Anne-Marie Dalin

Sperm morphology and chromatin integrity in Swedish warmblood stallions and their relationship to pregnancy rates

BackgroundArtificial 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.MethodsAliquots 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.ResultsRanges 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_DFI) 41.5–98.9, and mean of DNA fragmentation index (mean_DFI), 267.7–319.5. There was considerable variation among stallions, which was statistically significant for all these parameters except for mean_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_DFI (P < 0.001), and between normal morphology and mean_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.ConclusionEither 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.

Reproductive performance of purebred Landrace and Yorkshire sows in Thailand with special reference to seasonal influence and parity number.

The purpose of this study was to analyze reproductive performance in purebred Landrace and Yorkshire sows with special reference to seasonal influence and parity number, under tropical conditions where day length is almost constant throughout the year. Data from three purebred sow herds in Thailand during the period from 1993 to 1996 were analyzed. The two breeds were present in all three herds. The analysis comprised records of 3848 Landrace sow litters and 2033 Yorkshire sow litters. The statistical models included the fixed effects of month, year, parity, breed of the sow, herd, and two-way interactions of breed-parity, breed-herd, breed-month, breed-year, parity-month, month-herd, year-herd and month-year. The random effect of sow within breed was included in all models. Analysis of covariance was performed to analyze the effect of temperature, humidity and heat index on number of total born per litter (NTB), weaning to first service interval (WSI) and farrowing rate (FR). Landrace sows had significantly higher NTB (0.6 piglets), number of live born per litter (0.5 piglets), and average birth weight (0.13 kg) than Yorkshire sows (P<0.001). Farrowing rate was 3.9% higher in Landrace sows than in Yorkshire sows (P<0.01). However, Yorkshire sows had significantly shorter WSI (P<0.001) and significantly higher proportion of sows served within 7 days after weaning (P<0.01) than Landrace sows. No breed differences were found in number of stillborn per litter and weaning to conception interval. Parity had significant effect on all reproductive parameters analyzed. Number of total born and live born per litter was significantly lower for sows farrowing during the rainy season than in other seasons. Farrowing rate was low for sows mated during the hot and rainy season. Weaning to service interval and WSI7 were prolonged for sows weaned during the hot and rainy season. Reproductive performance was significantly unfavorably influenced by elevated temperature and heat index after mating (NTB and FR) or during lactation (WSI).

Post mortem findings in sows and gilts euthanised or found dead in a large Swedish herd.

BackgroundThe 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.MethodsA 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.ResultsThe 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.ConclusionThis 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.

A preliminary study on the induction of dioestrous ovulation in the mare – a possible method for inducing prolonged luteal phase

BackgroundStrong 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.MethodsIn 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 ≥ 30 mm was detected. A fifth mare included in this group was not treated due to no detectable dioestrous follicles ≥ 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.ResultsThree 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.ConclusionHCG treatment may be a method to induce prolonged luteal phases in the mare provided there is a dioestrous follicle ≥ 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.

Stallion spermatozoa selected by single layer centrifugation are capable of fertilization after storage for up to 96 h at 6°C prior to artificial insemination.

BackgroundOne 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.FindingsSixteen mares were inseminated with SLC-selected sperm doses that had been cooled and stored at 6°C for 48u2009h, 72u2009h or 96u2009h. Embryos were identified in 11 mares by ultrasound examination 16–18u2009days after presumed ovulation.ConclusionSLC-selected stallion spermatozoa stored for up to 96u2009h are capable of fertilization.

High Prevalence of Hepatitis E Virus in Swedish Moose – A Phylogenetic Characterization and Comparison of the Virus from Different Regions

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.

Reproductive characteristics in female Swedish moose (Alces alces), with emphasis on puberty, timing of oestrus, and mating

BackgroundThe 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.ResultsA 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.ConclusionsThe 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 14xa0days in southern Sweden.

Surveys on Coxiella burnetii infections in Swedish cattle, sheep, goats and moose

BackgroundQ 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.ResultsThe 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%.ConclusionsThe 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.

A 25 years experience of group-housed sows-reproduction in animal welfare-friendly systems.

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 4xa0weeks after service to 1xa0week 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 25xa0years 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.

Puberty in female wild boar (Sus scrofa) in Sweden

BackgroundSince 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 5–15xa0months, 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).ResultsIn total, 29 (16.6xa0%) animals were classified as post-pubertal. Field dressed weight ranged from 20.6 to 65.3xa0kg. 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.ConclusionsSeason, 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.