Exploring associations between the teat apex metagenome and Staphylococcus aureus intramammary infections in primiparous cows under organic directives.

The primary objective of this study was to identify associations between the prepartum teat apex microbiome and the presence of Staphylococcus aureus intramammary infections (IMI) in primiparous cows during the first 5 weeks after calving. We performed a case-control study using shotgun metagenomics of the teat apex and culture-based milk data collected longitudinally from 710 primiparous cows on five organic dairy farms. Cases had higher odds of having S. aureus metagenomic DNA on the teat apex prior to parturition compared to controls (OR = 38.9, 95% CI: 14.84-102.21). Differential abundance analysis confirmed this association, with cases having a 23.8 higher log fold change (LFC) in the abundance of S. aureus in their samples compared to controls. Of the most prevalent microorganisms in controls, those associated with a lower risk of post-calving S. aureus IMI included Microbacterium phage Min 1 (OR = 0.37, 95% CI: 0.25-0.53), Corynebacterium efficiens (OR = 0.53, 95% CI: 0.30-0.94), Kocuria polaris (OR = 0.54, 95% CI: 0.35-0.82), Micrococcus terreus (OR = 0.64, 95% CI: 0.44-0.93), and Dietzia alimentaria (OR = 0.45, 95% CI: 0.26-0.75). Genes encoding for Microcin B17 AMPs were the most prevalent on the teat apex of cases and controls (99.7% in both groups). The predicted abundance of genes encoding for Microcin B17 was also higher in cases compared to controls (LFC 0.26). IMPORTANCE: Intramammary infections (IMI) caused by Staphylococcus aureus remain an important problem for the dairy industry. The microbiome on the external skin of the teat apex may play a role in mitigating S. aureus IMI risk, in particular the production of antimicrobial peptides (AMPs) by commensal microbes. However, current studies of the teat apex microbiome utilize a 16S approach, which precludes the detection of genomic features such as genes that encode for AMPs. Therefore, further research using a shotgun metagenomic approach is needed to understand what role prepartum teat apex microbiome dynamics play in IMI risk.

Relationships of beta-casein genetics with production, fertility, and survival of purebred organic Holstein dairy cows.

The objective of this study was to compare ß-casein genotype of purebred certified-organic Holstein cows, and their effect on production, fertility, and survival. Holstein cows (n = 1,982) from 13 certified-organic dairy herds from the western, midwestern, and northeastern United States were genomically tested with CLARIFIDE Plus (Zoetis) for ß-casein genotype. Two hundred fourteen cows were A1A1 (11%), 848 cows were A1A2 (43%), and 920 cows were A2A2 (46%). In total, 2,249 lactation records, 1,025 from the first parity and 1,224 records during second and greater parities were used. Test-day milk, fat, and protein production (305-d) and somatic cell score were obtained from the Dairy Herd Improvement Association. A lower limit of 50 d for days open was applied, and cows with more than 250 d open had days open set to 250 d. Independent variables for statistical analysis were the fixed effects of herd, parity, ß-casein genotype (A1A1, A1A2, A2A2), and ß-casein genotype by parity interaction. Cow nested within parity was the random effect in the statistical models for fertility and production traits. Herd had a significant effect on all fertility, production, and survival variables. Parity affected the number of times bred per pregnancy and days open, milk, fat, and protein production, and somatic cell score. Beta-casein genotype and herd influenced the percentage of cows surviving to first and second lactation. Results indicate no difference in production and fertility regarding ß-casein genotype for organic dairy herds. Survival was biased against the A1 allele, which is indicated by lower survival rates during first lactation. These results may offer organic producers more flexibility in breeding and culling decisions to produce A2A2 milk.

Precision technologies to improve dairy grazing systems.

Pasture-based dairy herds continue to grow around the world as demand increases for sustainable farming practices. Grazing dairy farmers may benefit from the utilization of precision dairy technologies because these technologies have the potential to improve animal welfare, increase farm efficiency, and reduce costs. Precision dairy technologies have provided novel information about activity, rumination, and grazing behavior of various breeds in pasture-based systems. Previous research with wearable technologies has indicated that rumination, eating, and no activity have moderate to high correlations (r = 0.65 to 0.88) with visual observation; however, activity may be difficult to record in grazing herds. However, many grazing dairy farmers around the world are using activity monitors with generally positive success. Grazing is a complex behavior to define because cows may walk to an area and stop to eat or continuously walk and take bites of grass from the pasture. Wearable technologies can detect whether a cow is grazing with reasonable accuracy. However, the challenge is to determine pasture intake as bite rate and bite size because these can vary as the pasture is grazed to a low residual height. Nevertheless, grazing behavior data collected with wearable technologies was highly correlated (r = 0.92 to 0.95) with visual observations. Grazing is a behavior that should continue to be explored, especially with precision dairy technologies. As healthy and productive pastures are integral to grazing systems, accurate forage biomass measurements can improve efficiency and production of pastured dairy cows. However, few farms use technology to determine forage availability. Therefore, using dairy technologies to monitor forage dry matter from pasture may provide a potential benefit for grazing-based dairy farms. Current satellite technology with the normalized difference vegetation index and electronic rising plate meters may provide new technologies for farms to monitor forage biomass and fine-tune grazing within pastures. In the future, pasture-based dairy farms may rely on virtual fencing, drones to detect animal health issues and forage availability, and autonomous vehicles to move cattle and to detect weeds on pasture.

Effects of oral white willow bark (Salix alba) and intravenous flunixin meglumine on prostaglandin E2 in healthy dairy calves.

White willow bark (WWB) is commonly used in combination with other medicinal herbs and analgesics to alleviate inflammatory pain in disbudded calves under organic management, but there is no evidence to confirm an effect of WWB on inflammatory biomarkers in calves. The objective of this study was to determine whether WWB affects the inflammatory biomarker prostaglandin E2 (PGE2) in healthy dairy calves. A randomized crossover trial with 2 periods and 5 treatments was used for this experiment. A 7-d washout period was used to minimize carryover effects. The treatments were (1) 57.6 mg/kg oral WWB (low dose; L-WWB), (2) 115.1 mg/kg oral WWB (medium dose; M-WWB), (3) 230.3 mg/kg oral WWB (high dose; H-WWB), (4) 2.2 mg/kg i.v. flunixin meglumine (FM), or (5) no treatment (NT). Calves (n = 25) were randomly assigned to receive 1 of the 25 treatment sequences. Blood samples were collected at 1, 2, and 4 h after administration to determine PGE2 and salicylic acid plasma concentrations. The WWB had 2,171 µg/g (± 4.3% relative standard error) salicin (0.22%). On average, calves in the FM (721 ± 274 pg/mL) treatment had lower PGE2 than calves in all other treatments. Calves in the NT (2,606 ± 271 pg/mL), L-WWB (2,509 ± 276 pg/mL), M-WWB (2,343 ± 270 pg/mL), and H-WWB (3,039 ± 270 pg/mL) treatments had similar PGE2 averaged across sampling times. Calves in the L-WWB (23.4 ± 1.9 ng/mL), M-WWB (21.5 ± 1.9 ng/mL), and H-WWB (23.3 ± 1.9 ng/mL) treatments had similar maximum salicylic acid plasma concentrations. Results from this study indicate that the WWB doses used in this experiment were ineffective at achieving dose-dependent PGE2 and salicylic acid plasma concentration responses.

Evaluation of the RumiWatch system as a benchmark to monitor feeding and locomotion behaviors of grazing dairy cows.

Direct visual observation is a common method for validation of animal behavior technologies; however, visual observations are time consuming and subject to human error. The objective of this study was to evaluate the RumiWatch system (Itin and Hoch GmbH, Liestal, Switzerland), which is composed of a noseband sensor and a pedometer, for monitoring feeding and locomotion behaviors of grazing dairy cows, to determine its accuracy for use as a benchmark in validation studies. The study was conducted at the University of Minnesota West Central Research and Outreach Center in Morris, Minnesota, from May to June 2018. Two experiments were conducted and validated: (1) feeding and locomotion behaviors and (2) rumination cycle and grazing bites. Lactating crossbred dairy cows (n = 12) were offered pasture for 22 h/d, and cows were milked twice daily. Visual observations were recorded by 3 observers with the Pocket Observer app (Noldus Information Technology, Leesburg, VA). The first experiment determined agreement for visual observations and the RumiWatch noseband sensor and pedometer from 144 h of feeding and locomotion behaviors. The second experiment determined agreement for visual observations and the RumiWatch noseband sensor from 17.75 h of rumination cycle and grazing bites. Pearson correlations evaluated associations for visual observations, and the RumiWatch noseband sensor and pedometer and were 0.84 for rumination, 0.76 for grazing, 0.39 for drinking, 0.57 for other activities, 0.83 for standing, 0.91 for lying, and 0.38 for walking. Correlations for visual observations and rumination cycle and grazing bites were -0.13 and 0.47, respectively. The RumiWatch system evaluated rumination, grazing, standing, and lying behaviors with high precision and accuracy, and the RumiWatch system may be used as a benchmark instead of visual observation to validate animal behavior technologies.

Evaluation of solar photovoltaic systems to shade cows in a pasture-based dairy herd.

The combined use of solar photovoltaics and agriculture may provide farmers with an alternative source of income and reduce heat stress in dairy cows. The objective of this study was to determine the effects on grazing cattle under shade from a solar photovoltaic system. The study was conducted at the University of Minnesota West Central Research and Outreach Center in Morris, Minnesota on a grazing dairy. Twenty-four crossbred cows were randomly assigned to 2 treatment groups (shade or no shade) from June to September in 2019. The replicated (n = 4) treatment groups of 6 cows each were provided shade from a 30-kW photovoltaic system. Two groups of cows had access to shade in paddocks, and 2 groups of cows had no shade in paddocks. All cows were located in the same pasture during summer. Behavior observations and milk production were evaluated for cows during 4 periods of summer. Boluses and an eartag sensor monitored internal body temperature, activity, and rumination on all cows, respectively. Independent variables were the fixed effects of breed, treatment group, coat color, period, and parity, and random effects were replicate group, date, and cow. No differences in fly prevalence, milk production, fat and protein production, or drinking bouts were observed between the treatment groups. Shade cows had more ear flicks (11.4 ear flicks/30 s) than no-shade cows (8.6 ear flicks/30 s) and had dirtier bellies and lower legs (2.2 and 3.2, respectively) than no-shade cows (1.9 and 2.9, respectively). During afternoon hours, shade cows had lower respiration rates (66.4 breaths/min) than no-shade cows (78.3 breaths/min). From 1200 to 1800 h and 1800 to 0000 h, shade cows had lower body temperature (39.0 and 39.2°C, respectively) than no-shade cows (39.3 and 39.4°C, respectively). Furthermore, between milking times (0800 and 1600 h), the shade cows had lower body temperature (38.9°C) than no-shade cows (39.1°C). Agrivoltaics incorporated into pasture dairy systems may reduce the intensity of heats stress in dairy cows and increase well-being of cows and the efficiency of land use.

Growth, health, and economics of dairy calves fed organic milk replacer or organic whole milk in an automated feeding system.

The objective of the study was to investigate the growth, health, behavior, and economics of dairy calves fed organic milk replacer (n = 41) or organic whole milk (n = 40) in an automatic feeding system. Calves were fed either organic milk replacer or whole milk (assigned to treatment in birth order) during 2 seasons from March to July 2018 and from September to December 2018 at the University of Minnesota West Central Research and Outreach Center in Morris, Minnesota. The treatment groups were (1) pasteurized whole milk fed at 13% total solids of organic milk (WM), or (2) milk replacer fed at 150.98 g of dry replacer powder per liter of water (MR). Milk replacer was fed at 14.65% total solids based on the manufacturer's recommendation. Calves were introduced to the automated feeder at 5 d and allowed to drink up to 8 L/d at the maximum allowance. At 50 d, the allowance was reduced by 0.2 L/d and calves were weaned at 56 d. Milk feeding behavior (feeding station visit behaviors and drinking speeds) were collected from the automatic feeding system and analyzed by feeding group. Body weights were recorded at birth and then weekly until weaning (56 d). Health scores of calves were recorded twice a week. Data were analyzed using PROC MIXED (SAS Institute Inc.). Independent variables for analyses were the fixed effects of breed group, season of birth, and treatment group, and the interaction of season and treatment group along with pen as a random effect. No differences were found between treatment groups for average daily gain, weaning weight, hip height, or heart girth. Milk feeding behavior varied between the 2 feeding treatment groups. The WM calves had shorter visits to the feeding station (2.44 vs. 3.01 min, respectively) compared with MR calves. Overall drinking speeds of the WM calves were higher (1,301 mL/min) than those of the MR calves (581 mL/min). The MR calves had higher fecal scores than WM calves. The average cost per kilogram of gain was lower for WM calves ($6.35/kg) compared with MR calves ($8.82/kg). The results of this study indicate health and economic advantages to feeding organic dairy heifer calves whole milk during the preweaning period.

Linking routinely collected social work, education and health data to enable monitoring of the health and health care of school-aged children in state care ('looked after children') in Scotland: a national demonstration project.

BACKGROUND AND OBJECTIVES: Children in state care ('looked after children') have poorer health than children who are not looked after. Recent developments in Scotland and elsewhere have aimed to improve services and outcomes for looked after children. Routine monitoring of the health outcomes of looked after children compared to those of their non-looked after peers is currently lacking. Developing capacity for comparative monitoring of population-based outcomes based on linkage of routinely collected administrative data has been identified as a priority. To our knowledge there are no existing population-based data linkage studies providing data on the health of looked after and non-looked after children at national level. Smaller scale studies that are available generally provide very limited information on linkage methods and hence do not allow scrutiny of bias that may be introduced through the linkage process. STUDY DESIGN AND METHODS: National demonstration project testing the feasibility of linking routinely collected looked after children, education and health data. PARTICIPANTS: All children in publicly funded school in Scotland in 2011/12. RESULTS: Linkage between looked after children data and the national pupil census classified 10,009 (1.5%) and 1757 (0.3%) of 670,952 children as, respectively, currently and previously looked after. Recording of the unique pupil identifier (Scottish Candidate Number, SCN) on looked after children returns is incomplete, with 66% of looked after records for 2011/12 for children of possible school age containing a valid SCN. This will have resulted in some under-ascertainment of currently and, particularly, previously looked after children within the general pupil population. Further linkage of the pupil census to the National Health Service Scotland master patient index demonstrated that a safe link to the child's unique health service (Community Health Index) number could be obtained for a very high proportion of children in each group (94%, 95% and 95% of children classified as currently, previously, and non-looked after, respectively). In general, linkage rates were higher for older children and those living in more affluent areas. Within the looked after group, linkage rates were highest for children with the fewest placements and for those in permanent fostering. CONCLUSIONS: This novel data linkage demonstrates the feasibility of monitoring population-based health outcomes of school-aged looked after and non-looked after children using linked routine administrative data. Improved recording of the unique pupil identifier number on looked after data returns would be beneficial. Extending the range of personal identifiers on looked after children returns would enable linkage to health data for looked after children who are not in publicly funded schooling (i.e. those who are preschool or postschool, home schooled or in independent schooling).

New insights for identification of doping with recombinant human erythropoietin micro-doses after high hydration.

To minimize the chances of being caught after doping with recombinant human erythropoietins (rhEPO), athletes have turned to new practices using micro-doses and excess fluid ingestion to accelerate elimination and decrease the probability of detection. Our objective was to test the sensitivity of detection by validated methods (IEF: isoelectric focusing; SDS-PAGE: sodium dodecyl sulfate polyacrylamide gel electrophoresis) when such practices are used. First, after a three-week rhEPO boost period and 10 days of wash out, detection of a single 900 IU micro-dose of Eprex® was evaluated in healthy male subjects. After an injection in the evening, urine and plasma samples were collected the following morning. Half of the subjects then drank a bolus of water and new samples were collected 80 min later. Interestingly, rhEPO was detected in 100% of the samples even after water ingestion. A second similar protocol was then performed with a single injection of a micro-dose of rhEPO (500 IU or 900 IU), without a prior rhEPO boost. In addition, urine and plasma samples were also collected 15 and 20 h post rhEPO administration. Once again drinking water did not affect the rate of detection. Urine appeared a better matrix to detect micro-doses after 10 h, enabling between 92% and 100% of identification at that time. The rate of identification decreased rapidly thereafter, in particular for the 500 IU micro-dose. However IEF analysis still resulted in 71% identification of rhEPO in urine after 20 h. These results could help to define a better strategy for controlling and identifying athletes using rhEPO micro-doses. Copyright © 2016 John Wiley & Sons, Ltd.

Ductal Plate Malformation in the Liver of Boxer Dogs: Clinical and Histological Features.

Ductal plate malformations (DPMs) represent developmental biliary disorders with a wide phenotypic spectrum. This study characterizes DPM in 30 Boxer dogs. Median age was 1.5 (range, 0.3-10.0) years, with 12 dogs <1 year. Clinical features included increased serum levels of liver enzymes (28), gastrointestinal signs (16), poor body condition (14), abdominal effusion (9), and hepatic encephalopathy (2). Additional malformations included gallbladder atresia (8), atrophied left liver (2), absent quadrate lobe with left-displaced gallbladder (1), portal vasculature atresia (left liver, 1), intrahepatic portosystemic shunt (1), and complex intrahepatic arteriovenous malformation (1). All dogs had portal tracts dimensionally expanded by a moderate-to-severe multiple small bile duct phenotype embedded in abundant extracellular matrix; 80% displayed variable portal-to-portal bridging. Quantitative analysis confirmed significantly increased fibrillar collagen and a 3-fold increased portal tract area relative to 6 Boxer and 10 non-Boxer controls. Biliary phenotype was dominated by tightly formed CK19-positive ductules, typically 10 to 15 µm in diameter, with 3 to >30 profiles per portal tract, reduced luminal apertures, and negative Ki-67 immunoreactivity. CK19-positive biliary epithelium intersected directly with zone 1 hepatocytes as a signature feature when considered with other DPM characteristics. Phenotypic variation included a multiple small bile duct phenotype (all dogs), predominantly thin-walled sacculated ducts (4), well-formed saccular ducts (4), and sacculated segmental, interlobular, and intralobular ducts (Caroli malformation, 2 dogs, one with bridging portal fibrosis). Histologic evidence of portal venous hypoperfusion accompanied increased biliary profiles in every case. We propose that this spectrum of disorders be referred to as DPM with appropriate modifiers to characterize the unique phenotypes.

Laparoscopic kidney transplant by extra peritoneal approach: the safe transition from laboratory to the clinic.

The aim of this study is to develop a novel laparoscopic surgery by extra-peritoneal approach for kidney transplant and pave the way of safe transition from laboratory to the clinic. The study was established to explore the feasibility and safety of human laparoscopic kidney transplant. The experiment was first conducted on the deceased animals, then live animals and human cavader before human kidney transplant was approved. The study patient was a 49-year-old male who received the kidney for laparoscopic kidney transplant by extra-peritoneal approach. The control patient received the contralateral kidney for open kidney transplant. The estimated blood loss was minimal during surgery. Both kidneys experienced delayed graft function but the kidneys started function on Day 6 postoperation. The analgesia consumption was significantly less in the study patient. There is no surgical complication during 6-month follow-up. This study has developed a new technique for laparoscopic kidney transplant by extra-peritoneal approach. It has retained the advantages of open kidney transplant, which allows the graft located in the extra-peritoneal space without violating peritoneum. This study has also paved the way of safe transition for a novel laparoscopic surgery from laboratory to the clinic.

Psychological outcomes in critically ill patients receiving renal replacement therapy: a matched-pairs analysis.

We conducted a single-centre observational study over five years to assess the impact of renal replacement therapy on the psychological health of survivors of critical illness. We hypothesised that the added burden of renal replacement would increase the prevalence and severity of anxiety, depression and stress reactions in these patients, compared with matched pairs (matched for age, sex and APACHE II score) who did not receive renal replacement. Participants completed postal questionnaires. A total of 342 patients with acute kidney injury received renal replacement. One hundred and seventy-nine (52.3%) survived to hospital discharge, and 161 (47.1%) were alive at 90 days. Seventy-seven (47.8% of survivors) completed questionnaires. We found 77 matches for the Hospital Anxiety and Depression Scale analysis and 72 for the Impact of Events Scale analysis. Clinically relevant symptoms of psychiatric morbidity were common, with anxiety and depression affecting 49 (63.6%) patients and stress reactions affecting 24 (33.3%) patients. Mean scores (95% CI) were 11.4 (9.6-13.2) and 20.1 (15.7-24.6), respectively. On multivariate analysis, we found no significant differences between renal replacement patients and controls, in either the frequency or severity of these symptoms.

Detection of microdoses of rhEPO with the MAIIA test.

The detection of recombinant human erythropoietin (rhEPO) is difficult and becomes more challenging when only microdoses are administered intravenously. Twenty-three subjects were divided into two groups: EPO group (n = 7) and CONTROL group (n = 16). Seven urine and blood samples per subject were collected at least 5 days apart to determine within- and between-subject standard deviations in the percentage of migrating isoforms by the MAIIA test. Six injections of 50 IU/kg bw (boosting dosage) of epoetin beta (Neorecormon, Roche Diagnostics, Hvidovre, Denmark) were performed intravenously during a 3-week period, followed by two microinjections of only 10 IU/kg bw. Blood and urine samples were collected 2, 6, 12, and 72 h after the microinjection, as well as 72 h after the last boosting dose. Sensitivities and specificities of the MAIIA test were examined by absolute and passport thresholds. Sensitivity was 100% for at least 12 h after the microinjection, with ∼30% of plasma samples still exceeding the 99.9% passport threshold 72 h after a microinjection. The specificity was higher for the passport approach compared to the absolute approach, but there were no differences in sensitivities between approaches or between specimens (urine and plasma). We conclude that the MAIIA test shows potential for detecting very small doses of rhEPO.

Stability of athlete blood passport parameters during air freight.

INTRODUCTION: Fluctuations in ambient temperature and pressure, as well as physical jostling, may affect the stability of whole blood samples transported by air freight. The aim of this study was to characterize the stability of key blood variables during air freight and to investigate whether vibration or reduced pressure alone affected results. METHODS: Over a 72-h interval, we evaluated the stability of full blood count indices (plus reticulocytes) in tubes that were air-freighted a total of 2, 10 and 28 h. We also examined the impact of 24 h of reduced atmospheric pressure (750 hpa or approximately 2500 m.a.s.l) and vibration (5 Hz). Samples were measured on a Sysmex XT-2000i instrument. RESULTS: The two key variables in the context of antidoping (haemoglobin concentration, reticulocytes) remained stable over a 72-h period regardless of the duration of air freight. Atmospheric pressure and vibration had no discernible effect. CONCLUSION: Whole blood samples stored in NanoCool devices can be relied upon to remain stable for at least 72 h despite interim air freight.

A prospective evaluation of VEGF-targeted treatment cessation in metastatic clear cell renal cancer.

BACKGROUND: Vascular endothelial growth factor (VEGF)-targeted therapy is administered continuously until progression in metastatic clear cell renal cancer (mRCC). The role of intermittent therapy is under investigation. Preclinical data raise concerns about this approach. MATERIALS AND METHODS: This study combined the data from three similar phase II studies investigating VEGF-targeted therapy prior to planned nephrectomy for untreated mRCC (European Union Drug Regulating Authorities Clinical Trials 2006-004511-21, 2006-006491-38 and 2009-016675-29). The significance of progression during the planned treatment break (median 4.3 weeks) was assessed. RESULTS: Sixty-two patients had a structured treatment interruption for nephrectomy after achieving clinical benefit from treatment and restarted therapy. Twenty-three of these patients (37%) progressed (Response Evaluation Criteria In Solid Tumors v1.1) on the first scan after the treatment break. Subsequent stabilisation of disease occurred in 16 of the 23 (70%) progressing patients when the same VEGF tyrosine kinase inhibitor (TKI) was reintroduced. Baseline characteristics, such as the Memorial Sloan Kettering Cancer Centre prognostic score, did not predispose to the development of this progression. Progression during the treatment break was associated with an increased risk of death on multivariate analysis {hazard ratio (HR) 5.56; [95% confidence interval 2.29-13.5], P < 0.01}. Sequential fluorodeoxyglucose positron emission tomography showed a rebound in metabolic activity during the treatment break. CONCLUSIONS: Progression during planned VEGF TKI treatment interruptions is frequent and associated with a poor prognosis. Treatment cessation should be pursued with caution.

The effects of injury and illness on haemoglobin mass.

This study sought to quantify the effects of reduced training, surgery and changes in body mass on haemoglobin mass (Hbmass) in athletes. Hbmass of 15 athletes (6 males, 9 females) was measured 9±6 (mean±SD) times over 162±198 days, during reduced training following injury or illness. Additionally, body mass (n=15 athletes) and episodes of altitude training (n=2), iron supplementation (n=5), or surgery (n=3) were documented. Training was recorded and compared with pre-injury levels. Analysis used linear mixed models for ln(Hbmass), with Sex, Altitude, Surgery, Iron, Training and log(Body Mass) as fixed effects, and Athlete as a fixed and random effect. Reduced training and surgery led to 2.3% (p=0.02) and 2.7% (p=0.04) decreases in Hbmass, respectively. Altitude and iron increased Hbmass by 2.4% (p=0.03) and 4.2% (p=0.05), respectively. The effect of changes in body mass on Hbmass was not statistically significant (p=0.435).The estimates for the effects of surgery and altitude on Hbmass should be confirmed by future research using a larger sample of athletes. These estimates could be used to inform the judgements of experts examining athlete biological passports, improving their interpretation of Hbmass perturbations, which athletes claim are related to injury, thereby protecting innocent athletes from unfair sanctioning.

Stability of athlete passport parameters during extended storage.

INTRODUCTION: Extended intervals between sample collection and analyses render athlete's whole-blood specimens collected in the field for antidoping purposes susceptible to storage degradation. The aim of this study was to characterize the stability of key blood variables under different storage durations and temperatures. METHODS: We evaluated stability of full blood count indices (plus reticulocytes) in individual tubes left undisturbed during 36, 48, 72, 96, 120, 144 and 168 h of storage at approximately 4, 6 and 12 °C. Samples were measured on a Sysmex XT-2000i instrument. RESULTS: The two key variables in the context of antidoping (haemoglobin concentration, reticulocytes) were stable for at least 168 h, except under 12 °C (stable 48 h only). Volume-dependent variables changed in a predictable manner that enabled a nomogram to be generated to predict original values provided storage duration and temperature were known. CONCLUSION: Key blood results can be relied upon for at least 7 days if storage temperature is kept at 4-6 °C.

Within-patient variation of hemoglobin and reticulocytes: implications for evaluating ESA responsiveness in dialysis patients.

INTRODUCTION: Techniques that assess percent reticulocytes (%retics) or hemoglobin (Hb) to detect erythropoiesis-stimulating agents (ESA) use in athletes may be useful in evaluating ESA responsiveness in dialysis patients. However, within-patient variation, appropriate transformation, or the relationship between the blood draw interval length and analyte variation are untested. METHODS: In a prospective, single-arm trial, we determined Hb and %retics in 30 hemodialysis patients receiving stable ESA doses. Within-patient results were evaluated for variance homogeneity and normality with and without transformation. RESULTS: Square-root transformation (sqrt) of %retics produced the most constant variance (lowest r-value for correlation between sqrt|normalized residuals| and fitted values: 0.018 highest P-value 0.739) compared with log transformation (r = -0.198, P < 0.001) or no transformation (r = 0.215, P < 0.001) and showed the least departure from normality (highest P-value: 0.002 vs. < 0.001 vs. < 0.001, respectively). Hb results did not improve with transformation. Within-patient variance in both %retics and Hb increased with interval length between lab draws (P < 0.001). CONCLUSIONS: Initial assessment of anti-doping tool use in dialysis patient anemia management indicates square-root transformation of %retics and adjustment for time between lab draw intervals for both %retics and Hb will be required.

Quality control technique to reduce the variability of longitudinal measurement of hemoglobin mass.

The sensitivity of the athlete blood passport to detect blood doping may be improved by the inclusion of total hemoglobin mass (Hb(mass)), but the comparability of Hb(mass) from different laboratories is unknown. To optimize detection sensitivity, the analytical variability associated with Hb(mass) measurement must be minimized. The aim of this study was to investigate the efficacy of using quality controls to minimize the variation in Hb(mass) between laboratories. Three simulated laboratories were set up in one location. Nine participants completed three carbon monoxide (CO) re-breathing tests in each laboratory. One participant completed two CO re-breathing tests in each laboratory. Simultaneously, quality controls containing Low (1-3%) and High (8-11%) concentrations of percent carboxyhemoglobin (%HbCO) were measured to compare hemoximeters in each laboratory. Linear mixed modeling was used to estimate the within-subject variation in Hb(mass), expressed as the coefficient of variation, and to estimate the effect of different laboratories. The analytic variation of Hb(mass) was 2.4% when tests were conducted in different laboratories, which reduced to 1.6% when the model accounted for between-laboratory differences. Adjustment of Hb(mass) values using quality controls achieved a comparable analytic variation of 1.7%. The majority of between-laboratory variation in Hb(mass) originated from the difference between hemoximeters, which could be eliminated using appropriate quality controls.

Changing practice with changing research: results of two UK national surveys of intensive insulin therapy in intensive care patients.

We conducted two telephone surveys of all United Kingdom adult intensive care units in 2007/8 and 2010 to assess practice with regard to intensive insulin therapy for glycaemic control in critically ill patients, and to assess the change in practice following publications in 2008 and 2009 that challenged the evidence for this therapy. Of 243 units that had a written policy for intensive insulin therapy in 2007/8, 232 (96%) still had a policy in 2010. One hundred and six (46%) units had updated their policy in response to new evidence, whereas 126 (54%) stated that it had remained the same. Where intensive care units had changed their policy, we found a significant increase in target limits and a wider target range. Regional variations in practice were also seen. Across seven regions, the percentage of units where the glycaemic control policy had been updated since 2007/8 varied from nil to 78.9%.