Tracheostomy decannulation in children: a proposal for a structured approach on behalf of the working group chronic respiratory insufficiency within the German-speaking society of pediatric pulmonology.

The number of children with tracheostomies with and without home mechanical ventilation has grown continuously in recent years. For some of these children, the need for tracheostomy resolves and the child can be weaned from the tracheal cannula. Choosing the optimal time point for decannulation after elaborated prior diagnostic work-up needs careful consideration. The decannulation process requires an interdisciplinary team; however, these specialized structures for the experienced care of these children with tracheostomy are not available in all areas. The Working Group on Chronic Respiratory Insufficiency in the German Speaking Pediatric Pneumology Society (GPP) developed these recommendations to guide through a decannulation process. Initial evaluation of decannulation feasibility starts in the outpatient clinic with a detailed history, examination, and a speaking valve trial and is followed by an inpatient workup including sleep study, airway endoscopy and possibly modifications of the tracheal cannula. Downsizing the tracheal cannula allows a stepwise controlled weaning prior to removal of the tracheal cannula. After shrinking of the tracheostomy, the final surgical closure is performed.  Conclusion: An algorithm with diagnostic and therapeutic procedures for a safe and successful decannulation process is proposed. What is Known: • In children tracheostomy decannulation is a complex process that requires careful preparation and surveillance. What is New: • This statement of the German speaking society of pediatric pulmonology provides an expert practice guidance on the decannulation procedure and the value of one-way speaking valves.

Hepatitis E virus: Efficacy of pasteurization of plasma-derived VWF/FVIII concentrate determined by pig bioassay.

BACKGROUND: Hepatitis E virus (HEV) is the leading cause of acute hepatitis throughout the world. Increasing blood component transfusion-associated HEV infections highlight the need for reliable virus inactivation procedures for plasma derivatives from pooled plasma donations. STUDY DESIGN AND METHODS: An animal infection study was conducted to evaluate the efficiency of HEV inactivation by pasteurization during the manufacturing process of the von Willebrand Factor/Factor VIII (VWF/FVIII) concentrate Haemate P/Humate-P (CSL Behring, Marburg, Germany). For this purpose, groups of pigs were inoculated with stabilized VWF/FVIII intermediate spiked with HEV-positive liver homogenate and exposed to increasing incubation times of 0, 3, 6, and 10 h at 60°C. Animals were evaluated for virus replication over 27 days and in a subsequent trial over 92 days. RESULTS: Virus replication was detected in animals up to the 6-h pasteurization group. In contrast, pasteurization for 10 h did not reveal virus detection when the observation period was 27 days. In an additional experiment using the 10-h pasteurized material, two individuals started virus excretion and seroconverted when the observation period was extended to 92 days. Based on the total infection rate (2 of 12) of the animals inoculated with the sample pasteurized for 10 h, a virus reduction factor of at least 4.7 log10 is calculated. CONCLUSION: This study demonstrates that pasteurization at 60°C for 10 h of an HEV-positive plasma derivative leads to the effective reduction of infectivity, resulting in a VWF/FVIII product with an appropriate margin of safety for HEV.

Empowering Parents of Obese Children (EPOC): A randomized controlled trial on additional long-term weight effects of parent training.

Although inpatient lifestyle treatment for obese children and adolescents can be highly effective in the short term, long-term results are unconvincing. One possible explanation might be that the treatment takes place far from parents' homes, limiting the possibility to incorporate the parents, who play a major role in establishing and maintaining a healthy lifestyle in childhood and adolescence. The main goal was to develop a brief behaviorally oriented parent training program that enhances 'obesity-specific' parenting skills in order to prevent relapse. We hypothesized that the inclusion of additional parent training would lead to an improved long-term weight course of obese children. Parents of obese children (n = 686; 7-13 years old) either participated in complementary cognitive-behavioral group sessions (n = 336) or received written information only (n = 350) during the inpatient stay. Children of both groups attended multidisciplinary inpatient rehabilitation. BMI-SDS as a primary outcome was evaluated at baseline, post-intervention and at 6- and 12-month follow-up. Intention-to-treat (ITT) as well as per-protocol analyses (PPA) were performed. A significant within-group decrease of 0.24 (95% CI 0.18 to 0.30) BMI-SDS points from the beginning of the inpatient stay through the first year was found, but no group difference at the one-year follow-up (mean difference 0.02; 95% CI -0.04 to 0.07). We also observed an increase in quality of life scores, intake of healthy food and exercise for both groups, without differences between groups (ITT and PPA). Thus, while the inpatient treatment proved highly effective, additional parent training did not lead to better results in long-term weight maintenance or to better psychosocial well-being compared to written psycho-educational material. Further research should focus on subgroups to answer the question of differential treatment effects.

Looking deep inside: detection of low-abundance proteins in leaf extracts of Arabidopsis and phloem exudates of pumpkin.

The field of proteomics suffers from the immense complexity of even small proteomes and the enormous dynamic range of protein concentrations within a given sample. Most protein samples contain a few major proteins, which hamper in-depth proteomic analysis. In the human field, combinatorial hexapeptide ligand libraries (CPLL; such as ProteoMiner) have been used for reduction of the dynamic range of protein concentrations; however, this technique is not established in plant research. In this work, we present the application of CPLL to Arabidopsis (Arabidopsis thaliana) leaf proteins. One- and two-dimensional gel electrophoresis showed a decrease in high-abundance proteins and an enrichment of less abundant proteins in CPLL-treated samples. After optimization of the CPLL protocol, mass spectrometric analyses of leaf extracts led to the identification of 1,192 proteins in control samples and an additional 512 proteins after the application of CPLL. Upon leaf infection with virulent Pseudomonas syringae DC3000, CPLL beads were also used for investigating the bacterial infectome. In total, 312 bacterial proteins could be identified in infected Arabidopsis leaves. Furthermore, phloem exudates of pumpkin (Cucurbita maxima) were analyzed. CPLL prefractionation caused depletion of the major phloem proteins 1 and 2 and improved phloem proteomics, because 67 of 320 identified proteins were detectable only after CPLL treatment. In sum, our results demonstrate that CPLL beads are a time- and cost-effective tool for reducing major proteins, which often interfere with downstream analyses. The concomitant enrichment of less abundant proteins may facilitate a deeper insight into the plant proteome.

Seroprevalence and GIS-supported risk factor analysis of Fasciola hepatica infections in dairy herds in Germany.

A total of 20 749 bulk tank milk (BTM) samples was collected in November 2008 from all over Germany, corresponding to 20.9% of all German dairy herds. The BTM samples were analysed for antibodies against Fasciola hepatica using the excretory-secretory (ES) ELISA. A geospatial map was drawn to show herd prevalences per postal code area. Various spatial risk factors were tested for potential statistical associations with the ELISA results in logistic regression supported by a geographical information system (GIS). The mean seroprevalence was 23.6% and prevalences in different German federal states varied between 2.6% and 38.4%. GIS analysis revealed statistically significant positive associations between the proportion of grassed area and water bodies per postal code area and positive BTM ELISA results. This can be explained by the biology of the intermediate host, the amphibious snail Galba (Lymnea) truncatula and the pasture-borne nature of fasciolosis. The full logistic regression model had a Pseudo-R 2 of 22%, while the final model obtained by controlled stepwise model building revealed a Pseudo-R 2 of 14%, indicating that additional, unrecorded factors and random effects contributed substantially to the occurrence of positive ELISA results. Considering the high seroprevalences in some areas and the economic impact of fasciolosis, farmers and veterinarians should be strongly advised to implement effective liver fluke control programmes.

SARS-CoV-2 Associated Pediatric Inflammatory Multisystem Syndrome With a High Prevalence of Myocarditis - A Multicenter Evaluation of Clinical and Laboratory Characteristics, Treatment and Outcome.

Introduction: Pediatric inflammatory multisystem syndrome - temporally associated with SARS-CoV-2 infection (PIMS -TS) comprises a new disease entity having emerged after the COVID-19 outbreak in 2019. Materials and Methods: For this multicenter, retrospective study children between 0 and 18 years with PIMS-TS between March 2020 and May 2021 were included, before availability of vaccination for children. Frequent SARS-CoV-2 variants at that period were the wildtype virus, alpha, beta and delta variants. Inclusion criteria were according to the PIMS-TS criteria, proposed by the Royal College of Pediatrics and WHO. Study aim was to review their clinical, laboratory and echocardiographic data with a focus on cardiac involvement. Results: We report 45 patients, median age 9 years, 64% male. SARS-CoV-2 antibodies were positive in 35/41 (85%). PIMS occurrence followed local COVID-19 peak incidence periods with a time lag. The most common symptoms at presentation were fever (98%), abdominal pain (89%) and rash (80%). Fever history of > 5 days was associated with decreased left ventricular function (p = 0.056). Arterial hypotension and cardiac dysfunction were documented in 72% patients, increased brain natriuretic peptide in 96% and increased cardiac troponin in 64% of the children. Echocardiography revealed mitral valve regurgitation (64%), coronary abnormalities (36%) and pericardial effusions (40%). Increased NT-proBNP was significantly associated with the need of inotropics (p < 0.05), which were necessary in 40% of the patients. Treatment comprised intravenous immunoglobulin (93%), systemic steroids (84%) and acetylsalicylic acid (100%; 26/45 started with high dosages). For insufficient response to this treatment, five (11%) children received the interleukin-1 receptor antagonist anakinra. All patients were discharged with almost resolved cardiac signs. Conclusion: Our analysis of non-vaccinated children with PIMS-TS demonstrates that a considerable number have associated myocarditis requiring intensive care and inotropic support. Most children showed adequate response to intravenous immunoglobulin and steroids and good recovery. Further evaluation of pediatric patients with COVID-19 associated diseases is required to evaluate the impact of new virus variants.

Proteomic analysis of defense response of wildtype Arabidopsis thaliana and plants with impaired NO- homeostasis.

In recent years, nitric oxide (NO) has been recognized as a signalling molecule of plants, being involved in diverse processes like germination, root growth, stomatal closing, and responses to various stresses. A mechanism of how NO can regulate physiological processes is the modulation of cysteine residues of proteins (S-nitrosylation) by S-nitrosoglutathione (GSNO), a physiological NO donor. The concentration of GSNO and the level of S-nitrosylated proteins are regulated by GSNO reductase, which seems to play a major role in NO signalling. To investigate the importance of NO in plant defense response, we performed a proteomic analysis of Arabidopsis wildtype and GSNO-reductase knock-out plants infected with both the avirulent and virulent pathogen strains of Pseudomonas syringae. Using 2-D DIGE technology in combination with MS, we identified proteins, which are differentially accumulated during the infection process. We observed that both lines were more resistant to avirulent infections than to virulent infections mainly due to the accumulation of stress-, redox-, and defense-related proteins. Interestingly, after virulent infections, we also observed accumulation of defense-related proteins, but no or low accumulation of stress- and redox-related proteins, respectively. In summary, we present here the first detailed proteomic analysis of plant defense response.

Retrospective analysis of the oral immunisation of wild boar populations against classical swine fever virus (CSFV) in region Eifel of Rhineland-Palatinate.

In the present study the effect of control measures implemented during the classical swine fever (CSF) epidemic in wild boar in the Eifel region of the German federal state of Rhineland-Palatinate from 1999 to 2005 was assessed. During the first 3 years after official confirmation of virus detection these measures comprised intensive hunting, especially of young animals and hygiene measures. Subsequently oral immunisation (o.i.) using a modified live virus vaccine was introduced as an additional control tool. All shot wild boar from the restricted area were tested virologically and serologically for CSF. The laboratory results from over 110,000 animals accompanied by information about age, gender and geographical origin of the animals were collected in a relational database. In total about 82% of all virologically positive wild boars were piglets, thus confirming the importance of this age group in the perpetuation of the epidemic. An analysis of the hunting bag showed that piglets were underrepresented compared to older animals throughout the eradication programme. This finding indicated that hunters did not comply with the control strategy of intense targeting of young animals. Before as well as after the implementation of o.i. a significantly higher virological prevalence and a significantly lower serological prevalence were observed in piglets compared to yearlings and adults. Shortly after the beginning of the vaccination campaign in February 2002 CSFV prevalence decreased significantly whereas the serological prevalence increased markedly in all age classes. In order to test the influence of age and vaccination on the serological prevalence a logistic regression model was used. Our results strongly suggest that under the field conditions in the Eifel region vaccination against CSFV had a crucial influence on the increase of seroprevalence rate and the elimination of CSFV. The last virus-positive pig was found 13 months after start of o.i.

Comparison of different commercial DNA extraction kits and PCR protocols for the detection of Echinococcus multilocularis eggs in faecal samples from foxes.

Effective and sensitive methods for the molecular detection of Echinococcus multilocularis in faecal samples of final hosts are crucial for the prevention and control of human alveolar echinococcosis and for studies on the epidemiology of the parasite. Little is known about the suitability of commercial test kits for isolating DNA of E. multilocularis from fox faeces and the performance of standard Polymerase Chain Reaction (PCR) protocols in relation to the quality of DNA extracted by these kits. We compared four different kits: ZR Faecal DNA MiniPrep™ (Zymo Research), FastDNA® SPIN Kit for Soil (MP Biomedicals), QIAamp® Fast DNA Stool Mini Kit (QIAGEN) and NucleoSpin® Soil Kit (Macherey-Nagel) for the extraction of DNA from E. multilocularis eggs present in faeces of foxes. Negative faecal samples were spiked with 600, 300, 150, 75, 37, 18, 9, 5 or 2 E. multilocularis eggs, and each egg concentration was tested 10 times with each of the DNA extraction kits. Each extracted DNA sample was amplified using three PCR protocols: i. conventional PCR (cPCR, Platinum®Taq, Invitrogen), ii. qPCR with the iQ™ Supermix (Bio-Rad) and iii. qPCR with the QuantiTect® Multiplex-Master Mix (QIAGEN). The highest analytical sensitivities for molecular detection of E. multilocularis eggs in spiked fox faeces were observed when combining either the QIAamp® Fast DNA Stool Mini Kit or the ZR Faecal DNA MiniPrep™ kit with the qPCR using the QuantiTect® Multiplex-Master Mix (Sensitivities 97% and 94%, respectively). Combinations including the remaining test kits (NucleoSpin® Soil Kit and FastDNA® SPIN Kit for Soil) showed a markedly lower analytical sensitivity for PCR examinations. The results of the present study indicate that it is of utmost importance to select suitable DNA extraction kits in combination with robust PCR methods or reagents to achieve acceptable analytical sensitivity in the molecular detection of E. multilocularis eggs in fox faecal samples.

Strategies for the identification of disease-related patterns of volatile organic compounds: prediction of paratuberculosis in an animal model using random forests.

Modern statistical methods which were developed for pattern recognition are increasingly being used for data analysis in studies on emissions of volatile organic compounds (VOCs). With the detection of disease-related VOC profiles, novel non-invasive diagnostic tools could be developed for clinical applications. However, it is important to bear in mind that not all statistical methods are equally suitable for the investigation of VOC profiles. In particular, univariate methods are not able to discover VOC patterns as they consider each compound separately. The present study demonstrates this fact in practice. Using VOC samples from a controlled animal study on paratuberculosis, the random forest classification method was applied for pattern recognition and disease prediction. This strategy was compared with a prediction approach based on single compounds. Both methods were framed within a cross-validation procedure. A comparison of both strategies based on these VOC data reveals that random forests achieves higher sensitivities and specificities than predictions based on single compounds. Therefore, it will most likely be more fruitful to further investigate VOC patterns instead of single biomarkers for paratuberculosis. All methods used are thoroughly explained to aid the transfer to other data analyses.

Survey for zoonotic pathogens in Norway rat populations from Europe.

BACKGROUND: The Norway rat Rattus norvegicus is an important reservoir of various zoonotic pathogens, such as cowpox virus and Leptospira, but also for agents of no or unknown zoonotic potential. We describe a survey of 426 Norway rats originating from five European countries and different habitats for Leptospira spp., rickettsiae, orthopoxvirus (OPV), avian metapneumovirus subtypes A and B (aMPV) and rat polyomavirus (rat PyV). RESULTS: Leptospira DNA was detected in 60 out of 420 (14.3%) rats, and Rickettsia DNA was found in three out of 369 (0.8%) rats investigated. PCR-based typing resulted in the identification of L. interrogans sequence type 17, which corresponds to the serogroup Icterohaemorrhagiae, and Rickettsia helvetica respectively. Rat PyV DNA was detected in 103 out of 421 (24.5%) rats. OPV DNA and aMPV RNA were detected in none of the rats, but OPV-specific antibodies were detected in three out of 388 (0.8%) rats. The frequency of single Leptospira and rat PyV infections and coinfections was, independent of sex, greater for adults compared with juveniles/subadults and greater at rural sites compared with urban areas. CONCLUSIONS: Study results indicate a broad geographical distribution of Leptospira DNA in rats within Europe, underlining the need to investigate further the potential mechanisms leading to increased prevalence in rural habitats and to assess the relevance to public health. In contrast, rickettsia and OPV infections rarely occurred in wild rat populations. The potential influence of rat PyV on the susceptibility to infections with other pathogens should be investigated in future studies. © 2016 Society of Chemical Industry.

Reduction of Portion Size and Eating Rate Is Associated with BMI-SDS Reduction in Overweight and Obese Children and Adolescents: Results on Eating and Nutrition Behaviour from the Observational KgAS Study.

OBJECTIVE: Treatment of paediatric obesity focuses on changes of nutrition and eating behaviour and physical activity. The evaluation of the patient education programme by KgAS was utilised to analyse the association of changes of portion size, eating rate and dietary habits with BMI-SDS reductions. METHODS: Patients (n = 297) were examined at the beginning and at the end of treatment and after 1-year follow-up at different out-patient centres. Their parents completed questionnaires including estimation of children's portion size, eating rate and frequency of food intake. Associations of 1- and 2-year changes in BMI-SDS and behaviour were calculated for patients with complete data in BMI-SDS, portion size, eating rate, frequency of green, yellow and red food intake (n = 131) by multiple linear regression models. RESULTS: Significant changes were found in the desired direction for BMI-SDS, portion size, eating rate and the intake of unfavourable red food items both after 1 and 2 years as well as for the consumption of favourable green food items after 1 year. Significant positive associations with BMI-SDS reduction after 1 and 2 years were detected for portion size (Cohen's f2 0.13 and 0.09) and eating rate (Cohen's f2 0.20 and 0.10), respectively. CONCLUSION: Reduced portion sizes and eating rates are associated with BMI-SDS reduction after 1 and 2 years. These findings suggest to focus on appropriate portion sizes and reduced eating rates in patient education programmes.

Spatial and temporal expression patterns of Avr1b-1 and defense-related genes in soybean plants upon infection with Phytophthora sojae.

The Avr1b locus is required for avirulence of the oomycete pathogen Phytophthora sojae on soybeans carrying resistance gene Rps1b. One of the Avr genes of the locus (Avr1b-1) was shown to encode an elicitor. We have analyzed the spatial and temporal expression patterns of Avr1b-1 in comparison to defense-related genes induced in soybean. Avr1b-1 expression was detectable mainly in close proximity to the site of infection, in wound-inoculated hypocotyls as well as in roots infected with zoospores. Usually, in compatible interactions, higher expression levels of Avr1b-1 were observed in roots when compared with incompatible P. sojae-soybean interactions, whereas neither the timing nor the amount of transcript accumulation of defense-related genes showed cultivar-specific differences. In contrast, the PsojNIP gene encoding a proposed virulence factor was expressed only during the necrotrophic phase in the compatible interaction.

Occurrence and Spread of the Invasive Asian Bush Mosquito Aedes japonicus japonicus (Diptera: Culicidae) in West and North Germany since Detection in 2012 and 2013, Respectively.

The invasive Asian bush mosquito Aedes japonicus japonicus was first recognised as established in Germany in 2008. In addition to the first known and quickly expanding population in the southwestern part of the country, three separate populations were discovered in West, North and southeastern Germany in 2012, 2013 and 2015, respectively, by means of the 'Mueckenatlas', a German instrument of passive mosquito surveillance. Since the first findings of mosquito specimens in West and North Germany, these regions were checked annually for continuing colonisation and spread of the species. Both affected areas were covered by a virtual 10x10km2 grid pattern in the cells of which cemeteries were screened for immature stages of the mosquito. The cells were considered populated as soon as larvae or pupae were detected, whereas they were classified as negative when no mosquito stages were found in the cemeteries of at least three different towns or villages. Presence was also recorded when Ae. j. japonicus adults were submitted to the 'Mueckenatlas' from the respective cell or when there was evidence of local occurrence in localities other than cemeteries. Based on this approach, a significant expansion of the populated area was documented in West Germany since the first detection of Ae. j. japonicus in 2012 (increase in positive grid cells by more than 400%), while the North German population appears not to be expanding so far (reduction of positive grid cells by ca. 30% since 2013). As Ae. j. japonicus finds suitable climatic and ecological conditions in Germany, the differential expansion of the two populations might be attributed to the West German population being older and thus more firmly established than the closely related but younger North German population that might still be in its founder phase. However, geographic spread of all German populations in the future is anticipated. Continuous surveillance is recommended, as Ae. j. japonicus is a competent vector of several pathogens in the laboratory.

Spine malformation complex in 3 diverse syndromic entities: Case reports.

RATIONALE: Clinical and radiographic phenotypic characterizations were the base line tool of diagnosis in 3 syndromic disorders in which congenital cervico-thoracic kyphosis was the major deformity. PATIENTS CONCERNS: Directing maximal care toward the radiographic analysis is not only the axial malformation but also toward the appendicular abnormalities was our main concern. We fully documented the diversity of the spine phenotypic malformation complex via the clinical and radiographic phenotypes. DIAGNOSES: We established the diagnosis via phenotypic/genotypic confirmation in 3 diverse syndromic entities namely acampomelic campomelic dysplasia, Larsen syndrome and Morquio syndrome type A (mucopolysaccharidosis type IV A). INTERVENTIONS: Surgical interventions have been carried out in the Larsen syndrome and Morquio syndrome type A, resepectively. OUTCOMES: The earliest the diagnosis is, the better the results are. The necessity to diagnose children in their first year of life has many folds, firstly the management would be in favor of the child's growth and development and secondly, the prognosis could be clearer to the family and the medical staff as well. Our current paper is to sensitize paediatricians, physicians and orthopedic surgeons regarding the necessity to detect the aetiological understanding in every child who manifests a constellation of malformation complex. LESONS: Scoliosis and kyphosis/kyphoscoliosis are not a diagnosis in themselves. Such deformities are mostly a symptom complex correlated to dozens of types of syndromic associations. The rate curve progression and the final severity of congenital spine tilting are related to 3 factors: (a) the type of vertebral malformation present, (b) the patient's phenotype, and

[Detection of Coxiella burnetii in dairy cattle bulk tank milk and single tank milk samples by confirmatory testing]. / Nachweis von Coxiella burnetii beim Rind in Tank- und Einzelmilchproben im Rahmen von Abklärungsuntersuchungen.

Q fever is a worldwide zoonotic disease caused by the pathogen Coxiella (C.) burnetii. A wide range of animal species is susceptible to this intracellular bacterium with great importance in ruminants. Human infections occur mainly by airborne transmission. C burnetii was detected in animal products such as raw milk, raw-milk cheese and butter prepared from raw milk as well as in the meat of infected animals. In cattle milk, the pathogen was detected up to 13 months after calving. The risk of human foodborne C. Burnetii infection is still considered to be low, but cannot be completely ruled out and remains under discussion. The aim of this study was to compare different laboratory diagnostic methods for C. burnetii in milk sample. The bulk tank and individual milk samples were sent and studied at the National Reference Laboratory for Q-fever in the context of confirmatory laboratory testing after clinical suspicion or retesting of previously antibody detection was in the analysis of 888 individual milk samples a match of 93.3% (Cohen-kappa). A total of 173 bulk milk samples and 2,807 individual milk samples from bovine herds for the presence of C. burnetii DNA and antibodies were tested against the pathogen. The pathogen was detected in 62.5% of the bulk milk samples and up to 60% in individual milk samples. The highest proportion of positive bulk milks was determined as 68.3% in 2012. In individual milk samples, the highest proportion of seropositive samples was 62.2%.

Evaluation of an Erns-based enzyme-linked immunosorbent assay to distinguish Classical swine fever virus-infected pigs from pigs vaccinated with CP7_E2alf.

Infections with Classical swine fever virus (CSFV) are a major economic threat to pig production. To combat CSF outbreaks and to maintain trade, new marker vaccines were developed that allow differentiation of infected from vaccinated animals (DIVA principle). The chimeric pestivirus CP7_E2alf was shown to be safe and efficacious. Its DIVA strategy is based on the detection of CSFV E(rns)-specific antibodies that are only developed on infection. However, for the new marker vaccine to be considered a valuable control tool, a validated discriminatory assay is needed. One promising candidate is the already commercially available enzyme-linked immunosorbent assay, PrioCHECK CSFV E(rns) ELISA (Prionics BV, Lelystad, The Netherlands). Four laboratories of different European Union member states tested 530 serum samples and country-specific field sera from domestic pigs and wild boar. The ELISA displayed a good robustness. However, based on its reproducibility and repeatability, ranges rather than single values for diagnostic sensitivity and specificity were defined. The ELISA displayed a sensitivity of 90-98% with sera from CSFV-infected domestic pigs. A specificity of 89-96% was calculated with sera from domestic pigs vaccinated once with CP7_E2alf. The ELISA detected CSFV infections in vaccinated domestic pigs with a sensitivity of 82-94%. The sensitivity was lower with sera taken ≤21 days post-challenge indicating that the stage of CSFV infection had a considerable influence on testing. Taken together, the PrioCHECK CSFV E(rns) ELISA can be used for detection of CSFV infections in CP7_E2alf-vaccinated and nonvaccinated domestic pig populations, but should only be applied on a herd basis by testing a defined number of animals.

[Aspects seroepidemiological studies on Q fever in unvaccinated dairy cattle herds]. / Aspekte seroepidemiologischer Untersuchungen zum Q-Fieber in nicht geimpften Rinderbeständen.

Q fever is a zoonosis distributed worldwide and important in human as well as in veterinary medicine in Germany. In Baden-Wurttemberg, the pathogen is endemic. Usually Q fever is associated with infected sheep flocks. In contrast, however, in the animal disease reporting system (TSN) 88.1% of all listed Q fever infections during the last 12 years have been registered in cattle. Accordingly, in Baden-Württemberg and Freudenstadt 78.3 and, respectively, 62.5% of the Q fever cases were from cattle. Long term studies on appearance of Coxiella burnetii in normal herds of cattle are missing. Increasing vaccination of cattle herds against Q fever with the vaccine approved in Germany (no marker vaccine) complicates the future opportunities to gain data from serological studies. In the present study, a total of 1640 bovine sera taken from unvaccinated, clini- tion against C burnetii for analysis and comparison. The results show, depending on the test, a seroprevalence of 4.3% to 7.4%. Seasonal comparison revealed a significant increase of up to 9%.The month with the highest seroprevalence aver aged over three years was June with a prevalence of 24.7%. Overall, the findings of this study demonstrate that even the high number of entries does not fully capture the true prevalence of Q fever in cattle herds.

Lungworm Infections in German dairy cattle herds--seroprevalence and GIS-supported risk factor analysis.

In November 2008, a total of 19,910 bulk tank milk (BTM) samples were obtained from dairy farms from all over Germany, corresponding to about 20% of all German dairy herds, and analysed for antibodies against the bovine lungworm Dictyocaulus viviparus by use of the recombinant MSP-ELISA. A total number of 3,397 (17.1%; n = 19,910) BTM samples tested seropositive. The prevalences in individual German federal states varied between 0.0% and 31.2% positive herds. A geospatial map was drawn to show the distribution of seropositive and seronegative herds per postal code area. ELISA results were further analysed for associations with land-use and climate data. Bivariate statistical analysis was used to identify potential spatial risk factors for dictyocaulosis. Statistically significant positive associations were found between lungworm seropositive herds and the proportion of water bodies and grassed area per postal code area. Variables that showed a statistically significant association with a positive BTM test were included in a logistic regression model, which was further refined by controlled stepwise selection of variables. The low Pseudo R(2) values (0.08 for the full model and 0.06 for the final model) and further evaluation of the model by ROC analysis indicate that additional, unrecorded factors (e.g. management factors) or random effects may substantially contribute to lungworm infections in dairy cows. Veterinarians should include lungworms in the differential diagnosis of respiratory disease in dairy cattle, particularly those at pasture. Monitoring of herds through BTM screening for antibodies can help farmers and veterinarians plan and implement appropriate control measures.