The SWI/SNF chromatin remodelling complex regulates pancreatic endocrine cell expansion and differentiation in mice in vivo.

AIMS/HYPOTHESIS: Strategies to augment functional beta cell mass include directed differentiation of stem cells towards a beta cell fate, which requires extensive knowledge of transcriptional programs governing endocrine progenitor cell differentiation in vivo. We aimed to study the contributions of the Brahma-related gene-1 (BRG1) and Brahma (BRM) ATPase subunits of the SWI/SNF chromatin remodelling complex to endocrine cell development. METHODS: We generated mice with endocrine progenitor-specific Neurog3-Cre BRG1 removal in the presence of heterozygous (Brg1Δendo;Brm+/-) or homozygous (double knockout: DKOΔendo) BRM deficiency. Whole-body metabolic phenotyping, islet function characterisation, islet quantitative PCR and histological characterisation were performed on animals and tissues postnatally. To test the mechanistic actions of SWI/SNF in controlling gene expression during endocrine cell development, single-cell RNA-seq was performed on flow-sorted endocrine-committed cells from embryonic day 15.5 control and mutant embryos. RESULTS: Brg1Δendo;Brm+/- mice exhibit severe glucose intolerance, hyperglycaemia and hypoinsulinaemia, resulting, in part, from reduced islet number; diminished alpha, beta and delta cell mass; compromised islet insulin secretion; and altered islet gene expression programs, including reductions in MAFA and urocortin 3 (UCN3). DKOΔendo mice were not recovered at weaning; however, postnatal day 6 DKOΔendo mice were severely hyperglycaemic with reduced serum insulin levels and beta cell area. Single-cell RNA-seq of embryonic day 15.5 lineage-labelled cells revealed endocrine progenitor, alpha and beta cell populations from SWI/SNF mutants have reduced expression of Mafa, Gcg, Ins1 and Ins2, suggesting limited differentiation capacity. Reduced Neurog3 transcripts were discovered in DKOΔendo endocrine progenitor clusters, and the proliferative capacity of neurogenin 3 (NEUROG3)+ cells was reduced in Brg1Δendo;Brm+/- and DKOΔendo mutants. CONCLUSIONS/INTERPRETATION: Loss of BRG1 from developing endocrine progenitor cells has a severe postnatal impact on glucose homeostasis, and loss of both subunits impedes animal survival, with both groups exhibiting alterations in hormone transcripts embryonically. Taken together, these data highlight the critical role SWI/SNF plays in governing gene expression programs essential for endocrine cell development and expansion. DATA AVAILABILITY: Raw and processed data for scRNA-seq have been deposited into the NCBI Gene Expression Omnibus (GEO) database under the accession number GSE248369.

Mitochondrial genetic diversity and phylogenetic relationships of Echinococcus multilocularis in Europe.

The cestode Echinococcus multilocularis is the causative agent of alveolar echinococcosis, a fatal zoonotic parasitic disease of the northern hemisphere. Red foxes are the main reservoir hosts and, likely, the main drivers of the geographic spread of the disease in Europe. Knowledge of genetic relationships among E. multilocularis isolates at a European scale is key to understanding the dispersal characteristics of E. multilocularis. Hence, the present study aimed to describe the genetic diversity of E. multilocularis isolates obtained from different host species in 19 European countries. Based on the analysis of complete nucleotide sequences of the cob, atp6, nad2, nad1 and cox1 mitochondrial genes (4,968 bp), 43 haplotypes were inferred. Four haplotypes represented 62.56 % of the examined isolates (142/227), and one of these four haplotypes was found in each country investigated, except Svalbard, Norway. While the haplotypes from Svalbard were markedly different from all the others, mainland Europe appeared to be dominated by two main clusters, represented by most western, central and eastern European countries, and the Baltic countries and northeastern Poland, respectively. Moreover, one Asian-like haplotype was identified in Latvia and northeastern Poland. To better elucidate the presence of Asian genetic variants of E. multilocularis in Europe, and to obtain a more comprehensive Europe-wide coverage, further studies, including samples from endemic regions not investigated in the present study, especially some eastern European countries, are needed. Further, the present work proposes historical causes that may have contributed to shaping the current genetic variability of E. multilocularis in Europe.

Parasite Spillover from Domestic Sheep to Wild Reindeer-The Role of Salt Licks.

Attraction sites are important for environmental pathogen transmission and spillover. Yet, their role in wildlife disease dynamics is often poorly substantiated. Herein, we study the role of salt licks as potential attraction sites for the spillover of gastrointestinal parasites from domestic sheep to wild reindeer. Eggs from the introduced sheep nematode Nematodirus battus were found in faecal samples of both species, suggestive of spillover. DNA metabarcoding of soil, collected at salt licks, revealed that N. battus, in addition to Teladorsagia circumcincta, were the most frequently occurring parasitic nematodes, with a significantly higher prevalence of nematodal DNA in salt lick soil compared to soil from control sites nearby. The finding of similar DNA haplotypes of N. battus in sheep, reindeer, and salt lick soil supports the hypothesis of spillover to reindeer via salt licks. More detailed investigation of the genetic diversity of N. battus across these hosts is needed to draw firm conclusions. Infection with these sheep nematodes could potentially explain a recently observed decline in the calf recruitment rate of the Knutshø reindeer herd. This study also supports the hypothesized role of artificial salt licks as hot spots for the transmission of environmentally persistent pathogens and illustrates the importance of knowledge about such attraction points in the study of disease in free-roaming animals.

Dynamic regulation of pancreatic ß cell function and gene expression by the SND1 coregulator in vitro.

The pancreatic ß cell synthesizes, packages, and secretes insulin in response to glucose-stimulation to maintain blood glucose homeostasis. Under diabetic conditions, a subset of ß cells fail and lose expression of key transcription factors (TFs) required for insulin secretion. Among these TFs is Pancreatic and duodenal homeobox 1 (PDX1), which recruits a unique subset of transcriptional coregulators to modulate its activity. Here we describe a novel interacting partner of PDX1, the Staphylococcal Nuclease and Tudor domain-containing protein (SND1), which has been shown to facilitate protein-protein interactions and transcriptional control through diverse mechanisms in a variety of tissues. PDX1:SND1 interactions were confirmed in rodent ß cell lines, mouse islets, and human islets. Utilizing CRISPR-Cas9 gene editing technology, we deleted Snd1 from the mouse ß cell lines, which revealed numerous differentially expressed genes linked to insulin secretion and cell proliferation, including limited expression of Glp1r. We observed Snd1 deficient ß cell lines had reduced cell expansion rates, GLP1R protein levels, and limited cAMP accumulation under stimulatory conditions, and further show that acute ablation of Snd1 impaired insulin secretion in rodent and human ß cell lines. Lastly, we discovered that PDX1:SND1 interactions were profoundly reduced in human ß cells from donors with type 2 diabetes (T2D). These observations suggest the PDX1:SND1 complex formation is critical for controlling a subset of genes important for ß cell function and is targeted in diabetes pathogenesis.

Pharmacokinetics of a long-acting subcutaneous eprinomectin injection in semi-domesticated reindeer (Rangifer tarandus tarandus) - A pilot study.

Reindeer (Rangifer tarandus tarandus) are exposed to the pathogenic parasitic nematode Elaphostrongylus rangiferi during grazing. The severity of disease is dose-dependent. Prophylactic anthelmintic treatment is needed to improve animal health and reindeer herding sustainability. Herds are traditionally only gathered once during the summer, requiring a drug with a persistent effect. In this study we investigated the suitability of long-acting eprinomectin, given as a single subcutaneous injection at 1 mg/kg bodyweight in adult reindeer and calves. Plasma and faeces concentrations were determined using ultra-high performance liquid chromatography high resolution mass spectrometry (UHPLC-HRMS). Plasma concentrations remained above the presumed effect level of 2 ng/mL for 80 days, demonstrating the drug's potential. Pharmacokinetic parameters were compared to other species using allometric scaling. Calves and adults had slightly different profiles. No viable faecal nematode eggs were detected during treatment. Eprinomectin was measurable in the reindeer faeces up to 100 days, which is of environmental concern.

The Chd4 Helicase Regulates Chromatin Accessibility and Gene Expression Critical for ß-Cell Function In Vivo.

The transcriptional activity of Pdx1 is modulated by a diverse array of coregulatory factors that govern chromatin accessibility, histone modifications, and nucleosome distribution. We previously identified the Chd4 subunit of the nucleosome remodeling and deacetylase complex as a Pdx1-interacting factor. To identify how loss of Chd4 impacts glucose homeostasis and gene expression programs in ß-cells in vivo, we generated an inducible ß-cell-specific Chd4 knockout mouse model. Removal of Chd4 from mature islet ß-cells rendered mutant animals glucose intolerant, in part due to defects in insulin secretion. We observed an increased ratio of immature-to-mature insulin granules in Chd4-deficient ß-cells that correlated with elevated levels of proinsulin both within isolated islets and from plasma following glucose stimulation in vivo. RNA sequencing and assay for transposase-accessible chromatin with sequencing showed that lineage-labeled Chd4-deficient ß-cells have alterations in chromatin accessibility and altered expression of genes critical for ß-cell function, including MafA, Slc2a2, Chga, and Chgb. Knockdown of CHD4 from a human ß-cell line revealed similar defects in insulin secretion and alterations in several ß-cell-enriched gene targets. These results illustrate how critical Chd4 activities are in controlling genes essential for maintaining ß-cell function. ARTICLE HIGHLIGHTS: Pdx1-Chd4 interactions were previously shown to be compromised in ß-cells from human donors with type 2 diabetes. ß-Cell-specific removal of Chd4 impairs insulin secretion and leads to glucose intolerance in mice. Expression of key ß-cell functional genes and chromatin accessibility are compromised in Chd4-deficient ß-cells. Chromatin remodeling activities enacted by Chd4 are essential for ß-cell function under normal physiological conditions.

Systematic Review and Modelling of Age-Dependent Prevalence of Toxoplasma gondii in Livestock, Wildlife and Felids in Europe.

Toxoplasma gondii is a zoonotic parasite of importance to both human and animal health. The parasite has various transmission routes, and the meat of infected animals appears to be a major source of human infections in Europe. We aimed to estimate T. gondii prevalence in a selection of animal host species. A systematic literature review resulting in 226 eligible publications was carried out, and serological data were analyzed using an age-dependent Bayesian hierarchical model to obtain estimates for the regional T. gondii seroprevalence in livestock, wildlife, and felids. Prevalence estimates varied between species, regions, indoor/outdoor rearing, and types of detection methods applied. The lowest estimated seroprevalence was observed for indoor-kept lagomorphs at 4.8% (95% CI: 1.8-7.5%) and the highest for outdoor-kept sheep at 63.3% (95% CI: 53.0-79.3%). Overall, T. gondii seroprevalence estimates were highest within Eastern Europe, whilst being lowest in Northern Europe. Prevalence data based on direct detection methods were scarce and were not modelled but rather directly summarized by species. The outcomes of the meta-analysis can be used to extrapolate data to areas with a lack of data and provide valuable inputs for future source attribution approaches aiming to estimate the relative contribution of different sources of T. gondii human infection.

The Chd4 subunit of the NuRD complex regulates Pdx1-controlled genes involved in ß-cell function.

Type 2 diabetes (T2D) is associated with loss of transcription factors (TFs) from a subset of failing ß-cells. Among these TFs is Pdx1, which controls the expression of numerous genes involved in maintaining ß-cell function and identity. Pdx1 activity is modulated by transcriptional coregulators and has recently been shown, through an unbiased screen, to interact with the Chd4 ATPase subunit of the nucleosome remodeling and deacetylase complex. Chd4 contributes to the maintenance of cellular identity and functional status of numerous different cell types. Here, we demonstrated that Pdx1 dynamically interacts with Chd4 under physiological and stimulatory conditions within islet ß-cells and established a fundamental role for Chd4 in regulating insulin secretion and modulating numerous Pdx1-bound genes in vitro, including the MafA TF, where we discovered Chd4 is bound to the MafA region 3 enhancer. Furthermore, we found that Pdx1:Chd4 interactions are significantly compromised in islet ß-cells under metabolically induced stress in vivo and in human donor tissues with T2D. Our findings establish a fundamental role for Chd4 in regulating insulin secretion and modulating Pdx1-bound genes in vitro, and disruption of Pdx1:Chd4 interactions coincides with ß-cell dysfunction associated with T2D.

The Contribution of Transcriptional Coregulators in the Maintenance of ß-cell Function and Identity.

Islet ß-cell dysfunction that leads to impaired insulin secretion is a principal source of pathology of diabetes. In type 2 diabetes, this breakdown in ß-cell health is associated with compromised islet-enriched transcription factor (TF) activity that disrupts gene expression programs essential for cell function and identity. TF activity is modulated by recruited coregulators that govern activation and/or repression of target gene expression, thereby providing a supporting layer of control. To date, more than 350 coregulators have been discovered that coordinate nucleosome rearrangements, modify histones, and physically bridge general transcriptional machinery to recruited TFs; however, relatively few have been attributed to ß-cell function. Here, we will describe recent findings on those coregulators with direct roles in maintaining islet ß-cell health and identity and discuss how disruption of coregulator activity is associated with diabetes pathogenesis.

PCR detection of Angiostrongylus vasorum in faecal samples of dogs and foxes.

The cardiovascular nematode Angiostrongylus vasorum is spreading in the fox and dog populations of northern Europe. A. vasorum can result in severe clinical manifestations in dogs; therefore, specific diagnosis is crucial for assessing its prevalence. In the present study, faecal samples from foxes and domestic dogs were tested by a new polymerase chain reaction (PCR) targeting the second internal transcribed region of the ribosomal DNA (ITS2) of A. vasorum. Initial isolation of faecal larvae by sieving facilitated the processing of larger sample volumes and allowed for the recovery of dead larvae from frozen samples. The sieve-PCR method enabled the identification of a single larva per 2 g of faecal sample and did not amplify DNA of a range of canine helminths, thus presenting a non-invasive tool for wildlife surveillance and for confirmative diagnosis in dogs.

Infection with brainworm (Elaphostrongylus rangiferi) in reindeer (Rangifer tarandus ssp.) in Fennoscandia.

Sami reindeer herders have considerable traditional knowledge about a neurological reindeer disease resembling elaphostrongylosis, but the causative agent was not identified prior to the description of the brainworm Elaphostrongylus rangiferi in Russia in 1958. Elaphostrongylosis was quickly recognised as a serious cause of reindeer morbidity and mortality. The ecology, epidemiology and pathophysiology of the disease were studied in Sweden and Norway during the 1960s and in particular the 1970s to 1990s. In Finland, elaphostrongylosis was not recognised as an important disease for Finnish reindeer husbandry, even though the presence of brainworm infection has been documented. Brainworm has an indirect lifecycle with snail and slug intermediate hosts. The free-living L1 larvae have extremely good freeze tolerance and can survive > 360 days at - 80 °C in water (solid ice). Even though reindeer brainworm is clearly well adapted to the Arctic chill, the lifecycle stages outside the reindeer final host are sped up at warmer environmental temperatures. Arctic summer temperatures are close to the developmental threshold of the parasite in the intermediate gastropod hosts (8-10 °C), and the parasite has typically had a 2-year life cycle. Disease outbreaks generally occur during the winter following the infection of reindeer with infected snails and slugs during the summer and autumn. Warmer summers result in faster development of brainworm larvae in the intermediate hosts. Clinical symptoms have been seen reported as early as August, such as in the outbreak in Trøndelag, Norway in 2018. The reindeer brainworm is also a cause of conflict between reindeer herders and small ruminant farmers, because it can cause severe disease in goats and sheep, which share pasture with reindeer. Many knowledge gaps remain if we wish to successfully predict and mitigate for large-scale outbreaks in a future with a predicted warmer, wetter and wilder climate.

Modeling Thermal Suitability for Reindeer (Rangifer tarandus ssp.) Brainworm (Elaphostrongylus rangiferi) Transmission in Fennoscandia.

The brainworm, Elaphostrongylus rangiferi, is a nematode which causes neurological disorders (elaphostrongylosis) in reindeer (Rangifer tarandus ssp.). Favorable climatic conditions have been inferred as the cause of sporadic outbreaks of elaphostrongylosis in Norway, supported by positive associations between observed outbreaks/intensity of infection and summer temperatures in the previous years. Climate warming which results in increased transmission of E. rangiferi therefore presents a risk to the health of semi-domesticated and wild reindeer in Fennoscandia (Norway, Sweden, and Finland), the health of co-grazing small ruminants, and the livelihoods of indigenous Sámi herders. As a first step toward developing climate change impact assessments for E. rangiferi, a degree-day model was developed for larval development in a range of gastropod hosts and applied to historic weather data. Predictions were validated by statistical and qualitative comparison against historic parasitological and outbreak records. The model predicted an overall increase in thermal suitability for E. rangiferi, which was statistically significant in the north and along the Scandinavian mountain ranges, where reindeer density is highest. In these regions annual cumulative temperature conditions are suitable for larval development within a single year, potentially changing E. rangiferi epidemiology from a 2-year transmission cycle to a 1-year transmission cycle. This is the first mechanistic model developed for E. rangiferi and could be used to inform veterinary risk assessments on a broad spatial scale. Limitations and further developments are discussed.

The loss of STAT3 in mature osteoclasts has detrimental effects on bone structure.

Signal Transducer and Activator of Transcription 3 (STAT3) has recently been shown to be involved in bone development and has been implicated in bone diseases, such as Job's Syndrome. Bone growth and changes have been known for many years to differ between sexes with male bones tending to have higher bone mass than female bones and older females tending to lose bone mass at faster rates than older males. Previous studies using conditional knock mice with Stat3 specifically deleted from the osteoblasts showed both sexes exhibited decreased bone mineral density (BMD) and strength. Using the Cre-Lox system with Cathepsin K promotor driving Cre to target the deletion of the Stat3 gene in mature osteoclasts (STAT3-cKO mice), we observed that 8-week old STAT3-cKO female femurs exhibited significantly lower BMD and bone mineral content (BMC) compared to littermate control (CN) females. There were no differences in BMD and BMC observed between male knock-out and male CN femurs. However, micro-computed tomography (µCT) analysis showed that both male and female STAT3-cKO mice had significant decreases in bone volume/tissue volume (BV/TV). Bone histomorphometry analysis of the distal femur, further revealed a decrease in bone formation rate and mineralizing surface/bone surface (MS/BS) with a significant decrease in osteoclast surface in female, but not male, STAT3-cKO mice. Profiling gene expression in an osteoclastic cell line with a knockdown of STAT3 showed an upregulation of a number of genes that are directly regulated by estrogen receptors. These data collectively suggest that regulation of STAT3 differs in male and female osteoclasts and that inactivation of STAT3 in osteoclasts affects bone turnover more in females than males, demonstrating the complicated nature of STAT3 signaling pathways in osteoclastogenesis. Drugs targeting the STAT3 pathway may be used for treatment of diseases such as Job's Syndrome and osteoporosis.

Parasites in the changing world - Ten timely examples from the Nordic-Baltic region.

The world is changing, and parasites adapt. The Nordic-Baltic region in northern Europe - including the Nordic countries Denmark, Finland, Iceland, Norway and Sweden, and the Baltic States Estonia, Latvia and Lithuania - is facing new parasitological challenges due to changes in populations of parasites and their hosts and the spread of new parasites to the region due to climate change. Some changes can also be ascribed to increased awareness and detection. In this paper, we review and discuss a convenience selection of ten timely examples of recent observations that exemplify trends and challenges from different fields of parasitology, with particular focus on climate change and potential changes in epidemiology of pathogens in northern Europe. The examples illustrate how addressing parasitological challenges often requires both intersectoral and international collaboration, and how using both historical baseline data and modern methodologies are needed.

Serological detection of anti-Trichinella antibodies in wild foxes and experimentally infected farmed foxes in Norway.

Trichinella surveillance in wildlife has relied on the detection of muscle larvae using digestion techniques. Serology has been proposed as more suitable for large-scale epidemiological studies in wildlife. In this study, 328 individual sera from wild red foxes and 16 sera from experimentally infected farmed foxes were serologically tested with both excretory/secretory antigen (E/S) and the synthetic beta-tyvelose glycan antigen, in indirect ELISA tests. The wild red foxes (Vulpes vulpes) had previously been examined for muscle larvae, using muscle digestion, whilst the experimentally infected farmed foxes were inoculated per os with either a low dose, 500 larvae, or a high dose, 10,000 of Trichinella nativa muscle larvae. Western blot (WB) was carried out on all seropositive samples using crude larval antigen. The present study found both beta-tyvelose and E/S antigen suited for the detection of antibodies to Trichinella spp., and T. nativa in particular, in foxes. Both ELISA antigens performed well, although, the E/S antigen was superior to the beta-tyvelose antigen, with sera that had been stored at -20 degrees C for more than 10 years. Neither antigen, however, detected all of the samples proven seropositive by WB: E/S detected 21 of the 27 wild red fox sera positive by WB; beta-tyvelose detected 22 positive sera; and in total 24 of the 27 positive WB sera were identified using both antigens. Serology alone, without WB or muscle digestion, led to a two- to threefold higher seroprevalence estimate, respectively. The use of E/S antigen in conjunction with the WB was the method of choice for the screening of wild red fox populations for Trichinella. Antibody persistence to T. nativa was short in the low dose group where antibody levels were not different from background by 32 wpi. In total, 7.3% (24/328) of the wild red fox population had antibodies to Trichinella on ELISA and WB. Antibodies were identified in foxes from a further two regions in Norway compared to the original muscle digestion results.

Elaphostrongylus and Dictyocaulus infections in Norwegian wild reindeer and red deer populations in relation to summer pasture altitude and climate.

Nematodes of the genera Elaphostrongylus and Dictyocaulus are associated with disease in semi-domesticated tundra reindeer and farmed red deer whereas less knowledge exists in the wild. Their first stage larvae (L1) develop to the infective third stage (L3) in the environment; Elaphostrongylus spp. within intermediate gastropod hosts and Dictyocaulus spp. as free-living larvae. Larval development of Elaphostrongylus is highly temperature dependent with a developmental minimum of 9-10 °C. Larval development of Dictyocaulus spp. may occur at low temperatures (5 °C) but the larvae are sensitive to desiccation. We examined the prevalence and intensity of Elaphostrongylus spp. and Dictyocaulus spp. infections in six wild reindeer and two wild red deer populations in relation to altitude, temperature and rainfall in their respective main summer pasture area over the 5 summers prior to sampling. The parasitological examination was based upon morphological identification of L1 in the faeces of hunted animals. Altitude was calculated from animal position data and temperature and precipitation by means of a nationwide gridded data set. Temperature decreased with increasing altitude, from 13.3 °C for the lowest located red deer population (300 m) to 6.1 °C for the highest located reindeer population (1400 m). No significant relationship between altitude and rainfall was identified. Elaphostrongylus spp. infection decreased in prevalence with increasing altitude, being identified in 89% of investigated samples from the lowest located population and in 3% of samples from the highest. The prevalence of Dictyocaulus spp. infection varied between 28 and 80% and no relationship with altitude was found. The intensity of Elaphostrongylus spp. infection was low in reindeer and moderate in red deer whereas the intensity of Dictyocaulus spp. infection was moderate in both species. Our results indicated that the climatic conditions in all areas studied were suitable for Dictyocaulus spp., whereas summer temperature was a restrictive factor for Elaphostrongylus sp. in reindeer.

Long-term study of Sarcoptes scabiei infection in Norwegian red foxes (Vulpes vulpes) indicating host/parasite adaptation.

The red fox (Vulpes vulpes) population, in Norway, was naïve to Sarcoptes scabiei prior to the late 1970s when this parasite was first recorded and a still ongoing epidemic started. During the course of this protracted epidemic some degree of host/parasite adaptation, with the occurrence of healthy antibody positive foxes, might be expected. In the present study the prevalence of sarcoptic mange and serologically identified S. scabiei exposure was investigated in 363 Norwegian red foxes, shot by hunters during two different study periods (1994-1995 and 2002-2005). The sarcoptic mange diagnosis was based upon the presence of clearly visible lesions in the skin of the cadaver with confirmatory demonstration of S. scabiei. The serodiagnosis was based on an indirect-ELISA. There was a significant decrease in prevalence of both mange cases and seropositive animals from the first to the second study period. Whilst the mange prevalence fell more than threefold, from 30.0% to 6.6%, the seroprevalence dropped less dramatically from 53.3% to 19.1%. The smaller decrease in seroprevalence compared to mange cases reflected a significantly higher ratio of seropositive-mange negative versus seropositive-mange positive foxes, during the second study period, 40:18, compared to the first, 14:18. These findings indicate that the red fox population is adapting to live with the parasite and that low-grade or sub-clinical infections, and even recoveries, occur amongst exposed foxes. Mange positive foxes had significantly poorer body condition than those without sarcoptic mange. No significant difference in body condition was seen between seropositive-mange negative versus seronegative-mange negative foxes. The ELISA sensitivity was found to be 95% and proved a useful tool for investigating the exposure to S. scabiei in wild foxes. This study is believed to be the first pointing to a long-term Sarcoptes/fox adaptation, combining long-term prevalence studies of clinical sarcoptic mange and serological evidence of exposure to the parasite in the general fox population.

Prevalence of Trichinella larvae and extra-intestinal nematodes in Norwegian red foxes (Vulpes vulpes).

A survey of the parasitic fauna of the Norwegian red fox (Vulpes vulpes) population was carried out in 1994/1995 and 2002-2005. All foxes were killed during the licensed hunting season from October to April and, in total, 393 animals from all regions of the country were examined. The present study details the results of extra-intestinal nematode and Trichinella larvae examinations. All individuals were examined for Trichinella, using routine digestion methods. Parasitological examination of the internal organs of some of the foxes also identified a number of different extra-intestinal nematodes. The following prevalences were identified (number positive/number foxes examined): Trichinella larvae 19/393 (4.8%); Capillaria böhmi (C. böhmi) 88/174 (51%); Capillaria aerophila (C. aerophila) 160/181 (88%); Crenosoma vulpis (Cr. vulpis) 105/181 (58%) and Capillaria plica (C. plica) 81/154 (53%). No evidence of Angiostrongylus vasorum infection was found. The 19 different Trichinella isolates were species typed by PCR and sequence analysis; 18 isolates were identified as Trichinella nativa and one as Trichinella britovi. A wide geographical distribution of the parasites was seen. The following exceptions were recorded: C. böhmi, the prevalence of which was significantly lower in northern Norway (6%) compared to other regions (central Norway, eastern Norway and southern and western Norway; 52-57%). There was a significantly higher prevalence of Trichinella infection in eastern Norway (8.1%), when compared with the rest of the country (0.6%). Cr. vulpis prevalence was significantly higher in central Norway (83%) than in other regions (41-56%). There were no significant differences in age and sex distribution of the parasites with the exception of Cr. vulpis where juvenile foxes had a greater likelihood of infection. The data also indicated that adult foxes were more commonly infected with Trichinella larvae (5.8%) than juveniles (3.3%) (no statistical significance).

Present status, actions taken and future considerations due to the findings of E. multilocularis in two Scandinavian countries.

When Echinococcus (E.) multilocularis was first detected in mainland Scandinavia in Denmark in 2000, surveillance was initiated/intensified in Sweden, mainland Norway and Finland. After 10 years of surveillance these countries all fulfilled the requirements of freedom from E. multilocularis as defined by the EU, i.e. a prevalence in final hosts <1% with 95% confidence level. However, in 2011 E. multilocularis was detected in Sweden for the first time and surveillance was increased in all four countries. Finland and mainland Norway are currently considered free from E. multilocularis, whereas the prevalence in foxes in Sweden and Denmark is approximately 0.1% and 1.0%, respectively. E. multilocularis has been found in foxes from three different areas in Denmark: Copenhagen (2000), Højer (2012-14) and Grindsted (2014). Unlike Sweden, Norway and Finland, human alveolar echinococcosis (AE) is not notifiable in Denmark, and the number of human cases is therefore unknown. In Sweden, E. multilocularis has been found in foxes in four counties, Västra Götaland, Södermanland, Dalarna (2011) and Småland (2014). E. multilocularis has also been found in an intermediate host in Södermanland (2014). Two cases of AE have been reported in humans (2012), both infected abroad. No cases of E. multilocularis or AE have been reported in Finland and Norway. Recommendations and future considerations are discussed further.