CD2/CD21 index: a new marker to evaluate udder health in dairy cows.

Lymphocytes play a significant role in the immunological processes of the bovine mammary gland and were found to be the dominant cell population in the milk of healthy udder quarters. The objective of this study was to investigate the quantitative relationship between CD2(+) T and CD21(+) B lymphocytes using flow cytometry. In a first study, quarter foremilk samples from apparently healthy udder quarters [somatic cell counts (SCC) ≤100,000 cells/mL; n=65] were analyzed and compared with diseased quarters (SCC >100,000 cells/mL; n=15). Percentages of CD2(+) T cells were significantly higher in milk samples with SCC ≤100,000 cells/mL than in those with SCC >100,000 cells/mL, whereas percentages of CD21(+) B cells developed in the opposite direction. As a result of this opposing trend, a new variable, the CD2/CD21 index-representing the percentages of CD2(+) cells per CD21(+) cells-was defined. Although diseased quarters with SCC >100,000 cells/mL and the detection of major pathogens revealed generally CD2/CD21 indices <10, values >10 were observed in apparently healthy quarters. Hence, a CD2/CD21 index cutoff value of 10 may be suitable to aid differentiation between unsuspicious and microbiologically suspicious or diseased udder quarters. To test whether CD2/CD21 indices <10 were primarily related to pathogens, quarters with SCC ≤100,000 cells/mL and >100,000 cells/mL with different bacteriological status (culture negative, or minor or major pathogens) were selectively examined in a second biphasic study. In the first trial, 63 udder quarters were analyzed and 55 of these quarters were able to be sampled again in the second trial carried out 14 d later. In both trials, results of the first study were confirmed. Indeed, CD2/CD21 indices <10 were also found in quarters showing SCC ≤100,000 cells/mL and containing minor or major pathogens at the time of the current or previous bacteriological analysis. The results of our examinations indicated a clear relationship between the CD2/CD21 index and the bacteriological status of the mammary gland. In combination with SCC, it offers a new marker for quick differentiation of unsuspicious and microbiologically suspicious or diseased udder quarters.

Comparison of joining and constant κ-light chain regions in different cattle breeds.

A comparative transcription analysis of Ig κ-light chains (IGK) of the cattle breeds Holstein Friesian (HF), German Black Pied (GBP), German Simmental (GS) and Aubrac (A) revealed three alleles coding for two putative allotypic variants of IGKC. The amino acid residues p.Asp100Asn and p.Thr116Ala were located at the outer edge of the constant domain as demonstrated by homology-based modelling. Alleles were distributed in unequal frequencies within the breeds examined. While cattle breeds HF, GS, and A possessed all alleles and allotypic variants, GBP exhibited alleles encoding allotypic variant IGKC(a) . All three IGKJ segments were detected in 320 sequences analysed. IGKJ1 was combined predominantly with IGKC. The ORF2 of IGKJ2 was detected for the first time on transcriptional level.

Flow cytometric differential cell counts in milk for the evaluation of inflammatory reactions in clinically healthy and subclinically infected bovine mammary glands.

Somatic cell counts (SCC) are generally used as an indicator of udder health. In Germany, a cutoff value of 100,000 cells/mL is currently used to differentiate between healthy and diseased mammary glands. In addition to SCC, differential cell counts (DCC) can be applied for a more detailed evaluation of the udder health status. The aim of this study was to differentiate immune cells in milk of udder quarters classified as healthy based on SCC values of <100,000 cells/mL. Twenty cows were selected and 65 healthy udder quarters were compared with a control group of 15 diseased udder quarters (SCC>100,000 cells/mL). Cells were isolated from milk of all quarters to measure simultaneously percentages of lymphocytes, macrophages, and polymorphonuclear neutrophilic leukocytes (PMNL) by flow cytometric analysis. The bacteriological status of all 80 quarters was also determined. Differential cell count patterns of milk samples (n = 15) with extreme low SCC values of ≤ 6,250 cells/mL revealed high lymphocyte proportions of up to 88%. Milk cell populations in samples (n = 42) with SCC values from >6,250 to ≤ 25,000 cells/mL were also dominated by lymphocytes, whereas DCC patterns of 6 out of 41 milk samples with SCC values from ≥ 9,000 to ≤ 46,000 cells/mL indicated already inflammatory reactions based on the predominance of PMNL (56-75%). In 13 of 15 milk samples of the diseased udder quarters (SCC >100,000 cells/mL), PMNL were categorically found as dominant cell population with proportions of ≥ 49%. Macrophages were the second predominant cell population in almost all samples tested in relation to lymphocytes and PMNL. Further analysis of the data demonstrated significant differences of the cellular components between udder quarters infected by major pathogens (e.g., Staphylococcus aureus; n = 5) and culture-negative udder quarters (n = 56). Even the percentages of immune cells in milk from quarters infected by minor pathogens (e.g., coagulase-negative staphylococci; n = 19) differed significantly from those in milk of culture-negative quarters. Our flow cytometric analysis of immune cells in milk of udder quarters classified as healthy by SCC <100,000 cells/mL revealed inflammatory reactions based on DCC.

Refinement of quantitative trait loci on equine chromosome 10 for radiological signs of navicular disease in Hanoverian warmblood horses.

Navicular disease is characterized by a progressive degenerative alteration of the equine podotrochlea. In this study, we refined a previously identified quantitative trait locus (QTL) on horse chromosome 10 for the abnormal development of canales sesamoidales (DCS) of the navicular bone in Hanoverian warmblood horses. Genotyping was done in 192 Hanoverian warmblood horses from 17 paternal half-sib groups. The whole marker set comprised 45 markers including seven newly developed microsatellites and 13 single nucleotide polymorphisms (SNPs) within positional candidate genes. Chromosome-wide significant QTL were confirmed and refined for DCS on horse chromosome (ECA) 10 at 0.16-2.70 Mb and at 14.45-36.37 Mb. Nine microsatellites and three SNP markers reached the highest multipoint Zmeans and LOD scores at 19.34-20.38 Mb and at 23.17-30.73 Mb with genome-wide error probabilities of P<0.05. In addition, a significant association of a SNP within VSTM1 and a significant haplotype-trait association within IRF3 could be shown. These results support a possible role of the candidate genes VSTM1 and IRF3 within the QTL on ECA10 for DCS. This study is a further step towards the identification of the genes responsible for navicular disease in Hanoverian warmblood horses.

Somatic cell counts and bacteriological status in quarter foremilk samples of cows in Hesse, Germany--a longitudinal study.

Somatic cell counts (SCC) are generally used as an indicator of udder health. Currently in Germany, 100,000 cells/mL is the threshold differentiating infected and noninfected mammary glands. The aim of our study was the detailed analysis of udder health in a representative part of the dairy cow population in Hesse, Germany. Between 2000 and 2008, 615,187 quarter foremilk samples were analyzed. In addition to evaluation of distribution of SCC and prevalence of mastitis pathogens, pathogen prevalence was also calculated depending on SCC. The data indicated that 38% of all samples had SCC >100,000 cells/mL and 62% showed SCC ≤ 100,000 cells/mL; 31% of all samples revealed SCC ≤ 25,000 cells/mL. Coagulase-negative staphylococci were the dominant pathogens in the Hessian quarter foremilk samples (17.17% of all samples) followed by Corynebacterium spp. (13.56%), Streptococcus uberis (8.7%), and Staphylococcus aureus (5.01%). Mastitis pathogens were detected in 83% of all samples with SCC >100,000 cells/mL. However, the prevalence of mastitis pathogens in the SCC range from 1,000 to ≤ 100,000 cells/mL was 8.5% (5.51% minor pathogens, 2.01% major pathogens, and 0.98% other pathogens). For farms producing high quality milk, exceptional hygiene management is compulsory. One of the farms randomly selected showed clearly different results from the Hessian survey. Fifteen percent more samples lay in the SCC range ≤ 100,000 cells/mL with a lower prevalence of mastitis pathogens of 1.91% (1.03% minor pathogens, 0.83% major pathogens, and 0.05% other pathogens). Based on these results, inflammatory processes can obviously be detected in mammary glands of udder quarters healthy according to the current definitions. However, we argue that such inflammation can be detected by examination of the relationship of immune cells in milk.

Whole-genome scan for guttural pouch tympany in Arabian and German warmblood horses.

Equine guttural pouch tympany (GPT) is a hereditary disease in foals of several breeds, including thoroughbreds, Arabian, Quarter and warmblood horses. We performed a whole-genome scan for GPT in 143 horses from five Arabian and five German warmblood families and genotyped 257 microsatellites. Chromosome-wide significant linkage was detected on ECA2 and ECA15 using multipoint non-parametric linkage analyses. Analyses stratified by sex revealed chromosome-wide significant linkage on ECA2 for fillies and chromosome-wide significant linkage on ECA15 for colts. For Arabian colts, the quantitative trait locus (QTL) on ECA15 was genome-wide significant. Haplotypes including two to four microsatellites within the QTL on ECA2 and 15 in fillies and colts, respectively, were significantly associated with GPT for both breeds. Thus, our analysis indicated sex-specific QTL, a fact which is in agreement with a two- to fourfold higher incidence of GPT in females. This is the first report of QTL for equine GPT and a first step towards identifying genes responsible for GPT.

Fine mapping a quantitative trait locus on horse chromosome 2 associated with radiological signs of navicular disease in Hanoverian warmblood horses.

Navicular disease or podotrochlosis is one of the main causes of progressive forelimb lameness in warmblood horses. The objective of this study was to refine a quantitative trait locus on horse chromosome 2 for radiological alterations in the contour of the navicular bone (RAC) in Hanoverian warmblood horses. Genotyping was performed in 192 Hanoverian warmblood horses from 17 paternal half-sib groups. The marker set was extended to 58 informative microsatellites including nine newly developed microsatellites. QTL for RAC could be delineated at 32.50-43.13 Mb and a further new QTL for RAC could be identified at 59.08-65.14 Mb. The markers ABGe342 and ABGe343 reached the highest multipoint Z(mean) and LOD scores at 34.42 and 35.23 Mb with genome-wide error probabilities of P = 0.013 and P = 0.064. In addition, significant associations of markers and haplotypes within the QTL could be shown. The results support the location of the QTL on ECA2 associated with RAC. This work is a further step towards the development of a marker test for navicular disease in Hanoverian warmblood horses.

[Swinepox--skin disease with sporadic occurrence]. / Schweinepocken--eine sporadisch vorkommende Hauterkrankung.

Swinepox virus infection results in an acute, mild or subclinical course and is characterised by typical poxvirus skin lesions in affected pigs. Additionally, sporadic vertical swinepox virus transmission leads to congenital generalised infection and subsequent abortion or stillbirth. The present report describes the occurrence of epidermal efflorescences in two piglets after intrauterine natural suipoxvirus infection. No clinical abnormalities of the gilt and littermates as well as in other pigs from this herd were present. One of the affected piglets was stillborn and submitted for necropsy, the other animal was alive at birth, but died 3 days later. Histologically, a proliferative to ulcerative dermatitis with epithelial ballooning degeneration and characteristic intracytoplasmatic inclusion bodies was observed. The pathomorphological and histopathological suspected diagnosis of a poxvirus infection was confirmed by electron microscopy. Furthermore, the agent was identified as suipoxvirus by polymerase chain reaction. As demonstrated here, obvious skin lesions in suipoxvirus infection leads to a suspected diagnosis in newborn piglets on macroscopic examination. However, further post mortem examinations, including electron microscopy as well as molecular techniques are essential for the identification of the aetiology and the exclusion of differential diagnoses. Because the disease only affected two pigs there was only a small economic loss. A valid diagnostic plays an important role in advising farmers and for herd health monitoring.

Review of genetic aspects of radiological alterations in the navicular bone of the horse.

Navicular disease or podotrochlosis has long been known to cause forelimb lameness in horses. It had been proposed that the development of podotrochlosis has similarities to the human osteoarthritis (OA) complex. Alterations of the navicular bone can be made visible early in life only on the basis of radiographs. Reports on the prevalences of navicular disease indicate that radiological alterations in the navicular bone are present in different warmblood populations at frequencies of between 14.9% and 87.6%. Genetic factors play an important role in the development of the radiological signs. Estimates of heritability using animal threshold models range from h2 = 0.09 to h2 = 0.40. Estimated additive genetic correlations between radiological changes in the navicular bone and other orthopaedic health traits indicated that they mostly develop genetically independently of each other. There was a negative genetic correlation between radiological changes in the navicular bone and the number of tournament entries and placings. It has also been shown that reduction of radiological changes of navicular bones and improvement of breeding values for performance of riding horses can be achieved if selection is based on breeding values for these traits simultaneously. An optimised markerset was developed to detect quantitative trait loci (QTL) for pathologic changes in the navicular bone of Hanoverian warmblood horses. The horse genome was scanned using 214 highly polymorphic microsatellites chromosome-wide significant QTL were located on equine chromosomes (ECA) 2, 3, 4, 10, and 26. Genome-wide significant QTL were on ECA2 and on ECA10. Unravelling QTL associated with navicular disease will enhance selection progress for a healthy limb constitution in horses.