Hepatorenal dysfunction in patients with chronic thromboembolic pulmonary hypertension.

Background: Cardiac interactions with organs such as the liver or kidneys have been described in different cardiovascular diseases. However, the clinical relevance of hepatorenal dysfunction in chronic thromboembolic pulmonary hypertension (CTEPH) remains unclear. We determined the association of hepatorenal dysfunction (measured using the Model for End-stage Liver Disease Sodium [MELDNa] score) with right heart function and survival in patients with CTEPH. Methods: We analyzed all patients with CTEPH in the Giessen Pulmonary Hypertension Registry who had available MELDNa scores and were not taking vitamin K antagonists. The MELDNa score was calculated as MELD score - serum Na - (0.025 * MELD score * (140 - serum Na)) + 140; the MELD score was calculated as 10*(0.957*ln(creatinine)+0.378*ln(bilirubin)+1.12*ln(International Normalized Ratio))+6.43. Results: Seventy-two patients were included (74% female; median [Q1, Q3] MELDNa: 9 [6, 11]). MELDNa correlated well with right atrial and ventricular function and pulmonary hemodynamics. Forward regression analysis revealed that hepatorenal dysfunction mainly depends on right atrial strain and tricuspid regurgitation, but not right ventricular systolic dysfunction. Hepatorenal dysfunction predicted mortality at baseline and follow-up (adjusted hazard ratios [95% confidence intervals] per unit increase of MELDNa: 1.6 [1.1, 2.4] and 1.8 [1.1, 2.9], respectively). Changes in hepatorenal function also predicted mortality. Conclusion: Hepatorenal dysfunction in CTEPH is primarily associated with venous congestion rather than cardiac forward failure. As a surrogate parameter for hepatorenal dysfunction, MELDNa is a simple method to identify at-risk patients at baseline and follow-up.

Modelling the monthly abundance of Culicoides biting midges in nine European countries using Random Forests machine learning.

BACKGROUND: Culicoides biting midges transmit viruses resulting in disease in ruminants and equids such as bluetongue, Schmallenberg disease and African horse sickness. In the past decades, these diseases have led to important economic losses for farmers in Europe. Vector abundance is a key factor in determining the risk of vector-borne disease spread and it is, therefore, important to predict the abundance of Culicoides species involved in the transmission of these pathogens. The objectives of this study were to model and map the monthly abundances of Culicoides in Europe. METHODS: We obtained entomological data from 904 farms in nine European countries (Spain, France, Germany, Switzerland, Austria, Poland, Denmark, Sweden and Norway) from 2007 to 2013. Using environmental and climatic predictors from satellite imagery and the machine learning technique Random Forests, we predicted the monthly average abundance at a 1 km2 resolution. We used independent test sets for validation and to assess model performance. RESULTS: The predictive power of the resulting models varied according to month and the Culicoides species/ensembles predicted. Model performance was lower for winter months. Performance was higher for the Obsoletus ensemble, followed by the Pulicaris ensemble, while the model for Culicoides imicola showed a poor performance. Distribution and abundance patterns corresponded well with the known distributions in Europe. The Random Forests model approach was able to distinguish differences in abundance between countries but was not able to predict vector abundance at individual farm level. CONCLUSIONS: The models and maps presented here represent an initial attempt to capture large scale geographical and temporal variations in Culicoides abundance. The models are a first step towards producing abundance inputs for R0 modelling of Culicoides-borne infections at a continental scale.

Monthly variation in the probability of presence of adult Culicoides populations in nine European countries and the implications for targeted surveillance.

BACKGROUND: Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) are small hematophagous insects responsible for the transmission of bluetongue virus, Schmallenberg virus and African horse sickness virus to wild and domestic ruminants and equids. Outbreaks of these viruses have caused economic damage within the European Union. The spatio-temporal distribution of biting midges is a key factor in identifying areas with the potential for disease spread. The aim of this study was to identify and map areas of neglectable adult activity for each month in an average year. Average monthly risk maps can be used as a tool when allocating resources for surveillance and control programs within Europe. METHODS: We modelled the occurrence of C. imicola and the Obsoletus and Pulicaris ensembles using existing entomological surveillance data from Spain, France, Germany, Switzerland, Austria, Denmark, Sweden, Norway and Poland. The monthly probability of each vector species and ensembles being present in Europe based on climatic and environmental input variables was estimated with the machine learning technique Random Forest. Subsequently, the monthly probability was classified into three classes: Absence, Presence and Uncertain status. These three classes are useful for mapping areas of no risk, areas of high-risk targeted for animal movement restrictions, and areas with an uncertain status that need active entomological surveillance to determine whether or not vectors are present. RESULTS: The distribution of Culicoides species ensembles were in agreement with their previously reported distribution in Europe. The Random Forest models were very accurate in predicting the probability of presence for C. imicola (mean AUC = 0.95), less accurate for the Obsoletus ensemble (mean AUC = 0.84), while the lowest accuracy was found for the Pulicaris ensemble (mean AUC = 0.71). The most important environmental variables in the models were related to temperature and precipitation for all three groups. CONCLUSIONS: The duration periods with low or null adult activity can be derived from the associated monthly distribution maps, and it was also possible to identify and map areas with uncertain predictions. In the absence of ongoing vector surveillance, these maps can be used by veterinary authorities to classify areas as likely vector-free or as likely risk areas from southern Spain to northern Sweden with acceptable precision. The maps can also focus costly entomological surveillance to seasons and areas where the predictions and vector-free status remain uncertain.

Spatial and temporal variation in the abundance of Culicoides biting midges (Diptera: Ceratopogonidae) in nine European countries.

BACKGROUND: Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) are vectors of bluetongue virus (BTV), African horse sickness virus and Schmallenberg virus (SBV). Outbreaks of both BTV and SBV have affected large parts of Europe. The spread of these diseases depends largely on vector distribution and abundance. The aim of this analysis was to identify and quantify major spatial patterns and temporal trends in the distribution and seasonal variation of observed Culicoides abundance in nine countries in Europe. METHODS: We gathered existing Culicoides data from Spain, France, Germany, Switzerland, Austria, Denmark, Sweden, Norway and Poland. In total, 31,429 Culicoides trap collections were available from 904 ruminant farms across these countries between 2007 and 2013. RESULTS: The Obsoletus ensemble was distributed widely in Europe and accounted for 83% of all 8,842,998 Culicoides specimens in the dataset, with the highest mean monthly abundance recorded in France, Germany and southern Norway. The Pulicaris ensemble accounted for only 12% of the specimens and had a relatively southerly and easterly spatial distribution compared to the Obsoletus ensemble. Culicoides imicola Kieffer was only found in Spain and the southernmost part of France. There was a clear spatial trend in the accumulated annual abundance from southern to northern Europe, with the Obsoletus ensemble steadily increasing from 4000 per year in southern Europe to 500,000 in Scandinavia. The Pulicaris ensemble showed a very different pattern, with an increase in the accumulated annual abundance from 1600 in Spain, peaking at 41,000 in northern Germany and then decreasing again toward northern latitudes. For the two species ensembles and C. imicola, the season began between January and April, with later start dates and increasingly shorter vector seasons at more northerly latitudes. CONCLUSION: We present the first maps of seasonal Culicoides abundance in large parts of Europe covering a gradient from southern Spain to northern Scandinavia. The identified temporal trends and spatial patterns are useful for planning the allocation of resources for international prevention and surveillance programmes in the European Union.

Effects of temperature and photoperiod on the development of overwintering immature Culicoides chiopterus and C. dewulfi.

In areas with harsh winters, several studies have recorded low numbers - or even zero - adult Culicoides during the winter period followed by a population peak in the subsequent spring. This experimental study determined whether temperature, photoperiod, or the combination thereof affect the development of overwintering immature dung breeding Culicoides, resulting in this peak. Temperature had a significant impact on the development period of Culicoides chiopterus (Meigen), 1830 and C. dewulfi Goetghebuer, 1936. In treatments with constant high temperature, emergence occurred shortly after the beginning of the experiment (mean=day 9). In contrast, no individuals emerged from the other two treatments, as long as the temperatures were below 10°C. In these treatments, the emergence of Culicoides started when the temperature exceeded 20°C for some days (mean=day 33). There was no significant difference between the two photoperiods (February or April day length; 9 h:15 h [light:dark] vs. 13 h:11 h). Our results highlight the importance of temperature on the spring emergence of C. chiopterus and C. dewulfi, but the response to the four temperature-photoperiod treatments did not differ between the two species.

Physico-Chemical Characteristics of Culex pipiens sensu lato and Culex torrentium (Diptera: Culicidae) Breeding Sites in Germany.

Culex pipiens sensu lato (L.) and Culex torrentium Martini, 1925 are two widely distributed mosquito species in Europe. Due to difficulties in morphological discrimination, the current knowledge on differences in the breeding ecology is fragmentary. Therefore, this study evaluated the relation between the presence-absence of both species at various types of breeding habitats in response to physico-chemical parameters, using a recently developed molecular assay that allows reliable species-specific typing of larvae and pupae. The results revealed that the two species often occur in sympatry at the studied breeding sites, and there were no substantial differences concerning presence-absence of the two species with regard to the various environmental parameters investigated.

Topsoil conditions correlate with the emergence rates of Culicoides chiopterus and Culicoides dewulfi (Diptera: Ceratopogonidae) from cowpats.

Culicoides chiopterus (Meigen), 1830 and Culicoides dewulfi Goetghebuer, 1936 (Diptera: Ceratopogonidae) are considered to develop exclusively in dung, but do not necessarily show an equal distribution and abundance on livestock farms in Northern Europe. Recent modelling studies identified soil parameters to explain these differences. The present study addressed the question whether topsoil conditions underneath cowpats correlate with the number of emerging C. chiopterus and C. dewulfi. We recorded the emergence of biting midges from 24 cowpats over a period of 4 weeks and analysed samples from the topsoil. In agreement with species distribution models based on remote data, our results detected the correlation of soil moisture, organic matter and soil texture with the number of emerging C. chiopterus and C. dewulfi. With increasing soil moisture, the number of emerging adults increased for both species and the amount of organic matter was positively correlated with the number of emerging C. chiopterus. In contrast, soil textures showed conflicting results, i.e. a positive and negative relationship with the same variables. According to our results, soil underneath dung can explain the number of emerging Culicoides species. The knowledge of these effects might improve the interpretation of large-scaled distribution models for dung-breeding biting midges.

Culicoides biting midge density in relation to the position and substrate temperature in a cattle dung heap.

Dung heaps offer warm breeding sites, which might be suitable for a continuing development or even emergence at low air temperatures in winter. Therefore, this study collected substrate samples from the outer surface and core of a cattle dung heap at the beginning of the winter period. We aimed to analyze the density of immature Culicoides in relation to substrate position and temperature. We took samples from the outer layer and core of the dung heap at different heights. Floatation was used to extract Culicoides larvae from the dung heap samples. In order to rear larvae individually, we separated them in glass tubes. A total of 229 Culicoides larvae were extracted from the dung heap samples. Highest densities (99.1% of all larvae) were recorded for the outer layers of the dung heap but hardly any in the core (0.9% of all individuals). While the density of larvae was negatively correlated with increasing substrate temperatures, Culicoides larvae were found in a temperature range between 7.9 and 38.0 °C (mean 16.6 °C). Extracted larvae were reared to adults. All male individuals were identified as Culicoides obsoletus (Meigen), 1818 and all female individuals as C. obsoletus/Culicoides scoticus. It can be concluded that dung heaps offer temperature conditions, which allow the survival and probably also the development to adults for immature Culicoides also under harsh climate conditions in winter.

Impact of flooding on the immature stages of dung-breeding Culicoides in Northern Europe.

In Northern Europe, dung-breeding Culicoides (e.g., Culicoides chiopterus (Meigen 1830) and Culicoides dewulfi (Goetghebuer 1936)) are considered to be important vectors of the Bluetongue virus and Schmallenberg virus. The interpretation of their distribution is difficult due to the lack of knowledge about their ecology. Previously, soil moisture and especially flooding were identified as important factors that influence the development of several biting-midge species. Therefore, this experimental study addressed the question whether flooding has a negative impact on the development of immature stages of Obsoletus group species. Ten cowpats were collected, and each was divided into four quarters and kept at different moisture regimes in a greenhouse: (1) "dry" (no water added), (2) "control" (regularly moistened), (3) "alternately flooded" and (4) "permanently flooded", to compare Culicoides emergence. Flooding had a significant negative impact on the emergence of Culicoides. No individuals emerged from the "permanently flooded" treatment and only two individuals were sampled from the "alternately flooded" treatment. In contrast, the total emergence from the non-flooded samples in the "dry" (96 individuals, 38.6% of all Culicoides) and "control" (151 individuals, 60.6% of all biting midges) treatments was considerably higher. Biting midges were predominantly identified as C. dewulfi (161 individuals, 64.7% of all Culicoides) and C. chiopterus (63 individuals, 25.3% of all Culicoides). There were no significant differences in emergence between the "dry" and "control" treatments. Our results highlight the importance of soil moisture on the distribution of C. chiopterus and C. dewulfi. Regarding physiological and behavioural adaptations of other Culicoides species, we argue that pupae of C. chiopterus and C. dewulfi are in danger of drowning when breeding sites are flooded as they cannot float. On the contrary, our results indicate that desiccation might not be harmful to these species.

Field evaluation of four widely used mosquito traps in Central Europe.

BACKGROUND: To monitor adult mosquitoes several trapping devices are available. These are differently constructed and use various mechanisms for mosquito attraction, thus resulting in different trapping sensitivities and efficacies for the various species. Mosquito monitoring and surveillance programs in Europe use various types of mosquito traps, but only a few comparisons have been conducted so far. This study compared the performance of four commercial trapping devices, which are commonly used in Europe. METHODS: Four different traps, Biogents Sentinel trap (BG trap), Heavy Duty Encephalitis Vector Survey trap (EVS trap), Centres for Disease Control miniature light trap (CDC trap) and Mosquito Magnet Patriot Mosquito trap (MM trap) were compared in a 4 × 4 latin square study. In the years 2012 and 2013, more than seventy 24-hour trap comparisons were conducted at ten different locations in northern and southern Germany, representing urban, forest and floodplain biotopes. RESULTS: Per 24-hour trapping period, the BG trap caught the widest range of mosquito species, the highest number of individuals of the genus Culex as well as the highest number of individuals of the species Ochlerotatus cantans, Aedes cinereus/geminus, Oc. communis and Culex pipiens/torrentium. The CDC trap revealed best performance for Aedes vexans, whereas the MM trap was most efficient for mosquitoes of the genus Anopheles and the species Oc. geniculatus. The EVS trap did not catch more individuals of any genus or species compared to the other three trapping devices. The BG trap caught the highest number of individuals per trapping period in urban environments as well as in wet forest, while the CDC trap caught the highest number of individuals in the floodplain biotopes. Additionally, the BG trap was most efficient for the number of mosquito species in urban locations. CONCLUSION: The BG trap showed a significantly better or similar performance compared to the CDC, EVS or MM trap with regard to trapping efficacy for most common mosquito species in Germany, including diversity of mosquito species and number of mosquitoes per trapping period. Thus, the BG trap is probably the best solution for general monitoring or surveillance programs of adult mosquitoes in Central Europe.

The use of morphometric wing characters to discriminate female Culex pipiens and Culex torrentium.

The reliability of the length of wing radial vein r(2/3) as a character for the morphological discrimination of the two potential arbovirus vectors Culex pipiens s.s. and Cx. torrentium from Germany was reassessed, after this character had been neglected for more than 40 years. Additionally, multivariate morphometric analyses were applied to evaluate wing shape variation between both species. Although high-throughput molecular tools are now available to differentiate the two species, a simple, low-cost routine alternative may be useful in the absence of a molecular laboratory, such as under semi-field conditions. A thin-plate splines transformation confirmed that primarily the shrinkage of vein r(2/3) is responsible for the wing differences between the two species. In the bivariate analysis, the r(2/3)/r3 indices of Cx. pipiens s.s. and Cx. torrentium were 0.185 and 0.289, respectively, resulting in a correct classification of more than 91% of all tested specimens. Using the absolute length of vein r(2/3) alone still allowed for more than 90% accurate discrimination. Furthermore, classification accuracy of linear discriminant analysis exceeded 97%.

Impact of mechanical disturbance on the emergence of Culicoides from cowpats.

The outbreaks of Bluetongue virus and Schmallenberg virus between 2006 and 2012 highlighted the need for control methods for north-western Palaearctic biting midge species. There is especially a demand for alternative control methods without application of insecticides. Therefore, the objective of this paper was to evaluate the impact of mechanical disturbance as a control method for Culicoides biting midges in cowpats. We used a trash rake to destroy the compact structure of cowpats and spread the dung over the ground. We expected the disturbance to cause a faster alteration of physical characteristics and drying of the dung, resulting in lower emergence from disturbed cowpats. Emerging biting midges were collected with emergence traps. The number of emerged Culicoides was compared between treated and control cowpats. A total of 12,979 biting midges emerged, all belonging to the Obsoletus group. The majority of the 6,758 male individuals were identified as Culicoides chiopterus (Meigen, 1830) (91.4%), followed by Culicoides dewulfi (Goetghebuer, 1936) (6.5%), and a small number of Culicoides scoticus (Downes & Kettle, 1952) (1.2%). Our results showed no significant differences between the emergence of Culicoides (males and females) from disturbed and control cowpats. The lack of differences is discussed in light of the time period chosen for the experiment and the climatic conditions (especially low temperatures) during the study period. The climatic conditions during the study did not favour desiccation effects and therefore did not cause a severe alteration of the dung and the biotic parameters. In conclusion, immature Culicoides showed a high tolerance against mechanical disturbance.