Current Transition Practice for Primary Immunodeficiencies and Autoinflammatory Diseases in Europe: a RITA-ERN Survey.

BACKGROUND: Due to the absence of curative treatments for inborn errors of immunity (IEI), children born with IEI require long-term follow-up for disease manifestations and related complications that occur over the lifespan. Effective transition from pediatric to adult services is known to significantly improve adherence to treatment and long-term outcomes. It is currently not known what transition services are available for young people with IEI in Europe. OBJECTIVE: To understand the prevalence and practice of transition services in Europe for young people with IEI, encompassing both primary immunodeficiencies (PID) and systemic autoinflammatory disorders (AID). METHODS: A survey was generated by the European Reference Network on immunodeficiency, autoinflammatory, and autoimmune diseases Transition Working Group and electronically circulated, through professional networks, to pediatric centers across Europe looking after children with IEI. RESULTS: Seventy-six responses were received from 52 centers, in 45 cities across 17 different countries. All services transitioned patients to adult services, mainly to specialist PID or AID centers, typically transferring up to ten patients to adult care each year. The transition process started at a median age of 16-18 years with transfer to the adult center occurring at a median age of 18-20 years. 75% of PID and 68% of AID centers held at least one joint appointment with pediatric and adult services prior to the transfer of care. Approximately 75% of PID and AID services reported having a defined transition process, but few centers reported national disease-specific transition guidelines to refer to. CONCLUSIONS: Transition services for children with IEI in Europe are available in many countries but lack standardized guidelines to promote best practice.

Slaughterhouse Visual and Palpation Method for Estimating the Economic Damage of Porcine Proliferative Enteropathy (PPE).

BACKGROUND: Ileitis is a wasting disease of pigs. Clinical symptoms are diarrhea in growing pigs, wasting and reduced performance. Ileitis is ubiquitous in pig producing countries all around the world. It is estimated that the economic losses caused by the disease are USD 4.65 per fattening pig, and American pig farmers lose USD 56.1 million annually. It has become necessary to develop a slaughterhouse ileitis monitoring method that is simple, feasible to perform at modern slaughter lines, leads to immediate results and is cost effective. The practical experiences of applying the method are presented below. METHODS: Our studies were performed on pig herds and slaughterhouses in Central European countries (Hungary, Romania, Poland, Croatia, and Slovakia). Experiences were evaluated based on visual and palpation. The results of our investigations were evaluated by a scoring method. Authors made histological and immunohistochemical examinations of investigated ileums. The hypothetic economic losses due to the disease in each farm were determined by estimating the loss of profit according to Holtkamp's presentation in 2019. RESULTS: The essentials of the method we have developed are: it can be performed during slaughterhouse processing, it does not interfere with or make it impossible to carry out normal technological processes, and the results of the procedure are correlated with the results of other laboratory diagnostic tests for ileitis (histology, immunohistochemistry, herd serology, fecal PCR). It is noteworthy that the results of the method can be used to immediately estimate the impact of Lawsonia intracellularis infection on the performance of the herd from which the slaughter animals come. CONCLUSION: Using the results of the slaughtered pigs' visual and tactile examination at the slaughterhouse, the magnitude of the loss caused by Lawsonia intracellularis infection can be estimated quickly and accurately, and the return on investment of the strategy to be applied can be accurately planned.

Development and validation of a flow cytometry antibody test for Lawsonia intracellularis.

Lawsonia intracellularis is the etiologic agent of porcine proliferative enteropathy (PPE), an inflammatory bowel disease with a major economic impact on the pig industry. The serological diagnosis of PPE can be performed using Blocking or Indirect ELISA, Immunoperoxidase Monolayer Assay (IPMA) and Indirect Fluorescence Antibody Test (IFAT). Here, we designed a most sophisticated immunological method for the detection of porcine anti-L. intracellularis IgGs, named Flow Cytometry Antibody Test - FCAT. This assay uses whole, live-attenuated L. intracellularis bacteria derived from a commercial vaccine. For the assay, we set up the optimal antigen concentration (106 bacterium/assay), primary antibody dilution (1:100), time of incubation (20 min), antigen stability (15 days), precision (coefficient of variation - CV < 10%), reproducibility (CV ≤ 13%) and Receiver Operating Characteristic (ROC). When using a cut-off of >15.15% for FCAT, we determined that it showed a sensitivity of 98.8% and specificity of 100%. The rate of agreement with IPMA was 84.09% with a kappa index of 0.66. FCAT was used to screen 1,000 sera from non-vaccinated pigs housed in 22 different farms and we found that 730 pigs (73%) from 16 farms (72.7%) had L. intracellularis IgG. This high prevalence confirms that L. intracellularis is endemic on Brazilian pig farms. Finally, we determined that FCAT is an easy to perform diagnostic assay and we would highly recommend it for: i) seroepidemiological studies; ii) evaluation of infection dynamics; and iii) characterization of the humoral response profile induced by vaccines.

A mesoionic carbene complex of manganese in five oxidation states.

Reaction between a carbazole-based mesoionic carbene ligand and manganese(II) iodide results in the formation of a rare air-stable manganese(IV) complex after aerobic workup. Cyclic voltammetry reveals the complex to be stable in five oxidation states. The electronic structure of all five oxidation states is elucidated chemically, spectroscopically (NMR, high-frequency EPR, UV-Vis, MCD), magnetically, and computationally (DFT, CASSCF).

Correlation of Lawsonia intracellularis positivity in quantitative PCR and herd factors in European pig herds.

BACKGROUND: Lawsonia intracellularis is causing diarrhea, poor growth and sudden death in pigs. It can be found in most pig populations leading to large economic losses worldwide. Many potential risk factors for the occurrence of disease or seropositivity have been described. The current study therefore focused on herd characteristics in European countries associated with direct detection of the pathogen determined by quantitative polymerase chain reaction. RESULTS: A median number of less than 30 nursery pigs per pen was correlated to less positive nursery pigs (p < 0.01) and generally less samples positive per herd (p < 0.05) as well as a lower median of genome equivalents determined per herd (p < 0.05). Routine use of zinc oxide at/ around weaning, which was mentioned by 41.0% of all farmers, was correlated to higher number of positive nursery pigs (p < 0.01) as well as higher median genome equivalents determined per herd (p < 0.05). Slatted flooring of more than 78.0% of the surface in nursery units was correlated to lower number of positive animals (p < 0.05) and a lower median of genome equivalents per herd (p < 0.05). A weight of more than 7.8 kg at weaning was correlated to a higher number of positive growing pigs (p < 0.05) as well as general higher number of positive samples/ herd (p < 0.01). CONCLUSIONS: Weaning and subsequent accommodation of nursery pigs seem to be of particular importance in prevention of infection with Lawsonia intracellularis and the spread of the pathogen within the herd.

Prevalence of Lawsonia intracellularis in pig herds in different European countries.

BACKGROUND: Lawsonia intracellularis causes large economic losses in the pig industry worldwide. Pigs suffer from reduced daily weight gain, poor feed conversion ratio and increased mortality. The number of affected animals and herds in Europe remains unknown. This study will provide an overview of the prevalence of Lawsonia intracellularis in herds with a history of diarrhoea in different European countries and thereby identify country specific differences. RESULTS: Out of the 144 herds sampled in Germany, Denmark, Spain, the Netherlands and the United Kingdom, 90.3% (79.2-100.0%) contained at least one positive faecal sample on quantitative polymerase chain reaction (qPCR). Of the 6450 nursery, growing and finishing pigs of the previously mentioned herds, 26.2% (15.9-41.5%) of the animals were tested positive in faecal samples. Enzyme linked immunosorbent assay (ELISA) results of 60 herds were 91.7% (70-100%) positive. The percentage of positive samples in these 1791 blood samples was 31.6% (20.3-51.0%). Herd prevalence did not differ significantly by qPCR or ELISA. Significant differences between the countries were found regarding: Within-herd prevalence- qPCR: Samples from Denmark were more often positive than samples of Spain or the United Kingdom. Within-herd prevalence- ELISA: Samples from Denmark were more often positive than samples from Spain and the Netherlands. Affected age category- qPCR: Nursery pigs in Denmark were more often positive and shed more genome equivalents than nursery pigs in the other countries. Concentration of detected genome equivalents- qPCR: The concentration of genome equivalents from Lawsonia intracellularis in herds in Denmark was higher compared to all other countries. CONCLUSION: A widespread of Lawsonia intracellularis in the six European countries was confirmed, whereby a large part of the positive animals only excreted small amounts of genome equivalents. Country specific differences were found with Denmark in particular diagnosing more Lawsonia intracellularis then the other countries. Herd data collected in this study needs to be analysed to get more information about possible reasons for the differences found between the countries.

Occurrence and severity of lung lesions in slaughter pigs vaccinated against Mycoplasma hyopneumoniae with different strategies.

Different vaccination strategies against Mycoplasma hyopneumoniae have been adopted worldwide. Reports from the field indicate varying levels of protection among currently available vaccines. The goal of the present study was to compare the efficacies of three widespread commercial vaccination strategies against M. hyopneumoniae under field conditions. 20 farms were included. 14 farms used different single dose vaccines (vaccine 1 [V1], 8 herds; vaccine 2 [V2], 6 herds); another 6 farms (V3) used a two dose vaccination strategy. Gross lesions of 854 lungs and histopathology from 140 lungs were quantified, and a quantitative PCR was applied to detect M. hyopneumoniae and porcine circovirus 2 (PCV2) DNA in lung tissue (n=140). In addition, porcine reproductive and respiratory disease virus (PRRSV), swine influenza virus (SIV), Actinobacillus pleuropneumoniae, Haemophilus parasuis and Pasteurella multocida were tested by qualitative PCR. 53% of lungs were positive for M. hyopneumoniae. 55.9% of lungs showed macroscopic enzootic pneumonia (EP)-like lesions. Lung lesion scores (P<0.001) and M. hyopneumoniae-loads (P<0.008) differed significantly among the vaccination groups, with the most severe cases and highest amounts occurring in V1. Histological alterations differed (P<0.001) between V1 and V3. Lung lesion scores and histopathological changes were significantly correlated, with prevalence and load of M. hyopneumoniae indicating that the applied diagnostic tools are valuable in confirming the prevalence and severity of M. hyopneumoniae infections. Comparing different vaccination strategies against M. hyopneumoniae indicates varying levels of protection. M. hyopneumoniae is still a major problem despite the widely applied vaccination.

Analysis of bacterial load and prevalence of mixed infections with Lawsonia intracellularis, Brachyspira hyodysenteriae and/or Brachyspira pilosicoli in German pigs with diarrhoea.

Lawsonia (L.) intracellularis, Brachyspira (B.) hyodysenteriae and B. pilosicoli are important pathogens in domestic pig production world-wide, responsible for porcine intestinal adenomatosis, swine dysentery, and porcine intestinal spirochetosis, respectively. Conventional PCR is the major diagnostic tool in the detection of the three pathogens, but the sole detection of bacterial DNA might lead to misinterpretations of results with respect to their clinical relevance, especially with mixed infections. Thus, the present study targeted the detection and quantification of the three pathogens in samples from herds with a case history of diarrhoea. Herds and samples were selected by the practitioners on a voluntary basis. Results were based on 1176 individual samples from 95 herds from Southern Germany. The pathogens were detected simultaneously by multiplex real-time PCR. The overall prevalence for L. intracellularis, B. hyodysenteriae and B. pilosicoli was 12.6%, 8.4% and 3.2% in faecal samples and 48.4%, 24.2% and 31.6% in herds, respectively. Sixty one percent, 82.6%, and 73.4% of herds positive for L. intracellularis, B. hyodysenteriae, and B. pilosicoli, respectively, had mixed infections. Median log values of DNA equivalents/g of faeces for L. intracellularis, B. hyodysenteriae and B. pilosicoli were 3.3, 5.9 and 3.2, with maxima of 8.3, 8.0 and 6.3, respectively. Within herd prevalence of B. hyodysenteriae and B. pilosicoli as well as the load of B. hyodysenteriae were significantly associated with the severity of diarrhoea.

[Haemorrhagic septicaemia in a pig caused by extraintestinal pathogenic Escherichia coli (ExPEC) as a differential diagnosis in classical swine fever--case report and review of the literature]. / Durch Extraintestinal pathogene Escherichia coli (ExPEC) bedingte haemorrhagische Septikaemie beim Schwein als Differentialdiagnose zur Klassischen Schweinepest--Fallbericht und Literaturübersicht.

Domestic pig herds in some regions of Germany are permanently threatened by Classical Swine Fever. In the case of suspicion, a series of infectious and non infectious causes has to be excluded. The present paper describes a case of Escherichia coli septicaemia, with clinical and pathological symptoms that could not be differentiated from European or African Swine Fever. The E. coli strain could not be classified by standard serotyping. Virulence factors common for ETEC (enterotoxic E. coli) or EDEC (edema-disease E. coli) were not detected. Instead, we found P-fimbriae and aerobactin, thus characterising this strain as an extraintestinal pathogenic strain. Such strains have sporadicly been reported as the cause of septicaemia in piglets or weaners, but the present case is the first report of an E. coli-associated septicaemia in an adult pig. This case shows that extraintestinal pathogenic E. coli can be the cause of severe septicaemia and haemorrhagia. They thus have to be considered as a further differential diagnosis in swine fever.