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1.  Osteochondritis dessicans and subchondral cystic lesions in draft horses: a retrospective study. 
The Canadian Veterinary Journal  1998;39(10):627-633.
The clinical features, radiographic findings, treatment, and outcome in 51 draft horses with osteochondritis dessicans (OCD) or subchondral cystic lesions (SC) are reported. Clydesdale and Percheron were the most commonly affected breeds, and affected animals represented only 5% of the hospital population of draft horses. Horses were most frequently affected in the tibiotarsal joints and 73% (24 of 33 cases) of the horses with tibiotarsal effusion were affected bilaterally. Osteochondritis dessicans of the distal intermediate ridge was the most common lesion found in the tibiotarsal joint. The stifle was also frequently affected; 87% (13 of 15 cases) of horses with femoropatellar OCD only were lame, and lesions were most commonly located on the lateral trochlear ridge. Sixteen cases were managed conservatively, 30 received surgery, and 5 were euthanized. Lameness, effusion, or both clinical signs resolved in more than 50% of surgically treated cases, but clinical signs improved in 30% of conservatively-managed cases.
PMCID: PMC1539460  PMID: 9789673
2.  Comparative pathogenesis of type 1 (European genotype) and type 2 (North American genotype) porcine reproductive and respiratory syndrome virus in infected boar 
Virology Journal  2013;10:156.
Background
Porcine reproductive and respiratory syndrome virus (PRRSV) now has two main genotypes, genotype 1 (European) and genotype 2 (North American). There is a lack of data on the comparison of pathogenicity of the two genotypes in boars. The objectives of the present study were to evaluate the amount of PRRSV present in semen over time and compare the viral distribution and microscopic lesions of type 1 and type 2 PRRSV-infected boars.
Methods
Twenty-four 8-month-old PRRSV-naïve Duroc boars were randomly allocated to 3 treatment groups. The boars in groups 1 (n = 9) and 2 (n = 9) were intranasally inoculated with type 1 or type 2 PRRSV, respectively. The boars in groups 1 (n = 6) served as negative controls. Semen and blood samples were collected up to 35 days post-inoculation (dpi), and necropsies were performed on 14, 21, and 35 dpi.
Results
There were no significant differences in the genomic copy number of PRRSV, microscopic testicular lesion score, number of PRRSV-positive germ cells, or number of apoptotic cells between the type 1 and type 2 PRRSV-infected boars throughout the experiment. Histopathological changes were manifested by the desquamation of spermatocytes and the presence of multinucleated giant cells in seminiferous tubules of both type 1 and type 2 PRRSV-infected boars. The distribution of PRRSV-positive cells was focal; the virus was found in single germ cells or small clusters of germ cells, localized to the spermatogonia, spermatocytes, spermatids, and non-sperm cells in type 1 and type 2 PRRSV-infected boars.
Conclusions
The results of this study demonstrated that two genotypes of PRRSV do not have significantly different virulence toward the male reproductive system of pigs.
doi:10.1186/1743-422X-10-156
PMCID: PMC3663669  PMID: 23687995
Boar; Genotype; Male reproductive system; Porcine reproductive and respiratory syndrome virus; Semen
3.  An evaluation of chemical arthrodesis of the proximal interphalangeal joint in the horse by using monoiodoacetate. 
The use of monoiodoacetate (MIA) for arthrodesis of the proximal interphalangeal joint (PIJ) and the effect of exercise on the degree of fusion were investigated. Eight horses received 3 injections (Weeks 0, 3, 6) of MIA (2 mL; 60 mg/mL) into the right or left front PIJ. Peri-operatively, the horses received phenylbutazone, butorphanol, and abaxial sesamoidean nerve blocks to relieve pain. During the study, the horses were monitored for general health, lameness, and swelling around the injection area. Radiographs were taken biweekly to evaluate bony fusion. Horses were randomly divided into non-exercised and exercised groups. Exercise consisted of 20 minutes of trotting on a treadmill (4 m/s), 3 days per week for 13 weeks. The horses were euthanized at 24 weeks. Slab sections of the PIJ were evaluated grossly and radiographically for bony fusion. Histologic examinations were performed to evaluate articular cartilage. Three horses were excluded from the study after developing soft tissue necrosis around the injection site, septic arthritis, and necrotic tendinitis. The remaining horses remained healthy, developed a grade 1 to 4 lameness with minimal to severe swelling in the PIJ region. All 5 horses showed radiographic evidence of bony fusion, however, no fusion was present when injected joints were examined on postmortem examination. Histologic examination revealed thinning of the cartilage, diffuse necrosis of chondrocytes, with the calcified zone intact. Subjectively, exercise did not influence the degree of cartilage destruction. Based on this study, chemical arthrodesis cannot be advocated in clinical cases because of the high complication rate and lack of bony fusion.
Images
PMCID: PMC1189620  PMID: 11041498
4.  Joint disorder; a contributory cause to reproductive failure in beef bulls? 
The lame sire, unsound for breeding, can cause substantial economic loss due to reduced pregnancies in the beef-producing herd.
To test the hypothesis that joint disorder is a possible cause of infertility in beef sires, right and left hind limb bones from 34 beef sires were examined postmortem to identify lesions in the femorotibial, femoropatellar (stifle), tarsocrural, talocalcaneus, and proximal intertarsal (tarsal) joints. The bulls were slaughtered during or after the breeding season due to poor fertility results. Aliquots of the cauda epididymal contents taken postmortem from 26 bulls were used for sperm morphology evaluation. As a control, hind limbs (but no semen samples) from 11 beef bulls with good fertility results were included.
Almost all infertile bulls (30/34) had lesions in at least one joint. Twenty-eight bulls (28/30, 93%) had lesions in the stifle joint, and 24 (24/28, 86%) of these were bilateral. Fourteen bulls (14/30, 47%) had lesions in the tarsal joint, and 10 (10/14, 71%) of these were bilateral. Four bulls (4/34, 12%) had no lesions, three bulls (3/34, 9%) had mild osteoarthritis (OA), 5 (5/34, 15%) moderate OA, 17 (17/34, 50%) severe OA and 5 (5/34, 15%) deformed OA. Almost all OA lesions (97%) were characterized as lesions secondary to osteochondrosis dissecans. All the bulls with satisfactory sperm morphology (n = 12/34) had joint lesions, with mostly severe or deformed bilateral lesions (83%). Consequently, the most likely cause of infertility in these 12 bulls was joint disease. Almost all control bulls (10/11) had OA lesions, but most of them were graded as mild (55%) or moderate (36%). None of the control bulls had severe lesions or deformed OA.
We suggest that joint lesions should be taken into consideration as a contributory cause of reproductive failure in beef sires without symptoms of lameness.
doi:10.1186/1751-0147-49-31
PMCID: PMC2212630  PMID: 17983470
5.  Cartilage proteoglycans from normal and osteochondrotic porcine joints. 
Modern pigs grow fast but are highly susceptible to degenerative joint abnormalities, including osteochondrosis. Normal and osteochondrotic humeri and femurs were obtained from five normal and ten lame adolescent boars to study cartilage proteoglycans. Histological examination of joints indicated a locally-reduced intensity of proteoglycan staining by safranin-O in lesion areas of cartilage. Cartilage proteoglycans extracted with 4.0 M guanidinium chloride were studied using Sepharose 2B gel chromatography. The proteoglycans from severely osteochondrotic joints were less (P less than 0.05) aggregated and contained a greater (P less than 0.05) proportion of smaller monomers than those from normal joints. Loss or damage of core protein, including its hyaluronic acid-binding regions, may account for the greater proportion of small monomers. The results also indicated that the proportion of hyaluronic acid in the total glycosaminoglycan uronic acid fraction, estimated by Sephadex G-200 chromatography and cellulose acetate electrophoresis, was lower (P less than 0.05) for the extracted proteoglycans than for the residual or the whole cartilage proteoglycans in all joints studied.
PMCID: PMC1236153  PMID: 3926288
6.  Detection of porcine reproductive and respiratory syndrome virus in boar semen by PCR. 
Journal of Clinical Microbiology  1995;33(7):1730-1734.
Porcine reproductive and respiratory syndrome virus (PRRSV) causes a devastating disease in swine. The presence and transmission of PRRSV by boar semen has been demonstrated by using a swine bioassay. In this assay, 4- to 8-week-old pigs were inoculated intraperitoneally with semen from PRRSV-infected boars. Seroconversion of these piglets indicated the presence of PRRSV in semen. Seroconversion in gilts has also been demonstrated following artificial insemination with semen from PRRSV-infected boars. These methods of detecting PRRSV in boar semen are time-consuming, laborious, and expensive. The objective of this study was to develop a reliable and sensitive PCR assay to directly detect PRRSV in boar semen. Primers from open reading frames 1b and 7 of the PRRSV genome were used in nested PCRs. Virus was detected at concentrations as low as 10 infectious virions per ml in PRRSV-spiked semen. Specificity was confirmed by using a nested PCR and a 32P-labeled oligonucleotide probe. The primers did not react with related arteriviruses or other swine viruses. The PCR assay showed good correlation with the swine bioassay, and both methods were superior to virus isolation. To consistently identify PRRSV in boar semen, the cell fraction was separated by centrifugation at 600 x g for 20 min, a lysis buffer without a reducing agent (2-mercaptoethanol) was used, and nondiluted and 1:20-diluted cell fractions were evaluated by PCR. PRRSV was not reliably detected in the seminal plasma fraction of boar semen.
PMCID: PMC228258  PMID: 7665637
7.  A study of repair cartilage from osteochondrotic humeral condyles of swine: preliminary report. 
A total of 16 animals, including 12 lame and four normal boars, were used. All lame boars had severe osteochondrotic humeral condyles in which repair cartilage tissues originating from subchondral bone were observed. Quantitative chemical studies of repair cartilage and normal cartilage were carried out using humeral condyles from four selected animals (two lame and two normal boars, respectively). The repair cartilage contained a higher concentration of collagen and lower concentration of proteoglycan than did the normal cartilage, consistent with the light microscopic observation of fibrous and fibrocartilaginous tissues in the repair cartilage. The proportion of proteoglycan extractable with 4M-guanidine hydrochloride was less in the repair cartilage than the normal cartilage. The proportion of proteoglycan that can interact with hyaluronic acid was also less than normal in the repair cartilage.
PMCID: PMC1263573  PMID: 1477807
8.  Incidence of osteochondrosis (dissecans) in Dutch warmblood horses presented for pre-purchase examination 
Irish Veterinary Journal  2008;61(1):33-37.
Data are lacking in the literature regarding the incidence of osteochondrosis (dissecans) [OC(D)] in relation to lameness evaluation in Dutch Warmblood horses. The objective of this retrospective study was to assess the incidence of radiological abnormalities consistent with osteochondrosis or osteochondrosis dissecans in 1,231 sound Dutch Warmblood (DW) horses presented for pre-purchase examination. Standardised (Dutch) pre-purchase examination protocols were evaluated. The pre-purchase examination included a clinical, lameness and radiological evaluation, performed at a private equine clinic in the Netherlands. Radiographical examination included views of the distal (DIP) and proximal (PIP) interphalangeal, metacarpo- and metatarsophalangeal (MCP/MTP), tarsocrural (TC) and femoropatellar (FP) joints. Radiographical evidence of OC(D) was found in 44.3% of clinically sound DW horses. In this study, 443 horses (36%, n = 1,231) had evidence of OCD and 102 horses (8.3%, n = 1,231) had evidence of OC on pre-purchase radiographs. The results also indicated that the TC joints were significantly more likely to be affected. A considerable number of horses did not demonstrate any lameness, although radiographs revealed OC(D).
doi:10.1186/2046-0481-61-1-33
PMCID: PMC3113880  PMID: 21851701
Osteochondrosis; dissecans; lameness; pre-purchase; examination
9.  Symmetric dimethyl arginine and N-acetyl-β-D-glucosaminidase lysozimuria of proximal renal tubules as a target for nephrotoxicity in patients with rheumatoid arthritis treated with disease modifying antirheumatic drugs 
Journal of Nephropathology  2013;2(1):36-52.
Background
The aim of this study was to determine the effect of initial therapy with some disease modifying antirheumatic drugs (DMARDs) (Methotrexate and Ketoprofen) on glomerular and tubular integrity in patients with Rheumatoid arthritis (RA).
Objectives
Objectives: To determine whether there is a change in clinical and laboratory indicators of renal function in course of the follow up of treatment and whether that change correlates with the dynamics of the quantity of enzymes excreted in urine and reactants of the acute phase.
Materials and Methods
Using colorimetric method for determination of NAG, samples of 70 participants were examined (35 RA patients treated with Ketoprofen only, 35 RA patients treated with combined use of Methotrexate and Ketoprofen). The follow up was 5 time-intervals in the course of 24 weeks.
Results
There was moderate correlation between NAG and microalbuminuria (r=0,34) in the group of patients treated with Ketoprofen only, while statistically significant correlation (r=0,21) was seen in group of patients with combined use of Methotrexate and Ketoprofen. NAG enzymuria in size, number of patients registered, and time of appearance were greater and appears earlier in the group with the combined use of Methotrexate and Ketoprofen compared with the mono-therapy with Ketoprofen. Mean urinary NAG induction was increasing with the concomitant use of Methotrexate and Ketoprofen.
Conclusions
Methotrexate is more potent NAG inductor than Ketoprofen and provokes greater tubular enzymuria than Ketoprofen.
doi:10.5812/nephropathol.8989
PMCID: PMC3886170  PMID: 24475424
N-acetyl-D-glucosaminidase; Symmetric dimethyl arginine; Rheumatoid arthritis; Immunosuppressive drugs
10.  Identification of QTL with effects on intramuscular fat content and fatty acid composition in a Duroc × Large White cross 
BMC Genetics  2007;8:55.
Background
Improving pork quality can be done by increasing intramuscular fat (IMF) content. This trait is influenced by quantitative trait loci (QTL) sought out in different pig populations. Considering the high IMF content observed in the Duroc pig, it was appealing to determine whether favourable alleles at a major gene or QTL could be found. The detection was performed in an experimental F2 Duroc × Large White population first by segregation analysis, then by QTL mapping using additional molecular information.
Results
Segregation analysis provided evidence for a major gene, with a recessive Duroc allele increasing IMF by 1.8% in Duroc homozygous pigs. However, results depended on whether data were normalised or not. After Box-Cox transformation, likelihood ratio was indeed 12 times lower and no longer significant. The QTL detection results were partly consistent with the segregation analysis. Three QTL significant at the chromosome wide level were evidenced. Two QTL, located on chromosomes 13 and 15, showed a high IMF Duroc recessive allele with an overall effect slightly lower than that expected from segregation analysis (+0.4 g/100 g muscle). The third QTL was located on chromosome 1, with a dominant Large White allele inducing high IMF content (+0.5 g/100 g muscle). Additional QTL were detected for muscular fatty acid composition.
Conclusion
The study presented results from two complementary approaches, a segregation analysis and a QTL detection, to seek out genes involved in the higher IMF content observed in the Duroc population. Discrepancies between both methods might be partially explained by the existence of at least two QTL with similar characteristics located on two different chromosomes for which different boars were heterozygous. The favourable and dominant allele detected in the Large White population was unexpected. Obviously, in both populations, the favourable alleles inducing high IMF content were not fixed and improving IMF by fixing favourable alleles using markers can then be applied both in Duroc and LW populations. With QTL affecting fatty acid composition, combining an increase of IMF content enhancing monounsaturated fatty acid percentage would be of great interest.
doi:10.1186/1471-2156-8-55
PMCID: PMC1978499  PMID: 17705816
11.  Quantitative trait loci analysis for leg weakness-related traits in a Duroc × Pietrain crossbred population 
Background
Leg weakness issues are a great concern for the pig breeding industry, especially with regard to animal welfare. Traits associated with leg weakness are partly influenced by the genetic background of the animals but the genetic basis of these traits is not yet fully understood. The aim of this study was to identify quantitative trait loci (QTL) affecting leg weakness in pigs.
Methods
Three hundred and ten F2 pigs from a Duroc × Pietrain resource population were genotyped using 82 genetic markers. Front and rear legs and feet scores were based on the standard scoring system. Osteochondrosis lesions were examined histologically at the head and the condylus medialis of the left femur and humerus. Bone mineral density, bone mineral content and bone mineral area were measured in the whole ulna and radius bones using dual energy X-ray absorptiometry. A line-cross model was applied to determine QTL regions associated with leg weakness using the QTL Express software.
Results
Eleven QTL affecting leg weakness were identified on eight autosomes. All QTL reached the 5% chromosome-wide significance level. Three QTL were associated with osteochondrosis on the humerus end, two with the fore feet score and two with the rear leg score. QTL on SSC2 and SSC3 influencing bone mineral content and bone mineral density, respectively, reached the 5% genome-wide significance level.
Conclusions
Our results confirm previous studies and provide information on new QTL associated with leg weakness in pigs. These results contribute towards a better understanding of the genetic background of leg weakness in pigs.
doi:10.1186/1297-9686-43-13
PMCID: PMC3072315  PMID: 21418602
12.  Prevalence of lameness and claw lesions during different stages in the reproductive cycle of sows and the impact on reproduction results 
Animal  2013;7(7):1174-1181.
Lameness in sows is an emerging disease condition with major effects on animal welfare and economics. Yet the direct impact on reproduction results remains unclear. The present field study investigated the impact of lameness and claw lesions throughout the reproductive cycle on (re)production results of sows. In five farms, a total of 491 group-housed sows were followed up for a period of one reproductive cycle. Sows were assessed for lameness every time they were moved to another area in the farm. Claw lesions were scored at the beginning and at the end of the cycle. Reproduction results included the number of live-born piglets, stillborn piglets, mummified fetuses and crushed piglets, weaning-to-oestrus interval and the presence of sows not showing oestrus post weaning, returning to service and aborting. Sows that left the group were recorded and the reason was noted. A mean prevalence of lameness of 5.9% was found, although it depended on the time in the productive cycle. The highest percentage of lame sows (8.1%) was found when sows were moved from the post-weaning to the gestation stable. No significant associations were found between lameness and reproduction parameters with the exception of the effect on mummified foetuses. Wall cracks, white line lesions, heel lesions and skin lesions did have an effect on farrowing performance. Of all sows, 22% left the group throughout the study, and almost half of these sows were removed from the farm. Lameness was the second most important reason for culling. Sows culled because of lameness were significantly younger compared with sows culled for other reasons (parity: 2.6 ± 1.3 v. 4.0 ± 1.8). In conclusion, the present results indicate that lameness mainly affects farm productivity indirectly through its effect on sow longevity, whereas claw lesions directly affect some reproductive parameters. The high percentage of lame sows in the insemination stable indicate that risk factor studies should not only focus on the gestation stable, but also on housing conditions in the insemination stable.
doi:10.1017/S1751731113000232
PMCID: PMC3666190  PMID: 23714359
sow; lameness; reproduction; culling; claw lesions
13.  Pathological changes in the lumbar spine of boars. 
Lumbar vertebral columns from 73 boars were examined for pathological changes. While pathological changes were minimal at five months of age, several animals aged seven to 18 months of age had facet asymmetry and large osteophytes surrounding lumbar facets. In the adult group 36% had degeneration of intervertebral discs and 28% had ventrolateral vertebral osteophytes. Lesions characterized by irregular areas of cavitation in the annulus and adjacent vertebral epiphyses were also observed. Of animals submitted for necropsy with a history of clinical lameness. lesions included nonsuppurative polyarthritis, fracture of the sacrum and femoral head, osteochondrosis involving articular cartilage of the distal humerus and distal femur, vertebral osteomyelitis, meningeal abscess and ulnar osteomyelitis.
Images
PMCID: PMC1320094  PMID: 7448627
14.  Abrasions and lameness in piglets born in different farrowing systems with different types of floor 
Background
The quality of the floor is essential to the welfare of piglets as abrasions often are recorded in newborn piglets, and such lesions may lead to lameness. Apart from animal suffering, lameness contributes to losses in form of dead piglets, decreased growth, and increased use of antibiotics and manual labour.
Methods
In a herd with three different farrowing systems, 37 litters (390 piglets) were studied until the age of 3 weeks with respect to presence of skin wounds and abrasions. Lameness was registered until the age of 7 weeks. Eight lame piglets were sacrificed before medical treatment and subjected to necropsy including histopathological and microbiological examinations. Isolates of streptococci, staphylococci and E. coli were tested with respect to antimicrobial resistance. Mastitis was observed in ten sows.
Results
The most severe abrasions at carpus and soles were seen in the system with a new solid concrete floor with a slatted floor over the dunging area. The lowest magnitude was observed in the deep litter system with peat. Sole bruising was more common in the systems with concrete floor compared to the deep litter system with peat, and the differce in prevalence was significant at all examination days. The lesions decreased with time and about 75% of the treatments for lameness were performed during the first three weeks of life. The overall prevalence of lameness was highest in the system with new solid concrete floor with a slatted floor over the dunging area (9.4%) followed by the old solid concrete floor (7.5%). A lower (p < 0.05) prevalence was seen in the deep litters system with peat (3.3%). No significant relationship between mastitis and abrasions or lameness in the offspring was observed.
Conclusion
There were large differences in the prevalence of abrasions and lameness between the floor types. The deep litter system with peat provided a soft and good floor for piglets. The overall prevalence of lameness was only diagnosed in every fourth litter in that system compared to in every second litter in the systems with concrete floor. In contrast, the incidence of mastitis in the sows during the first week after farrowing was higher than in the systems with concrete floor.
doi:10.1186/1751-0147-50-37
PMCID: PMC2564923  PMID: 18816414
15.  Transcript profiling of candidate genes in testis of pigs exhibiting large differences in androstenone levels 
BMC Genetics  2010;11:4.
Background
Boar taint is an unpleasant odor and flavor of the meat and occurs in a high proportion of uncastrated male pigs. Androstenone, a steroid produced in testis and acting as a sex pheromone regulating reproductive function in female pigs, is one of the main compounds responsible for boar taint. The primary goal of the present investigation was to determine the differential gene expression of selected candidate genes related to levels of androstenone in pigs.
Results
Altogether 2560 boars from the Norwegian Landrace and Duroc populations were included in this study. Testicle samples from the 192 boars with most extreme high or low levels of androstenone in fat were used for RNA extraction, and 15 candidate genes were selected and analyzed by real-competitive PCR analysis. The genes Cytochrome P450 c17 (CYP17A1), Steroidogenic acute regulatory protein (STAR), Aldo-keto reductase family 1 member C4 (AKR1C4), Short-chain dehydrogenase/reductase family member 4 (DHRS4), Ferritin light polypeptide (FTL), Sulfotransferase family 2A, dehydroepiandrosterone-preferring member 1 (SULT2A1), Cytochrome P450 subfamily XIA polypeptide 1 (CYP11A1), Cytochrome b5 (CYB5A), and 17-beta-Hydroxysteroid dehydrogenase IV (HSD17B4) were all found to be significantly (P < 0.05) up-regulated in high androstenone boars in both Duroc and Landrace. Furthermore, Cytochrome P450 c19A2 (CYP19A2) was down-regulated and progesterone receptor membrane component 1 (PGRMC1) was up-regulated in high-androstenone Duroc boars only, while CYP21 was significantly down-regulated (2.5) in high-androstenone Landrace only. The genes Nuclear Receptor co-activator 4 (NCOA4), Sphingomyrlin phosphodiesterase 1 (SMPD1) and 3β-hydroxysteroid dehydrogenase (HSD3B) were not significantly differentially expressed in any breeds. Additionally, association studies were performed for the genes with one or more detected SNPs. Association between SNP and androstenone level was observed in CYB5A only, suggesting cis-regulation of the differential transcription in this gene.
Conclusion
A large pig material of highly extreme androstenone levels is investigated. The current study contributes to the knowledge about which genes that is differentially expressed regard to the levels of androstenone in pigs. Results in this paper suggest that several genes are important in the regulation of androstenone level in boars and warrant further evaluation of the above mentioned candidate genes, including analyses in different breeds, identification of causal mutations and possible gene interactions.
doi:10.1186/1471-2156-11-4
PMCID: PMC2823645  PMID: 20100319
16.  Incidence of lameness and abrasions in piglets in identical farrowing pens with four different types of floor 
Background
Lameness in piglets is a major animal welfare issue. Floor abrasiveness is a common cause of superficial injury in piglets in farrowing pens. The abrasion achieved may act as a gate for infections, which in turn may induce development of infectious arthritis. In this study, the influence of improvements of the floor quality and of increased ratios of straw in identical farrowing pens was measured.
Methods
The study was carried out at a herd with four identical farrowing units with solid concrete floor bedded with 1 kg chopped straw per sow and 1 hg per piglet and day. Nothing was changed in the management of the four identical farrowing units, but four experimental groups were created: Group I – control, Group II – the amount of bedding was doubled. The surface of the floor was repaired in two units, Group III – Piglet Floor®, Flowcrete Sweden AB, Perstorp, Sweden and Group IV – Thorocrete SL®, Växa Halland, Sweden. Three farrowing batches were studies in each unit. In total, 93 litters (1,073 piglets) were examined for foot and skin lesions until the age of 3 weeks. The occurrence of lameness was registered until weaning at an average age of 4.5 weeks. Twenty seven lame piglets were culled instead of medicinally treated and subjected to necropsy including histopathological and microbiological examinations. Isolates of streptococci, staphylococci and E. coli were tested with respect to antimicrobial resistance.
Results
Piglet born on the repaired floors had the lowest prevalences of abrasions at carpus. Also the doubled straw ration decreased the abrasions. Skin lesions at carpus decreased significantly in magnitude in all four systems from day 10. At day 3, the sole bruising scores of the control unit were greater than the other three units (p < 0.001). At day 10 and 17, sole bruising was less common in the units with repaired floors than in the control group and the group with doubled straw ration. In total 41 piglets were diagnosed as lame, corresponding to 3.8% of all live-born piglets (n = 1,073). Around 85% of these diagnoses took place during the first 3 weeks of life and the risk incidence of lameness decreased from 1.5% during the first week of life to 0.5% during the fourth week. The incidence of lameness was highest in the control unit and lowest in the units with repaired floors. Twenty lame piglets were confirmed to have bacterial growth in the joint. The causative agents were Streptococcus dysgalactiae subsp. equisimilis (60%), Staphylococcus hyicus subsp. hyicus (35%) and Escherichia coli (5%). These isolates were sensitive to all antibiotics included in the antimicrobial panels.
Conclusion
The results suggest that proper maintenance of the floor can prevent the degree of roughness and abrasiveness of the floors, which in turn can contribute significantly to prevention of abrasions, sole bruising and lameness in piglets. Maintaining the surface of concrete floors with two different commercially available solutions both decreased the incidence of abrasions and sole bruisings and thereby also of arthritis significantly. Also doubling the amount of chopped straw turned out to prevent development of skin lesions and sole bruisings to some extent, and subsequently also the incidence of arthritis.
doi:10.1186/1751-0147-51-23
PMCID: PMC2697160  PMID: 19463187
17.  Age-Related Toxoplasma gondii Seroprevalence in Dutch Wild Boar Inconsistent with Lifelong Persistence of Antibodies 
PLoS ONE  2011;6(1):e16240.
Toxoplasma gondii is an important zoonotic pathogen that is best known as a cause of abortion or abnormalities in the newborn after primary infection during pregnancy. Our aim was to determine the prevalence of T. gondii in wild boar to investigate the possible role of their meat in human infection and to get an indication of the environmental contamination with T. gondii. The presence of anti-T. gondii antibodies was determined by in-house ELISA in 509 wild boar shot in 2002/2003 and 464 wild boar shot in 2007. Most of the boar originated from the “Roerstreek” (n = 673) or the “Veluwe” (n = 241). A binormal mixture model was fitted to the log-transformed optical density values for wild boar up to 20 months old to estimate the optimal cut-off value (−0.685) and accompanying sensitivity (90.6%) and specificity (93.6%). The overall seroprevalence was estimated at 24.4% (95% CI: 21.1–27.7%). The prevalence did not show variation between sampling years or regions, indicating a stable and homogeneous infection pressure from the environment. The relation between age and seroprevalence was studied in two stages. Firstly, seroprevalence by age group was determined by fitting the binary mixture model to 200 animals per age category. The prevalence showed a steep increase until approximately 10 months of age but stabilized at approximately 35% thereafter. Secondly, we fitted the age-dependent seroprevalence data to several SIR-type models, with seropositives as infected (I) and seronegatives as either susceptible (S) or resistant (R). A model with a recovery rate (SIS) was superior to a model without a recovery rate (SI). This finding is not consistent with the traditional view of lifelong persistence of T. gondii infections. The high seroprevalence suggests that eating undercooked wild boar meat may pose a risk of infection with T. gondii.
doi:10.1371/journal.pone.0016240
PMCID: PMC3024411  PMID: 21283764
18.  Association between SNPs within candidate genes and compounds related to boar taint and reproduction 
BMC Genetics  2009;10:32.
Background
Boar taint is an unpleasant odour and flavour of the meat from some uncastrated male pigs primarily caused by elevated levels of androstenone and skatole in adipose tissue. Androstenone is produced in the same biochemical pathway as testosterone and estrogens, which represents a particular challenge when selecting against high levels of androstenone in the breeding programme, without simultaneously decreasing levels of other steroids. Detection of single nucleotide polymorphisms (SNPs) associated with compounds affecting boar taint is important both for gaining a better understanding of the complex regulation of the trait and for the purpose of identifying markers that can be used to improve the gain of breeding. The beneficial SNPs to be used in breeding would have the combinational effects of reducing levels of boar taint without affecting fertility of the animals. The aim of this study was to detect SNPs in boar taint candidate genes and to perform association studies for both single SNPs and haplotypes with levels of boar taint compounds and phenotypes related to reproduction.
Results
An association study involving 275 SNPs in 121 genes and compounds related to boar taint and reproduction were carried out in Duroc and Norwegian Landrace boars. Phenotypes investigated were levels of androstenone, skatole and indole in adipose tissue, levels of androstenone, testosterone, estrone sulphate and 17β-estradiol in plasma, and length of bulbo urethralis gland. The SNPs were genotyped in more than 2800 individuals and several SNPs were found to be significantly (LRT > 5.4) associated with the different phenotypes. Genes with significant SNPs in either of the traits investigated include cytochrome P450 members CYP2E1, CYP21, CYP2D6 and CYP2C49, steroid 5α-reductase SRD5A2, nuclear receptor NGFIB, catenin CTNND1, BRCA1 associated protein BAP1 and hyaluronoglucosaminidase HYAL2. Haplotype analysis provided additional evidence for an effect of CYP2E1 on levels of skatole and indole, and for BAP1, HYAL2 and SRD5A2 on levels of androstenone.
Conclusion
The findings in this study indicate that polymorphisms in CYP2E1, CYP21, CYP2D6, CYP2C49, NGFIB and CTNND1 might be used to reduce levels of boar taint without affecting levels of testosterone, estrone sulphate, 17β-estradiol or length of bulbo urethralis gland.
doi:10.1186/1471-2156-10-32
PMCID: PMC2723134  PMID: 19575819
19.  Genome-wide association scan and phased haplotype construction for quantitative trait loci affecting boar taint in three pig breeds 
BMC Genomics  2012;13:22.
Background
Boar taint is the undesirable smell and taste of pork meat derived from some entire male pigs. The main causes of boar taint are the two compounds androstenone and skatole (3-methyl-indole). The steroid androstenone is a sex pheromone produced in the testis of the boars. Skatole is produced from tryptophan by bacteria in the intestine of the pigs. In many countries pigs are castrated as piglets to avoid boar taint, however, this is undesirable for animal welfare reasons. Genetic variations affecting the level of boar taint have previously been demonstrated in many breeds. In the study presented in this paper, markers and haplotypes, which can be applied to DNA-based selection schemes in order to reduce or eliminate the boar taint problem, are identified.
Results
Approximately 30,000 SNPs segregating in 923 boars from three Danish breeds; Duroc, Landrace, and Yorkshire, were used to conduct genome wide association studies of boar taint compounds. At 46 suggestive quantitative trait loci (QTL), 25 haplotypes and three single markers with effects were identified. Furthermore, 40% of the haplotypes mapped to previously identified regions. Haplotypes were also analysed for effects of slaughter weight and meat content. The most promising haplotype was identified on Sus scrofa chromosome 1. The gain in fixed effect of having this haplotype on level of androstenone in Landrace was identified to be high (1.279 μg/g). In addition, this haplotype explained 16.8% of the phenotypic variation within the trait. The haplotype was identified around the gene CYB5A which is known to have an indirect impact on the amount of androstenone. In addition to CYB5A, the genes SRD5A2, LOC100518755, and CYP21A2 are candidate genes for other haplotypes affecting androstenone, whereas, candidate genes for the indolic compounds were identified to be SULT1A1 and CYP2E1.
Conclusions
Despite the small sample size, a total of 25 haplotypes and three single markers were identified including genomic regions not previously reported. The haplotypes that were analysed showed large effects on trait level. However, little overlap of QTL between breeds was observed.
doi:10.1186/1471-2164-13-22
PMCID: PMC3315726  PMID: 22244367
20.  Borrelia burgdorferi migrates into joint capsules and causes an up-regulation of interleukin-8 in synovial membranes of dogs experimentally infected with ticks. 
Infection and Immunity  1997;65(4):1273-1285.
Twenty 6-week-old specific-pathogen-free beagles were infected with Borrelia burgdorferi by tick challenge, and five uninfected dogs served as controls. During the study, all dogs were monitored for infection, clinical signs, and antibody response against B. burgdorferi. During episodes of lameness or postmortem, synovial fluids from each dog were examined for volume, cell number, polymorphonuclear leukocyte (PMN) content, cell viability, and chemotactic activity. Twenty-five tissues collected postmortem from each dog were tested for interleukin-8 (IL-8) mRNA, tumor necrosis factor alpha (TNF-alpha) mRNA, presence of live spirochetes, and histopathological changes. Thirteen infected dogs (group A), which seroconverted rapidly (maximum titers within 50 to 90 days), developed acute and severe mono- or oligoarthritis almost exclusively in the limb closest to the tick bite (median incubation period, 66 days). Synovial fluids of the arthritic joints collected during episodes of lameness had significantly elevated volume, cell count, PMN proportion, cell viability, and chemotactic activity for PMNs. The remaining joints of the same animals contained synovial fluids with elevated chemotactic activity and cell viability. Twelve dogs tested positive for IL-8 mRNA in multiple tissues (synovia, pericardium, and peritoneum), and 10 dogs expressed TNF-alpha mRNA, but only in the tributary lymph nodes of the inflamed joints. Histological examinations revealed severe poly- or oligoarthritis and moderate to severe cortical hyperplasia in draining lymph nodes of the inflamed joints in all 13 dogs. Seven infected dogs with mild or no clinical signs (group B) seroconverted slowly (peak titers after 90 days), and only some joint fluids showed chemotactic activity, which on average was lower than that in inflamed and noninflamed joints from dogs in group A. Four dogs expressed IL-8 mRNA (in the synovia and pericardium), and three dogs had TNF-alpha mRNA in tributary lymph nodes. Histologically, nonsuppurative arthritis was found in multiple joints, and mild to moderate cortical hyperplasia was found in draining lymph nodes. Five uninfected dogs without lameness (group C) had normal synovial fluids and tissues. In all infected dogs, live spirochetes were demonstrated more frequently in tissues of the somatic quadrant closest to the tick bite than in tissues further from the site of infection, suggesting that dissemination of B. burgdorferi occurs more by migration than by blood-borne spread. From these studies employing a canine model of B. burgdorferi infection, we conclude that IL-8 is involved in the pathogenesis of acute Lyme arthritis.
PMCID: PMC175128  PMID: 9119462
21.  Porcine sperm-zona binding ability as an indicator of fertility 
Animal reproduction science  2007;104(1-2):69-82.
The escalated use of artificial insemination in swine has increased the importance of determining fertility of a semen sample before it is used. Multiple laboratory assays have been developed to assess fertilizing potential but they have yielded inconsistent results. This experiment sought to determine the relationship between in vitro competitive zona binding ability and in vivo fertility based on heterospermic inseminations and paternity testing. The zona pellucida binding ability and fertility of sperm from 15 boars was assessed by comparing sperm from one boar with sperm from other individual boars in a pair-wise fashion using four ejaculates. The relationship of zona binding ability to the mean number of piglets sired per litter for each boar as well as historic fertility data (litter size and farrowing rate) was assessed. The in vitro competition assay consisted of labeling sperm from each boar of the pair with a different fluorophore and incubating an equal number of sperm from each boar in the same droplet with porcine oocytes. The competitive assay was highly effective in ranking boars by zona binding ability (R2 = 0.94). Paternity testing using microsatellite markers was used to determine the mean number of piglets sired per litter for each boar during heterospermic inseminations. The pairwise heterospermic insemination assay was effective in ranking boar fertility (R2 = 0.59). Using historical data from these boars, average litter size and farrowing rate were correlated (r = 0.81, p < 0.001). However, zona binding ability was not significantly correlated with historic farrowing rate data or historic average litter size. Boar sperm zona binding ability was also not correlated significantly with the mean number of piglets sired per litter following heterospermic insemination. But the number of piglets sired by each boar was related to a combination of zona binding ability, sperm motility, normal morphology, acrosomal integrity, and the presence of distal droplets (R2 = 0.70). These results suggest that zona binding ability is not an accurate predictor of fertilizing ability when used alone; however, when coupled with other sperm assessments, fertility may be predicted successfully.
doi:10.1016/j.anireprosci.2007.01.012
PMCID: PMC2243185  PMID: 17303353
Pig - Fertility; Fertilization; Heterospermic Insemination; Sperm; Zona Pellucida
22.  Exposure of Wild Boar to Mycobacterium tuberculosis Complex in France since 2000 Is Consistent with the Distribution of Bovine Tuberculosis Outbreaks in Cattle 
PLoS ONE  2013;8(10):e77842.
The Eurasian wild boar (Sus scrofa) is increasingly considered as a relevant actor in the epidemiology of animal tuberculosis (TB). Therefore, monitoring TB in wild boar becomes a key tool for establishing comprehensive control schemes for this disease. To estimate the exposure of free living wild boar to Mycobacterium tuberculosis complex (MTC) in France, a bovine-purified protein derivative based ELISA was used to test 2,080 archived serum samples of hunter-harvested animals in 58 French “départements”. Two cut-off values were used for diagnostic interpretation: 0.2, recommended by the manufacturer (specificity: 96.43%; sensitivity: 72.6%), and 0.5 (specificity: 100%; sensitivity: 64%). During the same period, at the 0.2 cut-off, global true seroprevalence was 5.9% (IC95%: 4.3%-7.7%) and 76% of the sampled “départements” had seropositive wild boar, including seven cattle TB-free “départements. At the 0.5 cut-off, global true seroprevalence was 2.2% (IC95%: 1.5-3.2) and positive wild boar belonged to 21% of the “départements”. All but one of these positive “départements” had reported at least one cattle TB outbreak since 2000. A good consistence between seropositive wild boar and TB outbreaks in cattle was found, especially at the 0.5 cut-off value (the mean distance to the nearest cattle TB outbreak was 13km and 27km for seropositive and seronegative wild boar, respectively; P<0.05). The use of an ELISA to detect MTC antibodies in wild boar has permitted the description of the geographic distribution of MTC contact in wild boar in France. Our results suggest that the ELISA could be used as a first screening tool to conduct TB surveillance in wild boar at a population level. High-risk wild boar populations (e.g. overabundant) could be tested and if identified positive by ELISA they should be surveyed in detail by combining pathology and culture.
doi:10.1371/journal.pone.0077842
PMCID: PMC3805591  PMID: 24167584
23.  Gene expression profiles in testis of pigs with extreme high and low levels of androstenone 
BMC Genomics  2007;8:405.
Background:
Boar taint is a major obstacle when using uncastrated male pigs for swine production. One of the main compounds causing this taint is androstenone, a pheromone produced in porcine testis. Here we use microarrays to study the expression of thousands of genes simultaneously in testis of high and low androstenone boars. The study allows identification of genes and pathways associated with elevated androstenone levels, which is essential for recognising potential molecular markers for breeding purposes.
Results:
Testicular tissue was collected from 60 boars, 30 with extreme high and 30 with extreme low levels of androstenone, from each of the two breeds Duroc and Norwegian Landrace. The samples were hybridised to porcine arrays containing 26,877 cDNA clones, detecting 563 and 160 genes that were differentially expressed (p < 0.01) in Duroc and Norwegian Landrace, respectively. Of these significantly up- and down-regulated clones, 72 were found to be common for the two breeds, suggesting the possibility of both general and breed specific mechanisms in regulation of, or response to androstenone levels in boars. Ten genes were chosen for verification of expression patterns by quantitative real competitive PCR and real-time PCR. As expected, our results point towards steroid hormone metabolism and biosynthesis as important biological processes for the androstenone levels, but other potential pathways were identified as well. Among these were oxidoreductase activity, ferric iron binding, iron ion binding and electron transport activities. Genes belonging to the cytochrome P450 and hydroxysteroid dehydrogenase families were highly up-regulated, in addition to several genes encoding different families of conjugation enzymes. Furthermore, a number of genes encoding transcription factors were found both up- and down-regulated. The high number of clones belonging to ferric iron and iron ion binding suggests an importance of these genes, and the association between these pathways and androstenone levels is not previously described.
Conclusion:
This study contributes to the understanding of the complex genetic system controlling and responding to androstenone levels in pig testis. The identification of new pathways and genes involved in the biosynthesis and metabolism of androstenone is an important first step towards finding molecular markers to reduce boar taint.
doi:10.1186/1471-2164-8-405
PMCID: PMC2204014  PMID: 17988377
24.  Influence of lameness on follicular growth, ovulation, reproductive hormone concentrations and estrus behavior in dairy cows 
Theriogenology  2011;76(4):658-668.
The objective of this study was to examine the effect of a chronic stressor, lameness, on reproductive parameters. Seventy cows 30–80 days post-partum were scored for lameness and follicular phases synchronized with GnRH followed seven days later by prostaglandin (PG). Fifteen Lame animals did not respond to GnRH ovarian stimulation. Milk progesterone for 5 days prior to PG was lower in the remaining Lame cows than Healthy herdmates. Fewer Lame cows ovulated (26/37 versus 17/18; P = 0.04) and the interval from PG to ovulation was shorter in Lame cows. In Subset 1 (20 animals), the LH pulse frequency was similar in ovulating animals (Lame and Healthy) but lower in Lame non-ovulators. An LH surge always preceded ovulation but lameness did not affect the interval from PG to LH surge onset or LH surge concentrations. Before the LH surge, estradiol was lower in non-ovulating cows compared to those that ovulated and estradiol concentrations were positively correlated with LH pulse frequency. In Subset 2 (45 cows), Lame ovulating cows had a less intense estrus than Healthy cows, although Lame cows began estrus and stood-to-be-mounted earlier than Healthy cows. In conclusion, we have identified several parameters to explain poor fertility in some chronically stressed animals. From 30 to 80 days post-partum, there was a graded effect that ranged from 29% Lame cows with absence of ovarian activity, whereas another 21% Lame cows failed to express estrus or ovulate a low estrogenic follicle; in 50% cows, many reproductive parameters were unaffected by lameness.
doi:10.1016/j.theriogenology.2011.03.019
PMCID: PMC3156299  PMID: 21601262
Lameness; Dairy cattle; Follicle; Ovulation; Estrus
25.  Large scale genome-wide association and LDLA mapping study identifies QTLs for boar taint and related sex steroids 
BMC Genomics  2011;12:362.
Background
Boar taint is observed in a high proportion of uncastrated male pigs and is characterized by an unpleasant odor/flavor in cooked meat, primarily caused by elevated levels of androstenone and skatole. Androstenone is a steroid produced in the testis in parallel with biosynthesis of other sex steroids like testosterone and estrogens. This represents a challenge when performing selection against androstenone in breeding programs, without simultaneously decreasing levels of other steroids. The aim of this study was to use high-density genome wide association (GWA) in combination with linkage disequilibrium-linkage analysis (LDLA) to identify quantitative trait loci (QTL) associated with boar taint compounds and related sex steroids in commercial Landrace (n = 1,251) and Duroc (n = 918) breeds.
Results
Altogether, 14 genome wide significant (GWS) QTL regions for androstenone in subcutaneous fat were obtained from the LDLA study in Landrace and 14 GWS QTL regions in Duroc. LDLA analysis revealed that 7 of these QTL regions, located on SSC 1, 2, 3, 7 and 15, were obtained in both breeds. All 14 GWS androstenone QTLs in Landrace are also affecting the estrogens at chromosome wise significance (CWS) or GWS levels, while in Duroc, 3 of the 14 QTLs affect androstenone without affecting any of the estrogens. For skatole, 10 and 4 QTLs were GWS in the LDLA analysis for Landrace and Duroc respectively, with 4 of these detected in both breeds. The GWS QTLs for skatole obtained by LDLA are located at SSC 1, 5, 6, 7, 10, 11, 13 and 14.
Conclusion
This is the first report applying the Porcine 60 K SNP array for simultaneous analysis of boar taint compounds and related sex hormones, using both GWA and LDLA approaches. Several QTLs are involved in regulation of androstenone and skatole, and most of the QTLs for androstenone are also affecting the levels of estrogens. Seven QTLs for androstenone were detected in one breed and confirmed in the other, i.e. in an independent sample, although the majority of QTLs are breed specific. Most QTLs for skatole do not negatively affect other sex hormones and should be easier to implement into the breeding scheme.
doi:10.1186/1471-2164-12-362
PMCID: PMC3149590  PMID: 21752240

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