Intramammary infections and risk factors in freshly calved heifers in Swedish dairy herds.

Subclinical mastitis can be common among freshly calved heifers (FCH), but the prevalence differs between herds, possibly due to variation in risk factors. The aims of this observational study were to identify differences in occurrence of intramammary infection (IMI) in FCH between herds with documented good or poorer first-parity udder health based on cow somatic cell count (CSCC) in early lactation, and to study herd differences in animal factors important for udder health, such as udder and hock skin lesions and animal cleanliness. Three groups of herds were included: those with high proportions of FCH with low CSCC (≤75,000 cells/mL) at the first 2 milk recordings after calving (LL), herds with high proportions of FCH with high CSCC (>100,000 cells/mL) at the first and low CSCC at the second recording (HL), and herds with high proportions of FCH with high CSCC at both recordings (HH). Thirty-nine herds (13 LL, 11 HL, 15 HH) were visited 3 times during a 12-mo period for observation of cleanliness and hock lesions, and sampling of udder and teat skin using swab cloths of milk-fed calves, early-pregnant heifers, and late-pregnant heifers. In 25 (9 LL, 9 HL, 7 HH) udder quarter samples from colostrum and milk on d 3 to 4 after calving were taken by the farmers from FCH during one year. The farmers also provided information on calving (individual or group), use of restraint and oxytocin at milking, and presence of teat and udder skin lesions. Bacterial growth in swab samples and quarter samples was investigated by culturing, and a selection of isolates was genotyped using whole-genome sequencing. Cleanliness, hock and udder skin lesions other than udder-thigh dermatitis, and growth of bacteria in swab samples did not differ between herd groups. It was more common that FCH from LL herds, compared with FCH in HH and HL herds, calved in a group of animals. Use of restraint at milking was more common in LL herds than in HH herds, whereas presence of udder-thigh dermatitis was lowest in LL herds. Specific infection was found in 14% of 5,593 quarter samples from 722 FCH. The most common IMI was Staphylococcus chromogenes. Growth of Staphylococcus simulans was more common in HH than in LL and HL herds. In colostrum samples, Staphylococcus haemolyticus was more common in HL and HH than in LL herds. The proportion of quarters with the same specific infection at both samplings was higher in HH than in LL herds and tended to be higher in HH than in HL herds. The proportion of quarters with Staph. chromogenes IMI at both samplings tended to differ between herd groups and was highest in HH herds. Whole-genome sequencing found the same sequence type of Staph. chromogenes and Staphylococcus aureus in both samples in almost all quarters with the same infection at both samplings. The differences in IMI between herd groups were in line with the higher somatic cell count in HH herds. The reasons for the predominance of Staph. chromogenes IMI in FCH need further studies.

Genotypic characterization of Staphylococcus chromogenes and Staphylococcus simulans from Swedish cases of bovine subclinical mastitis.

Staphylococcus chromogenes and Staphylococcus simulans are commonly found in intramammary infections (IMI) associated with bovine subclinical mastitis, but little is known about genotypic variation and relatedness within species. This includes knowledge about genes encoding antimicrobial resistance (AMR) and potential virulence factors (pVF). The aim of this study was therefore to investigate these aspects by whole-genome sequencing of milk isolates from Swedish dairy cows with subclinical mastitis in an observational study. We also wanted to study if specific genotypes were associated with persistent IMI and the inflammatory response at udder quarter level. In total, 105 and 118 isolates of S. chromogenes and S. simulans, respectively, were included. Isolates were characterized using a 7-locus multilocus sequence typing (7-MLST), core genome analysis and in-silico analysis of AMR and pVF genes. Forty-seven sequence types (ST) and 7 core genome clusters of S. chromogenes were identified, and the most common ST were ST-6 and ST-109, both belonging to cluster VII. A 7-locus MLST scheme for S. simulans was not available, but 3 core genome clusters and 5 subclusters were described. Overall, substantial variation in ST and clusters among cows and herds were found in both species. Some ST of S. chromogenes were found in several herds, indicating spread between herds. Moreover, within-herd spread of the same genotype was observed for both species. Only a few AMR genes [blaZ, strpS194, vga(A)] were detected in a limited number of isolates, with the exception of blaZ coding for ß-lactamase, which was identified in 22% of the isolates of S. chromogenes with ST-19, ST-102, and ST-103 more commonly carrying this gene compared with other ST. However, the blaZ gene was not identified in S. simulans. The average total number of pVF detected per isolate was similar in S. chromogenes (n = 30) and S. simulans (n = 33), but some variation in total numbers and presence of specific pVF or functional groups of pVF, was shown between ST/clusters within species. Differences in inflammatory response and potentially in persistent IMI at udder quarter level were found between S. chromogenes subtypes but not between S. simulans subtypes. In conclusion, the results from the present study generates new insight into the epidemiology of bovine S. chromogenes and S. simulans IMI, which can have implications for future prevention and antimicrobial treatment of infections related to these species.

Risk and success factors for good udder health of early lactation primiparous dairy cows.

We compared the management and housing of dairy heifers from calf to calving in herds that were very successful versus less successful in preventing mastitis in early lactation primiparous cows. This retrospective observational study included 170 Swedish dairy herds. Eligible herds were identified from the Swedish official milk recording scheme (SOMRS). Each herd had at least 60 cows per year, production data from 3 consecutive years, and at least 10 primiparous cows per year with their first milk recording 5 to 35 d after calving and their second milk recording 20 to 40 d after the first milk recording. In each herd, primiparous cows with a low (≤75,000 cells/mL) cow somatic cell count (CSCC) at both the first and second milk recording were categorized as low-low (LL); those with a high (>100,000 cells/mL) CSCC at both recordings were categorized as high-high (HH). Cows with high CSCC at the first recording and low at the second were categorized as high-low (HL). The annual proportions of LL, HL, and HH cows within each herd were calculated. Herds with an above-median proportion of LL, HL, or HH cows during the first year of a 3-yr selection period, and above the third quartile proportion of LL, HL, or HH cows, respectively, during the second and third year were identified. These herds (LL herds, n = 129; HL herds, n = 92; HH herds, n = 139) were contacted until a maximum of 60 herds per category had agreed to participate. Field technicians/veterinarians visited each herd once in the mid to late indoor season to collect data on housing and management of the heifers from birth to calving. Additional data were retrieved from the SOMRS. Associations between herd category (LL, HL, or HH) and variables collected were analyzed in 8 multivariable multinomial logistic regression submodels covering herd characteristics, milk-fed calves, heifers in early pregnancy, heifers in late pregnancy, calving and colostrum period, miscellaneous factors, summarized heifer housing data, and general health, culling, and fertility data. A final multivariable model was built from the results of the submodels and univariable analyses. The final model showed that having a standard operating procedure for colostrum feeding was more common in LL and HL herds than in HH herds; the mean bulk milk SCC and overall culling rate due to udder health was higher in HH herds than in LL and HL herds; and automatic milking was less common in LL herds than in HL and HH herds. Several herd and management variables differed between herd categories in the submodels. In conclusion, we identified several success factors for herds with good udder health among early lactation primiparous cows. This knowledge can be used to improve preventive measures in dairy herds to ensure sustainable and economic milk production.

Longitudinal study of Staphylococcus aureus genotypes isolated from bovine clinical mastitis.

Bovine clinical mastitis is an important problem for the dairy industry, and Staphylococcus aureus is a common mastitis-causing pathogen in many countries. Detailed knowledge on genetic variation of Staph. aureus strains within the bovine population, including changes over time, can be useful for mastitis control programs, because severity of disease and effects on milk production are at least partly strain-associated. Therefore, the major aim of this study was to compare sequence types of Staph. aureus isolated from cases of bovine clinical mastitis from 2002 to 2003 with sequence types of a more recent set of isolates collected from 2013 to 2018, using core genome multi-locus sequence typing (cgMLST). We also wanted to compare antibiotic resistance genes of isolates from the 2 sets, to identify changes that may have occurred over time in the Staph. aureus population. A total of 157 isolates of Staph. aureus, almost equally distributed between the 2 time periods, were subjected to high-throughput sequencing and cgMLST. The results showed that the most prevalent sequence types found among the 2002 to 2003 isolates belonged to the clonal complexes CC97, CC133, and CC151, and that those complexes still dominated among the isolates from 2013 to 2018. However, a population shift from CC133 to CC97 and CC151 over time was observed. Likewise, no important differences in prevalence of antibiotic resistance genes were found between the 2 sets of isolates. As expected, genes belonging to the major facilitator superfamily of transporter proteins, and multidrug and toxic compound extrusion transporters, were very common. Moreover, several genes and mutations conferring resistance to fosfomycin were present, but not in CC97 isolates. The ß-lactamase gene blaZ was found in only 3 out of 81 isolates from 2002 to 2003 and 1 out of 76 isolates in 2013 to 2018. In conclusion, the results indicate that mastitis-associated Staph. aureus strains circulating among dairy cows in Sweden exhibit a remarkable genotypic persistence over a time frame of close to 15 yr.

Incidence of udder cleft dermatitis (UCD) in dairy cows and risk factors for transitions to UCD.

Udder cleft dermatitis (UCD) is a common skin condition in Swedish dairy cows, affecting the anterior parts of the udder. The main objective of this study was to investigate incidence rate and duration of UCD in a 1-yr longitudinal study. Other objectives were to investigate risk factors for transitions from being healthy to having mild or severe UCD, and from having mild UCD to having severe UCD, and associations between UCD and clinical mastitis, somatic cell count (SCC) and hock lesions. Seven herds were included in the study and visited 9 times each at 6-wk intervals. At the visits, mild and severe UCD lesions, hock lesions, udder conformation traits, and hygiene scores were registered for each cow milked in the milking parlor. Information on breed, parity, days in milk (DIM), results from test milkings (milk production, SCC, and urea level), and veterinary treatments was also obtained. A UCD case was defined as one or several consecutive observations of UCD. The incidence and duration of UCD were described. Mixed-effect logistic regression models were used to analyze associations between potential risk factors and transitions to any type of UCD. Separate risk factor analyses were performed for transitions to mild and severe UCD. Associations with SCC, mastitis, and hock lesions were also analyzed with mixed-effect logistic regression models. The mean overall incidence of new UCD cases for all visits and herds was 0.5 cases per cow-year at risk. Risk factors associated with a higher risk of a transition to any type of UCD and mild UCD were breed (Swedish Red vs. Swedish Holstein), an indentation or fold at the fore udder attachment, and increasing DIM. In addition, a low milk urea level was associated with a lower risk of transition to any type of and mild UCD. Cows with previous mild UCD and high-yielding cows had increased risk for a transition to severe UCD. Cows that had an observed transition to severe UCD had an increased risk of veterinary-treated clinical mastitis within 6 wk after the UCD observation. No associations were found between UCD and SCC or hock lesions. The median observed duration of a UCD case was 12 wk, but most cases did not have an observed start or end during the study period. The observed duration of cases including severe UCD was longer than for cases involving only mild UCD. The high incidence and often long duration of UCD emphasize the need for preventive measures and treatment strategies.

Udder health of early-lactation primiparous dairy cows based on somatic cell count categories.

The aims were to investigate the prevalence of intramammary infection in early-lactation primiparous dairy cows (PC) in Sweden using milk recording cow composite somatic cell count (CSCC) categories based on classification of CSCC at the first 2 milk recordings after calving as low or high using cut-offs indicating intramammary infection. We also wanted to evaluate if herd-specific patterns in CSCC categories among PC can be identified to indicate success and problem herds as well as cow-level associations between CSCC categories and breed and sire, and herd-level associations between CSCC categories and herd size, milk production, production system, milking system, and year. A total of 1,597 dairy herds were included. Inclusion criteria were enrolment in the Swedish official milk recording scheme and having production data from at least 10 PC per year during 2014, 2015, and 2016. Herd (size, SCC, milk production, production system, milking system) and cow (breed, lactation number, calving date, CSCC, milk production) data were collected from the Swedish official milk recording scheme. Each PC was assigned a CSCC category (low-low, low-high, high-high, high-low, or inconclusive) based on the CSCC at the first 2 milk recordings using the following cut-offs. At each milk recording a CSCC ≤75,000 cells/mL was considered low and a CSCC >100,000 cells/mL was considered high, whereas a CSCC in between those values was considered inconclusive. Associations between CSCC categories and breed or sire of the PC were analyzed using multivariable multinomial logistic regression models. Associations at herd level between number of PC in a CSCC category and herd variables were analyzed using multivariable Poisson or negative binomial regression models. The annual proportions of CSCC categories for all PC were 51.3%, 5.5%, 15.5%, 13.7%, and 14.0% for the low-low, low-high, high-low, high-high, and inconclusive categories, respectively. The distribution of CSCC categories varied markedly between herds. Overall, the median herd prevalence was 50.2 to 54.2% and 11.7 to 13.2% per year for the low-low and high-high categories, respectively. At cow level, both breed and sire were significantly associated with CSCC categories, showing for example that a higher proportion of Jersey PC were categorized as high-high compared with Swedish Holstein (SH) and Swedish Red (SR) PC, and that PC of certain SH and SR sires more often were categorized as low-low or high-high cows than PC of other sires within each breed. All herd factors, except conventional and organic production, were significantly associated with the proportion of PC in a CSCC category at herd level. For example, the proportion of PC in the low-low category was significantly lower in larger herds (≥80 cows) compared with smaller herds (<80 cows) and significantly lower in automatic milking system herds than in herds with other milking systems, but significantly higher in herds with higher milk production. Overall, the results indicate a substantial need for prevention of subclinical mastitis in early-lactation PC as only 50% of these cows had low CSCC at both milk recordings after calving. Moreover, we conclude that CSCC categories may be a useful tool for identifying success and problem herds.

Microbiota of bovine milk, teat skin, and teat canal: Similarity and variation due to sampling technique and milk fraction.

The aim of this study was to evaluate the effect of sampling technique and milk fraction on bovine milk microbiota data and to compare the microbiota in milk to microbiota on the teat end and in the teat canal. Representative milk samples are highly important for assessment of bacteriological findings and microbiota in milk. Samples were obtained from 5 healthy lactating dairy cows at udder quarter level during 1 milking. Swab samples from the teat end and teat canal, and milk samples collected using different techniques and in different milk fractions were included. Milk was collected by hand stripping and through a teat canal cannula before and after machine milking, through a trans-teat wall needle aspirate after milking, and from udder quarter composite milk. The microbiota of the samples was analyzed with sequencing of the V1-V3 region of the 16S rRNA gene. In addition, somatic cell counts and bacterial cultivability were analyzed in the milk samples. Microbiota data were analyzed using multivariate methods, and differences between samples were tested using analysis of similarity (ANOSIM). Differences between samples were further explored via individual studies of the 10 most abundant genera. The microbiota on the teat end, in the teat canal, and in udder quarter composite milk, collected using a milking machine, differed in composition from the microbiota in milk collected directly from the udder quarter. No differences in milk microbiota composition were detected between hand-stripped milk samples, milk samples taken through a teat canal cannula, or milk samples taken as a trans-teat wall needle aspirate before or after milking. We conclude that for aseptic milk samples collected directly from the lactating udder quarter, sampling technique or milk fraction has minor effect on the microbiota composition.

Intramammary infections with different non-aureus staphylococci in dairy cows.

Subclinical mastitis causes an increase in milk somatic cell count (SCC) and can lead to reduced milk production and early culling. In many countries, non-aureus staphylococci (NAS) is the most common bacterial finding in subclinical mastitis of dairy cows. New methodology makes it possible to identify NAS species, but knowledge about the epidemiology is limited. The objective of this project was to improve advisory services for mastitis control by investigating associations between NAS and SCC, milk production, and persistence of intramammary infections (IMI). Farmers who had sent milk samples to the Swedish National Veterinary Institute (Uppsala, Sweden) were asked to participate if NAS was identified in the samples. Participating farmers were asked to resample all udder quarters of the cow once within 1 mo. Regression models were used to investigate associations between NAS and cow factors, udder quarter California mastitis test and SCC, and persistence of IMI. Associations with cow composite milk yield and SCC were also investigated. In total, 671 cows from 201 herds were enrolled in the study, and 19 NAS species were identified, of which the 4 most common were Staphylococcus epidermidis, Staphylococcus simulans, Staphylococcus chromogenes, and Staphylococcus haemolyticus. Persistent IMI was more common in udder quarters with Staphylococcus hyicus and S. simulans and less common in those with Staphylococcus saprophyticus IMI. ß-Lactamase production by the different NAS species varied from 0 to 100%. There was a significant association between NAS species and California mastitis test and SCC of udder quarters, and this varied depending on parity. The cow composite milk SCC at the test milking before the initial sample was taken differed significantly with NAS species, but not at the subsequent test milking. Milk yield-at the test milking before or after the initial sample-did not differ significantly for NAS species. There were no significant associations between milk yield or SCC and persistent NAS IMI. In conclusion, the NAS species affects SCC and persistent IMI differently but not milk yield.

Mild and severe udder cleft dermatitis-Prevalence and risk factors in Swedish dairy herds.

Udder cleft dermatitis (UCD) is an inflammatory skin condition affecting the anterior parts of the udder of dairy cows. The lesions may present as mild or severe skin lesions and have been associated with mastitis and digital dermatitis. The full etiology and pathogenesis are not understood and no large-scale studies have investigated prevalence and risk factors. Therefore, the main objectives of the study were to investigate the prevalence of mild and severe UCD in Swedish dairy herds and to identify risk factors associated with such lesions. We also wanted to investigate risk factors for all cases of UCD and to determine whether UCD increases the risk for mastitis and culling. A random sample of 100 freestall dairy herds were included in the study, and each herd was visited once. Cows were registered as having no, mild, or severe UCD. Additional cow and herd data were obtained via observations, interviews, and the Swedish Official Milk Recording Scheme. The data were analyzed using logistic regression models to identify risk factors for mild and severe UCD. In total, data from 3,479 cows in 99 herds were analyzed. The prevalence of mild and severe UCD was 19 and 9%, respectively. Lesions were found in 98 of 99 herds but the within-herd prevalence of mild (0-43%) and severe (0-33%) UCD varied notably between herds. Breed (Swedish Red compared with Swedish Holstein), certain udder conformation traits, and higher parity were risk factors associated with increased risk of UCD. In addition, cows with hock lesions and cows in herds with high incidence of culling due to hoof and leg diseases had a higher risk for mild UCD. More days in milk and high milk yield were cow-related risk factors associated with severe UCD. Three housing-related factors (shorter cubicles, mattress as cubicle base, and cubicles installed before 2001 compared with 2001-2005), a high incidence of veterinary-treated clinical mastitis and culling due to udder diseases, and a low incidence of culling of first-parity cows in early lactation were herd-related risk factors associated with increased risk for severe UCD. In addition, cows in herds with a high proportion of heifers older than 17 mo that were not inseminated were associated with lower risk of all UCD. Finally, UCD was not associated with the outcomes milk somatic cell count, veterinary-treated clinical mastitis, or culling in the multivariable analyses. The etiology of UCD is most likely multifactorial, involving udder conformation traits and other cow-related risk factors as well as herd-related risk factors. The high prevalence of severe UCD lesions in Swedish dairy cows emphasizes the need for preventive measures and efficient treatments.

Diagnostic test performance of somatic cell count, lactate dehydrogenase, and N-acetyl-ß-D-glucosaminidase for detecting dairy cows with intramammary infection.

The main objective of this study was to investigate the diagnostic test performance of somatic cell count (SCC), lactate dehydrogenase (LDH), and N-acetyl-ß-D-glucosaminidase (NAGase), analyzed in composite test milking samples, for detecting dairy cows with or without intramammary infection (IMI). A second objective was to investigate whether an adjustment of these udder health indicators according to their associations with different influential factors (i.e., parity, days in milk, and season) improved their test performance. Moreover, we wanted to investigate whether test performance of SCC improved if SCC results from previous adjacent test milkings were included in the model. Such test milking data were not available for LDH or NAGase. In this cross-sectional study, quarter milk samples for bacteriological examination were taken from almost 1,000 cows from 25 dairy herds during 3 consecutive days: the day before test milking, the day of test milking, and the day after test milking. From each cow, a composite test milking sample was analyzed for milk composition, SCC, LDH, and NAGase. Among the cows sampled, 485 were IMI negative and 256 were IMI positive in one or more udder quarters according to the definitions used. The remaining cows had inconclusive IMI status. To assess the test performance of SCC, LDH, and NAGase to identify IMI-negative and IMI-positive cows, univariable generalized estimating equation models were used with the udder health indicator of interest as outcome and IMI status as explanatory variable. From these models, receiver-operator characteristic curves were created and the area under cure (AUC) was calculated. From each model, a cut-off was chosen for calculations of the sensitivity (Se), specificity (Sp), positive predictive value (PPV), negative predictive value (NPV), and accuracy (ACC) for each udder health indicator. The AUC was similar for the adjusted SCC (0.84), nonadjusted SCC (0.83) and geometric mean SCC (0.80-0.81), but much lower for LDH (0.66) and NAGase (0.62). The highest Se, Sp, PPV, NPV, and ACC were obtained using SCC. Adjustment of the udder health indicators for influential factors (e.g., parity) did not improve the test performance markedly, whereas adding information about SCC from previous adjacent test milkings improved the test performance of SCC slightly. In conclusion, of the udder health indicators investigated, SCC had the best overall ability to correctly identify IMI-negative and IMI-positive dairy cows.

Udder infections with Staphylococcus aureus, Streptococcus dysgalactiae, and Streptococcus uberis at calving in dairy herds with suboptimal udder health.

Udder infections with Staphylococcus aureus, Streptococcus dysgalactiae, and Streptococcus uberis are common causes of bovine mastitis. To study these pathogens in early lactation, a 12-mo longitudinal, observational study was carried out in 13 herds with suboptimal udder health. The aims of the study were to investigate the occurrence of these pathogens and to identify if presence of the 3 pathogens, and of genotypes within the pathogens, differed with respect to herd, season, and parity. Quarter milk samples, collected at calving and 4 d in milk (DIM), were cultured for the 3 pathogens. Genotyping of staphylococcal and streptococcal isolates was performed using spa typing and pulsed-field gel electrophoresis, respectively. For each of the 3 pathogens, cows with an udder infection at calving or 4 DIM were allocated to 1 of 4 infection types: cleared (pathogen present only at calving), persistent (pathogen present in the same quarter at calving and 4 DIM), new (pathogen present only at 4 DIM), or cleared/new (pathogen present in 1 quarter at calving and in another quarter at 4 DIM). Associations between season or parity and overall occurrence of pathogens or infection types were determined using univariable mixed-effect logistic-regression models and the Fisher's exact test, respectively. The most commonly occurring pathogen was Staph. aureus, followed by Strep. dysgalactiae and Strep. uberis. Persistent infections were the most common infection type among Staph. aureus-infected cows, whereas cleared infections were the most common among Strep. dysgalactiae- and Strep. uberis-positive cows. The proportion of cows with persistent Staph. aureus infections and the proportion of cows having a Strep. uberis infection at calving or 4 DIM were higher in the multiparous cows than in primiparous cows. Infections with Strep. dysgalactiae were less common during the early housing season than during the late housing or pasture seasons, whereas persistent Strep. uberis infections were less common during the pasture season than during the late housing season. The relative occurrence of the 3 pathogens, infection types of each pathogen, and genotype diversity of each pathogen throughout the year or in different seasons and parities varied among the herds, indicating that underlying factors predisposing for udder infections at calving differ between herds. Genotyping of bacterial isolates gave important insight into how such infection patterns differed within and between herds. These findings emphasize the need to choose preventive strategies for each individual herd.

Yersinia enterocolitica infections associated with improperly pasteurized milk products: southwest Pennsylvania, March-August, 2011.

In July 2011, a cluster of Yersinia enterocolitica infections was detected in southwestern Pennsylvania, USA. We investigated the outbreak's source and scope in order to prevent further transmission. Twenty-two persons were diagnosed with yersiniosis; 16 of whom reported consuming pasteurized dairy products from dairy A. Pasteurized milk and food samples were collected from this dairy. Y. enterocolitica was isolated from two products. Isolates from both food samples and available clinical isolates from nine dairy A consumers were indistinguishable by pulsed-field gel electrophoresis. Environmental and microbiological investigations were performed at dairy A and pasteurization deficiencies were noted. Because consumption of pasteurized milk is common and outbreaks have the potential to become large, public health interventions such as consumer advisories or closure of the dairy must be implemented quickly to prevent additional cases if epidemiological or laboratory evidence implicates pasteurized milk as the outbreak source.

Prevalence and risk factors for udder cleft dermatitis in dairy cattle.

Udder cleft dermatitis (UCD) is a skin lesion in dairy cattle mostly located at the anterior junction between the udder and the abdominal wall or between the front quarters. Relatively little is known about causative factors for UCD, and few studies have investigated prevalence and risk factors of UCD. Therefore, the aim of this study was to investigate the prevalence of UCD in a random sample of dairy herds with freestalls and milking parlors in a county of Sweden. Thirty dairy herds participated in the study. Each herd was visited once at milking, when every third cow was investigated for presence of UCD. Associations between UCD and milk production, breed, parity, days in milk, claw health, and udder health on the herd and cow levels were also investigated. In addition, a case-control study was performed in 6 herds with a high prevalence of UCD to investigate associations between udder conformation or mange and UCD. Udder cleft dermatitis was found in 18.4% of the 1,084 cows included in the study. The within-herd cow prevalence varied between zero and 39%, with an average of 18.5%. Risk factors for UCD at the herd level were a high proportion of Swedish Red cows and a high production level. At the cow level, breed, parity, and production level were identified as risk factors. The highest risk of having UCD was found in high-producing Swedish Red cows that had calved at least 3 times. Veterinary-treated clinical mastitis was associated with UCD, but cow composite somatic cell count was not. A strong anterior udder attachment was a protective factor, but signs of mange had no association with UCD. The primary cause of UCD is still unclear, and more research is needed to identify the best ways to prevent the development of this animal welfare problem.

Status of vitamins E and A and ß-carotene and health in organic dairy cows fed a diet without synthetic vitamins.

Synthetic vitamin supplementation is not consistent with organic production, so it is important to investigate whether dairy cows can maintain their health and production without synthetic vitamins being added to their diet. In basic dairy cow diets, provitamin A (ß-carotene) and vitamin E are mainly found in pasture and in grass and legume silages, but the concentrations are highly variable. This study compared the vitamin status and health of cows without synthetic vitamin supplementation (NSV group) with control cows (CON group) fed synthetic vitamins according to Swedish recommendations (600 IU of vitamin E and 80,000 IU of vitamin A per cow per day) to investigate whether dairy cows can fulfill their requirements of vitamins A and E without supplementation with synthetic vitamins. Vitamin concentrations in blood plasma and milk, health, fertility, milk yield, and milk composition were measured in Swedish Holstein cows (n=28) during 2 complete lactations. All cows were fed a 100% organic diet containing grass-legume silage, cold-pressed rapeseed cake, peas, cereal grains, and minerals. Blood samples were collected from each cow 3 wk before expected calving, at calving, and 3 wk, 3 to 5 mo, and 7 to 9 mo after calving. Samples of colostrum were taken and milk samples were collected 4d after calving and at the same time as the 3 blood samplings after calving. The only difference in vitamin status between groups was found in colostrum in yr 1, when CON cows tended to have a higher concentration of α-tocopherol, and their ß-carotene concentration was higher compared with NSV cows. The NSV cows tended to have more cases of mastitis than CON cows in yr 2. Within the NSV group, fewer cows were healthy and more cases of mastitis were observed in yr 2 than in yr 1. The groups did not differ in production parameters. In conclusion, the vitamin status in blood and milk of the studied cows indicated that cows in organic dairy production can fulfill their requirements of vitamins A and E without any supplementation of synthetic vitamins, except at the time around calving, when the requirements are high. However, the impaired health of NSV cows in yr 2 may indicate a long-term negative health effect in cows fed no synthetic vitamins.

Associations of udder-health indicators with cow factors and with intramammary infection in dairy cows.

The objective of this study was to investigate if and how cow factors and intramammary infection (IMI) are associated with 4 different udder-health indicators in dairy cows as a first step in investigating whether the diagnostic performance of these indicators can be improved. The investigated indicators were somatic cell count (SCC), lactate dehydrogenase (LDH), N-acetyl-ß-d-glucosaminidase (NAGase), and alkaline phosphatase (AP) measured in milk. In this cross-sectional study, approximately 1,000 cows from 25 dairy herds were sampled for bacteriology (quarter milk samples) during 3 consecutive days: the day before test milking, at the day of test milking, and at the day after test milking. The whole-udder test milking sample was analyzed for milk composition, SCC, LDH, NAGase, and AP. Cow data (parity, breed, milk yield, percentage of milk fat and protein, milk urea concentration, and days in milk from the sampled test milking) were collected from the Swedish milk-recording scheme. Of the sampled cows 485 were considered IMI negative and were used in multivariable mixed-effect linear regression models to investigate associations between cow factors and the udder-health indicators. A second modeling including all cows, both IMI negative and IMI positive (256 cows), was also performed. The results showed that all udder-health indicators were affected by cow factors but that different cow factors were associated with different indicators. Intramammary-infection status was significantly associated with all udder-health indicators except AP. Parity and milk urea concentration were the only cow factors associated with all indicators in all models. The significant cow factors explained 23% of the variation in SCC and >30% of the variation in LDH, NAGase, and AP in IMI-negative cows, showing that LDH, NAGase, and AP are more affected than SCC by cow factors. The IMI status explained 23% of the variation in SCC in the model with all cows but only 7% of the variation in LDH and 2% of the variation in NAGase, indicating that SCC has the best potential as a diagnostic tool in finding cows with IMI. However, further studies are needed to investigate whether the diagnostic properties of these udder-health indicators will improve with adjustment according to their associations with different cow factors when used as a diagnostic tool for finding cows with IMI.

The impact of forced joint exercise on lubricin biosynthesis from articular cartilage following ACL transection and intra-articular lubricin's effect in exercised joints following ACL transection.

OBJECTIVE: To evaluate the impact of forced joint exercise following acute knee injury on lubricin metabolism and its relationship to cartilage degeneration and to assess chondroprotection of a single-dose purified human lubricin injection in exercised injured joints. METHODS: Anterior cruciate ligament transection (ACLT) was performed in rats with six experimental groups; 3-week post-ACLT, 3-week post-ACLT + exercise, 5-week post-ACLT, 5-week post-ACLT + exercise, and 5-week post-ACLT + exercise treated with intra-articular phosphate buffered saline (PBS) or lubricin. Joint exercise was achieved using a rotating cylinder at a speed of 6 rpm for 30 min daily, 5 days a week starting 1 week following surgery. Cartilage lubricin expression in injured joints was determined. Histological analyses included Safranin O/Fast Green, activated caspase-3, and lubricin mRNA in-situ hybridization. Assessment of cartilage damage was performed by osteoarthritis research society international (OARSI) modified Mankin scoring and urinary CTXII (uCTXII) levels. RESULTS: At 3 weeks, lubricin expression in exercised ACLT joints was significantly (P < 0.001) lower compared to ACLT joints. The OARSI scores were significantly (P < 0.001) higher in the ACLT + exercise animals compared to ACLT animals at 5 weeks. Compared to 3-week ACLT, 3-week ACLT + exercise cartilage showed increased caspase-3 staining. Compared to ACLT + exercise and PBS-treated ACLT + exercise, lubricin intra-articular treatment resulted in a significant increase (P < 0.001) in cartilage lubricin gene expression and a reduction (P < 0.05) in uCTXII levels. CONCLUSION: Joint exercise resulted in decreased lubricin cartilage expression, increased cartilage degeneration and reduced superficial zone chondrocyte viability in the ACLT joint. Intra-articular lubricin administration ameliorated cartilage damage due to exercise and preserved superficial zone chondrocytes' viability.

Amyloid deposition in 2 feline thymomas.

Two cases of feline thymoma with amyloid deposition were encountered between 1982 and 2010. Neoplastic cells were separated by abundant, pale eosinophilic, homogeneous material that was congophilic and birefringent. Ultrastructurally, the neoplastic cells were connected by desmosomes, and the extracellular deposits were composed of nonbranching, hollow-cored fibrils, 8-10 nm in diameter. In the case with sufficient archived tissue for additional sections, the amyloid remained congophilic following potassium permanganate incubation, and the neoplastic cells were immunoreactive for pancytokeratin. The histologic, histochemical, ultrastructural, and immunohistochemical features of both neoplasms are consistent with epithelial-predominant thymoma with the unusual feature of intratumoral amyloid deposition. The affinity of the amyloid for Congo red following potassium permanganate incubation is consistent with non-AA amyloid. The ultrastructural findings were consistent with amyloid production by the neoplastic epithelial cells.

Leisure-time physical activity and type 2 diabetes during a 28 year follow-up in twins.

AIMS/HYPOTHESIS: The study aimed to investigate whether baseline physical activity protects against the occurrence of type 2 diabetes during a 28 year follow-up, after controlling for childhood environment and genetic predisposition. METHODS: At baseline in 1975 same-sex twin pairs born in Finland before 1958 were sent a questionnaire including questions on physical activity. The participants (20,487 individuals, including 8,182 complete twin pairs) were divided into quintiles by leisure-time physical activity metabolic equivalent (MET) index (MET h/day). Type 2 diabetes was determined from nationwide registers for the follow-up period (1 January 1976-31 December 2004). Individual and pairwise Cox proportional hazard models were used. RESULTS: During follow-up, 1,082 type 2 diabetes cases were observed. Among all individuals, participants in MET quintiles (Q) III-V had significantly decreased risk for type 2 diabetes compared with sedentary individuals (QI). The pairwise analysis on pairs discordant for physical activity showed that participants in MET QII to V had significantly lower hazard ratios (0.61, 0.59, 0.61, 0.61) compared with sedentary participants. These findings from the pairwise analysis persisted after adjusting for BMI. In the pairwise analysis, the BMI-adjusted hazard ratio for type 2 diabetes was lower for physically active members of twin pairs (combined QII-V) than for inactive co-twins (HR 0.54; 95% CI 0.37-0.78). Similar results were obtained for both dizygotic and monozygotic pairs, as well as for the subgroup of twin pairs defined as free of co-morbidities in 1981 (HR 0.36; 95% CI 0.17-0.76). CONCLUSIONS/INTERPRETATION: Leisure-time physical activity protects from type 2 diabetes after taking familial and genetic effects into account.

An improved method to culture Staphylococcus aureus from bovine milk.

Staphylococcus aureus is an important udder pathogen often associated with subclinical mastitis in dairy cows. Identification of Staph. aureus-positive udder quarters and cows is an important part of control programs to reduce spread of Staph. aureus within and between dairy herds. Therefore, accurate and easy-to-perform culturing methods of Staph. aureus in milk are needed. In the present study, 8 methods for isolation of Staph. aureus in bovine milk samples were investigated. The methods involved different culturing volumes, enrichment, incubation, and freezing processes as well as sedimentation and use of the Mastistrip cassette (SVA, Uppsala, Sweden). Three different sets of milk samples were collected, and 6, 5, and 4 methods were used in each subset of samples. Our results indicate an increased probability of detecting Staph. aureus in milk samples when a simple incubation step (37 degrees C for 18 h) without additives was included before culturing. Using this incubation method, the number of Staph. aureus-positive udder quarters and cows increased by 50 and 29%, respectively, compared with using the standard method of direct culturing of 10 microL of milk. The improved method may be especially useful for detection of low concentrations of Staph. aureus in milk; for example, when screening herds for Staph. aureus.

Short communication: relationships between α-lactalbumin and quality traits in bulk milk.

The main objective of this study was to investigate whether the α-lactalbumin (α-LA) content of bulk milk is related with some known inflammatory markers and milk quality traits. An additional objective was to study whether combining α-LA, haptoglobin (Hp), and serum amyloid A (SAA) in an acute phase index (API) could be useful as an alternative marker for bulk milk quality. For the dairy industry, it is of great importance to receive high quality bulk milk for manufacture of liquid milk and dairy products. The somatic cell count (SCC) is currently used as an indirect marker for bulk milk quality, but because it is somewhat insensitive and unspecific, interest exists in alternative markers. Bulk milk samples were analyzed for α-LA, SCC, polymorphonuclear leukocyte count, Hp, SAA, fat, lactose, total protein and casein contents, casein number, protein composition, proteolysis, and coagulating properties. No significant differences were found in SCC, polymorphonuclear leukocyte count, Hp, or SAA between milk samples containing low, medium, or high concentrations of α-LA. Differences between α-LA groups were, however, found in some milk quality traits because high α-LA concentration was related to low concentrations of α(S1)-, α(S2)-, and ß-caseins and high concentrations of lactose and ß-lactoglobulin. A high API was related to low lactose content and casein number. Samples with high SCC contained less casein and had a lower casein number than milk with a low SCC, and proteolysis was significantly higher in high SCC milk than in low SCC milk. Neither α-LA nor API proved to be a better marker than SCC for the quality traits investigated, and α-LA was not considered to be a useful inflammatory marker in bulk milk.