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Traditional methods for age determination of wildlife include either slicing thin sections off or grinding a tooth, both of which are laborious and invasive. Especially when it comes to ancient and valuable museum samples of rare or extinct species, non-invasive methods are preferable. In this study, X-ray micro-computed tomography (µ-CT) was verified as an alternative non-invasive method for age determination of three species within the order of Carnivora and suborders Odontoceti. Teeth from 13 red foxes (Vulpes vulpes), 2 American mink (Neogale vison), and 2 harbor porpoises (Phocoena phocoena) of known age were studied using µ-CT. The number of visible dental growth layers in the µ-CT were highly correlated with true age for all three species (R2 = 96%, p < 0.001). In addition, the Bland–Altman plot showed high agreement between the age of individuals and visible dental layers represented in 2D slices of the 3D µ-CT images. The true age of individuals was on average 0.3 (±0.6 SD) years higher than the age interpreted by the µ-CT image, and there was a 95% agreement between the true age and the age interpreted from visible dental layers in the µ-CT.

Background: Diagnosing and treatment of diseases in pigs are important to maintain animal welfare, food safety and productivity. At the same time antimicrobial resistance is increasing, and therefore, antibiotic treatment should be reserved for individuals with a bacterial infection. The aim of the study was to investigate gross and histological lesions and related pathogens in pigs that died during the nursery period in five Danish farms. In addition, high throughput, real-time qPCR monitoring of specific porcine pathogens in fecal sock and oral fluid samples were carried out to investigate the between-farm and between-batch variation in the occurrence of pathogens. Results: Twenty-five batches of nursery pigs from five intensive, indoor herds were followed from weaning (approximately four weeks) to the end of nursery (seven to eight weeks post weaning). Gross and histological evaluation of 238 dead and 30 euthanized pigs showed the highest prevalence of lesions in the skin, respiratory system, gastrointestinal tract, and joints. Gross and histological diagnoses of lung and joint lesions agreed in 46.5% and 62.2% of selected pigs, respectively. Bacteriological detection of Escherichia coli, Streptococcus suis or Staphylococcus aureus infections in joints, lungs and livers was confirmed as genuine infection on immunohistochemical staining in 11 out of 70 tissue sections. The real-time qPCR analysis of pooled samples showed that most pathogens detected in feces and in oral fluid in general followed the same shedding patterns in consecutive batches within herds. Conclusions: Gross assessment should be supplemented with a histopathological assessment especially when diagnosing lesions in the lungs and joints. Moreover, microbiological detection of pathogens should optimally be followed up by in situ identification to confirm causality. Furthermore, routine necropsies can reveal gastric lesions that may warrant a change in management. Real-time qPCR testing of fecal

Diet studies of carnivores are often based on analysis of prey remains found in scats or stomachs. In these analyses differences in decomposition rate and the degree of digestion of the prey item must be taken into consideration. In studies of raccoon dog diets, eggshells are rarely found. This may be due to three possible scenarios, which may act in combination: 1) raccoon dogs rarely eat bird´s eggs in the wild, 2) raccoon dogs eat eggs without swallowing shell parts, 3) rapid digestion of swallowed eggshells, so that they are only present briefly in stomach contents. In this study, the feeding behaviour of raccoon dogs on eggs of different sizes was observed under semi-natural conditions in an enclosure and the dissolution time of eggshells was examined. Dissolution time of eggshells of different thicknesses was tested experimentally using different concentrations of hydrochloric acid. The study showed a negative relationship between egg size/shell thickness and the amount of shell ingested by the raccoon dog. No eggshell, independent of bird species, was degraded beyond detectability after 12 hours in hydrochloric acid at pH 3.3. A correction factor estimating the mass of eggshell (g) ingested by the raccoon dog was calculated from the dry mass of eggshell found in the stomach or scat. Also the amount of eggshell estimated to be degraded by gastric acid is given. This correction factor ranged from 14 to 282 depending on eggshell thickness and egg size. From this study, it can be concluded that raccoon dogs do ingest some eggshell and that it is possible to detect eggshells in stomachs and scats of raccoon dogs, suggesting that the lack of eggshell in diet samples probably reflects a low proportion of eggs in the diet and/or preying more heavily on large eggs that are cracked open and have their contents eaten, rather than small eggs that are eaten shell-and-all.

The feral mink population in Denmark consists of two groups of animals: mink born in the wild and mink that have recently escaped from farms. The aims of this study were to: (1) estimate the reproductive performance and mortality of the Danish mink born in the wild (wild-born) and mink escaped from farms (captive-born); (2) discuss the likelihood of a self-sustaining population of wild-born mink in Denmark; and (3) model the relationship between the pulp cavity width and the age of mink. During 2018, 247 wild caught mink were sent for necropsy at the Danish National Veterinary Institute. Based on body length, 112 were determined as captive-born and 96 as wild-born. The mean litter size ± SE of wild-born females was 7.6 ± 0.9 (range: 5–11 kits) and for captive-born females 5.9 ± 0.9 (range: 1–10 kits). The relationship between age (in months) of mink and pulp cavity width was highly significant. Individuals with a pulp cavity width of >35% were younger than one year. Based on fecundity, the turnover of the mink population was estimated to be 66%, and the yearly mortality was estimated at 69%. Hence, the population is slightly declining. In conclusion, a feral reproducing mink population in Denmark persists without a continuous influx of captive-born mink from farms.

Recent data suggest that organic broilers often score worse on footpad lesions than conventional broilers but also that the current scoring of organic broiler feet may be misleading. In order to characterise footpad lesions in organic broilers, this study assessed and compared footpad lesions in a sample of 2987 conventional and 3578 organic broiler feet obtained from a large Danish abattoir during summer and winter. The feet were scored according to two scoring systems: the modified Danish surveillance scoring system and a histopathology-based new scoring system specifically developed to target the ability to differentiate between broiler feet with hyperkeratosis and ulcers. For both systems, all broiler feet with visible lesions were cross-sectionally incised. Significant differences between the two production systems were found for both scoring systems (χ2 = 710; P < 0.001 and χ2 = 247; P < 0.001 for the new and the surveillance systems, respectively), showing that a larger proportion of the organic feet compared to conventional feet - summer and winter - exhibited signs of hyperkeratosis. In addition, a smaller fraction of the organic feet than of the conventional feet were given the outermost scores, that is, normal or ulcerated; 13.4% v. 25.3% broiler feet were given score 0 for organic v. conventional production systems, respectively (χ2 = 152; P < 0.001), and 18.4% v. 23.8% feet were given score 4 for organic v. conventional production systems, respectively (χ2 = 308; P < 0.001). Thus, the results suggest that surveillance scoring systems such as the one used in Denmark are useful for the examination of footpad lesions in broilers from both types of production systems. However, the results have also raised attention to a typical characteristic of the feet of organic broilers, that is, profound hyperkeratosis, which may underlie potential misclassifications in surveillance scoring systems like the one used in Denmark. Among the possible solutions

Abortion is a major source of economic losses in cattle breeding. Abortion occurs due to a wide range of causes, but infections are the most frequently diagnosed. However, establishing an aetiological diagnosis remains challenging due to the large variety of bacteria, protozoa, viruses, and fungi that have been associated with abortion in cattle. Economic restraints limit the range of diagnostic methods available for routine diagnostics, and decomposition of the conceptus or lack of proper fetal and/or maternal samples further restrict the diagnostic success. In this study, we report recent diagnostic findings from bovine abortions in Denmark, a country that has a large dairy sector and is free from most infectious agents causing epizootic abortion in cattle. The aims of the study were: (i) to identify infectious causes of bovine abortion in Denmark, (ii) to categorise the diagnostic findings based on the level of diagnostic certainty, and (iii) to assess the diagnostic rate. Due to economic restraints, only a limited panel of routine diagnostic methods were available. Placentas and/or fetuses from mid- to late-term abortions and stillbirths (n = 162) were submitted to the Danish National Veterinary Institute between January 2015 and June 2017. The aborted materials were examined macroscopically, histologically, and by bacterial culture. Maternal blood samples were tested for bovine viral diarrhoea virus (BVDV) antibodies. The likely aetiology of the abortion was diagnosed in 52 cases, resulting in a diagnostic rate of 33%. The most common cause was protozoal infection (19%) followed by infection with Trueperella pyogenes (3%), Staphylococcus aureus (2%), and non-haemolytic Escherichia coli (2%). Lesions in fetuses with a protozoal infection were consistent with neosporosis. In many cases (38%), inflammatory changes were found in the placenta and/or fetal organs but no specific aetiology was identified. Neither infection with Brucella spp. nor maternal BVDV antibodi

In Denmark, American mink (Neovison vison) have been bred for their fur since the mid-1920s. Mink escaping from farms may supply the feral population. Often, it is of biological and management interest to separate the population of feral mink (i.e. mink caught in the wild) in two groups: 1) mink born on farms i.e., escapees, and 2) mink born in the wild. In this study, two methods were used for separating feral mink into the two groups: a) Comparison of body length of farmed mink and feral mink, and b) Presence of a biomarker (tetracycline: an oral antibiotic used on mink farms). A total of 367 wild caught mink (from the mainland of Denmark and the island of Bornholm), and 147 mink from farms, collected during the period 2014–2018, were used for the analysis of body length. For the testing of tetracycline (TC) as a biomarker, 78 mink from farms where there was knowledge about TC treatment (with or without) were examined for fluorescent markings in the canine teeth. Results from both univariate analyses and Gaussian mixture model analysis demonstrated clear divisions between the mean body length (mean ± S.E., range) of farmed males (52.1 cm ± 0.4, 48–68) and farmed females (mean 44.0 ± 0.2, 40–50), and between farmed mink and wild caught mink. Mixture analysis identified two groups within each sex of the wild caught mink, one assigned to farmed mink (born in captivity) and another group of smaller mink suspected of being born in the wild. On Bornholm, the mean (±SD, range) length of males born in the wild was 43.7cm (± 0.3, 36–57) and for females 37.5cm (± 0.3, 32–45). The mean length (±SD, range) of males born in the wild in the mainland of Denmark was 42.5cm (± 2.3, 36–46) and for females 36.1cm (± 1.0, 34–37). Among the feral mink from mainland Denmark, 28.4% of males and 21.6% of females were identified as escapees, while 0% of the males and 1% of the females were identified as escapees among the wild caught mink on Bornholm. Eight percent of mink from farms usin

Background: Vaccines may have non-specific effects, affecting resistance to heterologous pathogens. Veterinary vaccines have seldom been investigated for their non-specific effects. However, recent observational studies suggest that an inactivated paratuberculosis vaccine reduced all-cause mortality in goats and cattle. Aim: We tested if vaccination with a killed mycobacterial vaccine may have heterologous effects in swine (Sus domesticus), specifically on the pathogenic and clinical effects of a heterologous challenge with Actinobacillus pleuropneumoniae in young pigs. Methods: Newborn piglets were randomized to vaccination s.c. with the inactivated paratuberculosis vaccine Gudair (Zoetis Inc.) (n = 17) or no vaccine (n = 16). At 4-5 weeks after vaccination, all piglets were challenged intra-nasally with a high (Gudair: n = 8; control: n = 8) or a low (Gudair: n = 9; control: n = 8) dose of the gram-negative bacterium A. pleuropneumoniae causing acute porcine pleuropneumonia. The effect and severity of pathogen challenge was evaluated by measuring acute phase proteins C-reactive protein, haptoglobin and Porcine alpha 1-acid glycoprotein, and by gross pathology 1 day post challenge. Specific and non-specific in vitro cytokine responses to vaccination were evaluated in whole blood before bacterial challenge. Results: The vaccine was immunogenic in the pigs as evidenced by increased IFN-gamma responses to purified protein derivative of Mycobacterium paratuberculosis. However, Gudair vaccine did not affect IL-6 responses. The gross pathology of the lungs as well as the acute phase protein responses after the high A. pleuropneumoniae dose challenge was slightly increased in the vaccinated animals compared with controls, whereas this was not seen in the animals receiving the low-dose bacterial challenge. Conclusion: The inactivated paratuberculosis vaccine exacerbated the pathological and inflammatory effects of an experimental A. pleuropneumoniae infection in young pigs

A decline in the Danish population of red foxes Vulpes vulpes due to an outbreak of canine distemper (CDV) in 2012 gave us the opportunity to test the hypothesis that the reproductive performance of foxes increases when the population density declines. The reproductive performance of 280 female foxes from two periods (mainly shot or road killed) in 1997-2000 and 2012-16, were compared. Game Bag Records of Jutland (GBRJ) were used as an estimate of population density. After a drop in GBRJ in 2013 due to the CDV epidemic, the mean litter size (based on dark placental scars from partum to oestrus) became significantly larger than in previous years; F=4.3, p>0.03, Hc=8.1, p<0.02. In 2015-2016, after population decline the mean litter size was 8.2 (±2.5 SD) and in the breeding seasons before population decline in 1997-1999 and 2012-2013 the mean litter size was 5.6 (±2.1 SD) and 5.7 (±2.0 SD), respectively. During the period 1997-1999, barrenness was relatively high especially in yearlings, and the reproducing yearling foxes made up only 6% of the breeding females compared to 2012-13 and 2015-16 where breeding yearling females made up 53% and 61%, respectively. Age related differences in litter size and productivity were found in years with a relatively high population density, when older females in their third and fourth breeding seasons had the largest litter sizes and highest productivity. This was in contrast to the years with low population size, when no age-related reproduction was found, and when young females had relatively large litter sizes and high productivity. Rump fat thickness (RFT) of the breeding females was significantly higher in breeding females than in barren females, and the RFT was positively correlated to the number of embryos (R2=41%). This study confirms that the number of barren females drops, and the proportion of yearling females and litter size increase with reduced population density. Hence, culling or epidemics in fox populations inc

For the first time, temporal variability in body size and sexual dimorphism is revealed in foxes Vulpes vulpes from the same geographical area at over time. The weights and lengths of 552 Danish foxes were documented during three different periods: 1965–1977, 2012–2014 and the winter of 2015/2016. During the first and the third periods, the fox population was below the carrying capacity due to hunting pressure and canine distemper, respectively. Adult males were significantly (p < 0.01) heavier (mean weight: 7.7 kg and 7.5 kg respectively) in periods of low population density, i.e. 1965–1977 and compared to 2015/2016, compared to 2012–2014, when population density was high (the mean weight: 6.8 kg). However, no significant differences were found in the weight of females. Hence, sexual dimorphism ranged from 7.6 to 3.6 in adult foxes in low and high-density periods, respectively. During the winters of 2012–2014, no difference in body fat measured by rump fat thickness (RFT) was found between age groups and genders in contrast to 2015/2016, when RFT was significantly (p < 0.001) larger in adult females (mean RFT: 0.77 cm) than in adult males (mean RFT = 0.58cm).

Advanced animal models, such as minipigs, are needed for the development of a globally requested human Chlamydia vaccine. Previous studies have shown that vaginal inoculation of sexually mature Göttingen minipigs with Chlamydia trachomatis resulted in an infection lasting only 3-5 days. The aim of this study was to evaluate the effect of targeting the upper porcine genital tract by transcervical and transabdominal intrauterine inoculation, compared to previously performed vaginal inoculation. Furthermore, we investigated the effect of the hormonal cycle, estrus vs. diestrus, on the establishment of a C. trachomatis infection in the minipig. Targeting the upper genital tract (transcervical inoculation) resulted in a longer lasting infection (at least 7 days) compared to vaginal inoculation (3-5 days). When comparing intrauterine inoculation during estrus and diestrus, inoculation during diestrus resulted in a longer lasting infection (at least 10 days) compared to estrus (3-5 days). Furthermore, we found a significant C. trachomatis specific IFN-γ response in pigs inoculated during estrus correlating with the accelerated clearance of infection in these pigs. These findings suggest that for implementation of an optimal model of C. trachomatis in minipigs, inoculation should bypass the cervix and preferable be performed during diestrus.

I perioden april til oktober 2016, blev fæces fra mink (Neovison vison) på 30 danske minkgårde undersøgt med henblik på at fastlægge forekomsten og arterne af Eimeria hos danske mink (Neovison vison). På basis af morfologisk undersøgelse af sporulerede oocyster, blev oocyster som lignede tidligere beskrevet art, kaldet E. vison identificeret. Den fundne art var dog en smule mindre end E. vison. Efterfølgende blev oocysterne forsøgt identificeret molekylært og sammenlignet med sekvenser i Genbank. Desværre var der ikke tilgængelige mink sekvenser i GenBank, og den nyligt fundne Eimeria art kaldes derfor for Eimeria vison-like. I alt fandtes 2,6% (108/4.141) af de underøgte fæcesprøver positive for E. vison-like oocyster ved mikroskopi, svarende til at 23,9% (78 mink) af minkene udskilte E. vison-like oocyster mindst én gang i undersøgelsesperioden.

Infectious skin disorders are not uncommon in mink. Such disorders are important as they have a negative impact on animal health and welfare as well as on the quality and value of the fur. This study presents the isolation of Arcanobacterium phocae from mink with severe skin lesions and other pathological conditions, and from wild seals and otters. In 2015, A. phocae was isolated for the first time in Denmark from outbreaks of dermatitis in mink farms. The outbreaks affected at least 12 farms. Originating from these 12 farms, 23 animals cultured positive for A. phocae. The main clinical findings were necrotizing pododermatitis or dermatitis located to other body sites, such as the lumbar and cervical regions. A. phocae could be isolated from skin lesions and in nine animals also from liver, spleen and lung, indicating a systemic spread. The bacterium was also, for the first time in Denmark, detected in dead seals (n = 9) (lungs, throat or wounds) and otters (n = 2) (throat and foot). An infectious skin disorder in mink associated with A. phocae has started to occur in Danish farmed mink. The origin of the infection has not been identified and it is still not clear what the pathogenesis or the port of entry for A. phocae infections are.

Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is the cause of severe reproductive and respiratory disease in swine worldwide. In Denmark, both PRRSV-1 and PRRSV-2 are circulating and approximately 35% of pig herds are seropositive for PRRSV. In November 2010, a pig herd in the Northern part of Denmark experienced an infection with PRRSV-2 with clinical signs that were much more severe than normally reported from current Danish PRRSV-2 affected herds. Due to the clinical observations of reproductive failure in sows and high mortality in piglets, it was speculated that a new, more pathogenic or vaccine evading PRRSV strain had emerged in Denmark. The overall aim of the present study was to perform a genetic and biological characterization of the virus isolated from the diseased herd. Complete genome sequencing of isolates from this herd revealed that although the case strain had some unique genetic features including a deduced 3 amino acid deletion, it was in overall very similar to the other PRRS-2 viruses circulating in Denmark. In an experimental trial in growing pigs, no overt clinical signs or pathology were observed following intranasal inoculation with the new virus isolate. Virus shedding, acute phase protein responses and serological responses were comparable to those seen after experimental challenge with a Danish PRRSV-2 reference strain isolated in 1997. Vaccination with a commercial modified live PRRSV-2 vaccine had a clear reducing effect on virus shedding, magnitude, and duration of viremia and viral load in the lungs. Overall, the results indicate that the severe disease observed in the field was contributed by additional factors in combination with the PRRS virus infection.