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1.  Single layer centrifugation-selected boar spermatozoa are capable of fertilization in vitro 
Background
Good quality spermatozoa are important to achieve fertilization, viable embryos and offspring. Single Layer Centrifugation (SLC) through a colloid (Androcoll-P) selects good quality spermatozoa. However, it has not been established previously whether porcine spermatozoa selected by this method maintain their fertility.
Methods
The semen was prepared either by SLC or by standard centrifugation (control) and used for in vitro fertilization (IVF) at oocyte:spermatozoa ratios of 1:50; 1:100 and 1:300 (or 4 x 103, 8 x 103 and 24 x 103 spermatozoa/ml) to evaluate their subsequent ability to generate blastocysts. In addition, sperm motility was assessed by computer assisted sperm motility analysis.
Results
Total and progressive motility were significantly higher in sperm samples prepared by SLC compared to uncentrifuged samples. Sperm binding ability, polyspermy, cleavage and blastocyst rates were affected by the oocyte:sperm ratio, but not by sperm treatment.
Conclusion
The use of SLC does not adversely affect the in vitro fertilizing and embryo-generating ability of the selected spermatozoa compared to their unselected counterparts, but further modifications in the IVF conditions would be needed to improve the monospermy in IVF systems. Since SLC did not appear to have a negative effect on sperm fertilizing ability, and may in fact select for spermatozoa with a greater potential for fertilization, an in vivo trial to determine the usefulness of this sperm preparation technique prior to artificial insemination is warranted.
doi:10.1186/1751-0147-55-20
PMCID: PMC3602148  PMID: 23497680
IVF; Pig; Semen; Sperm selection
2.  Supplementation of a grape seed and grape marc meal extract decreases activities of the oxidative stress-responsive transcription factors NF-κB and Nrf2 in the duodenal mucosa of pigs 
Background
In pigs, enteric infections and the development of gut disorders such as diarrhoea are commonly observed, particularly after weaning. The present study investigated the hypothesis that feeding a grape seed and grape marc extract (GSGME) as a dietary supplement has the potential to suppress the inflammatory process in the small intestine of pigs by modulating the activities of NF-κB and Nrf2 due to its high content of flavonoids.
Methods
Twenty-four crossbred, 6 weeks old pigs were randomly assigned to 2 groups of 12 animals each and fed nutritionally adequate diets without or with 1% GSGME for 4 weeks.
Results
Pigs administered GSGME had a lower transactivation of NF-κB and Nrf2 and a lower expression of various target genes of these transcription factors in the duodenal mucosa than control pigs (P < 0.05). Concentrations of α-tocopherol and thiobarbituric acid reactive substances (TBARS) in liver and plasma and total antioxidant capacity of plasma and relative mRNA abundances of NF-κB and Nrf2 target genes in the liver did not differ between the two groups. However, the ratio of villus height:crypt depth and the gain:feed ratio was higher in the pigs fed GSGME than in control pigs (P < 0.05).
Conclusions
This study shows that dietary supplementation of a polyphenol rich GSGME suppresses the activity of NF-κB in the duodenal mucosa of pigs and thus might provide a useful dietary strategy to inhibit inflammation in the gut frequently occurring in pigs. Feeding GSGME did not influence vitamin E status and the antioxidant system of the pigs but improved the gain:feed ratio. In overall, the study suggests that polyphenol-rich plant extracts such GSGME could be useful feed supplements in pig nutrition, in order to maintain animal health and improve performance.
doi:10.1186/1751-0147-55-18
PMCID: PMC3599961  PMID: 23453040
NF-κB; Nrf2; Polyphenol; Pig; Intestine
3.  Effects on boar semen quality after infection with porcine reproductive and respiratory syndrome virus: a case report 
The effect of porcine reproductive and respiratory syndrome virus (PRRSV) on semen quality was examined in a group of 11 spontaneously infected boars in a commercial boar stud. Semen samples were collected 4 weeks prior to 4 weeks post-infection (wpi). Infection with PRRSV of the European genotype subtype 1 (EU-1) was verified by specific quantitative real-time polymerase chain reaction (RT-PCR) in 36% of the serum samples. All boars seroconverted before 4 wpi and remained in normal condition throughout the study. Comparison of the percentage of morphologically intact spermatozoa revealed an increase of acrosome-defective spermatozoa (P = 0.012) between −4 and 4 wpi. Significant deleterious effects on semen quality were detected for membrane integrity when semen had been stored for 2 days after sampling. Analysis of sperm subpopulations in a thermoresistance test on day 7 after sampling revealed alterations in the percentage of circular, progressively motile spermatozoa (P = 0.013), in the percentage of non-linear, progressively motile spermatozoa (P = 0.01), and on the amplitude of lateral sperm head displacement (P = 0.047). There was no difference in the incidence of mitochondrially active spermatozoa (P = 0.075). Investigation of routine production data between pre- and post-infection status showed no differences on ejaculate volume (P = 0.417), sperm concentration (P = 0.788), and percentage of motile spermatozoa (P = 0.321). This case report provides insights into a potential control strategy for PRRSV outbreaks in boar studs.
doi:10.1186/1751-0147-55-16
PMCID: PMC3599264  PMID: 23442207
PRRSV; Boar; CASA; Semen quality; Spermatozoa
4.  Coxiella burnetii associated reproductive disorders in domestic animals-a critical review 
The bacterium Coxiella burnetii has been detected in the fetal membranes, birth fluids and vaginal mucus, as well as in the milk and other excretions of several domestic mammals. The finding of C. burnetii in association with abortion, parturition and in the postpartum period has led to the hypothesis that C. burnetii causes a range of reproductive diseases. This review critically evaluates the scientific basis for this hypothesis in domestic mammals.
The review demonstrates a solid evidence for the association between C. burnetii infection and sporadic cases of abortion, premature delivery, stillbirth and weak offspring in cattle, sheep and goats. C. burnetii induced in-herd epidemics of this complete expression of reproductive failure have been reported for sheep and goats, but not for cattle. The single entities occur only as part of the complex and not as single events such as generally increased stillbirth rate. Studies show that C. burnetii initially infects the placenta and that subsequent spread to the fetus may occur either haematogenous or by the amniotic-oral route. The consequences for the equine, porcine, canine and feline conceptus remains to the elucidated but that infection of the conceptus may occur is documented for most species. There is no solid evidence to support a hypothesis of C. burnetii causing disorders such as subfertility, endometritis/metritis, or retained fetal membranes in any kind of domestic animal species.
There is a strong need to validate non-pathology based methods such as polymerase chain reaction for their use in diagnostic and research in relation to establishing C. burnetii as the cause of abortion and to adapt an appropriate study design and include adequate control animals when linking epidemiological findings to C. burnetii or when evaluating effects of vaccination in production herds.
doi:10.1186/1751-0147-55-13
PMCID: PMC3577508  PMID: 23419216
Coxiella burnetii; Q fever; Reproduction; Abortion; Cattle; Sheep; Goat; Buffalo; Pig; Dog; Cat
5.  A polymorphism in the insulin-like growth factor 1 gene is associated with postpartum resumption of ovarian cyclicity in Holstein-Friesian cows under grazing conditions 
Background
Insulin-like growth factor 1 (IGF-1) gene is considered as a promising candidate for the identification of polymorphisms affecting cattle performance. The objectives of the current study were to determine the association of the single nucleotide polymorphism (SNP) IGF-1/SnaBI with fertility, milk production and body condition traits in Holstein-Friesian dairy cows under grazing conditions.
Methods
Seventy multiparous cows from a commercial herd were genotyped for the SNP IGF-1/SnaBI. Fertility measures evaluated were: interval to commencement of luteal activity (CLA), calving to first service (CFS) and calving to conception (CC) intervals. Milk production and body condition score were also evaluated. The study period extended from 3 wk before calving to the fourth month of lactation.
Results and discussion
Frequencies of the SNP IGF-1/SnaBI alleles A and B were 0.59 and 0.41, respectively. Genotype frequencies were 0.31, 0.54 and 0.14 for AA, AB and BB, respectively. Cows with the AA genotype presented an early CLA and were more likely to resume ovarian cyclicity in the early postpartum than AB and BB ones. No effect of the SNP IGF-1/SnaBI genotype was evidenced on body condition change over the experimental period, suggesting that energy balance is not responsible for the outcome of postpartum ovarian resumption in this study. Traditional fertility measures were not affected by the SNP IGF-1/SnaBI.
Conclusion
To our knowledge this is the first report describing an association of the SNP IGF-1/SnaBI with an endocrine fertility measure like CLA in cattle. Results herein remark the important role of the IGF-1gene in the fertility of dairy cows on early lactation and make the SNP IGF-1/SnaBI an interesting candidate marker for genetic improvement of fertility in dairy cattle.
doi:10.1186/1751-0147-55-11
PMCID: PMC3599257  PMID: 23409757
IGF-1; Polymorphism; Commencement of luteal activity; Cattle
6.  Aleutian mink disease virus in furbearing mammals in Nova Scotia, Canada 
Background
Aleutian mink disease virus (AMDV) is widespread among ranched and free-ranging American mink in Canada, but there is no information on its prevalence in other wild animal species. This paper describes the prevalence of AMDV of 12 furbearing species in Nova Scotia (NS), Canada.
Methods
Samples were collected from carcasses of 462 wild animals of 12 furbearing species, trapped in 10 NS counties between November 2009 and February 2011. Viral DNA was tested by PCR using two primer pairs, and anti-viral antibodies were tested by counterimmunoelectrophoresis (CIEP) on spleen homogenates.
Results
Positive PCR or CIEP samples were detected in 56 of 60 (93.3%) American mink, 43 of 61 (70.5%) short-tailed weasels, 2 of 8 (25.0%) striped skunks, 2 of 11 (18.2%) North American river otters, 9 of 85 (10.6%) raccoons, and 2 of 20 (10.0%) bobcats. Samples from six fishers, 24 coyotes, 25 red foxes, 58 beavers, 45 red-squirrels and 59 muskrats were negative. Antibodies to AMDV were detected by CIEP in 16 of 56 (28.6%) mink and one of the 8 skunks (12.5%). Thirteen of the mink were positive for PCR and CIEP, but three mink and one skunk were CIEP positive and PCR negative. Positive CIEP or PCR animals were present in all nine counties from which mink or weasel samples were collected.
Conclusions
The presence of AMDV in so many species across the province has important epidemiological ramifications and could pose a serious health problem for the captive mink, as well as for susceptible wildlife. The mechanism of virus transmission between wildlife and captive mink and the effects of AMDV exposure on the viability of the susceptible species deserve further investigation.
doi:10.1186/1751-0147-55-10
PMCID: PMC3602201  PMID: 23394546
Aleutian mink disease virus; American mink; Carnivores; Furbearing species; Mustelidae; Rodents
7.  Structural changes in femoral bone tissue of rats after subchronic peroral exposure to selenium 
Background
The role of selenium (Se) on bone microarchitecture is still poorly understood. The present study aims to investigate the macroscopic and microscopic structures of femoral bone tissue in adult male rats after subchronic peroral administration of Se.
Methods
Twenty one-month-old male Wistar rats were randomly divided into two experimental groups. In the first group (Se group) young males were exposed to 5 mg Na2SeO3/L in drinking water, for 90 days. Ten one-month-old males without Se administration served as a control group. At the end of the experiment, macroscopic and microscopic structures of the femurs were analysed using analytical scales, sliding instrument, and polarized light microscopy.
Results
The body weight, femoral length and cortical bone thickness were significantly decreased in Se group rats. These rats also displayed different microstructure in the middle part of the femur, both in medial and lateral views, where vascular canals expanded into the central area of the bone while, in control rats, these canals occurred only near the endosteal surfaces. Additionally, a smaller number of primary and secondary osteons was identified in Se group rats. Histomorphometric analyses revealed significant increases for area, perimeter, maximum and minimum diameters of primary osteons’ vascular canals but significant reductions for all measured variables of Haversian canals and secondary osteons.
Conclusions
Se negatively affected the macroscopic and microscopic structures of femoral bone tissue in adult male rats. The results contribute to the knowledge on damaging impact of Se on bone.
doi:10.1186/1751-0147-55-8
PMCID: PMC3598879  PMID: 23369508
8.  Methicillin-resistant Staphylococcus aureus containing mecC in Swedish dairy cows 
Background
Hitherto, methicillin-resistant Staphylococcus aureus (MRSA) has not been detected in Swedish cattle. However, due to the report of mecC, a novel homologue to the mecA gene, there was reason to re-evaluate susceptibility results from strain collections of Staphylococcus aureus and test suspected isolates for the presence of mecC.
Findings
Bovine isolates of S. aureus with elevated minimum inhibitory concentrations of beta-lactams were retrospectively tested for presence of mecC. In four of the isolates mecC was detected.
Conclusion
In Sweden, this is the first finding of MRSA in cattle and the first detection of MRSA harbouring mecC of domestic animal origin. MRSA in animal populations has implications as a potential reservoir with risk for spread to humans. Occurrence of MRSA among Swedish cattle appears still very limited.
doi:10.1186/1751-0147-55-6
PMCID: PMC3563487  PMID: 23369449
MRSA; Staphylococcus aureus; mecC; Mastitis; Dairy cows
9.  Malignant peripheral nerve sheath tumor as a cause of chronic cardiac insufficiency in cattle 
Chronic cardiac insufficiency was associated with a malignant peripheral nerve sheath tumor in a cow. An eight-year-old cow developed a progressive condition (over a period of three months) characterized by an enhanced abdominal volume, reluctance to move, a positive jugular pulse, watery diarrhea and death. At necropsy, moderate subcutaneous edema and an enhanced hepatic lobular pattern were observed. A 23x20x11 cm firm, grayish-white mass adhered to and infiltrated the right atrium. Multiple firm, yellowish-white nodules of 0.5 to 12 cm in diameter were diffusely scattered in the epicardium and parietal pericardium. Histologically, the tumor was poorly circumscribed with foci of infiltration of the myocardium. The neoplastic cells had two major histologic patterns, Antoni types A and B. Within occasional foci, pleomorphic cells with an epithelioid appearance were present in addition to multinucleated cells with periodic acid Schiff (PAS)-positive cytoplasmic globules. Foci of cartilaginous and granular differentiations were interspersed among the neoplastic cells. Multiple vessels presented wall hyalinization and tumoral embolus. Large necrotic foci with mineralization and cholesterol clefts were also observed. Immunohistochemically, the tumor was positive for S100 protein, vimentin and neuron-specific enolase labeling.
doi:10.1186/1751-0147-55-7
PMCID: PMC3577581  PMID: 23369465
Cattle; Immunohistochemical procedures; Neoplasm; Malignant schwannoma
10.  An evaluation of the ability of Dichelobacter nodosus to survive in soil 
Background
Dichelobacter nodosus is the causative agent of footrot in sheep. The survival of the bacterium in soil is of importance for the epidemiology of the disease. The investigation evaluates the survival of D. nodosus in soil with and without added hoof powder stored under different temperatures.
Results
An experimental setup was used with bacteriological culture and real-time polymerase chain reaction (PCR), and the results indicate that the bacteria can survive in soil for longer time than previously expected. The survival time was found to be dependent on temperature and the addition of hoof powder to the soil, with the longest survival time estimated to be 24 days in soil samples with hoof powder stored at 5°C.
Conclusion
Our findings indicate that the survival time of D. nodosus and its ability to infect susceptible sheep on pasture under different climatic conditions should be studied further.
doi:10.1186/1751-0147-55-4
PMCID: PMC3561160  PMID: 23343097
Dichelobacter nodosus; Soil; Real-time PCR; Survival; Bacteriological culture
11.  Spread of extended-spectrum beta-lactamase producing Escherichia coli isolates in Swedish broilers mediated by an incI plasmid carrying blaCTX-M-1 
Background
The already high and increasing occurrence of extended-spectrum beta-lactamases (ESBL) producing Escherichia coli in European broiler populations is of concern due to the fact that third and fourth generation cephalosporins are deemed critically important in human medicine. In Sweden 34% of the broilers carry ESBL/pAmpC producing E. coli in their gut, despite the absence of a known selection pressure such as antimicrobial usages. The aim of the current study was to characterise a selection of E. coli strains carrying the blaCTX-M-1, to determine if the spread was due to a specific clone.
Findings
Ten isolates carrying blaCTX-M-1 from Swedish broilers belonged to eight different multi-locus sequence types with three isolates belonging to ST155. The ST155 isolates were identical as assessed by PFGE. The blaCTX-M-1 was in all isolates carried on a plasmid of replicon type incI, which also transferred resistance to tetracycline and sulfamethoxazole.
Conclusion
The occurrence of ESBL-producing E. coli in the Swedish broilers is not due to the emergence of a single clone, but rather the spread of a specific incI plasmid carrying blaCTX-M-1.
doi:10.1186/1751-0147-55-3
PMCID: PMC3557190  PMID: 23336334
Extended spectrum betalactamases; ESBL; Broiler; CTX-M-1; Escherichia coli
12.  Assay dependence of Brucella antibody prevalence in a declining Alaskan harbor seal (Phoca vitulina) population 
Background
Brucella is a group of bacteria that causes brucellosis, which can affect population health and reproductive success in many marine mammals. We investigated the serological prevalence of antibodies against Brucella bacteria in a declining harbor seal population in Glacier Bay National Park, Alaska.
Results
Prevalence ranged from 16 to 74 percent for those tests detecting antibodies, indicating that harbor seals in Glacier Bay have been exposed to Brucella bacteria. However, the actual level of serological prevalence could not be determined because results were strongly assay-dependent.
Conclusions
This study reinforces the need to carefully consider assay choice when comparing different studies on the prevalence of anti–Brucella antibodies in pinnipeds and further highlights the need for species- or taxon-specific assay validation for both pathogen and host species.
doi:10.1186/1751-0147-55-2
PMCID: PMC3577514  PMID: 23324565
Brucella; Harbor seals; Alaska; Assay dependence
13.  Ultrasonographic examination of the forestomachs and the abomasum in ruminal drinker calves 
Background
The study investigated the ultrasonographic appearance of the reticulum, rumen, omasum and abomasum of calves with ruminal drinking syndrome.
Methods
In ten milk-fed calves with ruminal drinking syndrome the reticulum, rumen, omasum and abomasum were examined by ultrasonography using a 5-MHz linear transducer before, during and after the ingestion of milk.
Results
The reticulum could be imaged in eight of ten calves before feeding. The reticular wall appeared as an echoic line, similar to mature cattle, and reticular folds were seen in eight calves. The reticular content appeared as echoic heterogeneous fluid. Reticular contractions were biphasic with 1.0 ± 0.38 contractions per minute. The rumen had a mean wall thickness of 2.1 mm dorsally, 3.5 mm at the level of the longitudinal groove, and 3.2 mm ventrally. The ventral sac of the rumen of all calves contained echoic heterogeneous liquid. During feeding the milk entering the rumen could be seen as hyperechoic liquid in five calves. The omasum was seen on the right side as a crescent-shaped line medial to the liver in seven calves. Only the omasal wall closest to the transducer was seen as an echoic line with a mean thickness of 2.7 mm. The ultrasonographic appearance of the omasum did not change during or after feeding. The abomasum was seen immediately caudal to the xyphoid on both sides of the midline before feeding. The mean length at the ventral midline was 22.2 cm. The ingesta were heterogeneous in all calves and the abomasal folds were distinct in eight. The mean lateral expansion of the abomasum from the ventral midline to the left and right varied from 8.7 to 13.8 cm and from 4.3 to 11.3 cm. The milk entering the abomasum was observed in all calves, and signs of milk clotting were seen in all calves 15 minutes after feeding.
Conclusion
This study showed that ultrasonography is useful for detecting milk in the reticulum and rumen of calves with ruminal drinking syndrome.
doi:10.1186/1751-0147-55-1
PMCID: PMC3599229  PMID: 23298472
14.  Physiological evaluation of free-ranging moose (Alces alces) immobilized with etorphine-xylazine-acepromazine in Northern Sweden 
Background
Evaluation of physiology during capture and anesthesia of free-ranging wildlife is useful for determining the effect that capture methods have on both ecological research results and animal welfare. This study evaluates capture and anesthesia of moose (Alces alces) with etorphine-xylazine-acepromazine in Northern Sweden.
Methods
Fifteen adult moose aged 3–15 years were darted from a helicopter with a combination of 3.37 mg etorphine, 75 mg xylazine, and 15 mg acepromazine. Paired arterial blood samples were collected 15 minutes apart with the first sample at 15–23 minutes after darting and were analyzed immediately with an i-STAT®1 Portable Clinical Analyzer.
Results
All animals developed hypoxemia (PaO2 <10 kPa) with nine animals having marked hypoxemia (PaO2 5.5-8 kPa). All moose were acidemic (ph<7.35) with nine moose having marked acidemia (pH<7.20). For PaCO2, 14 moose had mild hypercapnia (PaCO2 6-8 kPa) and two had marked hypercapnia (PaCO2>8 kPa). Pulse, respiratory rate, pH and HCO3 increased significantly over time from darting whereas lactate decreased.
Conclusions
The hypoxemia found in this study is a strong indication for investigating alternative drug doses or combinations or treatment with supplemental oxygen.
doi:10.1186/1751-0147-54-77
PMCID: PMC3541067  PMID: 23276208
Alces; Moose; Anesthesia; Etorphine; Xylazine; Immobilization
15.  Effect of propofol on oxidative stress status in erythrocytes from dogs under general anaesthesia 
Background
Alterations of the normal redox balance might be attributed to increase of plasma free-radical concentration and a disruption of the antioxidant defense system. One of the adverse effects of general anaesthetics is the exogen sources of reactive oxygen radicals that are responsible for several diseases. The purposes of the current study were to evaluate the effect of propofol on oxidative stress and to compare the differences between propofol induction only and induction plus continuous infusion on antioxidant status in dogs.
Findings
Beagle dogs were evaluated in the present study. The dogs were assigned randomly to receive three treatments in a crossover model. The three treatments were: group 1 (n = 9), 2% isoflurane; group 2 (n = 9), anaesthesia induced with an intravenous (IV) bolus dose of 6 mg/kg propofol and maintained with 1.5–2% isoflurane; group 3 (n = 9), total IV anaesthesia (induction with 6 mg/kg propofol, infusion with 0.6 mg/kg/min propofol). The results of this study show that dogs exposed to isoflurane had decreased antioxidant enzymes activities, whereas dogs injected with propofol had increased antioxidant enzymes activities.
Conclusions
The results of this study showed that an infusion dose of propofol has antioxidant effects in dogs. These effects may be beneficial to patients in whom free radicals play a role in oxidative stress, such as those with ischemia. Further studies are needed to evaluate whether these antioxidant effects of the anaesthetic are of clinical value.
doi:10.1186/1751-0147-54-76
PMCID: PMC3542034  PMID: 23268830
Anaesthesia; Antioxidant effects; Dogs; Oxidative stress; Propofol
16.  Seroprevalence of Toxoplasma gondii infection in Norwegian dairy goats 
Background
Toxoplasma gondii is a major problem for the sheep industry as it may cause reproduction problems. The importance of T. gondii in Norwegian goat herds is uncertain, but outbreaks of toxoplasmosis in dairy goat farms have been recorded. The aim of this study was to describe the prevalence of T. gondii infection in Norwegian dairy goats by using serology.
Findings
Goat serum originally collected as part of two nationwide surveillance and control programmes between 2002 and 2008 were examined for T. gondii antibodies by using direct agglutination test. In total, 55 of 73 herds (75%) had one or more serologically positive animals, while 377 of 2188 (17%) of the individual samples tested positive for T. gondii antibodies.
Conclusions
This is the first prevalence study of T. gondii infection in Norwegian goats. The results show that Norwegian goat herds are commonly exposed to T. gondii. Nevertheless, the majority of goat herds have a low prevalence of antibody positive animals, which make them vulnerable to infections with T. gondii during the gestation period.
doi:10.1186/1751-0147-54-75
PMCID: PMC3538615  PMID: 23259528
Toxoplasma gondii; Goats; Norway; Prevalence; Serology
17.  Changes in mean scrotal circumference in performance tested Swedish beef bulls over time 
Background
There is a growing interest in beef cattle breeding in Sweden. The majority of the females are bred naturally, which is why it is important to choose healthy fertile bulls to obtain good reproduction and profitability. The breeding soundness evaluation includes measurement of scrotal circumference (SC). Our aim was to analyze if the SC of performance tested beef bulls has changed over the years. In total, 1332 bulls (Angus, Charolais, Hereford and Simmental) from 13 batches (1997-2010) were included in the study. Case book entries from final evaluation of the bulls, 11-13 months old, were compiled and analyzed.
Results
An overall mean SC of 34.7 cm independent of breed and age was found which is above the set minimum level. Only eleven bulls did not reach the minimum level. An increase in SC of 0.06-0.07 cm/year was shown for all breeds. In all (1997-2010), the increase of the average SC (independent of breed and age) was approximately 1 cm. The positive trend was apparent for all breeds but only statistically significant for the Charolais breed.
Conclusions
In conclusion, our results indicated an increase in the SC over time, which improves the possibilities to obtain performance tested beef sires in Sweden with the potential for achieving better fertility results.
doi:10.1186/1751-0147-54-74
PMCID: PMC3538610  PMID: 23241352
Andrology; Cattle; Genetics; Testicles; Scrotal circumference; Beef bull
18.  Muscle reorganisation through local injection of stem cells in the diaphragm of mdx mice 
Background
The diaphragm is the major respiratory muscle affected by Duchenne muscular dystrophy (DMD) and is responsible for causing 80% of deaths. The use of mechanical forces that act on the body or intermittent pressure on the airways improves the quality of life of patients but does not prevent the progression of respiratory failure. Thus, diseases that require tissue repair, such as DMD, represent a group of pathologies that have great potential for cell therapy. The application of stem cells directly into the diaphragm instead of systemic application can reduce cell migration to other affected areas and increase the chances of muscle reorganisation. The mdx mouse is a suitable animal model for this research because its diaphragmatic phenotype is similar to human DMD. Therefore, the aim of this study was to assess the potential cell implantation in the diaphragm muscle after the xenotransplantation of stem cells.
Methods
A total of 9 mice, including 3 control BALB/Cmice, 3 5-month-old mdx mice without stem cell injections and 3 mdx mice injected with stem cells, were used. The animals injected with stem cells underwent laparoscopy so that stem cells from GFP-labelled rabbit olfactory epithelium could be locally injected into the diaphragm muscle. After 8 days, all animals were euthanised, and the diaphragm muscle was dissected and subjected to histological and immunohistochemical analyses.
Results
Both the fresh diaphragm tissue and immunohistochemical analyses showed immunopositive GFP labelling of some of the cells and immunonegativity of myoblast bundles. In the histological analysis, we observed a reduction in the inflammatory infiltrate as well as the presence of a few peripheral nuclei and myoblast bundles.
Conclusion
We were able to implant stem cells into the diaphragm via local injection, which promoted moderate muscle reorganisation. The presence of myoblast bundles cannot be attributed to stem cell incorporation because there was no immunopositive labelling in this structure. It is believed that the formation of the bundles may have been stimulated by cellular signalling mechanisms that have not yet been elucidated.
doi:10.1186/1751-0147-54-73
PMCID: PMC3537552  PMID: 23231953
Muscular dystrophy; Mice; Stem cell; Animal model
19.  The effect of two pre-cryopreservation single layer colloidal centrifugation protocols in combination with different freezing extenders on the fragmentation dynamics of thawed equine sperm DNA 
Background
Variability among stallions in terms of semen cryopreservation quality renders it difficult to arrive at a standardized cryopreservation method. Different extenders and processing techniques (such us colloidal centrifugation) are used in order to optimize post-thaw sperm quality. Sperm chromatin integrity analysis is an effective tool for assessing such quality. The aim of the present study was to compare the effect of two single layer colloidal centrifugation protocols (prior to cryopreservation) in combination with three commercial freezing extenders on the post-thaw chromatin integrity of equine sperm samples at different post-thaw incubation (37°C) times (i.e., their DNA fragmentation dynamics).
Results
Post-thaw DNA fragmentation levels in semen samples subjected to either of the colloidal centrifugation protocols were significantly lower (p<0.05) immediately after thawing and after 4 h of incubation at 37°C compared to samples that underwent standard (control) centrifugation. The use of InraFreeze® extender was associated with significantly less DNA fragmentation than the use of Botu-Crio® extender at 6 h of incubation, and than the use of either Botu-Crio® or Gent® extender at 24 h of incubation (p<0.05).
Conclusions
These results suggest that single layer colloidal centrifugation performed with extended or raw semen prior to cryopreservation reduces DNA fragmentation during the first four hours after thawing. Further studies are needed to determine the influence of freezing extenders on equine sperm DNA fragmentation dynamics.
doi:10.1186/1751-0147-54-72
PMCID: PMC3599590  PMID: 23217215
Colloidal centrifugation; Equine sperm; DNA fragmentation; Cryopreservation; Equipure®
20.  Increases in the completeness of disease records in dairy databases following changes in the criteria determining whether a record counts as correct 
Background
The four Nordic countries: Denmark (DK), Finland (FIN), Norway (NO) and Sweden (SE), all have national databases in which mainly records of treated animals are maintained. Recently, the completeness of locomotor disorder records in these databases has been evaluated using farmers’ recordings as a reference level. The objective of the present study was to see how previous estimates of completeness figures are affected by the criteria determining whether a recording in the database is to be judged correct. These demands included date of diagnosis and disease classification. In contrast with the previous study, a period of time between the date of disease recording in the database and by the farmer was allowed. Further, the calculations were brought to bear on individual locomotor diagnoses instead of a common locomotor disease complex code.
Methods
Randomly selected dairy herds (≥ 15 cows) were invited to participate. During two 2-month periods in 2008 the farmers recorded the diseases they observed on the farm and their recordings constituted a farmer database (FD). These recordings were compared to disease recordings in the National Databases (ND). Earlier calculations of completeness for locomotor complex cases assuming an exact match on date were compared with ±7 day and ±30 day discrepancies calculated in this study.
Results
The farmers in DK, FIN, NO and SE recorded 426, 147, 97 and 193 locomotor disorders, respectively. When a window of ±7 days was allowed there was a relative increase in completeness figures lying in the range of 24–100%. Further increases were minor, or non-existent, when the window was expanded to ±30 days. The same trend was seen for individual diagnoses.
Conclusion
In all four of the Nordic countries a common pattern can be observed: a further increase in completeness occurs when individual locomotor diagnoses recorded by the farmer are permitted to match any locomotor diagnosis recorded in the ND. Completeness increased when both time span and different diagnoses within the locomotor complex were allowed.
doi:10.1186/1751-0147-54-71
PMCID: PMC3538600  PMID: 23206729
Nordic dairy disease databases; Validation; Completeness; Farmer disease recording; Locomotor disorders
21.  Investigation of the presence of human or bovine respiratory syncytial virus in the lungs of mink (Neovison vison) with hemorrhagic pneumonia due to Pseudomonas aeruginosa 
Background
Hemorrhagic pneumonia is a disease of farmed mink (Neovison vison) caused by Pseudomonas aeruginosa. The disease is highly seasonal in Danish mink with outbreaks occurring almost exclusively in the autumn. Human respiratory syncytial virus (RSV) has been shown to augment infection with P. aeruginosa in mice and to promote adhesion of P. aeruginosa to human respiratory cells.
Findings
We tested 50 lung specimens from mink with hemorrhagic pneumonia for bovine RSV by reverse transcriptase polymerase chain reaction (PCR) and for human RSV by a commercial real-time PCR. RSV was not found.
Conclusions
This study indicates that human and bovine RSV is not a major co-factor for development of hemorrhagic pneumonia in Danish mink.
doi:10.1186/1751-0147-54-70
PMCID: PMC3533935  PMID: 23181342
Hemorrhagic pneumonia; Mink; Pseudomonas aeruginosa; Respiratory syncytial virus
22.  Genetic basis of penicillin resistance of S. aureus isolated in bovine mastitis 
Background
The blaZ gene encoding penicillin resistance can be located either chromosomally or on plasmids. The aim of this study was to investigate the genetic relationships and to determine the location of the blaZ gene in S. aureus isolated in bovine mastitis in Finland and Sweden.
Methods
Seventy-eight β-lactamase positive S. aureus isolates from bovine mastitis (34 from Finland and 44 from Sweden) were included in the study. The localization of blaZ gene was determined by Southern blotting. The blaZ genes of the isolates were sequenced and the sequences were translated to beta-lactamase proteins and further grouped as different protein signatures. The isolates and, as control, 33 Swedish and 36 Finnish beta-lactamase negative isolates were typed with pulsed-field gel electrophoresis (PFGE).
Results
In 26 out of 34 Finnish isolates (76.5%) and in 25 out of 44 Swedish isolates (56.8%) the blaZ gene was localized on a plasmid. Six different protein signatures were found. One signature was found only in four Swedish isolates, but all other signatures were found both in Finnish and Swedish isolates. The PFGE results revealed a diversity of S. aureus clones. The protein signatures were not clearly associated with certain pulsotypes.
Conclusions
The plasmid location of the blaZ gene was not statistically significantly more common in Finland than in Sweden, and hence does not explain the higher proportion of penicillin-resistant isolates of S. aureus causing bovine mastitis in Finland compared to Sweden.
doi:10.1186/1751-0147-54-69
PMCID: PMC3575348  PMID: 23176642
Mastitis; blaZ; Staphylococcus aureus; Beta-lactamases; Penicillin-resistance Correspondence
23.  Outside enclosure and additional enrichment for dairy goats – a preliminary study 
Background
Dairy goats are commonly housed at a space allowance of 0.7 – 0.8 m2/goat in commercial Norwegian goat herds, which is very low compared to regulations and recommendations in other European countries. One easy and cheap way to increase space allowance is to allow the animals’ access to outdoor area. The aim of this study was to investigate the effect of access to an outside enclosure and environmental enrichment for dairy goats kept in slatted floor pens with low space allowance on their activity pattern and social behaviour.
Methods
A group of 82 dairy goats on a commercial Norwegian dairy farm were kept inside during the winter period from October to April. In April the goats were given access to an outside enclosure for 8 hours per day. After having access to the enclosure for another for two days, enrichment (branches) was provided, and after 19 days the enrichment were removed. The goats were observed for 5 hours per day for the two last days before they got access to the outside enclosure, the two days in the enclosure, the two first and the two last days with enrichment and for the following two days without enrichment by two trained observers.
Results
When allowed access to the enclosure, the goats spent nearly 50% of the time outside, and later the time spent outside was reduced to less than 40% (P < 0.0001), but there was no clear effect of enrichment. All the goats appeared to have a regular use of the enclosure. Time spent resting decreased 59.2% to only 25.2% when the goats first got access to the enclosure, but then started to increase again (P < 0.0001). Initially time spent exploring and chewing the branches was 20%, but this was reduced to around 12% in the last part of the ENRICH period (P < 0.0001). Number of aggressive interactions tended to increase when the goats were allowed access to the outdoor enclosure whereas play behaviour was only observed in the outside enclosure (P < 0.05).
Conclusions
In conclusion, the goats preferred to use the outside enclosure when being active, and branches were perceived as an attractive enrichment.
doi:10.1186/1751-0147-54-68
PMCID: PMC3538566  PMID: 23173769
Goat; Outside enclosure; Environmental enrichment
24.  Toxocara cati larva migrans in domestic pigs - detected at slaughterhouse control in Norway 
Routine Trichinella meat inspection at the slaughterhouse detected one larva in a pooled batch of 100 pig samples. The larva was sent to the Norwegian Veterinary Institute (NVI) for species identification.
Morphological examination revealed that the larva was not Trichinella spp. Molecular analysis was performed. PCR and sequencing of 5S/ITS identified the larva as Toxocara cati. A second round of digests was carried out at the meat inspection laboratory, in smaller batches to try to identify the infected animal. No further larvae were detected and it was not possible to identify which of the 100 animals the larva had come from. This is the first time that Toxocara cati has been reported in slaughterhouse pigs in Norway.
Although the infected individual could not be identified, the meat originated from one of six potential farms. A small survey regarding rodent control and cats was sent to each of these farms. Cats had restricted access to food storage areas (two farms reported that cats had access) whilst none of the farms allowed cats into the production housing. Cats were, however, present on all the farms (mostly stray cats of unknown health status). Half of the farms also reported seeing rodents in the pig housing during the previous six months and half reported finding rodents in the feed and straw storage areas. We were unable to narrow down the source of infection – however contamination of food or bedding material, with cat faeces or infected rodents, in addition to the presence of infected rodents in pig housing remain potential routes of infection.
doi:10.1186/1751-0147-54-66
PMCID: PMC3537544  PMID: 23171732
Fattening pigs; Internal transcribed spacer; Larva migrans; Meat inspection; Toxocara cati; Zoonosis; 5S ribosomal DNA
25.  Efficacy of single dose of an inactivated porcine circovirus type 2 (PCV2) whole-virus vaccine with oil adjuvant in piglets 
Background
Post-weaning multisystemic wasting syndrome (PMWS) associated with PCV2 is one of the most costly diseases currently faced by the swine industry. The development of effective vaccines against PCV2 infection has been accepted as an important strategy in the prophylaxis of PMWS.
Methods
In the present study, a PK-15 cell-adapted formalin-inactivated prototype vaccine candidate was prepared using a strain of PCV2 from China. Inactivation of the virus was accomplished using a standard formalin inactivation protocol. The protective properties of the inactivated PCV2 vaccine were evaluated in piglets. Ten 28-day-old pigs were randomly assigned to two groups, each with five. Group 1 was vaccinated intramuscularly with the inactivated virus preparation; Group 2 received sterile PBS as a placebo. By 28 days post-vaccination (DPV), Groups 1 and 2 were challenged intranasally and intramuscularly with 5 × 107 TCID50 of a virulent PCV2 isolate.
Results
The vaccinated pigs seroconverted to PCV2 and had high levels of serum antibodies to PCV2 at 28 days after vaccination, whereas the control pigs remained seronegative. No significant signs of clinical disease were recorded following the challenge with PCV2, but moderate amounts of PCV2 antigen were detected in most lymphoid organs of the control pigs. PCV2 was detected in two out of the five vaccinated pigs. Furthermore, pathological lesions and viremia were milder in the vaccinated group.
Conclusions
The obtained results indicate that the inactivated PCV2 virus vaccine with an oil adjuvant induce an immunological response in pigs that appears to provide protection from infection with PCV2. The vaccine, therefore, may have the potential to serve as a vaccine aimed to protect pigs from developing PMWS.
doi:10.1186/1751-0147-54-67
PMCID: PMC3539982  PMID: 23171757
Porcine circovirus type 2; Post-weaning multisystemic wasting syndrome; Single-dose immunization; Formalin-inactivated vaccine

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