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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.  Consequences of the natural course of articular osteochondrosis in pigs for the suitability of computed tomography as a screening tool 
BMC Veterinary Research  2014;10(1):212.
A significant heritability has been documented for articular osteochondrosis. Selection against osteochondrosis has historically been based on macroscopic evaluation, but as computed tomography (CT) now is used to select boars with optimal body composition it can potentially also be used to screen for osteochondrosis. False negative diagnosis will occur if defects have not developed or have resolved prior to screening at a single time point.
The aim of the current study was to assess the suitability of the use of CT at a single point in time as a screening tool in piglets for articular osteochondrosis, which is known to be a highly dynamic condition in which lesions develop and resolve over time.
Male Landrace piglets (n = 18) were serial CT scanned from 2–8 times at biweekly intervals from 70–180 days of age. At each interval, 1–2 piglets were euthanased and the left distal femur processed for histological validation.
A total of 795 defects were identified in the 112 available CT scans. Within the hind and fore limbs, the incidence of defects was highest in the stifle (n = 321) and elbow joints (n = 110), respectively. Ninety-eight per cent of the defects in the stifle and elbow joints had developed by the 7th examination interval when the piglets were a mean age of 159 days old. The proportion of defects that resolved was lowest in the stifle joint at 51% and highest in the elbow joint at 69%.
Scanning of the current piglets at an age of 159 days resulted in detection of 98% of the total number of defects that developed up to the maximum age of 180 days. The proportion of defects that resolved ranged from 51–69% for different joints, but may not adversely affect prevalence as this category of false negative diagnosis will result in selection of pigs that are disposed for healing. Optimally timed CT is a powerful screening tool for osteochondrosis.
PMCID: PMC4172834  PMID: 25199991
Articular osteochondrosis; Leg weakness; Pig; CT; Histology; Development; Screening; Selection; Breeding
3.  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.
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.
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.
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.
The results of this study demonstrated that two genotypes of PRRSV do not have significantly different virulence toward the male reproductive system of pigs.
PMCID: PMC3663669  PMID: 23687995
Boar; Genotype; Male reproductive system; Porcine reproductive and respiratory syndrome virus; Semen
4.  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.
PMCID: PMC1189620  PMID: 11041498
5.  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.
PMCID: PMC2212630  PMID: 17983470
6.  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
7.  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
8.  Pressure mat analysis of naturally occurring lameness in young pigs after weaning 
BMC Veterinary Research  2014;10:193.
Lameness is a common problem in modern swine husbandry. It causes welfare problems in affected pigs as well as financial problems for farmers. To minimize these negative consequences of lameness, new treatment and prevention strategies need to be developed and validated using objective and quantitative measurement techniques. An example of such a putative diagnostic tool is the use of a pressure mat. Pressure mats are able to provide both objective loading (kinetic) as well as objective movement (kinematic) information on pig locomotion.
In this study, pressure mat analysis was used to assess compensatory force redistribution in lame pigs; in particular a predefined set of four pressure mat parameters was evaluated for its use to objectively distinguish clinically lame from sound pigs. Kinetic data from 10 clinically lame and 10 healthy weaned piglets were collected. These data were analyzed to answer three research questions. Firstly the pattern of compensatory weight distribution in lame animals was studied using the asymmetry indices (ASI) for several combinations of limbs. Secondly, the correlation between total left-right asymmetry index and visual scores of lameness was assessed. Thirdly, by using receiver-operated curve (ROC) analysis, optimal cutoff values for these ASIs were then calculated to objectively detect lame pigs.
Lame animals generally showed a shift in loading towards their diagonal and contralateral limbs, resulting in a clear left-right asymmetry. The degree of lameness as graded by visual scoring correlated well with the total left-right ASIs. Lame pigs could be objectively distinguished from sound pigs based on clear cutoff points calculated by ROC analysis for the complete set of four evaluated parameters.
The gait of lame pigs is asymmetric, due to the unloading of the affected limb and concomitant weight redistribution towards other limbs. This asymmetry objectively expressed as total left-right asymmetry, correlates well with the subjective visual lameness scoring and can be used to objectively distinguish lame from sound pigs. Pressure mat gait analysis of pigs, therefore, appears to be a promising and useful tool to objectively quantify and possibly early detect lameness in pigs.
PMCID: PMC4236822  PMID: 25139245
Kinetics; Gait analysis; Porcine; Symmetry; Redistribution; Loading
9.  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
10.  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.
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: 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.
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.
Methotrexate is more potent NAG inductor than Ketoprofen and provokes greater tubular enzymuria than Ketoprofen.
PMCID: PMC3886170  PMID: 24475424
N-acetyl-D-glucosaminidase; Symmetric dimethyl arginine; Rheumatoid arthritis; Immunosuppressive drugs
11.  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).
PMCID: PMC3113880  PMID: 21851701
Osteochondrosis; dissecans; lameness; pre-purchase; examination
12.  Identification of QTL with effects on intramuscular fat content and fatty acid composition in a Duroc × Large White cross 
BMC Genetics  2007;8:55.
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.
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.
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.
PMCID: PMC1978499  PMID: 17705816
13.  Quantitative trait loci analysis for leg weakness-related traits in a Duroc × Pietrain crossbred population 
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.
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.
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.
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.
PMCID: PMC3072315  PMID: 21418602
14.  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.
PMCID: PMC3666190  PMID: 23714359
sow; lameness; reproduction; culling; claw lesions
15.  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.
PMCID: PMC1320094  PMID: 7448627
16.  Effect of rider experience and evaluator expertise on subjective grading of lameness in sound and unsound sports horses under saddle 
The primary objective of this study was to investigate whether rider experience influences the assessment and grading of lameness in horses based on under-saddle gait analysis. Thirteen adult sports horses in active training were included in the study. After a baseline lameness and neurologic examination by the principal investigators, horses were videotaped while being ridden by an experienced and a less experienced rider. A 3-minute video was made for each horse and rider and 26 videos were randomly ordered and compiled on a DVD. Veterinarians with different levels of experience in evaluating lameness and veterinary students viewed the DVD and assigned a lameness score to each horse/rider combination. In a model accounting for the expertise of the evaluator, there was no difference in overall lameness scores between experienced and less experienced riders. This result was consistent for both sound and unsound horses. The overall lameness scores reported by specialists and students, however, differed significantly. The lameness score reported by the study participants while the horse was ridden was significantly associated with the subjective baseline lameness assessment reported by the principal investigators for the same limb when the horse was not under saddle. Additional work is necessary to determine whether riders with even lower skill levels would further alter the balance and motion pattern of the horse and have more influence on subjective grading of lameness.
PMCID: PMC3962283  PMID: 24688169
17.  Abrasions and lameness in piglets born in different farrowing systems with different types of floor 
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.
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.
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.
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.
PMCID: PMC2564923  PMID: 18816414
18.  Transcript profiling of candidate genes in testis of pigs exhibiting large differences in androstenone levels 
BMC Genetics  2010;11:4.
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.
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.
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.
PMCID: PMC2823645  PMID: 20100319
19.  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.
PMCID: PMC3024411  PMID: 21283764
20.  Incidence of lameness and abrasions in piglets in identical farrowing pens with four different types of floor 
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.
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.
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.
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.
PMCID: PMC2697160  PMID: 19463187
21.  Genome-wide association scan and phased haplotype construction for quantitative trait loci affecting boar taint in three pig breeds 
BMC Genomics  2012;13:22.
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.
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.
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.
PMCID: PMC3315726  PMID: 22244367
22.  Association between SNPs within candidate genes and compounds related to boar taint and reproduction 
BMC Genetics  2009;10:32.
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.
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.
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.
PMCID: PMC2723134  PMID: 19575819
23.  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.
PMCID: PMC2243185  PMID: 17303353
Pig - Fertility; Fertilization; Heterospermic Insemination; Sperm; Zona Pellucida
24.  Genetic structure of wild boar (Sus scrofa) populations from East Asia based on microsatellite loci analyses 
BMC Genetics  2014;15:85.
Wild boar, Sus scrofa, is an extant wild ancestor of the domestic pig as an agro-economically important mammal. Wild boar has a worldwide distribution with its geographic origin in Southeast Asia, but genetic diversity and genetic structure of wild boar in East Asia are poorly understood. To characterize the pattern and amount of genetic variation and population structure of wild boar in East Asia, we genotyped and analyzed microsatellite loci for a total of 238 wild boar specimens from ten locations across six countries in East and Southeast Asia.
Our data indicated that wild boar populations in East Asia are genetically diverse and structured, showing a significant correlation of genetic distance with geographic distance and implying a low level of gene flow at a regional scale. Bayesian-based clustering analysis was indicative of seven inferred genetic clusters in which wild boars in East Asia are geographically structured. The level of genetic diversity was relatively high in wild boars from Southeast Asia, compared with those from Northeast Asia. This gradient pattern of genetic diversity is consistent with an assumed ancestral population of wild boar in Southeast Asia. Genetic evidences from a relationship tree and structure analysis suggest that wild boar in Jeju Island, South Korea have a distinct genetic background from those in mainland Korea.
Our results reveal a diverse pattern of genetic diversity and the existence of genetic differentiation among wild boar populations inhabiting East Asia. This study highlights the potential contribution of genetic variation of wild boar to the high genetic diversity of local domestic pigs during domestication in East Asia.
PMCID: PMC4112609  PMID: 25034725
Microsatellites; East Asia; Genetic diversity; Genetic structure; Wild boar
25.  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

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