates. The results show that qPCR with a Ct cut-off at 37 had a higher Se than BC for composite DHI samples and it is therefore more suitable as a routine screening test for Staph. aureus. However, BC on quarter samples gave the highest Se and Sp and should therefore be used for confirmatory testing.

Prevalence of abomasal lesions in Danish Holstein cows at the time of slaughter.

Abomasal lesions in cattle are challenging to diagnose because the clinical signs are often subtle and nonspecific. An increasing number of studies suggests that abomasal lesions are commonly found in cattle of all ages, but the number of recent prevalence studies in dairy cows is limited. The main objective of this study was to estimate the prevalence of abomasal lesions in a population of Danish Holstein dairy cattle in Denmark. An abattoir survey of 1,327 dairy cows apparently suitable for slaughter was performed in 2016 and 2017. Abomasal lesions are usually classified using a standardized protocol with the following types: abomasal ulcer type I: erosions and nonperforating ulcers; type II: ulcers with arterial bleeding; type III: perforating ulcer with localized peritonitis; and type IV: perforating ulcer with diffuse peritonitis. A further subdivision of the nonperforating abomasal ulcers type I were classified as subtype Ia: erosions; subtype Ib: small ulcers with localized hemorrhage; subtype Ic: ulcers with a crater-like appearance; and subtype Id: retention of the mucosa due to tissue loss with either radial wrinkles converging at a central point or perforations of the spiral folds. Type I abomasal lesions were found in 84% of the examined cows. No ulcers with arterial bleeding (type II) or perforating ulcers with diffuse peritonitis (type IV) were found, but one perforating abomasal ulcer (type III) was observed. The total number of lesions found was 7,418 and when the lesion subtypes were evaluated individually the majority of subtypes Ia and Ic were found in the pyloric area (85 and 94%, respectively), whereas lesion subtypes Ib and Id dominated the fundus/corpus area (71 and 67%, respectively). When considering lesion subtypes according to parity, there was a high prevalence of subtype Ib across the 3 groups of parity (51, 58, and 55% for parity 1, 2, and ≥3, respectively). The prevalence of lesion subtype Id seemed to increase with increasing parity, and the prevalence of lesion subtype Ia seemed to decrease with increasing parity. The prevalence of abomasal lesions was higher than that shown in similar studies of cattle at the time of slaughter. However, the expectedly most painful cases were rare, with only one perforating and no arterial bleeding ulcers found. The etiology of the different subtype I lesions is still unknown, and further investigations are needed to establish the possible influence these lesions might have on welfare and production.

Ultrasonographical examination of bovine claws through the sole horn on weight-bearing claws.

Claw horn disruptions in the bovine claw are believed to be a consequence of pressure on the sole corium from the third phalanx, which may be caused by a weakening of the suspensory apparatus in the claw. We aimed to develop an ultrasonographic method that would make it possible to measure the thickness of the soft tissue between the third phalanx and the sole horn on a weight-bearing claw. A device was developed to record the sole horn and soft tissue thickness indirectly through a polyethylene plate, and 52 feet from slaughtered cows were examined using ultrasonography, both directly and indirectly. Soft tissue and sole horn thickness measurements in the apex and the plantar part of the sole were compared with anatomical measurements of transected claws. To assess the method on weight-bearing versus non-weight-bearing claws, we examined the hind claws of 10 live cattle without transection. We found a weak correlation between the soft tissue thickness measured by ultrasound and anatomical measurements. A strong correlation was observed between the direct ultrasound approach and the developed indirect method. There was a considerable difference between weight-bearing and non-weight-bearing claws, signifying a weak or nonexistent correlation. However, this part of the study was only done on 10 live cows and the results should be interpreted with caution. We concluded that it would be possible to measure the soft tissue using an indirect ultrasound approach on a weight-bearing-claw standing on a polyethylene plate. The major difference between the results of weight-bearing versus non-weight-bearing claws suggests that future studies of the suspensory apparatus could focus on weight-bearing claws.

Knowledge gaps that hamper prevention and control of Mycobacterium avium subspecies paratuberculosis infection.

In the last decades, many regional and country-wide control programmes for Johne's disease (JD) were developed due to associated economic losses, or because of a possible association with Crohn's disease. These control programmes were often not successful, partly because management protocols were not followed, including the introduction of infected replacement cattle, because tests to identify infected animals were unreliable, and uptake by farmers was not high enough because of a perceived low return on investment. In the absence of a cure or effective commercial vaccines, control of JD is currently primarily based on herd management strategies to avoid infection of cattle and restrict within-farm and farm-to-farm transmission. Although JD control programmes have been implemented in most developed countries, lessons learned from JD prevention and control programmes are underreported. Also, JD control programmes are typically evaluated in a limited number of herds and the duration of the study is less than 5 year, making it difficult to adequately assess the efficacy of control programmes. In this manuscript, we identify the most important gaps in knowledge hampering JD prevention and control programmes, including vaccination and diagnostics. Secondly, we discuss directions that research should take to address those knowledge gaps.

A radiological study on intra- and extra-cranial calcifications in adults with X-linked hypophosphatemia and associations with other mineralizing enthesopathies and childhood medical treatment.

OBJECTIVES: 1) The objective of this study was to explore radiological signs of intracranial and nuchal ligament calcifications in adult patients with X-linked hypophosphatemia (XLH) compared with controls and 2) to correlate signs of cranial calcifications in XLH patients with the presence of other extra-cranial enthesopathies, with the severity of skeletal XLH impact and with medical treatment during childhood. SETTING AND SAMPLE POPULATION: Lateral and postero-anterior cephalograms from 36 adult XLH patients and 49 adult controls and X-rays from spine, pelvis, knees and ankles from 31 of the 36 XLH patients. METHODS: Radiological signs of intracranial and nuchal ligament calcifications in XLH patients were compared with controls by Fischer's exact test. In XLH patients, the presence of cranial calcifications was correlated with the presence of other enthesopathies, with the severity of skeletal XLH impact and with medical treatment by Fischer's exact or chi-squared test. RESULTS: Six (17%) XLH patients revealed major signs of intracranial calcifications. Nuchal ligament calcifications were common in XLH patients compared with controls (p = 0.018). Enthesopathy was present at 0-24 sites per XLH patient (median 2). Intracranial calcifications trended to correlate positively with vertebral enthesopathies (p = 0.059). Nuchal calcifications correlated positively with the severity of skeletal XLH impact (p = 0.040). Vertebral enthesopathies correlated negatively with medical treatment (p = 0.008). CONCLUSION: More XLH patients than controls showed nuchal ligament calcifications, and some XLH patients showed intracranial calcifications. Severely affected XLH patients often had nuchal ligament calcifications. Medically treated XLH patients had few vertebral enthesopathies.

Morphological characteristics of frontal sinus and nasal bone focusing on bone resorption and apposition in hypophosphatemic rickets.

OBJECTIVES: To characterize the size and the morphology of the frontal sinus (i.e., structure evolved by bone resorption) and the nasal bone (i.e., structure evolved by bone formation) in adults with hypophosphatemic rickets (HR) compared with controls. SETTING AND SAMPLE POPULATION: Thirty-six patients with HR (12 males and 24 females) aged 21-74 years were included. The control group comprised 49 healthy individuals (23 males and 26 females) aged 20-79 years. MATERIAL AND METHODS: Profile cephalograms were obtained and the following measurements were included: height and width of the frontal sinus; length, width, and area of the nasal bone. The morphology of the nasal bone was assessed. Linear regression analyses were used to compare HR patients with controls. RESULTS: In HR patients, the size of the frontal sinus was unaffected (p = 0.406 to p = 0.862). The proximal width of the nasal bone, and the ratio between the proximal width and the axial length of the nasal bone were increased in HR patients (p < 0.05). CONCLUSIONS: The size of the frontal sinus was unaffected, indicating a normal ability of bone resorption within the bone. The morphology of the nasal bone was abnormal indicating a disturbance in bone formation during growth. The disturbances in nasal bone modeling were mainly expressed in the proximal part supported by structures of cartilaginous origin.

Effect of presampling procedures on real-time PCR used for diagnosis of intramammary infections with Staphylococcus aureus in dairy cows at routine milk recordings.

Aseptic procedures for milk sample collection are considered crucial for bacterial culture to avoid misdiagnosis and subsequently unnecessary treatment or culling. The objective of this field study was to investigate the effect of presampling procedures on the PCR-positivity at cycle threshold value ≤ 37 of real-time PCR assay to detect Staphylococcus aureus from composite milk samples at routine milk recordings while accounting for known cow-level risk factors. A total of 1,199 dairy cows from 6 herds with conventional milking parlors were sampled and tested by PCR in 2011. Following the farmers' routine premilking preparations, 624 cows of the 1,199 cows were randomly selected for bacterial culture preceded by presampling procedures. These procedures were: cleaning of udder teats, removing the first streams of milk, and 70% alcohol teat disinfection. Data on parity, somatic cell counts, days in milk, daily milk yield, fat %, and protein % were extracted from the Danish Cattle Database, whereas energy-corrected milk was calculated based on the latter 3. The within-herd prevalence of intramammary infections with Staph. aureus was 31%, ranging from 16 to 48% in the 6 herds. Univariable analysis showed that the presampling procedures, somatic cell counts, energy-corrected milk, and days in milk were significantly associated with PCR-positivity, whereas parity was not significant. A multivariable model with herd as random effect showed that presampling procedures decreased the chance of being PCR-positive to 0.75 (95% CI; 0.58-0.97) compared with cows where the presampling procedures were not carried out. In conclusion, presampling procedures decreased the cow's chance of being PCR-positive to Staph. aureus. Presampling procedures appeared to improve the specificity of PCR for detection of Staph. aureus by reducing false positives through destruction of Staph. aureus bacteria colonizing or contaminating the teat skin, orifice, and canal. Random herd effects accounted for only 8.9%, indicating that the cluster effect due to herd management on the PCR positivity to Staph. aureus was virtually negligible.

High bone mineral apparent density in children with X-linked hypophosphatemia.

UNLABELLED: Bone mineral apparent density (BMAD) in children with X-linked hypophosphatemia (XLH) was evaluated, as they are unlikely to have extra-skeletal ossifications contributing to the elevated bone mineral density of the spine in adult patients. Children with XLH also had significantly higher BMAD of the spine compared to femoral neck. INTRODUCTION: BMAD obtained by dual-energy X-ray absorptiometry scans in children with XLH was evaluated, as they are unlikely to have the extra-skeletal ossifications contributing to the elevated bone mineral density of the spine in adult patients. METHODS: A total of 15 children with biochemically and genetically verified XLH were recruited. Anthropometric measurements were performed, and to correct for the short stature (small bones), the BMAD of the spine and the femoral neck was evaluated. RESULTS: Z-scores of BMAD of the spine (mean (95 % CI); 2.0 (1.3-2.7); p < 0.001) were significantly elevated compared to reference children. Z-scores of the femoral neck (1.0 (-0.0 to 2.1); p = 0.059) tended to be elevated. Spine Z-scores were significantly higher than the Z-scores of the femoral neck, (paired t test, p = 0.02). BMAD of the spine was evaluated according to the Molgaard's approach; XLH children had normal bone size of the spine for age due to a normal sitting height Z-score of -0.4 (-1.0 to 0.1); p = 0.1. Z-scores of bone mineral content (BMC) of the spine for bone area were elevated (1.4 (0.8-2.1); p < 0.001). No reference data were available to allow evaluation of the BMAD of the femoral neck by the Molgaard's approach. CONCLUSIONS: Children with XLH have an increased BMAD and a high BMC for bone area at the lumbar spine, and this was due to causes other than extra-skeletal ossifications and corrected for bone size. The BMAD of the spine was significantly higher compared to the femoral neck.

Parity and tanned white skin as novel predictors of vitamin D status in early pregnancy: a population-based cohort study.

CONTEXT: In pregnancy, vitamin D insufficiency and deficiency, defined as serum 25-hydroxyvitamin D (25(OH)D) <50 nM, and <25 nM, respectively, may have adverse effects for both mother and child. Prevalence estimates, and identification of subgroups at special risk, may be useful for the planning of preventive strategies. OBJECTIVE: To study the prevalence and risk factors of hypovitaminosis D in early pregnancy. DESIGN AND METHODS: In a cross-sectional study of 1348 women in early pregnancy from the Odense Child Cohort, Denmark, 25(OH)D was determined and correlated to demographic and lifestyle variables (age, nationality, skin tone, parity, prepregnancy body mass index (BMI), smoking and sun exposure), using multiple linear and logistic regression analyses for all year, or stratified for summer and winter. The risk of vitamin D insufficiency was expressed as odds ratios (OR) with 95% confidence intervals in brackets. RESULTS: The prevalence of vitamin D insufficiency and deficiency was estimated to 27·8% and 3·5% respectively. In adjusted analyses, vitamin D insufficiency was directly associated with winter season, OR = 1·89 (1·35-2·63); increasing prepregnancy BMI, OR = 1·06 (1·03-1·10); and smoking, OR = 2·7 (1·34-5·41); but was less frequent in nulliparous, OR = 0·47 (0·33-0·68) and tanned Caucasians, OR = 0·63 (0·41-0·97). Season-specific associations having parental origin from outside Europe in summer, OR = 4·13 (1·41-12·13); in winter smoking, OR = 3·15 (1·19-8·36); and prepregnancy BMI, OR = 1·12 (1·06-1·18). CONCLUSIONS: Vitamin D insufficiency was widespread in early pregnancy. Associations to smoking, prepregnancy BMI and origin outside Europe varied with season. Multiparity and not being tanned in Caucasians represent new risk factors of vitamin D insufficiency.

Salmonella prevalence among reptiles in a zoo education setting.

Clinically healthy reptiles may shed Salmonella and therefore act as a potential zoonotic threat. Most people in Northern European countries are rarely exposed to reptiles, but many zoos have education departments where children have direct contact with this group of animals. The objectives of this study were to determine the prevalence and serotype distribution of Salmonella among reptiles in the Education Department (n = 55) at Copenhagen Zoo and compare it to the Zoo's main reptile collection (n = 145) to evaluate the zoonotic risk. Salmonella was isolated from cloacal swabs by selective enrichment, and a single isolate from each positive sample was further identified by biochemical tests and serotyped. The overall prevalence was 35% (69/200) with significant difference between the Education Department (64%, 35/55) and the main reptile collection (23%, 34/145). A total of 28 serotypes were detected. Ten serotypes were isolated from more than one specimen and four from more than one species. Salmonella enterica subsp. enterica serovar Eastbourne was the predominant serotype (32%, 22/69) and was also the serotype isolated from most reptile species (n = 7). Transmission of serotypes from one department to another was very limited indicated by the serotype distribution. Despite the relative high prevalence observed among the reptiles in the Zoo's Education Department compared to the reptiles in the Zoo's main reptile collection, no Salmonella cases have been linked to the Zoo, and Salmonella ser. Eastbourne is very rarely isolated from humans in Denmark. Simple hygienic procedures such as hand washing which is consistently carried out following handling of reptiles at the Education Department may reduce the risk and therefore contribute to this low prevalence.

Sample size estimation to substantiate freedom from disease for clustered binary data with a specific risk profile.

Disease cases are often clustered within herds or generally groups that share common characteristics. Sample size formulae must adjust for the within-cluster correlation of the primary sampling units. Traditionally, the intra-cluster correlation coefficient (ICC), which is an average measure of the data heterogeneity, has been used to modify formulae for individual sample size estimation. However, subgroups of animals sharing common characteristics, may exhibit excessively less or more heterogeneity. Hence, sample size estimates based on the ICC may not achieve the desired precision and power when applied to these groups. We propose the use of the variance partition coefficient (VPC), which measures the clustering of infection/disease for individuals with a common risk profile. Sample size estimates are obtained separately for those groups that exhibit markedly different heterogeneity, thus, optimizing resource allocation. A VPC-based predictive simulation method for sample size estimation to substantiate freedom from disease is presented. To illustrate the benefits of the proposed approach we give two examples with the analysis of data from a risk factor study on Mycobacterium avium subsp. paratuberculosis infection, in Danish dairy cattle and a study on critical control points for Salmonella cross-contamination of pork, in Greek slaughterhouses.

High-throughput biological small-angle X-ray scattering with a robotically loaded capillary cell.

With the rise in popularity of biological small-angle X-ray scattering (BioSAXS) measurements, synchrotron beamlines are confronted with an ever-increasing number of samples from a wide range of solution conditions. To meet these demands, an increasing number of beamlines worldwide have begun to provide automated liquid-handling systems for sample loading. This article presents an automated sample-loading system for BioSAXS beamlines, which combines single-channel disposable-tip pipetting with a vacuum-enclosed temperature-controlled capillary flow cell. The design incorporates an easily changeable capillary to reduce the incidence of X-ray window fouling and cross contamination. Both the robot-control and the data-processing systems are written in Python. The data-processing code, RAW, has been enhanced with several new features to form a user-friendly BioSAXS pipeline for the robot. The flow cell also supports efficient manual loading and sample recovery. An effective rinse protocol for the sample cell is developed and tested. Fluid dynamics within the sample capillary reveals a vortex ring pattern of circulation that redistributes radiation-damaged material. Radiation damage is most severe in the boundary layer near the capillary surface. At typical flow speeds, capillaries below 2 mm in diameter are beginning to enter the Stokes (creeping flow) regime in which mixing due to oscillation is limited. Analysis within this regime shows that single-pass exposure and multiple-pass exposure of a sample plug are functionally the same with regard to exposed volume when plug motion reversal is slow. The robot was tested on three different beamlines at the Cornell High-Energy Synchrotron Source, with a variety of detectors and beam characteristics, and it has been used successfully in several published studies as well as in two introductory short courses on basic BioSAXS methods.

Periapical and endodontic status of permanent teeth in patients with hypophosphatemic rickets.

Hypophosphatemic rickets (HR) is a rare hereditary disease in which dental problems in terms of spontaneous periapical infections are frequently reported. Most previous reports have been based on a small number of HR patients and have been published before the disease could be confirmed genetically. The aim of the present study was to describe the periapical and endodontic status of permanent teeth in patients with genetically and/or biochemically confirmed HR. The patients were recruited from a medical study on HR patients. The patients underwent a dental examination including a digital panoramic radiograph, which was scored for endodontically affected teeth (i.e. teeth with periapical radiolucencies and/or endodontically treated teeth). A total of 52 patients (age range: 5·7-74·5 years; 17 males and 35 females) were included. HR patients were characterised by a high number of endodontically affected teeth (mean: 4·2; s.d.: 5·0). The number of affected teeth rose significantly with age (P < 0·01), and no statistically significant gender difference was found. The relative distribution of endodontically affected teeth in the three tooth groups (incisors and canines, premolars, and molars) varied according to age. In the youngest age group, only incisors and canines were affected, while the relative proportion of affected premolars and molars increased with age. Endodontically affected teeth are common in HR patients, and the number of affected teeth increased significantly with age. Hence, the need for endodontic treatment among HR patients is comprehensive.

Stochastic models to simulate paratuberculosis in dairy herds.

Stochastic simulation models are widely accepted as a means of assessing the impact of changes in daily management and the control of different diseases, such as paratuberculosis, in dairy herds. This paper summarises and discusses the assumptions of four stochastic simulation models and their use in the design of certification, surveillance, and control strategies for paratuberculosis in cattle herds. A detailed comparison is made between the Dutch JohneSSim and the Danish PTB-Simherd, using the same context of a set of control strategies in a typical Dutch/Danish herd. The conclusion is that while the models are somewhat different in their underlying principles and do put slightly different values on the different strategies, their overall findings are similar. Therefore, simulation models may be useful in planning paratuberculosis strategies in dairy herds, although as with all models caution is warranted when interpreting and generalising the results.