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The Swedish wolf population is closely monitored and managed to keep the population at a sustainable level while avoiding conflicts. Detailed knowledge about reproduction is crucial for estimates of population size and the reproductive potential of a population. Post-mortem evaluation of reproductive organs can be used as a complementary tool to field monitoring for evaluation of cyclicity and previous pregnancy, including litter size. Therefore, we evaluated reproductive organs from 154 female wolves that were necropsied during the period 2007-2018. The reproductive organs were weighed, measured, and inspected according to a standardised protocol. Presence of placental scars was evaluated for estimates of previous pregnancy and litter size. Data about individual wolves were also obtained from national carnivore databases. Body weight increased during the first year of life before levelling out. There was evidence of cyclicity the first season after birth in 16.3 % of the 1-year-old females. No females < 2 years had evidence of a previous pregnancy. Pregnancy rates were significantly lower in 2- and 3-year old females than in older females. Mean uterine litter size was 4.9 ± 2.3, and did not differ significantly between age groups. Our data supports earlier field data that female wolves usually start to reproduce at the earliest at 2-years of age but that they occasionally start to cycle one season earlier. All females ≥ 4 years of age had reproduced. Pathological findings of the reproductive organs were rare, indicating that reproductive health of female wolves is not a limiting factor for population growth., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interests., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

The recently identified causative agent of congenital tremor in domestic piglets, atypical porcine pestivirus (APPV), was detected in serum from Swedish wild boar. A previous study from Sweden described APPV in domestic piglets suffering from congenital tremor, but the APPV situation in the wild boar population was unknown. In this study, 595 serum samples from wild boar originating from 13 counties in the south and central parts of Sweden, collected between 2000 and 2018, were analysed for the presence of the APPV-genome and for antibodies against the APPV-glycoprotein E rns . The results revealed that APPV is highly abundant in the Swedish wild boar population; 12% (73/595) were APPV-genome positive in serum and 72% (433/595) of the tested wild boars displayed APPV-specific antibodies. The present study also shows that APPV has been present in the Swedish wild boar population since at least the year 2000. The viral sequences obtained from the wild boars were highly similar to those obtained from Swedish domestic pigs positive for APPV and suffering from congenital tremor, suggesting a viral exchange between wild boars and domestic pigs. The high proportion of viraemic and seropositive wild boar is indicative of wild boar being an important reservoir for APPV., (© 2021 The Authors. Transboundary and Emerging Diseases published by Wiley-VCH GmbH.)

Objective: Insemination with spermatozoa, seminal plasma and extender, cause a rapid inflammatory response in pig endometrium, characterized by an influx of neutrophils into the uterus. The transient inflammatory response to semen involves cytokine induction. Potential functions for Interleukin-23 (IL-23) in the inflammatory response to different insemination treatments were examined by studying mRNA expression and immunostaining in gilt oviduct and endometrium 35-40 h after insemination. Insemination was performed with seminal plasma (SP), spermatozoa (SPZ) without SP in the extender Beltsville thawing solution (BTS), or BTS alone. In control gilts an insemination catheter was inserted without anything being inseminated., Results: Results showed that IL-23 mRNA was expressed in oviduct and endometrium after insemination regardless of treatment. There was an approximate two- to fourfold increase in expression of IL-23 mRNA in catheter-insertion control compared with SPZ, SP and BTS treatment groups. IL-23 immunolabelling was detected in a small number of separate cells and in the sub-epithelial connective tissue of the endometrium, the endosalpinx of isthmus and infundibulum., Conclusion: In conclusion, insemination with SP, SPZ in BTS, and BTS alone decreased the expression of IL-23 mRNA in the endometrium compared to catheter-insertion control, indicating a possible role for IL-23 in the inflammatory response after insemination in gilts.

Improved knowledge about reproductive patterns and potential in male wolves (i.e., testicular development and size relative to age, pubertal age, and seasonal effects) is needed for evaluation and monitoring of reproductive outcomes in populations. Reproductive organs from 215 male wolves, culled as a result of licensed hunting, protective culling or from carcasses found were examined. The testes and epididymis were weighed and measured. There were biopsy samples collected from the testes and the cauda epididymis for histological determinations if there were spermatozoa in tissues collected. There were reproductive tissue analyses of 197 males while there were separate evaluations of tissues from ten cryptorchid animals. Juvenile wolves (< 1 year, n = 47) had a lesser body mass and mean testes mass than subadult (1-2 years, n = 71) and adult (>2 years, n = 79) males. Season also affected testicular characteristics of structures evaluated with subadult and adult males having a lesser mass during summer months (May-August). Of the 197 males, 70 % had spermatozoa in the seminiferous tubules and the cauda epididymis and were classified as being 'potentially fertile' when tissues were collected, while 22 % were classified as being non-fertile (no spermatozoa, including males that were pre-pubertal) and tissues of 8% could not be evaluated. When testes mass was greater, there was a greater likelihood that spermatozoa were present. There were seven of the ten cryptorchid males of the unilateral type. These testicular and epididymal findings will be useful for evaluating the reproductive potential and management of wolves in Scandinavia., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Knowledge on dental disorders in commercial sows is limited although such conditions may have important animal welfare implications. In a pilot study, the dental and periodontal health of 58 sows (Landrace*Yorkshire-crosses) from 8 Swedish commercial pig herds, slaughtered at one abattoir, were investigated. The oral cavity was inspected and abnormalities were recorded on a dental chart modified for pigs. Dental abnormalities, absence of teeth, supernumerary teeth, tooth fractures, signs of caries, and malalignment were recorded. The study revealed that 19% of the sows had supernumerary teeth and 59% of the sows missed at least one tooth. Periodontitis, calculus and malalignment were observed in 33%, 45% and 17%, respectively. Tooth wear was very common both in incisors (total 83%) and in premolars/molars (total 84%). One or more tooth fractures (between 1 and 6 per sow) was found in 41%. Signs of caries was found in 9%. In order to assess oral health, three indices were used: calculus index (CI), periodontal index (PDI) and tooth wear index (TWI). Severe periodontitis, tooth wear in incisors and tooth wear in premolars/molars were found in 7%, 34% and 35%, respectively. With respect to animal welfare, the etiology and the effects of the disorders on health, stress and pain need to be investigated.

Tularaemia is a zoonotic disease, in Europe caused by Francisella tularensis subsp. holarctica. Many lagomorphs and a variety of small rodents are wildlife species prone to develop clinical disease, while predators and scavengers are relatively resistant and may serve as sentinels. Blood samples from 656 Swedish wild predators and scavengers were serologically investigated using slide agglutination and microagglutination. In the slide agglutination test, 34 seropositive animals were detected, and they were found among all species investigated: brown bear (Ursus arctos), Eurasian lynx (Lynx lynx), raccoon dog (Nyctereutes procyonoides), red fox (Vulpes vulpes), wild boar (Sus scrofa), wolf (Canis lupus) and wolverine (Gulo gulo). Due to haemolysis the microagglutination test was more difficult to read at low titres, and only 12 animals were classified as seropositive. F. tularensis subsp. holarctica was detected by a polymerase chain reaction in lymphatic tissues of the head in one brown bear, one red fox and one wolf. The significance of this finding regarding possible latency of infection is not clear. In conclusion, the results of this study indicate that all predator and scavenger species included in this study may serve as sentinels for tularaemia in Sweden. Their role as reservoirs is unclear.

Climate change, with warmer temperatures and altered precipitation patterns, has affected the distribution of vectors and vector-borne diseases. In the northern hemisphere, vectors are spreading north, and with them, pathogens of zoonotic and animal health impact. Eurasian moose (Alces alces alces) are physiologically and anatomically adapted for cold climate, and are rarely considered ideal hosts of vectors, apart from deer keds (Lipoptena cervi). To investigate the presence of vector-borne pathogens, spleen samples from 615 moose were collected in southern Sweden from 2008 to 2015. The samples were analyzed with a high-throughput PCR method for 24 bacterial, and 12 parasitic pathogens. Anaplasma (82%), Borrelia (3%), Babesia (3%), and Bartonella (1%) DNA was found, showing that moose are exposed to, and can act as hosts of some of these pathogens, which can have an impact of both animal and human health. These results show that Swedish moose are exposed to pathogens that in some instances are more commonly found in regions with warmer climate, and highlights the importance of also considering moose as sentinels of vector-borne pathogens. Further research is needed to understand the effect of these pathogens on the health of individual moose and to elucidate whether climate change and moose population density interact to create the pattern observed.

The wild boar ( Sus scrofa) population has increased markedly during the last three decades in Sweden and in other parts of Europe. This population growth may lead to increased contact between the wild boar and the domestic pig ( Sus scrofa scrofa), increasing the risk of transmission of pathogens. The objective of our study was to estimate the seroprevalence of selective pathogens, known to be shared between wild boars and domestic pigs in Europe, in three wild boar populations in Sweden. In total, 286 hunter-harvested female wild boars were included in this study. The sera were analyzed for antibodies against nine pathogens using different commercial or in-house enzyme-linked immunosorbent assays. Antibodies were detected against porcine parvovirus (78.0%), porcine circovirus type 2 (99.0%), swine influenza virus (3.8%), Erysipelothrix rhusiopathiae (17.5%), Mycoplasma hyopneumoniae (24.8%), and Toxoplasma gondii (28.6%). No antibodies were detected against porcine respiratory and reproductive syndrome virus, Brucella suis, or Mycobacterium bovis. Our results highlight the potential importance of the wild boar as a reservoir for pathogens potentially transmissible to domestic pigs and which also may affect human health.

Background: Sarcoptic mange has recently emerged in wild boar in Switzerland, raising the question of the origin of the infection. The main aim of this study was to assess the extent of exposure of the wild boar populations to Sarcoptes scabiei in Switzerland, prior to and after the detection of mange cases, to determine whether the mite has been recently introduced into the populations concerned. We performed a serological survey using a commercially available ELISA and 1056 archived blood samples of free-ranging wild boar from Switzerland. To facilitate the interpretation of the obtained data, we additionally estimated seroprevalence in wild boar populations of four other European countries (1060 samples), both from areas with confirmed clinical cases of mange and from areas without reported cases in wild boar. Lastly, we revised the evaluation of the commercial ELISA when used with wild boar sera., Results: Seropositive reactions were observed for samples from all five countries and from 15 of the 16 study areas. The obtained apparent seroprevalences ranged from 0.0% (0/82; 95% confidence interval [CI]: 0.0-4.4) to 17.4% (8/46; 95% CI: 7.8-31.4). Wild boar from study areas with known clinical cases and those ≤60 kg were four times more likely to be seropositive than wild boar from areas without reported cases and > 60 kg, respectively. Optical density values did not differ between the two types of study areas among seropositive samples but were significantly lower among seronegative samples from areas without than from areas with clinical cases. No difference was observed between the two sampling periods in Switzerland. The revised ELISA specificity was 96.8% (984/1017; 95% CI: 95.5-97.7) when wild boar from areas without history of mange were considered truly negative., Conclusions: Seropositivity to S. scabiei is more frequent and occurs over a larger geographic range than expected. Data suggest that the parasite is endemic within the wild boar populations of Switzerland and other European countries but that its presence is not necessarily associated with disease occurrence. Extrinsic factors which trigger disease emergence in infected populations remain to be investigated. The applied ELISA represents a promising tool for future studies.

The moose (Alces alces) is a ruminant that harvests energy from fiber-rich lignocellulose material through carbohydrate-active enzymes (CAZymes) produced by its rumen microbes. We applied shotgun metagenomics to rumen contents from six moose to obtain insights into this microbiome. Following binning, 99 metagenome-assembled genomes (MAGs) belonging to 11 prokaryotic phyla were reconstructed and characterized based on phylogeny and CAZyme profile. The taxonomy of these MAGs reflected the overall composition of the metagenome, with dominance of the phyla Bacteroidetes and Firmicutes. Unlike in other ruminants, Spirochaetes constituted a significant proportion of the community and our analyses indicate that the corresponding strains are primarily pectin digesters. Pectin-degrading genes were also common in MAGs of Ruminococcus, Fibrobacteres and Bacteroidetes and were overall overrepresented in the moose microbiome compared with other ruminants. Phylogenomic analyses revealed several clades within the Bacteriodetes without previously characterized genomes. Several of these MAGs encoded a large numbers of dockerins, a module usually associated with cellulosomes. The Bacteroidetes dockerins were often linked to CAZymes and sometimes encoded inside polysaccharide utilization loci, which has never been reported before. The almost 100 CAZyme-annotated genomes reconstructed in this study provide an in-depth view of an efficient lignocellulose-degrading microbiome and prospects for developing enzyme technology for biorefineries.

In recent decades, wild boars (Sus scrofa) have increased in numbers and distribution in Europe. Compared to other wild ungulates of similar body size, wild boars have a high reproductive capacity. To increase the knowledge of wild boar reproduction, the objective of this study was to investigate characteristics of reproductive organs, and to provide information on the occurrence of abnormalities in reproductive organs from free-ranging female wild boars. Between December 2011 and December 2015, reproductive organs from female wild boars (>30 kg body weight), were collected during hunting in four Swedish counties at estates where supplementary feeding was applied. The organs were macroscopically examined and measured. The stage of the reproductive cycle was defined according to the ovarian structures and in relation to uterus characteristics. Observed abnormalities were noted. The results from 569 animals that met the requirements to be included in this study showed significant differences in weight and length of the uterus between the various reproductive stages. Sampling region had significant effect on these differences. Abnormalities in the reproductive organs were present in approximately 10% of the examined animals. The prevalence of abnormalities increased significantly with age and was significantly affected by sampling region., (© 2017 Blackwell Verlag GmbH.)

Background: The number and spatial distribution of wild boars (Sus scrofa) has increased remarkably in Sweden as well as in other European countries. To understand the population dynamics of the wild boar, knowledge of its reproductive period, oestrus cycle and reproductive success is essential. The aim of this study was therefore to describe the seasonal reproductive pattern and reproductive potential of a wild boar population in Sweden. The study was based on findings from macroscopic examinations of the reproductive organs from 575 hunter-harvested female wild boars (>30 kg body weight). Samples were collected between December 2011 and December 2015 in the southern and middle parts of Sweden. The age of the sampled animals was determined and dressed weight was noted. The stage of the reproductive cycle was defined according to ovarian structures and in relation to the appearance of/and findings in the uterus. The crown-rump length (CRL) of the embryos/foetuses was used to calculate the oestrus/mating month and month for the expected farrowing., Results: The macroscopic examination revealed a seasonal variation of reproductive stages, although cyclic and pregnant females were found in all seasons. Moreover, the estimated oestrus/mating and farrowing months based on the CRL showed that mating and farrowing may occur 'off-season'. The average litter size (no. of embryos or foetuses) per pregnant female was 5.4. Sow weight and age had significant effect on both the reproductive potential (ovulation rate and litter size) and pregnancy rate, respectively., Conclusions: The reproductive potential in the studied wild boar population was high compared to studies from other countries and farrowing may occur 'off-season'. This suggests that the environmental conditions in Sweden, including supplemental feeding, are favourable for wild boar reproduction.

Background: Schmallenberg virus (SBV) first emerged in Europe in 2011, and in Sweden in late 2012. The virus was still circulating in parts of Europe in 2015. In recent testing, the virus has not been detected in Swedish domestic animals, indicating that it is no longer circulating in Sweden. It is not known if the virus has circulated and is still circulating in Swedish wild cervid populations and whether wildlife can act as virus reservoirs. The aim of this study was to investigate whether SBV has circulated, and is still circulating among wild cervids in Sweden., Results: Ninety-two sera from moose (Alces alces, n = 22), red deer (Cervus elaphus, n = 15), fallow deer (Dama dama, n = 44), and roe deer (Capreolus capreolus, n = 11) were collected and analyzed for antibodies against SBV. The sampling occurred in the southern and middle part of Sweden during three time periods: 1) before the vector season in 2012, 2) after the vector season in 2012, and 3) after the vector season in 2015. Animals from periods 1 and 2 were of varying ages, whereas animals collected in period 3 were born after the vector season 2013. Animals from period 1 (n = 15) and 3 (n = 47) were seronegative, but, 53% (16 of 30) of animals from period 2 were seropositive, determined by SBV competitive ELISA. Samples from period 2 were additionally analyzed for SBV-neutralizing antibodies. Such antibodies were detected in 16/16 SBV-N-antibody-positive, 3/12 negative and 2/2 doubtful sera. The two tests were in accordance at SBV-neutralizing antibody titers of 1:32 or higher., Conclusion: Our results show that SBV circulated among wild cervids during the vector season of 2012. Three years later, no SBV-antibodies were detected in animals born after the vector season 2013. The likely absence of SBV circulation in Sweden, in contrast to other parts of Europe, might be explained by the annual occurrence of a vector-free season due to climate conditions. Interpretations are limited by the small sample-size, but the results suggest that the SBV competitive ELISA has high specificity but might have slightly lower sensitivity compared to a seroneutralization assay, when using samples from wild cervids.

Lipopolysaccharides (LPS) from Gram negative bacteria are involved in the pathogeny of uterine diseases in cows. This study aimed to investigate LPS effects on the growth of bovine endometrial epithelial cells (bEEC) and relationships between LPS response and tissue characteristics. Uteri from 35 females were characterized for parity and stage of oestrous cycle. Densities of glandular tissue (dGT), CD11b+ cells and Ki67+ cells were measured in the endometrial tissue. Cells from 13 dioestrus cows were exposed to 0, 2, 4, 8, 12, 16 or 24μg/mL LPS. Effects of parity and stage of the oestrous cycle on tissue characteristics and effects of LPS dosage, cow and tissue characteristics on changes in cell numbers were analyzed by ANOVA. The dGT was higher in metoestrus and dioestrus samples than in pro-oestrus ones whereas densities of CD11b+ and Ki67+ cells were higher at pro-oestrus (p<0.05-p<0.01). LPS influenced bEEC populations in a dose related manner. An increase in number of live cells was observed for dosages ranging from 2 to 12μg/mL LPS (p<0.0001 vs controls). No effect was found on numbers and frequencies of dead cells. With higher dosages, the numbers of live cells did not increase but the numbers of dead did increase. No relationships were observed between cow or tissue characteristics and growth patterns or frequencies of viable bEEC in controls nor in the response to LPS. To conclude this model is suitable for further studies on dysregulations induced by LPS in endometrial tissue., (Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Since the re-appearance of wild boars in Sweden in the 1970s, the population has increased. Besides having large litter sizes, puberty at an early age is considered as an important factor contributing to the high reproductive potential of wild boar. Although controversial, supplemental feeding is applied to varying extent throughout the wild boar range in Sweden, and its effect on wild boar reproduction is debated. The aim of this study was to investigate the proportion of post-pubertal female wild boar gilts in a population subjected to supplemental feeding, in relation to age, weight, and season. Also, the effect of another definition of puberty (based upon follicular size) on the outcome of the proportion of female wild boar gilts considered to be able to reproduce in a population was illustrated. Between Jan 2013 and Dec 2015 reproductive organs from 592 female wild boars, were collected. Ovaries and uterus were macroscopically examined, and reproductive stage was determined by the presence of ovarian structures and uterus characteristics. Age was estimated using tooth eruption and tooth replacement, and weight was noted. A subset of 175 female wild boars, aged 5-15 months, was included in this study. An animal was considered to be post-pubertal if the ovaries contained one or more corpora lutea (CL) or if the uterus showed signs of previous pregnancy (presence of enlarged blood vessels in the cut surface between the mesometric ligament and the uterus)., Results: In total, 29 (16.6 %) animals were classified as post-pubertal. Field dressed weight ranged from 20.6 to 65.3 kg. Season, weight, and age class significantly influenced the variation in proportion of post-pubertal females. Post-pubertal animals were found in autumn, winter, and spring, but not in the summer season. Another definition of puberty, based on follicle size, lead to different result on proportions of animals considered to have passed puberty., Conclusions: Season, weight, and age significantly influenced the variation in proportion of post-pubertal wild boar females. The proportion of post-pubertal animals increased with age and weight. However, weight is probably a better proxy for puberty than age group is. The proportion of post-pubertal females also increased from summer to spring suggesting a seasonal reproductive pattern. Different definitions of puberty will result in various outcomes, which high-lights the importance of using adequate definition of puberty.

Both Rickettsia helvetica and Anaplasma phagocytophilum are common in Ixodes ricinus ticks in Sweden. Knowledge is limited regarding different animal species' competence to act as reservoirs for these organism. For this reason, blood samples were collected from wild cervids (roe deer, moose) and domestic mammals (horse, cat, dog) in central Sweden, and sera were tested using immunofluorescence assay to detect antibodies against spotted fever rickettsiae using Rickettsia helvetica as antigen. Sera with a titer ≥1:64 were considered as positive, and 23.1% (104/450) of the animals scored positive. The prevalence of seropositivity was 21.5% (23/107) in roe deer, 23.3% (21/90) in moose, 36.5% (23/63) in horses, 22.1% (19/90) in cats, and 17.0% (17/100) in dogs. PCR analysis of 113 spleen samples from moose and sheep from the corresponding areas were all negative for rickettsial DNA. In roe deer, 85% (91/107) also tested seropositive for A. phagocytophilum with a titer cutoff of 1:128. The findings indicate that the surveyed animal species are commonly exposed to rickettsiae and roe deer also to A. phagocytophilum.

Between January and December 2013, the dental and periodontal health of 99 Swedish wild boars (Sus scrofa) was investigated. Sampling occurred in conjunction with routine hunting at six large estates in the southern and middle parts of Sweden. All six of the estates use supplemental feeding. The weight of the animals, their sex and their dates of death were noted. Age was estimated using tooth eruption and tooth replacement patterns. The oral cavity was inspected and abnormalities were recorded on a dental chart modified for wild boars. The findings included supernumerary teeth, absence of teeth, mild class II malocclusion, severe tooth wear, periodontitis, calculus, caries, tooth fractures and the presence of enamel defects. Swedish wild boars suffer from different dental lesions and the impact of supplemental feeding on dental and periodontal health is still to be investigated., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Background: Hepatitis E virus (HEV) infects a range of species, including humans, pigs, wild boars and deer. Zoonotic transmission may contribute to the high HEV seroprevalence in the human population of many countries. A novel divergent HEV from moose (Alces alces) in Sweden was recently identified by partial genome sequencing. Since only one strain was found, its classification within the HEV family, prevalence in moose and zoonotic potential was unclear. We therefore investigated samples from 231 moose in seven Swedish counties for HEV, and sequenced a near complete moose HEV genome. Phylogenetic analysis to classify this virus within the family Hepeviridae and to explore potential host specific determinants was performed., Methods and Findings: The HEV prevalence of moose was determined by PCR (marker for active infection) and serological assays (marker of past infection) of sera and 51 fecal samples from 231 Swedish moose. Markers of active and past infection were found in 67 (29%) animals, while 34 (15%) were positive for HEV RNA, 43 (19%) were seropositive for anti-HEV antibodies, and 10 (4%) had both markers. The number of young individuals positive for HEV RNA was larger than for older individuals, and the number of anti-HEV antibody positive individuals increased with age. The high throughput sequenced moose HEV genome was 35-60% identical to existing HEVs. Partial ORF1 sequences from 13 moose strains showed high similarity among them, forming a distinct monophyletic clade with a common ancestor to HEV genotype 1-6 group, which includes members known for zoonotic transmission., Conclusions: This study demonstrates a high frequency of HEV in moose in Sweden, with markers of current and past infection demonstrated in 30% of the animals. Moose is thus an important animal reservoir of HEV. The phylogenetic relationship demonstrated that the moose HEV belonged to the genotype 1-6 group, which includes strains that also infect humans, and therefore may signify a potential for zoonotic transmission of this HEV.

Knowledge of the reproductive biology of game species is vital for sustainable management. In moose (Alces alces), research in reproductive characteristics has focused on the female, whereas there are few studies in male moose. The aim of the present study was to investigate sperm morphology and chromatin integrity (SCSA), and their relationships with testicular and epididymal features, as well as temporal aspects with respect to the hunting season. In total, 143 male moose aged 1.5-11.5 years were sampled from 2008 to 2011. The proportion of normal spermatozoa (PNS) ranged from 1.5% to 82.0%, with a mean of 51%, and the %DFI (DNA fragmentation index) ranged from 2.5% to 36.7% (mean 9.5). PNS decreased temporally, and was positively associated with carcass and testes weight. Body weight and testes weight had positive effect on PNS regardless of age. No effect of any explanatory variables was observed on the DFI. The testis/body weight ratio of moose (0.033%) is among the lowest reported among mammals, indicating a less polygynous mating system than in roe deer and red deer. For reproduction success in moose, a high body weight in males is favorable, as is a balanced sex ratio. Thus, males should not be harvested prior to the time when the majority of females have passed their first oestrus of the season., (Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.)

A successful outcome after artificial insemination with cooled semen is dependent on many factors, the sperm quality of the ejaculate being one. Previous studies have shown that spermatozoa with good motility, normal morphology, and good chromatin integrity can be selected by means of colloid centrifugation, particularly single layer centrifugation (SLC) using species-specific colloids. The purpose of the present study was to conduct an insemination trial with spermatozoa from "normal" ejaculates, i.e., from stallions with no known fertility problem, to determine whether the improvements in sperm quality seen in SLC-selected sperm samples compared with uncentrifuged controls in laboratory tests are reflected in an increased pregnancy rate after artificial insemination. In a multicentre study, SLC-selected sperm samples and uncentrifuged controls from eight stallions were inseminated into approximately 10 mares per treatment per stallion. Ultrasound examination was carried out approximately 16 days after insemination to detect an embryonic vesicle. The pregnancy rates per cycle were 45% for controls and 69% for SLC-selected sperm samples, which is statistically significant (P < 0.0018). Thus, the improvement in sperm quality reported previously for SLC-selected sperm samples is associated with an increase in pregnancy rate, even for ejaculates from stallions with no known fertility problem., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Background: Q fever is a zoonotic disease caused by the bacterium Coxiella burnetii. Prevalence data in ruminant species are important to support risk assessments regarding public and animal health. The aim was to investigate the presence of or exposure to C. burnetii in cattle, sheep, goats and moose, and to compare two enzyme-linked immunosorbent assays (ELISAs). National surveys of antibodies against C. burnetii were performed for dairy cattle (n=1537), dairy goats (n=58) and sheep (n=518). Bovine samples consisted of bulk milk, caprine of pooled milk, and ovine of pooled serum. Antibodies were investigated in moose samples (n=99) from three regions. A one-year regional cattle bulk milk survey was performed on the Isle of Gotland (n=119, four occasions). Cattle, sheep and goat samples were analysed with indirect ELISA and moose samples with complement fixation test. For the sheep, goat, and parts of the cattle survey, samples were run in parallel by ELISAs based on antigens from infected ruminants and ticks. Bulk milk samples from the regional cattle survey and vaginal swabs from a subset of the sheep herds (n=80) were analysed for the agent by polymerase chain reaction. Spatial clustering was investigated in the national cattle survey., Results: The prevalence of antibodies in dairy herds was 8.2% with large regional differences. High risk clusters were identified in the southern regions. The prevalence among dairy herds on the Isle of Gotland varied from 55.9% to 64.6% and 46.4% to 58.9.0% for antibodies and agent, respectively, overall agreement between agent and antibodies was 85.2%. The prevalence of antibodies in sheep was 0.6%, the agent was not detected the vaginal swabs. Antibodies were not detected in goats or moose, although parts of the moose samples were collected in an area with high prevalence in cattle. The overall agreement between the two ELISAs was 90.4%., Conclusions: The prevalence of antibodies against C. burnetii in dairy cattle in Sweden shows large regional differences. The results suggest that C. burnetii is a rare pathogen among Swedish moose, dairy goat and sheep. ELISAs based on ruminant and tick antigen performed in a similar manner under Swedish conditions.

Since January 1 2013, group housing of sows has been compulsory within the European Union (EU) in all pig holdings with more than ten sows. Sows and gilts need to be kept in groups from 4 weeks after service to 1 week before the expected time of farrowing (Article 3(4) of Directive 2008/120/EC on the protection of pigs). The legislation regarding group housing was adopted already in 2001 and a long transitional period was allowed to give member states and producers enough time for adaptation. Even so, group housing of sows still seems to be uncommon in the EU, and is also uncommon in commercial pig farming systems in the rest of the world. In this review we share our experience of the Swedish 25 years of animal welfare legislation stipulating that sows must be loose-housed which de facto means group housed. The two most important concerns related to reproductive function among group-housed sows are the occurrence of lactational oestrus when sows are group-housed during lactation, and the stress that is associated with group housing during mating and gestation. Field and clinical observations in non-lactating, group-housed sows in Sweden suggest that by making basic facts known about the pig reproductive physiology related to mating, we might achieve application of efficient batch-wise breeding without pharmacological interventions. Group housing of lactating sows has some production disadvantages and somewhat lower productivity would likely have to be expected. Recordings of behavioural indicators in different housing systems suggest a lower welfare level in stalled animals compared with group-housed ones. However, there are no consistent effects on the reproductive performance associated with different housing systems. Experimental studies suggest that the most sensitive period, regarding disturbance of reproductive functions by external stressors, is the time around oestrus. We conclude that by keeping sows according to the pig welfare-friendly Directive 2008/120/EC, it is possible to combine group-housing of sows with good reproductive performance and productivity. However, substantially increased research and development is needed to optimize these systems.

Summary: The occurrence of Anaplasma phagocytophilum was investigated in spleen and serum samples from Swedish moose (Alces alces) in southern Sweden (island and mainland). Samples were analysed for presence of A. phagocytophilum DNA by real-time PCR (n = 263), and for Anaplasma antibodies with ELISA serology (n = 234). All serum samples had antibodies against A. phagocytophilum. The mean DNA-based prevalence was 26·3%, and significant (P < 0·01) temporal, and spatial variation was found. Island moose had significantly (P < 0·001) higher prevalence of A. phagocytophilum DNA than moose from the mainland areas. Two samples were sequenced to determine genetic variation in the 16S rRNA and groESL genes. Genetic sequence similarity with the human granulocytic anaplasmosis agent, equine granulocytic ehrlichiosis agent, and different wildlife-associated A. phagocytophilum variants were observed in the 16S rRNA and groESL genes. Our study shows that moose are exposed to A. phagocytophilum in Sweden, and represent a potential wildlife reservoir of the pathogen.

Background: The moose (Alces alces) is an intensively managed keystone species in Fennoscandia. Several aspects of reproduction in moose have not been fully elucidated, including puberty, timing of mating and oestrus, and the length of the oestrus period. These aspects are relevant for an adaptive management of moose with respect to harvest, population size, demography and environmental conditions. Therefore, an investigation of female moose reproduction was conducted during the moose-hunting period in southern Sweden from 2008 to 2011., Results: A total of 250 reproductive organs and information on carcass weight and age was collected from four different hunting areas (provinces of Öland, Småland, Södermanland, and Västergötland) in southern Sweden. The results showed that puberty in female moose varied with carcass weight, age, and time of season. The period for oestrous/mating lasted from about mid September to the beginning of November., Conclusions: The oestrus period (predominantly for heifers) is longer than previously reported and was not finished when the hunting period started. Sampling the uterine cervix to detect spermatozoa was a useful method to determine if mating had occurred. To avoid hunting of moose during oestrus, we suggest that the hunting period should be postponed by at least 14 days in southern Sweden.

Knowledge on reproductive success is vital for successful management of large ungulates and is often measured by means of observing surviving offspring. In harvested ungulates, postmortem investigations of reproductive organs are used to estimate reproductive potential by obtaining ovulation rates and fetus numbers. However, there are differences in numbers of offspring observed, fetal/embryo counts, and ovulation rates. We hypothesize that the discrepancy between estimated reproductive potential and reproductive outcome in large ungulates is not only due to ova loss but also due to embryonic mortality. We investigated reproductive status in early pregnancy by sampling hunter-harvested moose ( Alces alces ) in southern Sweden from 2007 to 2011. In all, 213 reproductive organs were examined postmortem, and in confirmed pregnant moose ( n  = 53), 25 % (19 of 76) embryos were nonviable and 6 % of ova was unfertilized. The discrepancy between the ovulation rate of all pregnant moose (1.49) and the number of expected offspring per pregnant female, when embryonic mortality and unfertilized oocytes were accounted for (1.08), was 27.5 %. An association between inflammation of the inner mucous membrane (endometritis) of the moose's uterus and embryonic mortality was observed. This is the first comprehensive report of embryonic mortality and endometritis in moose. The observed discrepancy between ovulation rates and early embryonic development/survival shows that ovulation rates are indicative but not accurate estimates of moose reproductive rate. The use of ovulation rates as a sole estimator of future offspring rates may lead to an overharvest of a managed moose population.

There is controversy about whether the presence of some seminal plasma (SP) in an equine insemination dose is necessary for promoting fertility. A new technique for improving stallion sperm quality, single layer centrifugation (SLC) using a species-specific colloid, Androcoll-E, selects a sperm subpopulation that is highly motile with normal morphology, intact membranes and good chromatin integrity from the rest of the ejaculate and removes SP. The present study was designed to investigate the effect of restoring homologous SP (5% and 10%) on the progressive motility, velocity, and chromatin integrity of SLC-selected stallion spermatozoa in 44 semen samples over time. Sperm progressive motility (P<0.01) and the proportion with class A velocity (>50 μm/sec) were increased in samples where SP was restored, whereas the proportion with class B velocity (10 to 50 μm/sec) was decreased compared with SLC samples. However, after 24 h cold storage of treated samples, progressive motility was not different for the SP-treated groups compared with SLC, whereas chromatin damage DNA fragmentation index (%DFI) was higher. In contrast, adding SP to untreated 24 h-stored SLC samples did not affect progressive motility although it did increase the proportion of spermatozoa with class A velocity. There was individual variation between stallions whether 5% or 10% SP produced a greater increase in progressive motility. In conclusion, 5% to 10% SP can be added back to SLC-selected samples if considered necessary to optimize fertility. However, it should be added immediately before insemination rather than before storage of the sperm dose, to benefit from the transient increase in sperm progressive motility and avoid increased chromatin damage., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Background: One of the challenges faced by equine breeders is ensuring delivery of good quality semen doses for artificial insemination when the mare is due to ovulate. Single Layer Centrifugation (SLC) has been shown to select morphologically normal spermatozoa with intact chromatin and good progressive motility from the rest of the ejaculate, and to prolong the life of these selected spermatozoa in vitro. The objective of the present study was a proof of concept, to determine whether fertilizing ability was retained in SLC-selected spermatozoa during prolonged storage., Findings: Sixteen mares were inseminated with SLC-selected sperm doses that had been cooled and stored at 6°C for 48 h, 72 h or 96 h. Embryos were identified in 11 mares by ultrasound examination 16-18 days after presumed ovulation., Conclusion: SLC-selected stallion spermatozoa stored for up to 96 h are capable of fertilization.

Levels of the cytokines transforming growth factor (TGF)-β1, interleukin (IL)-10 and IL-6 in the boar seminal plasma (SP) as well as TGF-β1 level in different fractions of ejaculate were studied. These cytokines was chosen because of their expected effect on tissue immune response, i.e. suppressive (TGF-β1 and IL-10) and pro-inflammatory (IL-6). Three whole ejaculates from five boars A-E, (n=15) were sampled weekly to evaluate the levels of seminal plasma TGF-β1, IL-10 and IL-6 as well as their fluctuations over time. The effect of different storage temperatures, -20°C or -80°C, on the level of seminal plasma TGF β1 was also tested (three boars, two fractions in one ejaculate). In addition, in 4 different fractions of ejaculates: the pre-sperm-rich (Pre-SRF), first 10 ml of sperm-rich (10SRF), the rest of the sperm-rich fraction (Rest-SRF) and the rest of the ejaculate (RE) fraction, were collected from three boars (A-C) on four different occasions for TGF-β1 evaluation. In the whole ejaculates (n=15), a wide range in the concentration of the cytokines TGF-β1 (20.4 - 766.5 pg/mL) and IL-10, (73.7 - 837.3 pg/mL), was found. For IL-6, the concentration was low (range 11.5 - 30.9 pg/ml) and only detected in four out of 15 collections (from two boars). The mean levels of TGF-β1 and IL-10 between individual boars varied but were not statistical different. The level of TGF-β1 in Pre-SRF, Rest-SRF and RE fractions was significantly lower in boar A than the other boars. A significantly higher concentration of TGF-β1 was found in the 10SRF than in the other fractions. Different storage temperatures (-20°C or -80°C) did not affect the seminal plasma TGF-β1 level after one year of storage. To conclude: Boar seminal plasma contained TGF- β1 and IL-10 but with high individual variation. IL-6 was low or undetectable. The TGF- β1 level was highest in the first 10 mL of the sperm-rich fraction of the ejaculate. Further studies are needed on the role of different levels of cytokine in boar semen on porcine female reproductive tissue, especially for TGF- β1., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The effects of semen components or extender alone on the expression of selected cytokines [interleukine (IL)-1β, IL-6, granulocyte-macrophage colony stimulating factor (GM-CSF), IL-10 and transforming growth factor (TGF)-β1] on the porcine endometrium were studied, as well as the presence of polymorphonuclear neutrophilic granulocytes (PMNs). In experiment (Exp) I, groups of gilts were sampled at 5-6h after insemination with fresh semen in extender (Beltsville thawing solution, BTS), spermatozoa in extender (Spz), seminal plasma (SP), or only BTS (control). In Exp II, gilts were sampled 35-40h after insemination with Spz, SP, BTS or only catheter inserted (as control). Immunohistochemical (IHC) labelling of IL-6, IL-10 and TGF-β1 was evident, especially in surface and glandular epithelia of the porcine endometrium. There were no consistent differences in IHC-labelling of the cytokines in relation to different treatments. However, the scores for IL-6 and IL-10 in surface epithelium and sub-epithelial connective tissue compartments were higher at 35-40h than shortly (5-6h) after treatment. Cytoplasmic labelling in the sub-epithelial connective tissue was observed in scattered individual cells but not in PMNs. Shortly (5-6h) after insemination, there were no differences between animals inseminated with BTS (control) and the semen components for any of the cytokine mRNAs. Later however, at 35-40h, lower endometrial expression of TGF-β1 mRNA was observed in the Spz and BTS groups compared with the control (catheter only). The same pattern was found for IL-10 (NS). The mRNA expression of IL-6 in the BTS inseminated group was higher compared to the control group. Insemination with SP resulted in significantly lower PMN cell infiltration in the sub-epithelial connective tissue compared with Spz or BTS groups shortly (5-6h) after insemination. Later (35-40h), a significant difference was found between SP (lower) and the control group (only catheter). To conclude, our results show that insemination and/or inseminated components modulated cytokine expression in the gilt endometrium. The semen extender BTS stimulated immune reactivity, as shown by down-regulation of the suppressive cytokine TGF-β1. Insemination with solely SP clearly decreased PMN cell infiltration of the gilt endometrium. However, no clear relation between the cytokines studied and PMN cell presence was found., (Copyright © 2010 Elsevier B.V. All rights reserved.)

This study was designed to evaluate the effect of single layer centrifugation (SLC) and subsequent cold storage on stallion sperm capacitation-like status and acrosome reaction. Three stallions were included in the study, with three ejaculates per stallion. The samples were examined 4, 24 and 72 h after collection, extension and SLC, with storage at 6°C. Sperm capacitation-like status was investigated using the fluorescent dye chlortetracycline (CTC). There was no difference in capacitation-like status between colloid-selected and non-selected spermatozoa. Sperm motility decreased significantly during cold storage, whereas the proportion of apparently capacitated spermatozoa increased. There was no change in the proportion of acrosome-reacted spermatozoa. In conclusion, SLC through Androcoll™-E does not adversely affect the capacitation-like status of stallion spermatozoa, although it did increase with time during cold storage., (© 2010 Blackwell Verlag GmbH.)

Effects of semen components [fresh semen in extender, spermatozoa in extender (Spz), seminal plasma (SP)], or extender alone (Beltsville thawing solution, BTS) on the expression of selected cytokines [interleukin (IL)-1beta, IL-6, IL-10 and transforming growth factor (TGF)-beta1)] as well as the presence of cells positive for CD8 or CD25 were studied in the pig oviduct. In addition, cytokines in SP and oviductal flushings were analyzed. In experiment (Exp) I, groups of gilts were sampled at 5-6h after insemination with SP, Spz, fresh semen in BTS or only BTS (control). In Exp II, gilts were sampled 35-40 h after insemination with SP, Spz, BTS or only catheter insertion (control). Most oviductal flushing samples were positive (> or =detectable limits) for IL-10 and TGF-beta1 but only few for IL-6. The IHC-labelling of IL-6, IL-10 and TGF-beta1 was evident, especially in the epithelial cells of the isthmus and infundibulum as well as in the cells of the regional (mesometrial) lymph node. Cilia of the epithelium were positive for IL-6 (strongest in the infundibulum) and TGF-beta1 (strongest in the isthmus) but negative for IL-10. There were no consistent differences in IHC-labelling of the cytokines in relation to different treatments, except at 35-40 h after insemination (Exp II), when IL-6 was slightly higher in epithelium of the SP group and IL-10 in the infundibular connective tissue was higher in the SP and Spz groups. In the isthmus and infundibulum, there were no differences between animals inseminated with BTS (control) and the semen components for any of the cytokine mRNAs at 5-6h after insemination (Exp I). However, later (35-40 h, Exp II), insemination with SP, Spz and BTS alone appeared to up-regulate TGF-beta1 mRNA expression compared with the control group (without any fluid infused). In all treatment groups, the mRNA level for TGF-beta1 was higher than for IL-1beta, IL-6 and IL-10. Higher mRNA levels of all cytokines were found in the isthmus compared with the infundibulum. Numbers of CD8-positive cells (both in epithelium and connective tissue) appeared higher in the infundibulum compared with the isthmus and were mostly higher shortly (Exp I) after treatment with SP, SPZ and BTS than later (Exp II) in both segments. CD25-positive cells were few and found solely in the sub-epithelial connective tissue. The results indicate that in the porcine oviduct, IL-6, IL-10 and TGF-beta1 are endogenous produced and that TGF-beta1 may have a more important role for immunomodulation than the other cytokines, especially in isthmus. Differences between isthmus and infundibulum in cytokine mRNA expression and in presence of CD8-positive cells indicate different patterns of immune reactivity in the upper and lower parts of the oviduct., (Copyright (c) 2010 Elsevier B.V. All rights reserved.)

On-stud assessment of stallion sperm quality can be problematic. A new instrument, the Nucleocounter SP-100, was validated for measuring stallion sperm concentration and viability. It was subsequently used to evaluate sperm viability in Kenney's extender and INRA96. There was a strong correlation between sperm concentrations measured by the Nucleocounter SP-100 and by the Bürker counting chamber (r = 0.84; P < .001). Similarly, there was a good correlation between sperm viability results from the Nucleocounter SP-100 and flow cytometric results (r = 0.73; P < .001). Sperm viability at 24 hours was significantly better for samples extended in INRA96 than in Kenney's extender (P < .001). Furthermore, sperm kinematics were better for stored samples in INRA96 than in Kenney's extender. Single Layer Centrifugation selected spermatozoa that maintained their viability better during storage for 24 hours than the uncentrifuged samples. In conclusion, the type of semen extender used and Single Layer Centrifugation were found to influence both the kinematics and viability of stallion spermatozoa. The Nucelocounter-SP100 was considered to be a useful instrument for rapidly measuring stallion sperm concentration and viability.

The objective was to investigate the changes in stallion sperm quality (sperm motility, viability, membrane integrity and chromatin integrity) occurring during cool storage, and to study the effect of sperm selection by single layer colloidal centrifugation on these parameters of sperm quality. Spermatozoa from 3 stallions (10 ejaculates, 3-4 per stallion) were selected by centrifugation through a single layer of colloid (SLC). The resulting sperm preparations and the control samples (extended but unselected semen samples) were stored at 5 degrees C for 48h. Assessments of sperm quality, such as sperm motility, viability (SYBR-14/PI staining), membrane stability (Annexin-V/PI staining) and chromatin integrity, were performed on aliquots of the selected sperm preparations and unselected samples on the day of collection (3h) and after 24 and 48h of storage. In the SLC-selected sperm samples, sperm motility, sperm viability, proportions of spermatozoa with normal morphology and with intact chromatin were significantly better than in unselected samples (motility: 77+/-4% vs. 64+/-8% at 3h; P<0.001; viability: 79.5+/-9% vs. 64.7+/-9%, P<0.001; normal morphology 89+/-6% vs. 69+/-9%; chromatin integrity DFI 11.3+/-5% vs. 22.1+/-10%). Membrane stability, however, was not different in the SLC-selected and unselected samples (74.6+/-8% vs. 69.3+/-8%). The deterioration seen in sperm quality in the unselected samples was prevented by SLC, so that sperm viability, membrane stability and chromatin integrity were unchanged in the selected samples by 48h compared to 3h (P<0.001), whereas the unselected samples were significantly worse by 48h (P<0.001). Furthermore, it should be possible to send an aliquot of a normal insemination dose (i.e. unselected spermatozoa) overnight to a reference laboratory for analysis of both plasma membrane and chromatin integrity. In conclusion, centrifugation of stallion spermatozoa through a single layer of colloid is a useful technique for selecting the best spermatozoa from an ejaculate and, moreover, sperm quality is maintained during storage.

The objective of the current study was to optimize the volumes of Androcoll-E and sperm sample used in various sizes of centrifuge tube to scale up single-layer centrifugation (SLC) for routine use in the field. Although sperm suspensions of equivalent quality were produced using Androcoll-E in small and large tubes, the sperm yield was much lower in the latter (P<0.001). In contrast, in 200-mL tubes (XL), the yields were approximately 25% higher than those for the small tubes. An increased volume (4.5 mL) of extended ejaculate in small tubes (SLC-Inc) or 15 to 18 mL extended ejaculate on 15 mL of colloid of a reduced density, Androcoll-E-Large (SLC-Large), in 50-mL tubes were both found to give similar yields of motile spermatozoa as that of the SLC-Small method (SLC-Small, 49.7+/-18.6%; SLC-Inc, 53.3+/-17.1%; SLC-Large, 44.9+/-18.3%) and were found to be equivalent in quality (motility: 88.0+/-8.8%, 84.0+/-3.5%, 90.0+/-5.4%; normal morphology: 69.4+/-17.0%, 69.4+/-12.7%, 63.9+/-15.6%; viability: 78+/-16.7%, 83.8+/-12.5%, 80.05+/-14.6%; DNA fragmentation index: 14.7+/-10.9%, 12.8+/-8.1%, 11.6+/-7.6%, respectively). The processing of an "average" stallion Equus caballus ejaculate in approximately twenty-seven 10-mL tubes (SLC-Inc) or eight 50-mL tubes (SLC-Large) is feasible, the latter being considered more practical for on-stud use.

Background: Sex steroid hormone receptors have been identified in the adrenal gland of rat, sheep and rhesus monkey, indicating a direct effect of sex steroids on adrenal gland function., Methods: In the present study, immunohistochemistry using two different mouse monoclonal antibodies was employed to determine the presence of oestrogen receptor alpha (ERalpha) and progesterone receptor (PR) in the mare adrenal gland. Adrenal glands from intact (n = 5) and ovariectomised (OVX) (n = 5) mares, as well as uterine tissue (n = 9), were collected after euthanasia. Three of the OVX mares were treated with a single intramuscular injection of oestradiol benzoate (2.5 mg) 18-22 hours prior to euthanasia and tissue collection (OVX+Oe). Uterine tissue was used as a positive control and showed positive staining for both ERalpha and PR., Results: ERalpha staining was detected in the adrenal zona glomerulosa, fasciculata and reticularis of all mare groups. Ovariectomy increased cortical ERalpha staining intensity. In OVX mares and one intact mare, positive ERalpha staining was also detected in adrenal medullary cells. PR staining of weak intensity was present in a low proportion of cells in the zona fasciculata and reticularis of all mare groups. Weak PR staining was also found in a high proportion of adrenal medullary cells. In contrast to staining in the adrenal cortex, which was always located within the cell nuclei, medullary staining for both ERalpha and PR was observed only in the cell cytoplasm., Conclusion: The present results show the presence of ERalpha in the adrenal cortex, indicating oestradiol may have a direct effect on mare adrenal function. However, further studies are needed to confirm the presence of PR as staining in the present study was only weak and/or minor. Also, any possible effect of oestradiol treatment on the levels of steroid receptors cannot be determined by the present study, as treatment time was of a too short duration.

The aim of the present study was to investigate differences in the expression of mRNAs for ERalpha, ERbeta and PR in the sow uterus at different stages of the estrous cycle as well as in inseminated sows at estrus and during early pregnancy by use of solution hybridization and in relation to plasma levels of estradiol and progesterone. Uterine samples were collected at different stages of the estrous cycle and after insemination/early pregnancy. In the endometrium, the expression of ERalpha mRNA and PR mRNA was similar for cyclic and early pregnant groups. Both were highest at early diestrus/70 h after ovulation and ERalpha mRNA was lowest at late diestrus/d 19 while PR mRNA was lowest at diestrus and late diestrus/d 11 and d 19. The expression of endometrial ERbeta was constantly low during the estrous cycle but higher expression was found in inseminated/early pregnant sows at estrus and 70 h after ovulation. In the myometrium, high expression of ERalpha mRNA and PR mRNA was observed at proestrus and estrus in cyclic sows and at estrus in newly inseminated sows. Higher expression of myometrial ERbeta mRNA was found in inseminated/early pregnant sows compared with cyclic sows, although significant only at estrus. In conclusion, the expression of mRNAs for ERalpha, ERbeta and PR in the sow uterus differed between endometrium and myometrium as well as with stages of the estrous cycle and early pregnancy. In addition to plasma steroid levels, the differences between cyclic and inseminated/early pregnant sows suggest that other factors, e.g. insemination and/or the presence of embryos, influence the expression of these steroid receptor mRNAs in the sow uterus.

The aim of this study was to estimate genetic parameters for longevity from Swedish crossbred sows to investigate the possibilities of selecting for this trait. Data were collected from 16 commercial piglet-producing herds, on crossbred (Landrace × Yorkshire) sows farrowing in the period 1 January 2001 to 31 December 2004. The data set with records on 10 373 sows was split into two sets according to the breed of the sire, i.e. Landrace sires (LS) or Yorkshire sires (YS). Removal hazard during productive life (PL) was analysed with survival analysis, using a sire model. Stayability from first to second litter (STAY12), stayability from first to third litter (STAY13), length of productive life (LPL) and lifetime production (LTP) were analysed with linear models, using an animal model. Females after the worst sire had 1.7 times higher (progeny of LS) and 2.4 times higher (progeny of YS) risk of removal than females after the best sire. Heritability for PL was estimated at 0.06 (LS) and 0.12 (YS). The heritabilities for the linear longevity traits ranged from 0.03 to 0.08. Genetic correlations between the four linear longevity traits were all high and positive (0.6 to 1.0), as were the phenotypic correlations (0.5 to 0.8). The correlations (Spearman rank) between the sire's estimated breeding values for all the five longevity traits were all significant (P < 0.001) and moderate to strong in both data sets. Estimated breeding value (EBV) correlations between the five longevity traits and traits included in the present Swedish breeding evaluation (Quality Genetics (QG)) were significant in a few cases. Significant and favourable EBV correlations were found between age at first farrowing and both STAY12 and STAY13 (-0.20 and -0.31), as well as between litter weight at 3 weeks and LPL and LTP (0.13 to 0.20). Significant and unfavourable EBV correlations were found between age at 100 kg and STAY12 (0.32), as well as between the exterior conformation score from testing station and PL (-0.20). The level of the estimated heritabilities for longevity indicates that genetic improvement of sow longevity would be possible. However, overall, there was no strong indirect selection for sow longevity with the current Swedish breeding evaluation (QG).

The objective was to investigate whether it is possible to improve the quality of stallion semen, with respect to sperm morphology and chromatin integrity, both of which have been linked to fertility, using either density gradient centrifugation (DGC) or a new method, hereby named single layer centrifugation (SLC). The two methods of colloidal centrifugation were evaluated using 38 ejaculates from 10 stallions. Sperm morphology, subjective motility and sperm chromatin integrity were compared in uncentrifuged samples and in centrifuged sperm preparations. The proportion of morphologically normal spermatozoa varied between stallions (p < 0.001) and was increased by both methods of colloidal centrifugation (median value before centrifugation 67.5%; after SLC 78%; after DGC 77%; p < 0.001). The incidence of certain abnormalities was reduced, e.g. proximal cytoplasmic droplets were reduced from 12.9% to 8.8% (p < 0.001), and mid-piece defects from 5.3% to 1.4% (p < 0.05). Similarly, sperm motility and chromatin integrity were significantly improved (p < 0.001), with no difference between the two centrifugation methods. Centrifugation through colloids can enrich the proportions of stallion spermatozoa with normal morphology and normal chromatin structure in sperm preparations. The new method, SLC, was as effective as DGC in selecting motile stallion spermatozoa with normal morphology and intact chromatin. SLC, being simpler to use than DGC, would be appropriate for routine use by stud personnel to improve stallion sperm quality in insemination doses.

The present study investigated daily sperm output and sperm morphology of fresh semen in eight Swedish Warmblood stallions aged 5-8 years. They were used for artificial insemination, and their fertility during the breeding season of semen collection exceeded 60% per cycle. One ejaculate of semen was collected daily for 10 consecutive days from each stallion. The gel-free volume was measured, and the sperm concentration was assessed with a Bürker chamber. The volume of gel-free fraction was multiplied by the sperm concentration to give the total number of spermatozoa (TSN). Sperm morphology was examined in ejaculates collected on days 2, 5 and 10. An aliquot from each ejaculate was fixed in 1 ml formol-saline immediately after collection and examined under a phase-contrast microscope (magnification 1000x) to assess morphological abnormalities. Furthermore smears were prepared and stained according to Williams (carbolfuchsin-eosin) for a more detailed examination of the sperm heads under a light microscope (magnification 1000x). Analysis of variance was applied to data. Total spermatozoa number decreased progressively during the first 8 days of collection, and daily sperm output (DSO) was calculated as mean TSN of collections on days 8-10, being 6.4 x 10(9) spermatozoa. The overall percentages of morphologically normal spermatozoa in ejaculates collected on days 2, 5 and 10 were above 70%, being significantly lower in ejaculate 2 (68.6%) compared with ejaculates 5 and 10 (72.9% respectively 75.3%).

Reasons for Performing Study: A new, simpler, technique of colloidal centrifugation has recently been developed, designated single layer centrifugation (SLC). This technique requires evaluation by comparison with a density gradient for its ability to select the best quality spermatozoa and its practicality of use on studfarms., Objective: To compare the effect of 2 methods of colloidal centrifugation, density gradient centrifugation and single layer centrifugation, on stallion sperm motility, yield and survival, using freshly collected extended stallion semen., Methods: Aliquots of extended stallion semen from 10 stallions (38 ejaculates) were processed by the 2 methods of colloidal centrifugation. For both uncentrifuged and centrifuged samples, sperm yield was calculated and subjective sperm motility assessed over several days to provide an estimate of sperm survival. Some stored semen samples, held at 4 degrees C overnight, were also available for testing., Results: For fresh, extended semen, a similar recovery yield of motile spermatozoa was seen for the 2 methods of preparation for single layers and density gradients, respectively. Sperm motility and survival rate were significantly improved by colloidal centrifugation compared to unprocessed ejaculate, without any significant difference between methods (SLC vs. gradient). However, the yield was reduced by 18-20% when cold-stored semen was used for centrifugation compared to fresh semen; and more variation between ejaculates was observed than for fresh ejaculates. Again, sperm motility and sperm survival were improved in the centrifuged sperm preparations compared to stored, unprocessed ejaculates., Potential Relevance: The 2 colloid centrifugation techniques produce equivalent sperm preparations in terms of sperm quality. However, the SLC method would be more practical and convenient for use in the field.

Background: The aim of this study was to get information on post mortem diagnoses of sows found dead or euthanised and to understand the diagnoses aetiology (causative background). Moreover, the study was to evaluate the association between the clinical symptoms observed on farm and post mortem findings., Methods: A large Swedish herd was studied from January to September 2006. During the 32-week period 3.9% of the removed sows and gilts (old enough to be mated) were found dead, 12.0% were euthanised and the rest were sent to slaughter. Of 32 sows/gilts found dead 17 (53%) were post mortem examined, and of 98 sows euthanised 79 (81%) were examined. The 96 examined carcasses were after 70 sows and 26 gilts. The findings at examination were together with data from the herd monitoring program PigWin Sugg the base for the descriptive statistics presented., Results: The average parity number at removal was 2.8 for those found dead and 2.1 for those euthanised. The highest number euthanised and found dead was in parity 0 (gilts). The main proportion of post mortem examinations was made on sows being in the period = 28 d of gestation at death (37.5%), followed by weaning to next service period (24.0%). Arthritis, with an incidence of 36.4% was the most common main finding of pathological-anatomical diagnosis (PAD). Of sows/gilts found dead were circulatory/cardiac failure (23.5%) and trauma related injuries (23.5%) most common PAD. The most commonly observed clinical symptom and reason for euthanasia of the sows/gilts was lameness. Notably, in 43% of the cases with PAD arthritis, the clinical symptoms suggested it being a fracture. Further one or more abscesses (38.5%) and teeth injuries (31.0%) were common findings when also incidental findings were included., Conclusion: This post mortem study based on carcasses from sows/gilts found dead or euthanised showed that arthritis was a significant problem in the studied herd and that post mortem examination was important to get proper diagnosis.

The objective of this study was to investigate factors that might influence the length of productive life in Swedish crossbred (Landrace x Yorkshire) sows. The data set consisted of 20,310 sows farrowing between 2001 and 2004 in 21 commercial piglet-producing herds. Productive life (PL) was defined as the number of days between first farrowing and removal or termination of data collection. In addition to the overall risk analysis of PL, another 4 longevity traits were analyzed (competing risk analyses): reproductive disorder-determined length of PL (RPL), udder problem-determined length of PL (UPL), lameness-determined length of PL (LPL), and mortality-determined length of PL (MPL). Analyses were performed by using survival analysis, applying a Weibull model with 6 time-dependent and 1 time-independent variable (age at first farrowing). The factor with the largest contribution to the likelihood function for PL was days after farrowing, followed by parity, the herd x year combination, the total number of piglets born, days between weaning and next farrowing, farrowing month, and age at first farrowing. For all 4 competing risk traits, the factors contributing most to the likelihood function were days after farrowing, the herd x year combination, and parity, with a varied order between traits. The hazard for removal was greatest 30 to 40 d after farrowing (after weaning) for PL, UPL, and LPL (P < 0.001). However, for MPL the hazard was greatest just after farrowing (0 to 10 d), and for RPL the hazard peaked at 70 to 100 d after farrowing. The hazard for removal was, compared with parity 1, less in parities 2 to 7 and greater from parity 8 for PL (P < 0.001). The hazard was greatest in parity 1 (P < 0.01) for RPL, UPL, and LPL, whereas for MPL the hazard increased with greater parity number and was markedly greater from parity 9 (P < 0.001). Sows with litters of 9 piglets or less had a greater hazard for removal than sows with litters of 12 to 13 piglets (P < 0.001). Intervals between 120 and 122 d from weaning to the next farrowing showed the lowest hazard for removal (P < 0.001). The influence of farrowing month displayed no clear pattern for PL. Sows of 14 mo or older at their first farrowing had a 20% greater hazard for removal than younger sows (P < 0.001). The hazard for removal was greater for smaller litters in all parities but was more accentuated in greater parities. Overall, days after farrowing was the main risk factor for sow removal. Removal hazard was greatest shortly after weaning, and this peak increased with greater parity number.

Background: Artificial insemination is not as widely used in horses as in other domestic species, such as dairy cattle and pigs, partly because of the wide variation in sperm quality between stallion ejaculates and partly due to decreased fertility following the use of cooled transported spermatozoa. Furthermore, predictive tests for sperm fertilising ability are lacking. The objective of the present study was to assess sperm morphology and chromatin integrity in ejaculates obtained from 11 warmblood breeding stallions in Sweden, and to evaluate the relationship of these parameters to pregnancy rates to investigate the possibility of using these tests predictively., Methods: Aliquots from fortyone ejaculates, obtained as part of the normal semen collection schedule at the Swedish National Stud, were used for morphological analysis by light microscopy, whereas thirtyseven were used for chromatin analysis (SCSA) by flow cytometry. The outcome of inseminations using these ejaculates was made available later in the same year., Results: Ranges for the different parameters were as follows; normal morphology, 27-79.5%; DNA-fragmentation index (DFI), 4.8-19.0%; standard deviation of DNA fragmentation index (SD&#95;DFI) 41.5-98.9, and mean of DNA fragmentation index (mean&#95;DFI), 267.7-319.5. There was considerable variation among stallions, which was statistically significant for all these parameters except for mean&#95;DFI (P < 0.001, P < 0.01, P < 0.001 and P < 0.2 respectively). There was a negative relationship between normal morphology and DFI (P < 0.05), between normal morphology and SD&#95;DFI (P < 0.001), and between normal morphology and mean&#95;DFI (P < 0.05). For specific defects, there was a direct relationship between the incidence of pear-shaped sperm heads and DFI (P < 0.05), and also nuclear pouches and DFI (P < 0.001), indicating that either morphological analysis or chromatin analysis was able to identify abnormalities in spermiogenesis that could compromise DNA-integrity. A positive relationship was found between normal morphology and pregnancy rate following insemination (r = 0.789; P < 0.01) and a negative relationship existed between DFI and pregnancy rate (r = -0.63; P < 0.05). Sperm motility, assessed subjectively, was not related to conception rate., Conclusion: Either or both of the parameters, sperm morphology and sperm chromatin integrity, seem to be useful in predicting the fertilising ability of stallion ejaculates, particularly in determining cases of sub-fertility.

Ovariectomized mares and mares with inactive ovaries may show signs of estrus. The reason behind this phenomenon is not clear; however, steroid hormones of adrenal origin have been suggested. Moreover, aberrant adrenal hormone production has been implied as a reason why some intact mares may change behavior. In the present study, the effect of ACTH on plasma levels of cortisol, progesterone, androstenedione and testosterone was investigated in intact mares with normal estrous behavior ('controls', n=5) and intact mares that according to their owners showed deviant estrous behavior ('problem' mares, n=7). Blood samples were collected hourly from 12:00 h until 14:00 h the following day (half-hourly between 14:00 and 17:00 h) on two occasions (at two estruses), with saline or ACTH treatment (tetracosactide) at 14:00 h (saline treatment day or ACTH treatment day). ACTH treatment caused a significant increase in plasma levels of cortisol, progesterone, androstenedione and testosterone in all mares (P<0.05). An overall significant difference in cortisol response to ACTH was found (P<0.05), with 'problem' mares showing a significantly lower increase in cortisol levels 30 min to 3h post ACTH treatment (P<0.001). The 'problem' mares also showed a significantly higher increase than controls in progesterone levels in the same time period (P<0.05). The reason for the reduced adreno-cortical reactivity, with a low cortisol response to the ACTH treatment, in the 'problem' mares is unknown, but may indicate a difference in adrenal function as compared to control mares.

The mare is the only non-primate species known to display estrous signs after ovariectomy and adrenal hormones have been implicated as a possible cause. Moreover, in several species, estradiol seems to have a stimulatory effect on the hypothalamic-pituitary-adrenal axis. The aim of the present study was to compare the effect of ACTH (tetracosactide) on pertinent hormones [cortisol, progesterone, androstenedione, testosterone (intact and ovariectomized mares) and estradiol (ovariectomized mares only)] in intact mares in estrus with the same mares after ovariectomy (n=5). Blood samples were collected hourly from 12:00 until 14:00 h the following day (half-hourly between 14:00 and 17:00 h) on two occasions, with saline or ACTH treatment at 14:00 h (saline treatment day or ACTH treatment day). The mares, both when intact and after ovariectomy, showed a significant increase in all measured hormones, except estradiol (not measured in intact mares), after ACTH treatment, lasting at least 3h post-treatment (P<0.001). On the saline treatment day, cortisol levels in ovariectomized mares were lower than in intact mares in the evening (18:00-23:00 h), but higher at night (24:00-05:00 h). No differences in cortisol response between mares, when intact and after ovariectomy, were found after ACTH treatment (P=0.3). Androstenedione levels were lower (P<0.001) and increased less after ACTH treatment in ovariectomized mares, as compared to when intact (P<0.05). Progesterone concentrations were lower in the ovariectomized mares at night (24:00-05:00 h) on the saline treatment day and at all times on the ACTH treatment day (P<0.05). Testosterone concentrations were lower in ovariectomized mares on both treatment days, as compared to when intact (P<0.001). It was concluded that ovariectomy affected basal cortisol pattern. Ovarian androstenedione and testosterone contributed to the basal circulating levels and, in the case of androstenedione, was stimulated by ACTH. Endogenous estradiol did not act stimulatory on adrenal gland hormone production in the mare.

The aim of this study was to investigate the influence of pre- and post-ovulatory insemination and early pregnancy on the distribution of immune cells in the oviduct. Eighteen sows were pre-ovulatory and sixteen sows were post-ovulatory inseminated and slaughtered at different times, 5-6 h after insemination, 20-25 h and approximately 70 h after ovulation, day 11 and day 19. Immediately after slaughter, oviductal samples of three different segments (isthmus, ampulla and infundibulum) were fixed, embedded in plastic resin and stained with toluidine blue or cryofixed and stored in a freezer at -70 degrees C until analysed by immunohistochemistry (pre-ovulatory inseminated sows) with an avidin-biotin peroxidase method. Quantitative and qualitative examinations of oviductal epithelium and subepithelial connective tissue were performed by light microscopy. After pre- or post-ovulatory insemination, neutrophils were not observed in the oviductal epithelium from any of the segments or groups. The numbers of intraepithelial lymphocytes of all sows as well as CD2- and CD3-positive cells of the pre-ovulatory inseminated sows were higher in the infundibulum than in the other segments (p < or = 0.001). In the subepithelial connective tissue of the pre-ovulatory inseminated sows, significantly higher numbers of lymphocytes (p < or = 0.001) and plasma cells (p < or = 0.001) were found in infundibulum than in isthmus. Neutrophils were found mainly in infundibulum, the number approximately 40 h after pre-ovulatory insemination was significantly higher (p < or = 0.05) than in the other groups and segments. Significantly higher numbers of CD2 than CD3-positive cells were found for all groups and segments. In the subepithelial connective tissue of post-ovulatory inseminated sows, the numbers of lymphocytes was higher (p < or = 0.001) at day 19 than up to 50 h after insemination and lower (p < or = 0.001) in isthmus than in ampulla and infundibulum. Neutrophils were found in infundibulum in almost all groups and the number was significantly higher (p < or = 0.05) in the infundibulum up to 50 h after insemination than in other segments. In the oviductal epithelium, no influence of insemination was found on the presence of phagocytes, i.e. neutrophils and macrophages, but on lymphocytes. In the infundibular connective tissue, pre-ovulatory insemination had an effect on neutrophil distribution, indicating an active immune response to insemination in the upper segment. Post-ovulatory insemination changed the oviductal immune cell pattern.

Background: Strong oestrous symptoms in the mare can cause problems with racing, training and handling. Since long-acting progesterone treatment is not permitted in mares at competition (e.g. according to FEI rules), there is a need for methods to suppress unwanted cyclicity. Spontaneous dioestrous ovulations in the late luteal phase may cause a prolongation of the luteal phase in mares., Methods: In this preliminary study, in an attempt to induce ovulation during the luteal phase, human chorionic gonadotropin (hCG) (3000 IU) was injected intramuscularly in four mares (experimental group) in the luteal phase when a dioestrous follicle > or = 30 mm was detected. A fifth mare included in this group was not treated due to no detectable dioestrous follicles > or = 30 mm. Four control mares were similarly injected with saline. The mares were followed with ultrasound for 72 hours post injection or until ovulation. Blood samples for progesterone analysis were obtained twice weekly for one month and thereafter once weekly for another two to four months., Results: Three of the hCG-treated mares ovulated within 72 hours after treatment and developed prolonged luteal phases of 58, 68 and 82 days respectively. One treated mare never ovulated after the hCG injection and progesterone levels fell below 3 nmol/l nine days post treatment. Progesterone levels in the control mares were below 3 nmol/l within nine days after saline injection, except for one mare, which developed a spontaneously prolonged luteal phase of 72 days., Conclusion: HCG treatment may be a method to induce prolonged luteal phases in the mare provided there is a dioestrous follicle > or = 30 mm that ovulates post-treatment. However, the method needs to be tested on a larger number of mares to be able to draw conclusions regarding its effectiveness.

The aim of this study was to investigate the distribution of immune cells in the endometrium of anoestrous female pigs, five sows in anoestrus by lactation and five pre-pubertal gilts (Swedish Landrace x Swedish Yorkshire). Uterine samples, taken immediately after slaughter, were fixed, embedded in plastic resin and stained with toluidine blue or cryo fixed and stored in a freezer at -70 degrees C until analysed by immunohistochemistry with an avidin-biotin peroxidase method. Immune cells in the surface (luminal) and the glandular epithelium as well as the subepithelial and the glandular connective tissue layers were counted using light microscopy. In the surface (luminal) and the glandular epithelia of gilts and sows, lymphocytes were the predominant immune cells found. There were no significant differences between gilts and sows. Macrophages were detected in the glandular epithelium of sows but not in gilts. In the subepithelial and the glandular connective tissue layers of both gilts and sows, lymphocytes were also the most common immune cells found. The numbers of lymphocytes and macrophages were significantly higher in the sows than in the gilts (p