Intramammary antimicrobial treatment of subclinical mastitis and cow performance later in lactation.

The aim of this study was to evaluate long-term therapeutic effects of antimicrobial treatment of recently acquired subclinical mastitis (RASCM) during lactation. Quarter-level clinical mastitis (CM) follow-up, composite somatic cell counts (SCC), and cow-level milk yield later in lactation were evaluated using follow-up data from 2 previously published linked randomized field trials. The first trial randomly assigned antimicrobial treatment with any intramammary product or negative control to culture-positive quarters of cows having a first elevated composite SCC after 2 consecutive low composite SCC measurements. Untreated cows that had a second elevated composite SCC at the next measurement and were staphylococci-positive (i.e., Staphylococcus aureus or non-aureus staphylococci) were randomly assigned to treatment or control. Quarter-level CM cases were reported by the participating herd personnel, and milk yield and composite SCC data were obtained from the regular test-day recording. Frailty survival models were used to evaluate the long-term therapeutic effects of antimicrobial treatment of RASCM on quarter-level CM follow-up. Mixed linear regression models were applied to quantify the effect on milk yield and composite SCC. Data of 638 quarters from 486 cows in 38 herds were available for statistical analyses, of which 229 quarters of 175 cows received antimicrobial treatment for RASCM. Antimicrobial treatment culminated in reduced composite SCC levels later in lactation but did not result in different milk yield levels or CM follow-up compared with control cows. Antimicrobial treatment of cows with RASCM should therefore only be considered in exceptional situations given the current focus on antimicrobial usage reduction in animal husbandry.

Prevalence and risk factors for brucellosis in goats in areas of Mexico with and without brucellosis control campaign.

Brucellosis is a major constraint for small-scale goat farming systems in Mexico. This study estimated the prevalence of testing positive to brucellosis and identified and quantified risk factors in goats from small-scale farms of Michoacán that had participated in a brucellosis campaign (i.e. vaccination, serological testing, culling and awareness) and of Jalisco that had negligible brucellosis campaign participation. A cross-sectional serological survey was conducted among 1,713 goats of 83 flocks. The prevalence of testing positive to brucellosis was higher (38%) in Jalisco than in Michoacán (11%). Logistic regression analysis indicated that goats from Michoacán had lower odds to test positive for brucellosis (odds ratio (OR) = 0.32, 95% confidence interval (CI) 0.21-0.48) compared to goats from Jalisco. Goats in zero-grazing systems had lower odds than goats in grazing systems (OR = 0.22, 95% CI 0.09-0.57). When goats were kept in pens with low density (0.002 to 0.22 goat/m(2)), odds was lower (OR = 0.44, 95% CI 0.28-0.67) compared to goats kept in pens with higher density (0.23 to 1 goat/m(2)). Odds was higher for testing positive when farmers bought goats from goat traders (OR = 1.82, 95% CI 1.15-2.87) compared to farmers who did not. If scavenger poultry had access to goat pens, the odds was half (OR = 0.52, 95% CI 0.33-0.83) of those where poultry had no access. Regular disinfection of the pen reduced the odds (OR = 0.66, 95% CI 0.44-0.99) compared to where disinfection was not regular. The brucellosis control campaign was effective in reducing brucellosis seropositivity.

A randomized controlled pilot study into the effects of a restricted elimination diet on family structure in families with ADHD and ODD.

BACKGROUND: Behavioural improvements of children with attention-deficit hyperactivity disorder (ADHD) and oppositional defiant disorder (ODD) following a restricted elimination diet (RED), may be due to concurrent changes in family environment. METHODS: Twenty-four children with ADHD, were randomized to either a 5-week RED intervention, or a control intervention consisting of healthy food advices in a pilot study. RESULTS: No differences in family environment were found, neither at baseline nor when comparing the start and end measurements of both groups. CONCLUSIONS: In this pilot study, the effects of an RED on ADHD and ODD are not mediated by improvement of family environment in families motivated to follow an RED. Replication of this preliminary study in larger groups of children is advised.

Effects of a restricted elimination diet on the behaviour of children with attention-deficit hyperactivity disorder (INCA study): a randomised controlled trial.

BACKGROUND: The effects of a restricted elimination diet in children with attention-deficit hyperactivity disorder (ADHD) have mainly been investigated in selected subgroups of patients. We aimed to investigate whether there is a connection between diet and behaviour in an unselected group of children. METHODS: The Impact of Nutrition on Children with ADHD (INCA) study was a randomised controlled trial that consisted of an open-label phase with masked measurements followed by a double-blind crossover phase. Patients in the Netherlands and Belgium were enrolled via announcements in medical health centres and through media announcements. Randomisation in both phases was individually done by random sampling. In the open-label phase (first phase), children aged 4-8 years who were diagnosed with ADHD were randomly assigned to 5 weeks of a restricted elimination diet (diet group) or to instructions for a healthy diet (control group). Thereafter, the clinical responders (those with an improvement of at least 40% on the ADHD rating scale [ARS]) from the diet group proceeded with a 4-week double-blind crossover food challenge phase (second phase), in which high-IgG or low-IgG foods (classified on the basis of every child's individual IgG blood test results) were added to the diet. During the first phase, only the assessing paediatrician was masked to group allocation. During the second phase (challenge phase), all persons involved were masked to challenge allocation. Primary endpoints were the change in ARS score between baseline and the end of the first phase (masked paediatrician) and between the end of the first phase and the second phase (double-blind), and the abbreviated Conners' scale (ACS) score (unmasked) between the same timepoints. Secondary endpoints included food-specific IgG levels at baseline related to the behaviour of the diet group responders after IgG-based food challenges. The primary analyses were intention to treat for the first phase and per protocol for the second phase. INCA is registered as an International Standard Randomised Controlled Trial, number ISRCTN 76063113. FINDINGS: Between Nov 4, 2008, and Sept 29, 2009, 100 children were enrolled and randomly assigned to the control group (n=50) or the diet group (n=50). Between baseline and the end of the first phase, the difference between the diet group and the control group in the mean ARS total score was 23·7 (95% CI 18·6-28·8; p<0·0001) according to the masked ratings. The difference between groups in the mean ACS score between the same timepoints was 11·8 (95% CI 9·2-14·5; p<0·0001). The ARS total score increased in clinical responders after the challenge by 20·8 (95% CI 14·3-27·3; p<0·0001) and the ACS score increased by 11·6 (7·7-15·4; p<0·0001). In the challenge phase, after challenges with either high-IgG or low-IgG foods, relapse of ADHD symptoms occurred in 19 of 30 (63%) children, independent of the IgG blood levels. There were no harms or adverse events reported in both phases. INTERPRETATION: A strictly supervised restricted elimination diet is a valuable instrument to assess whether ADHD is induced by food. The prescription of diets on the basis of IgG blood tests should be discouraged. FUNDING: Foundation of Child and Behaviour, Foundation Nuts Ohra, Foundation for Children's Welfare Stamps Netherlands, and the KF Hein Foundation.

Genome-wide association study of insect bite hypersensitivity in two horse populations in the Netherlands.

BACKGROUND: Insect bite hypersensitivity is a common allergic disease in horse populations worldwide. Insect bite hypersensitivity is affected by both environmental and genetic factors. However, little is known about genes contributing to the genetic variance associated with insect bite hypersensitivity. Therefore, the aim of our study was to identify and quantify genomic associations with insect bite hypersensitivity in Shetland pony mares and Icelandic horses in the Netherlands. METHODS: Data on 200 Shetland pony mares and 146 Icelandic horses were collected according to a matched case-control design. Cases and controls were matched on various factors (e.g. region, sire) to minimize effects of population stratification. Breed-specific genome-wide association studies were performed using 70 k single nucleotide polymorphisms genotypes. Bayesian variable selection method Bayes-C with a threshold model implemented in GenSel software was applied. A 1 Mb non-overlapping window approach that accumulated contributions of adjacent single nucleotide polymorphisms was used to identify associated genomic regions. RESULTS: The percentage of variance explained by all single nucleotide polymorphisms was 13% in Shetland pony mares and 28% in Icelandic horses. The 20 non-overlapping windows explaining the largest percentages of genetic variance were found on nine chromosomes in Shetland pony mares and on 14 chromosomes in Icelandic horses. Overlap in identified associated genomic regions between breeds would suggest interesting candidate regions to follow-up on. Such regions common to both breeds (within 15 Mb) were found on chromosomes 3, 7, 11, 20 and 23. Positional candidate genes within 2 Mb from the associated windows were identified on chromosome 20 in both breeds. Candidate genes are within the equine lymphocyte antigen class II region, which evokes an immune response by recognizing many foreign molecules. CONCLUSIONS: The genome-wide association study identified several genomic regions associated with insect bite hypersensitivity in Shetland pony mares and Icelandic horses. On chromosome 20, associated genomic regions in both breeds were within 2 Mb from the equine lymphocyte antigen class II region. Increased knowledge on insect bite hypersensitivity associated genes will contribute to our understanding of its biology, enabling more efficient selection, therapy and prevention to decrease insect bite hypersensitivity prevalence.

Physical Complaints Decrease after Following a Few-Foods Diet in Children with ADHD.

Attention-deficit hyperactivity disorder (ADHD) symptoms may significantly decrease after following a few-foods diet (FFD). The results of a small randomised controlled trial (RCT) showed that co-occurring physical complaints in children with ADHD decreased as well. To further investigate the effect of an FFD on physical complaints, we analysed unpublished data from previously published studies (i.e., 'Impact of Nutrition on Children with ADHD' [INCA], an RCT, and 'Biomarker Research in ADHD: the Impact of Nutrition' [BRAIN], an open-label trial). In both trials, the association between an FFD, ADHD, and 21 individual physical complaints was assessed. Children either followed a 5-week FFD (the INCA FFD group and BRAIN participants) or received healthy food advice (the INCA control group). The ADHD rating scale and a physical complaint questionnaire were filled in at the start and end of the trials. The INCA results showed, for 10 of 21 complaints, a clinically relevant reduction in the FFD group compared to the control group. The open-label BRAIN results confirmed the outcomes of the FFD group. No association was detected between the decrease in physical complaints and the decrease in ADHD symptoms. The results point toward an association between the FFD and a decrease in thermoregulation problems, gastrointestinal complaints, eczema, and sleep problems.

Revisiting the relative effectiveness of slaughterhouses in Ireland to detect tuberculosis lesions in cattle (2014-2018).

Slaughterhouse or meat factory surveillance to detect factory lesions (FL) at slaughter is an important part of the bovine tuberculosis (bTB) eradication program in Ireland. The objective of this study was to quantify the effectiveness of Irish slaughterhouses or factories in submitting FL and the proportion of those submitted FL confirmed as being due to bTB in slaughtered cattle, and to identify and quantify the association of risk factors at animal, herd, and factory level with FL submission and confirmation. The data consisted of 6,611,854 animals slaughtered in Irish factories from 2014 to 2018 obtained from the Department of Agriculture, Food and Marine (DAFM), Ireland. Selected risk factors for this study included factory, year and month of slaughter, age, sex, breed, animal movement, ever inconclusive in the standard or severe skin test, herd type, herd size, and bTB history. The association of each risk factor on the FL submission and confirmation risk were analysed with univariable followed by a multivariable logistic regression with herd as random effect. Factories were ranked and compared based on the odds ratio (OR) obtained from the univariable (crude OR) and multivariable (adjusted OR) analysis. The average submission risk of all factories was 20 per 10,000 animals slaughtered, ranging from 1 to 42 per 10,000 animals slaughtered, and the average confirmation risk over all factories was 40.72%, ranging from 0.00 to 61.84%. The odds of submitting and confirming FL as bTB positive were higher in animals over eight years old compared to animals 1-2 years old (OR = 1.91, 95 CI 95% 1.77-2.06 and OR = 4.05, 95% CI 3.17-5.18, respectively), and were higher in animals that ever had inconclusive skin result based on severe interpretation (OR = 2.83, 95% CI 2.44-3.27 and OR = 4.48, 95% CI 2.66-7.54, respectively), animals originating from sucklers herds (OR = 1.08, 95% CI 1.02-1.14 and OR = 1.31, 95% CI 1.11-1.55, respectively), or herds with bTB history in the past three years (OR = 4.46, 95% CI 4.28-4.66 and OR = 319.90, 95% CI 237.98-430.04, respectively). The odds of FL submission and confirmation decreased as the herd size increased (OR = 0.95, 95% CI 0.93-0.96 and OR = 0.82, 95% CI 0.78-0.86, respectively). An inverse relationship of FL submission and confirmation was present for variable sex and inconclusive skin result with standard interpretation, where submission odds were higher in males (OR = 1.05, 95% CI 1.00-1.10) and ever inconclusive animals (OR = 74.24, 95% CI 69.39-79.43), although the confirmation odds were lower (males OR = 0.66, 95% CI 0.56-0.76; ever inconclusive OR = 0.44, 95% CI 0.36-0.54). The crude and adjusted ranking of factories did not differ greatly for FL submission, indicating that factory-related factors may contribute significantly to the submission variation between factories. However, a substantial difference between crude and adjusted confirmation ranking was present which may indicate that animal and herd-related factors were associated to variation in confirmation risk between factories.

Estimation of the likelihood of fecal-oral HEV transmission among pigs.

Sources for human hepatitis E virus (HEV) infections of genotype 3 are largely unknown. Pigs are potential animal reservoirs for HEV. Intervention at pig farms may be desired when pigs are confirmed as a source for human infections, requiring knowledge about transmission routes. These routes are currently understudied. The current study aims to quantify the likelihood of pig feces in causing new HEV infections in pigs due to oral ingestion. We estimated the daily infection risk for pigs by modeling the fate of HEV in the fecal-oral (F-O) pathway. Using parameter values deemed most plausible by the authors based on current knowledge the daily risk of infection was 0.85 (95% interval: 0.03-1). The associated expected number of new infections per day was ∼4 (2.5% limit 0.1, the 97% limit tending to infinity) compared to 0.7 observed in a transmission experiment with pigs, and the likelihood of feces causing the transmission approached 1. In alternative scenarios, F-O transmission of HEV was also very likely to cause new infections. By reducing the total value of all explanatory variables by 2 orders of magnitude, the expected numbers of newly infected pigs approached the observed number. The likelihood of F-O transmission decreased by decreasing parameter values, allowing for at most 94% of infections being caused by additional transmission routes. Nevertheless, in all scenarios F-O transmission was estimated to contribute to HEV transmission. Thus, despite the difficulty in infecting pigs with HEV via oral inoculation, the F-O route is likely to cause HEV transmission among pigs.

Risk factors for digital dermatitis in free-stall-housed, Canadian dairy cattle.

BACKGROUND: A comprehensive analysis of the relation between digital dermatitis (DD) and cow and herd characteristics in Canadian dairies is currently lacking. METHODS: A multilevel logistic regression analysis was performed using 12,260 cow records from 62 dairy farms to assess association between 27 cow and herd-level variables, and presence of DD. RESULTS: The odds for a cow to have at least 1 DD lesion were higher in first-parity cows and those in later lactation (≥45 days in milk). Housing cows on a concrete base was associated with higher odds (OR 2.24) for DD when bedding was added once a week or less. Bedding the concrete base more frequently reduced odds for DD. Wood shavings or other bedding types were more positively associated with DD (OR 2.31 and 1.87, respectively) compared to sawdust. Also, the odds of DD were lower on farms with a scraping manure frequency of every 2 h compared to less frequent scraping (OR 0.54). CONCLUSION: Nine risk factors for DD were identified and quantified, with stall base, bedding type, and manure scraping frequency associated with lower odds of DD. DD prevalence could be reduced by implementing management practices for first-parity cows, as they had higher odds of DD.

Correlation between brain function and ADHD symptom changes in children with ADHD following a few-foods diet: an open-label intervention trial.

Research into the effect of nutrition on attention-deficit hyperactivity disorder (ADHD) in children has shown that the few-foods diet (FFD) substantially decreases ADHD symptoms in 60% of children. However, the underlying mechanism is unknown. In this open-label nutritional intervention study we investigated whether behavioural changes after following an FFD are associated with changes in brain function during inhibitory control in 79 boys with ADHD, aged 8-10 years. Parents completed the ADHD Rating Scale before (t1) and after the FFD (t2). Functional magnetic resonance imaging (fMRI) scans were acquired during a stop-signal task at t1 and t2, and initial subject-level analyses were done blinded for ARS scores. Fifty (63%) participants were diet responders, showing a decrease of ADHD symptoms of at least 40%. Fifty-three children had fMRI scans of sufficient quality for further analysis. Region-of-interest analyses demonstrated that brain activation in regions implicated in the stop-signal task was not associated with ADHD symptom change. However, whole-brain analyses revealed a correlation between ADHD symptom decrease and increased precuneus activation (pFWE(cluster) = 0.015 for StopSuccess > Go trials and pFWE(cluster) < 0.001 for StopSuccess > StopFail trials). These results provide evidence for a neurocognitive mechanism underlying the efficacy of a few-foods diet in children with ADHD.

Effects of food on physical and sleep complaints in children with ADHD: a randomised controlled pilot study.

Attention deficit/hyperactivity disorder (ADHD), a common behavioural disorder in children, may be associated with comorbid physical and sleep complaints. Dietary intervention studies have shown convincing evidence of efficacy in reducing ADHD symptoms in children. In this pilot study, we investigated the effects of an elimination diet on physical and sleep complaints in children with ADHD. A group of 27 children (3.8-8.5 years old), who all met the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria for ADHD, were assigned randomly to either a diet group (15/27) or a control group (12/27). The diet group followed a 5-week elimination diet; the control group adhered to their normal diet. Parents of both groups had to keep an extended diary and had to monitor the behaviour and the physical and sleep complaints of their child conscientiously. The primary endpoint was the clinical response, i.e. a decrease of physical and sleep complaints, at the end of the trial, based on parent ratings on a Physical Complaints Questionnaire. The number of physical and sleep complaints was significantly decreased in the diet group compared to the control group (p < 0.001), with a reduction in the diet group of 77% (p < 0.001, effect size = 2.0) and in the control group of 17% (p = 0.08, effect size = 0.2). Specific complaints that were significantly reduced were in three domains: headaches or bellyaches, unusual thirst or unusual perspiration, and sleep complaints. The reduction of complaints seemed to occur independently of the behavioural changes (p = 0.1). However, the power of this comparison was low. A positive correlation existed between the reduction of physical and behavioural symptoms (p < 0.01). The reduction did not differ between children with or without an atopic constitution (p = 0.7). An elimination diet may be an effective instrument to reduce physical complaints in children with ADHD, but more research is needed to determine the effects of food on (functional) somatic symptoms in children with and without ADHD. This trial was registered as an International Standard Randomised Controlled Trial, ISRCTN47247160.

Retrospective Outcome Monitoring of ADHD and Nutrition (ROMAN): The Effectiveness of the Few-Foods Diet in General Practice.

INTRODUCTION: Double-blind placebo-controlled studies investigating the effect of a few-foods diet (FFD) on attention-deficit/hyperactivity disorder (ADHD) have provided consistent evidence that ADHD can be triggered by foods, indicating the existence of a food-induced ADHD subtype. In 2001 the "few-foods" approach was included in an ADHD treatment protocol. This approach consists of (a) determining, by means of an FFD, whether food is a trigger of ADHD; (b) reintroducing, in FFD responders, foods to assess which foods are incriminated; (c) finally composing a personalised diet eliminating the involved foods only. In the Netherlands the few-foods approach is applied in practice. We aimed to retrospectively assess its effectiveness on ADHD and oppositional defiant disorder (ODD) in real life. METHODS: Data from all children who started the few-foods approach in three specialised healthcare facilities during three consecutive months were included. Behavior was assessed at start and end of the 5-week FFD, using the ADHD Rating Scale and a structured psychiatric interview. Clinical responders (behavioral improvements ≥40%) proceeded with the reintroduction phase. RESULTS: Data of 57 children, 27 taking medication and 15 following some elimination diet at start, were available. No differences were noted between parental scores of children with and without medication or some elimination diet at start. 21/27 (78%) children stopped taking medication during the FFD. 34/57 (60%) children were ADHD responders, 20/29 (65%) children meeting ODD criteria were ODD responders. 26/34 (76%) ADHD responders started the reintroduction phase; 14/26 (54%) still participated at six months. Teacher data were available of 18/57 (32%) children. 9/18 (50%) children were ADHD responders. CONCLUSION: The FFD, if applied by trained specialists, may lead to clinically relevant reduction of ADHD and ODD symptoms in general practice, and a concomitant decrease of ADHD medication. These results corroborate the existence of an ADHD subgroup with food-induced ADHD. Defining and eliminating the incriminated foods, i.e. the underlying causal triggers, may result in secondary prevention of food-induced ADHD. Research into underlying mechanism(s) is of vital importance: finding an easier method or biomarkers for diagnosing food-induced ADHD and ascertaining the incriminated foods may lead to redundancy of the few-foods approach.

The course of hepatitis E virus infection in pigs after contact-infection and intravenous inoculation.

BACKGROUND: Worldwide, hepatitis E virus (HEV) genotype 3 is observed in pigs and transmission to humans is implied. To be able to estimate public health risks from e.g. contact with pigs or consumption of pork products, the transmission routes and dynamics of infection should be identified. Hence, the course of HEV-infection in naturally infected pigs should be studied. RESULTS: To resemble natural transmission, 24 HEV-susceptible pigs were infected either by one-to-one exposure to intravenously inoculated pigs (C1-pigs; n = 10), by one-to-one exposure to contact-infected pigs (C2-pigs: n = 7; C3-pigs: n = 5) or due to an unknown non-intravenous infection route (one C2-pig and one C3-pig). The course of HEV-infection for contact-infected pigs was characterized by: faecal HEV RNA excretion that started at day 7 (95% confidence interval: 5-10) postexposure and lasted 23 (19-28) days; viremia that started after 13 (8-17) days of faecal HEV RNA excretion and lasted 11 (8-13) days; antibody development that was detected after 13 (10-16) days of faecal HEV RNA excretion. The time until onset of faecal HEV RNA excretion and onset of viremia was significantly shorter for iv-pigs compared to contact-infected pigs, whereas the duration of faecal HEV RNA excretion was significantly longer. At 28 days postinfection HEV RNA was detected less frequently in organs of contact-infected pigs compared to iv-pigs. For contact-infected pigs, HEV RNA was detected in 20 of 39 muscle samples that were proxies for pork at retail and in 4 of 7 urine samples. CONCLUSION: The course of infection differed between infection routes, suggesting that contact-infection could be a better model for natural transmission than iv inoculation. Urine and meat were identified as possible HEV-sources for pig-to-pig and pig-to-human HEV transmission.

A randomised controlled trial into the effects of food on ADHD.

The aim of this study is to assess the efficacy of a restricted elimination diet in reducing symptoms in an unselected group of children with Attention deficit/hyperactivity disorder (ADHD). Dietary studies have already shown evidence of efficacy in selected subgroups. Twenty-seven children (mean age 6.2) who all met the DSM-IV criteria for ADHD, were assigned randomly to either an intervention group (15/27) or a waiting-list control group (12/27). Primary endpoint was the clinical response, i.e. a decrease in the symptom scores by 50% or more, at week 9 based on parent and teacher ratings on the abbreviated ten-item Conners Scale and the ADHD-DSM-IV Rating Scale. The intention-to-treat analysis showed that the number of clinical responders in the intervention group was significantly larger than that in the control group [parent ratings 11/15 (73%) versus 0/12 (0%); teacher ratings, 7/10 (70%) versus 0/7 (0%)]. The Number of ADHD criteria on the ADHD Rating Scale showed an effect size of 2.1 (cohen's d) and a scale reduction of 69.4%. Comorbid symptoms of oppositional defiant disorder also showed a significantly greater decrease in the intervention group than it did in the control group (cohens's d 1.1, scale reduction 45.3%). A strictly supervised elimination diet may be a valuable instrument in testing young children with ADHD on whether dietary factors may contribute to the manifestation of the disorder and may have a beneficial effect on the children's behaviour.

Opportunities for Brucellosis Control in Mexico: Views Based on the Sustainable Livelihoods Perspective.

Caprine brucellosis is a main constraint for small-scale goat husbandry systems in the Global South, as it negatively affects production parameters and can be transmitted to humans. The aim of this manuscript is to point out opportunities for brucellosis control in a resource poor area. The present paper draws from previous research in two Mexican states, Jalisco and Michoacán, both within the Bajío region. Main opportunities for brucellosis control are discussed within the "Sustainable Livelihoods Perspective." Goat farming gives farmers a number of livelihoods benefits: food, cash, prestige, and a job. Goat farming is also a reason for some farmers to stay in their villages rather than to migrate to the US. This livelihood strategy, however, is threatened by brucellosis, which is endemic in the goat population of the region. Brucellosis control, however, offers an opportunity for small-scale goat farmers to enhance health and welfare. The socio-economic context is very important in planning a successful brucellosis control campaign. Control strategies should be planned considering the local goat farming husbandry and the views of the farmers.

Biomarker Research in ADHD: the Impact of Nutrition (BRAIN) - study protocol of an open-label trial to investigate the mechanisms underlying the effects of a few-foods diet on ADHD symptoms in children.

INTRODUCTION: Attention deficit hyperactivity disorder (ADHD) is the most common childhood behavioural disorder, causing significant impediment to a child's development. It is a complex disorder with numerous contributing (epi)genetic and environmental factors. Currently, treatment consists of behavioural and pharmacological therapy. However, ADHD medication is associated with several side effects, and concerns about long-term effects and efficacy exist. Therefore, there is considerable interest in the development of alternative treatment options. Double-blind research investigating the effects of a few-foods diet (FFD) has demonstrated a significant decrease in ADHD symptoms following an FFD. However, an FFD requires a considerable effort of both child and parents, limiting its applicability as a general ADHD treatment. To make FFD intervention less challenging or potentially obsolete, we need to understand how, and in which children, an FFD affects ADHD behaviour and, consequently, the child's well-being. We hypothesise that an FFD affects brain function, and that the nutritional impact on ADHD is effectuated by a complex interplay between the microbiota, gut and brain, that is, the microbiota-gut-brain axis. METHODS AND ANALYSIS: The Biomarker Research in ADHD: the Impact of Nutrition (BRAIN) study is an open-label trial with researchers blinded to changes in ADHD symptoms during sample processing and initial data analyses. ETHICS AND DISSEMINATION: The Medical Research and Ethics Committee of Wageningen University has approved this study (NL63851.081.17, application 17/24). Results will be disseminated through peer-reviewed journal publications, conference presentations, (social) media and the BRAIN study website. A summary of the findings will be provided to the participants. TRIAL REGISTRATION NUMBER: NCT03440346. STUDY DATES: Collection of primary outcome data started in March 2018 and will be ongoing until 100 children have participated in the study. Sample data analysis will start after all samples have been collected.

A New Model to Calibrate a Reference Standard for Bovine Tuberculin Purified Protein Derivative in the Target Species.

Since 1986, use of a Bovine International Standard (BIS) for bovine tuberculin has been required to ensure national and international uniformity regarding the potency designation of bovine tuberculin Purified Protein Derivative (PPDb) preparations produced by multiple manufacturers. The BIS is the unique golden standard in the guinea pig potency assay, representing 100% potency, where potencies of production batches are calculated as relative potencies in comparison with the potency of the BIS which was set at 32,500 international Unit (IU) per mg. The stock supply and lifetime of the BIS is limited.The aim of this study was to develop a model to determine the potency of a newly produced in-house Reference Standard (RS) for PPDb with great accuracy in the target species (cattle) and to prove its precision and accuracy in the guinea pig potency test. First simulations were done to estimate the required number of cattle needed. Then, 30 naturally bTB infected cattle were subjected to a tuberculin skin test using multiple injections of both the RS and the BIS. Both were applied randomly in the same volume and concentration (1 dose). The potency of the RS against the BIS was directly derived from the least square means (LSMEANS) and was estimated as 1.067 (95% CI: 1.025-1.109), equal to a potency of 34,700 ± 1,400 IU/mg. In six guinea pig potency assays the RS was used to assign potencies to production batches of PPDb. Here, precision and accuracy of the RS was determined according to the parallel-line assay. Relative potencies were estimated by exponentiation of the common slope. The corresponding 95% confidence intervals were obtained according to Fieller's theorem. In sensitized guinea pigs, the relative potency of the RS against the BIS was 1.115 (95% CI: 0.871-1.432), corresponding to an absolute potency of 36,238 IU/mg (95% CI: 28,308-46,540).In conclusion: the method used to determine the potency of the RS against the BIS in naturally bTB infected cattle, resulted in a highly accurate potency estimate of the RS. The RS can be used in the guinea pig test to assign potencies to PPDb production batches with high precision and accuracy.

Seroprevalence and risk factors of lumpy skin disease in Ethiopia.

Lumpy skin disease (LSD) is an acute or inapparent viral disease of cattle which is endemic in many African and Middle East countries. LSD is one of the major transboundary livestock diseases in Ethiopia. A cross-sectional study using multistage cluster sampling was undertaken in central and north-western parts of Ethiopia with the objectives to estimate seroprevalence and to identify and quantify risk factors contributing to the occurrence of the disease. A total of 2386 cattle sera were sampled from 605 herds and 30 clusters (kebeles) located in 10 districts and tested for presence of LSD virus antibodies using virus neutralization test. All the serum samples were collected from cattle having no history of LSD vaccination. The overall animal level and herd level apparent seroprevalences were 25.4% (95% CI: 23.7-27.2) and 48.9% (95% CI: 44.9-52.9), respectively and varied significantly between districts. The true animal level and herd level prevalences were estimated as 26.5% (95% CI: 24.7-28.3) and 52.6% (95% CI: 48.3-56.9), respectively. At animal level, adult age (OR = 2.44 (95% CI: 1.67-3.55) compared to calf), contact with other animals (OR = 0.41 (95% CI: 0.23-0.74), compared to no contact) and presence of water bodies (OR = 1.61 (95% CI: 1.03-2.52), compared to no such bodies) were identified as the most important risk factors in relation to testing LSD positive. The putative risk factors altitude, breed, sex, and presence of animal trade route showed no significant association with LSD sero-status. Generally, cattle population with many adult animals and that live in wet areas are at highest risk, whereas cattle in frequent contact with other animals and animal species have lower risk, potentially due to a dilution effect of vectors.

Economic impact of lumpy skin disease and cost effectiveness of vaccination for the control of outbreaks in Ethiopia.

Lumpy skin disease (LSD), an infectious viral disease of cattle, causes considerable financial losses in livestock industry of affected countries. A questionnaire survey with the objectives of determining direct economic losses of LSD (mortality loss, milk loss, draft loss) and treatment costs (medication and labour cost) per affected herd, and assessing the cost effectiveness of vaccination as a means for LSD control was carried out in the central and north-western parts of Ethiopia. From a total of 4430 cattle (in 243 herds) surveyed, 941 animals (in 200 herds) were reported to be infected. The overall morbidity and mortality at animal level were 21.2% and 4.5%, and at herd level these were 82.3% and 24.3%. There was a significant difference in animal level morbidity and mortality between categories of animals. Over 94% of the herd owners ranked LSD as a big or very big problem for cattle production. A large proportion (92.2%) of the herd owners indicated that LSD affects cattle marketing. A median loss of USD 375 (USD 325 in local Zebu and USD 1250 in Holstein-Friesian local Zebu cross cattle) was estimated per dead animal. Median losses per affected lactating cow were USD 141 (USD 63 in local Zebu cows and USD 216 in Holstein-Friesian local Zebu cross cows) and, USD 36 per affected ox. Diagnosis and medication cost per affected animal were estimated at USD 5. The median total economic loss of an LSD outbreak at herd level was USD 1176 (USD 489 in subsistence farm and USD 2735 in commercial farm). At herd level, the largest component of the economic loss was due to mortality (USD 1000) followed by milk loss (USD 120). LSD control costs were the least contributor to herd level losses. The total herd level economic losses in the commercial farm type were significantly higher than in the subsistence farm type. The financial analysis showed a positive net profit of USD 136 (USD 56 for subsistence farm herds and USD 283 for commercial herds) per herd due to LSD vaccine investment. It should be noted that only the noticeable direct costs and treatment costs associated with the disease were considered in the study. Generally, vaccination is economically effective and should be encouraged.

Field study on the use of vaccination to control the occurrence of lumpy skin disease in Ethiopian cattle.

The current study was carried out in central and North-western parts of Ethiopia to assess the efficacy of Kenyan sheep pox virus strain vaccine (KS1 O-180) against natural lumpy skin disease (LSD) infection under field conditions by estimating its effect on the transmission and severity of the disease. For this study, an LSD outbreak was defined as the occurrence of at least one LSD case in a specified geographical area. An observational study was conducted on a total of 2053 (1304 vaccinated and 749 unvaccinated) cattle in 339 infected herds located in 10 sub-kebeles and a questionnaire survey was administered to 224 herd owners. Over 60% of the herd owners reported that the vaccine has a low to very low effect in protecting animals against clinical LSD; almost all of them indicated that the vaccine did not induce any adverse reactions. In the unvaccinated group of animals 31.1% were diagnosed with LSD while this was 22.5% in the vaccinated group (P<0.001). Severity of the disease was significantly reduced in vaccinated compared to unvaccinated animals (OR=0.68, 95% CI: 0.49; 0.96). Unvaccinated infected animals were more likely (predicted fraction=0.89) to develop moderate and severe disease than vaccinated infected animals (predicted fraction=0.84). LSD vaccine efficacy for susceptibility was estimated to be 0.46 (i.e. a susceptibility effect of 0.54) while the infectiousness effect of the vaccine was 1.83. In other words, the vaccine reduces the susceptibility by a factor of two and increases infectiousness by approximately the same amount. LSD transmission occurred in both vaccinated and unvaccinated animals, the estimated reproduction ratio (R) was 1.21 in unvaccinated animals compared to 1.19 in vaccinated ones, and not significantly different. In conclusion, KS1 O-180 vaccination, as applied currently in Ethiopia, has poor efficacy in protecting cattle populations against LSD, neither by direct clinical protection nor by reducing transmission, and this signifies the urgent need to either improve the quality of the vaccine or to develop potent alternative vaccines that will confer good protection against LSD.