A Disease Outbreak in Beef Cattle Associated with Anaplasma and Mycoplasma Infections.

An outbreak of disease in a Swedish beef cattle herd initiated an in-depth study to investigate the presence of bacteria and viruses in the blood of clinically healthy (n = 10) and clinically diseased cattle (n = 20) using whole-genome shotgun sequencing (WGSS). The occurrence of infectious agents was also investigated in ticks found attached to healthy cattle (n = 61) and wild deer (n = 23), and in spleen samples from wild deer (n = 30) and wild boars (n = 10). Moreover, blood samples from 84 clinically healthy young stock were analysed for antibodies against Anaplasma phagocytophilum and Babesia divergens. The WGSS revealed the presence of at least three distinct Mycoplasma variants that were most closely related to Mycoplasma wenyonii. Two of these were very similar to a divergent M. wenyonii variant previously only detected in Mexico. These variants tended to be more common in the diseased cattle than in the healthy cattle but were not detected in the ticks or wild animals. The DNA of A. phagocytophilum was detected in similar proportions in diseased (33%) and healthy (40%) cattle, while 70% of the deer, 8% of ticks collected from the cattle and 19% of the ticks collected from deer were positive. Almost all the isolates from the cattle, deer and ticks belonged to Ecotype 1. Based on sequencing of the groEL-gene, most isolates of A. phagocytophilum from cattle were similar and belonged to a different cluster than the isolates from wild deer. Antibodies against A. phagocytophilum were detected in all the analysed samples. In conclusion, uncommon variants of Mycoplasma were detected, probably associated with the disease outbreak in combination with immune suppression due to granulocytic anaplasmosis. Moreover, A. phagocytophilum was found to be circulating within this cattle population, while circulation between cattle and deer occurred infrequently.

Herd Routines and Veterinary Advice Related to Dry-Cow Therapy and Treatment with Internal Teat Sealants in Dairy Cows.

Dry-cow therapy with antibiotics (DCT) and treatment with internal teat sealants (ITS) are often used to control mastitis in dairy cows. However, the knowledge on farmer and veterinary compliance with recommendations for DCT and ITS is scarce. Thus, the main aim was to collect information on farmer routines and veterinary advice for such treatments. Associations with herd and veterinary variables were also studied. Web-based questionnaires including questions on demographics and the use of DCT and ITS were sent to 2472 farmers and 517 veterinarians in Sweden. The answers were summarized descriptively, and associations with demographics were evaluated using univariable regression models. The response rate was 14% for farmers and 25% for veterinarians. Among the farmers, 81% used selective DCT (SDCT), 3% used blanket DCT (BDCT), and 16% did not use DCT. Almost all (93%) veterinarians prescribed DCT and among those most recommended SDCT while 8% recommended BDCT. Eighty-two percent of the farmers did not use ITS and 45% of the veterinarians never prescribed ITS. Milking system and milk production, and post-graduate training and number of mastitis cases per month were associated with the largest numbers of farmer and veterinary answers, respectively. In conclusion, many farmer routines and veterinary advice complied with the recommendations available at the time, but a clear need for more education was also identified. The results also indicated that an up-date of the national recommendations was warranted.

Microbial Aetiology, Antibiotic Susceptibility and Pathogen-Specific Risk Factors for Udder Pathogens from Clinical Mastitis in Dairy Cows.

Mastitis is one of the most important infectious diseases and one of the diseases that causes the greatest use of antibiotics in dairy cows. Therefore, updated information on the bacteria that cause mastitis and their antibiotic susceptibility properties is important. Here, for the first time in over 10 years, we updated the bacterial findings in clinical mastitis in Swedish dairy cows together with their antibiotic resistance patterns and risk factors for each bacterial species. During the period 2013-2018, samples from clinical mastitis were collected, together with information on the cows and herds of origin. The samples were cultured, and a total of 664 recovered bacterial isolates were subjected to susceptibility testing. Staphylococcus aureus (S. aureus) was the most common pathogen and accounted for 27.8% of diagnoses, followed by Streptococcus dysgalactiae (S. dysgalactiae) (15.8%), Escherichia coli (E. coli) (15.1%), Streptococcus uberis (S. uberis) (11.4%), Trueperella pyogenes (T. pyogenes) (7.7%), non-aureus staphylococci (NAS) (2.8%), Klebsiella spp. (2.7%), Enterococcus spp. (1.3%), and Streptococcus agalactiae (S. agalactiae) (1.2%). Various other bacteria accounted for 2.6%. Staphylococci were, in general, susceptible to most antibiotics, but 2.6% of S. aureus and 30.4% of NAS were resistant to penicillin. No methicillin-resistant staphylococci were found. All S. agalactiae were susceptible to penicillin. Bimodal and trimodal MIC distributions for penicillin in S. dysgalactiae and S. uberis, respectively, indicate acquired reduced susceptibility in some isolates. The mostly unimodal MIC distributions of T. pyogenes indicate that acquired resistance does usually not occur in this species. Among E. coli, 14.7% were resistant to at least one antibiotic, most often ampicillin (8.7%), streptomycin (7.8%), or sulphamethoxazole (6.9%). Klebsiella spp. had low resistance to tetracycline (9.1%) but is considered intrinsically resistant to ampicillin. Pathogen-specific risk factors were investigated using multivariable models. Staphylococcus aureus, S. dysgalactiae, and T. pyogenes were more common, while E. coli was less common in quarters with more than one pathogen. S. aureus and T. pyogenes were mostly seen in early lactation, while E. coli was more common in peak to mid lactation and S. dysgalactiae in early to peak lactation. Trueperella pyogenes and Klebsiella spp. were associated with a previous case of clinical mastitis in the current lactation. Staphylococcus aureus was associated with tie stalls and T. pyogenes with loose housing. All pathogens except E. coli and S. dysgalactiae had a seasonal distribution. In conclusion, the aetiological agents for clinical bovine mastitis have remained relatively stable over the last 10-15 years, S. aureus, S. dysgalactiae, E. coli and S. uberis being the most important. Resistance to penicillin among Gram-positive agents was low, and in general, antibiotic resistance to other compounds was low among both Gram-positive and Gram-negative agents.

Prudent Use of Antibiotics in Dairy Cows: The Nordic Approach to Udder Health.

Global concerns regarding bacterial antibiotic resistance demand prudent use of antibiotics in livestock production. Dairy production in the Nordic countries has a low consumption of antibiotics, while animal health, productivity and milk quality are at high levels. Here, we describe the basis of Nordic mastitis control and treatment strategies, as a model for production of high-quality milk with prudent use of antibiotics. We hope this will be beneficial for dairy producers and advisors in other countries and regions that consider limiting antibiotic use in cattle herds. In this perspectives paper we describe the dairy sector in the Nordic countries, and present regulatory aspects of antibiotic use, diagnostics and current guidelines for treatment of clinical mastitis as well as dry cow therapy. We also show summary statistics of udder health indicators in Denmark, Finland, Norway and Sweden, to illustrate the effects of the implemented udder health management practices.

Recovery from udder cleft dermatitis in dairy cows.

Udder cleft dermatitis (UCD) is a common skin problem in dairy cows, localized at the fore udder attachment or between the udder halves. The main objectives of this study were to investigate spontaneous recovery from UCD in a longitudinal study and to investigate the treatment effect on UCD of a topical spray containing chelated copper and zinc in an additional treatment trial. Spontaneous recovery was investigated during a 1-yr longitudinal study in which 7 Swedish dairy herds were visited 9 times each, at 6-wk intervals. During the visits, all cows milked in the milking parlor were examined for mild and severe UCD, udder conformation traits, hygiene, and hock lesions. Additional cow and herd data were obtained from the Swedish official milk recording scheme. Recovery was defined as 2 consecutive observations of no UCD after a UCD case (1 or more consecutive observations of UCD). The recovery rate was calculated and factors associated with recovery were analyzed using discrete-time survival analysis. A treatment study was conducted in 4 of the herds after completion of the longitudinal study. Cows with UCD were allocated to treatment or a control group, and cows in the treatment group were treated once every day for 14 to 28 d. At follow-up visits on d 14, 28, and 56, all cows were scored for UCD. To investigate the effect of treatment, an ordered logistic regression model was used, with UCD score on d 56 (no, mild, or severe) as the outcome. Other potential explanatory variables were also included in the model. A total of 38% (126 of 329) of the cows in the longitudinal study made a spontaneous recovery from UCD. A long duration of UCD, severe lesions and higher parity reduced the chance of recovery. Among the recovered cows, 47% (59 of 126 cows) had a recurrent case of UCD after the recovery. In the treatment study, there was no difference in UCD score on d 56 between the treatment and control groups. Factors that affected the UCD score on d 56 were the duration of UCD before the treatment study started, UCD score on d 1 and udder conformation. Our findings emphasize the importance of further studies to identify effective treatment regimens as well as further studies to unravel the pathophysiology of UCD to improve the recommendations on how to prevent these lesions.

A shotgun metagenomic investigation of the microbiota of udder cleft dermatitis in comparison to healthy skin in dairy cows.

Udder cleft dermatitis (UCD) is a skin condition affecting the fore udder attachment of dairy cows. UCD may be defined as mild (eczematous skin changes) or severe (open wounds, large skin changes). Our aims were to compare the microbiota of mild and severe UCD lesions with the microbiota of healthy skin from the fore udder attachment of control cows, and to investigate whether mastitis-causing pathogens are present in UCD lesions. Samples were obtained from cows in six dairy herds. In total, 36 UCD samples categorized as mild (n = 17) or severe (n = 19) and 13 control samples were sequenced using a shotgun metagenomic approach and the reads were taxonomically classified based on their k-mer content. The Wilcoxon rank sum test was used to compare the abundance of different taxa between different sample types, as well as to compare the bacterial diversity between samples. A high proportion of bacteria was seen in all samples. Control samples had a higher proportion of archaeal reads, whereas most samples had low proportions of fungi, protozoa and viruses. The bacterial microbiota differed between controls and mild and severe UCD samples in both composition and diversity. Subgroups of UCD samples were visible, characterized by increased proportion of one or a few bacterial genera or species, e.g. Corynebacterium, Staphylococcus, Brevibacterium luteolum, Trueperella pyogenes and Fusobacterium necrophorum. Bifidobacterium spp. were more common in controls compared to UCD samples. The bacterial diversity was higher in controls compared to UCD samples. Bacteria commonly associated with mastitis were uncommon. In conclusion, a dysbiosis of the microbiota of mild and severe UCD samples was seen, characterized by decreased diversity and an increased proportion of certain bacteria. There was no evidence of a specific pathogen causing UCD or that UCD lesions are important reservoirs for mastitis-causing bacteria.

Microbiota data from low biomass milk samples is markedly affected by laboratory and reagent contamination.

Discoveries of bacterial communities in environments that previously have been described as sterile have in recent years radically challenged the view of these environments. In this study we aimed to use 16S rRNA sequencing to describe the composition and temporal stability of the bacterial microbiota in bovine milk from healthy udder quarters, an environment previously believed to be sterile. Sequencing of the 16S rRNA gene is a technique commonly used to describe bacterial composition and diversity in various environments. With the increased use of 16S rRNA gene sequencing, awareness of methodological pitfalls such as biases and contamination has increased although not in equal amount. Evaluation of the composition and temporal stability of the microbiota in 288 milk samples was largely hampered by background contamination, despite careful and aseptic sample processing. Sequencing of no template control samples, positive control samples, with defined levels of bacteria, and 288 milk samples with various levels of bacterial growth, revealed that the data was influenced by contaminating taxa, primarily Methylobacterium. We observed an increasing impact of contamination with decreasing microbial biomass where the contaminating taxa became dominant in samples with less than 104 bacterial cells per mL. By applying a contamination filtration on the sequence data, the amount of sequences was substantially reduced but only a minor impact on number of identified taxa and by culture known endogenous taxa was observed. This suggests that data filtration can be useful for identifying biologically relevant associations in milk microbiota data.

Hock lesions in dairy cows in freestall herds: a cross-sectional study of prevalence and risk factors.

BACKGROUND: Hock lesions (HL) in dairy cows are a common animal welfare problem in modern dairy production with freestall housing systems, but there are no large-scale studies addressing its epidemiology in Sweden. The aims of this cross-sectional study were to investigate the prevalence of HL of different severity in 100 Swedish freestall dairy herds, and to identify cow- and herd-related risk factors. Associations between HL and mastitis as well as culling were also investigated. RESULTS: In total, 3217 cows from 99 herds were included in the statistical analyses. The overall cow prevalence of hair loss on the hock (mild HL) was 68% and the prevalence of ulceration or evident swelling of the hock, with or without hair loss, (severe HL) was 6%. The within-herd prevalence varied among herds, between 23 and 100% for mild HL, and between 0 and 32% for severe ones. Breed (higher risk for Swedish Holstein than for Swedish Red) and days in milk (higher risk at 181-305 days than at 0-90 days) were cow-related risk factors associated with both types of lesions, whereas higher parity and cleaner cows were associated only with increased risk of severe HL. A reduced risk for mild HL was seen in cows housed on mattresses compared to rubber mats, and in cows housed on peat compared to other bedding materials. Also, cows in herds with a high proportion of not yet inseminated heifers older than 17 months had a lower risk of mild HL than cows in herds with a low proportion. Risk for severe HL was lower when cubicles were of recommended width compared to under recommendation, for organic production compared to conventional, and when teat dip or no treatment after milking was used, compared to teat spray. For both mild and severe HL, herringbone milking parlors were associated with higher risk than tandem parlors. We found no significant associations between HL and mastitis or culling. CONCLUSIONS: The prevalence of HL is high in Swedish dairy herds, although most lesions are mild. Several cow- and herd-related risk factors were identified and the results can be used to improve recommendations for the prevention of HL in Swedish freestall dairy herds.

An unusual presentation of pseudocowpox associated with an outbreak of pustular ulcerative vulvovaginitis in a Swedish dairy herd.

Species Pseudocowpox virus (PCPV; family Poxviridae) is known to cause pustular cutaneous disease in cattle. We describe an outbreak of pseudocowpox with an unusual clinical picture in a free-stall dairy herd of ~80 cows. Approximately 90% of the cows had vesicles, erosions, papules, and scabs on the vulva and vaginal mucosa. Histologic analysis of biopsy tissues indicated a primary, although not specified, viral infection. Transmission electron microscopy revealed parapoxvirus particles in both tissue and vesicular materials. Deep sequencing analysis of extracted DNA from swabbed vesicle areas gave a contig of nearly 120,000 nucleotides, matching the PCPV strain VR 634 with 100% identity. Analyses confirmed the absence of other potential causes of pustular vulvovaginitis such as bovine herpesvirus 1 and Ureaplasma diversum. A rolling cow brush was suspected to be the fomite.

Occurrence and Spread of Quinolone-Resistant Escherichia coli on Dairy Farms.

UNLABELLED: Quinolone-resistant Escherichia coli (QREC) is common in feces from young calves, but the prevalence and genetic diversity of QREC in groups of cattle of other ages and the farm environment are unknown. The aims of the study were to obtain knowledge about the occurrence of QREC on dairy farms, the genetic diversity of QREC within and between farms, and how these relate to the number of purchased animals and geographic distances between farms. We analyzed the within-sample prevalence of QREC in individual fecal samples from preweaned dairy calves and postpartum cows and in environmental samples from 23 Swedish dairy farms. The genetic diversity of the QREC isolates on 10 of these farms was assessed. In general, QREC was more prevalent in the dairy farm environment and in postpartum cows if QREC was commonly found in calves than if QREC was rare in calves. In particular, we found more QREC organisms in feed and water troughs and in environments that may come into contact with young calves. Thus, the results suggest that QREC circulates between cattle and the farm environment and that calves are important for the maintenance of QREC. Some genotypes of QREC were widespread both within and between farms, indicating that QREC has spread within the farms and likely also between farms, possibly through purchased animals. Farms that had purchased many animals over the years had greater within-farm diversity than farms with more closed animal populations. Finally, animals on more closely located farms were more likely to share the same genotype than animals on farms located far apart. IMPORTANCE: This study investigates the occurrence of a specific type of antimicrobial-resistant bacterium on dairy farms. It contributes to increased knowledge about the occurrence and spread of these bacteria, and the results pave the way for actions or further studies that could help mitigate the spread of these bacteria on dairy farms and in the community as a whole.

Udder health in beef cows and its association with calf growth.

BACKGROUND: Studies outside the Nordic countries have indicated that subclinical mastitis (measured by milk somatic cell count or the California Mastitis Test), intramammary infections (IMI), or blind quarters in beef cows may have negative effects on beef calf growth. Knowledge on prevalence of such udder health problems in Swedish beef cows is scarce. Therefore, the main aim of this study was to investigate subclinical mastitis, IMI and udder conformation in a number of beef cow herds. Production of ß-lactamase in staphylococci was also investigated. Associations between certain cow factors and subclinical mastitis and IMI, and associations between cow and calf factors and 200 day calf weaning weight were also studied. The herds were visited once within a month after calving and once at weaning. Udder examination and quarter milk sampling, for somatic cell count and bacteriology, were performed in 8 to 12 cows per herd and occasion. RESULTS: Approximately 50%, 40% and 10% of the cows had subclinical mastitis, IMI, and at least one blind quarter, respectively, but the prevalence varied markedly between herds. Intramammary infections (mainly due to staphylococci) were identified in 13-16% of the milk samples. Less than 5% of the staphylococcal isolates produced ß-lactamase. Approximately 11% of the cows sampled twice had the same IMI (mostly Staphylococcus aureus) at both samplings. Cow factors of importance for subclinical mastitis and/or IMI were teat and udder shape, breed, parity, presence of blind quarters, and cow hygiene. No significant associations were found between udder health parameters studied and calf weaning weights. CONCLUSIONS: Subclinical mastitis and IMI, but not blind quarters, were common in beef cows, but the prevalence varied markedly between herds. Most IMI were caused by staphylococci and more than 95% of those were sensitive to penicillin. Cows with large funnel-shaped teats or pendulous udder after calving, and cows with blind quarters were at risk of having subclinical mastitis and/or IMI. Poor hygiene was also a risk factor for udder health problems. No significant associations were found between udder health and calf weaning weight. More studies on risk factors are warranted to improve advisory services on awareness and prevention of mastitis in beef cows.

Relationship between somatic cell count, polymorphonuclear leucocyte count and quality parameters in bovine bulk tank milk.

The somatic cell count (SCC) in bovine bulk tank milk is presently used as an indicator of raw milk quality, reflecting the udder health status of the herd. During mastitis, SCC increases, mostly owing to an influx of polymorphonuclear leucocytes (PMN) from blood into milk, with a concomitant change in milk composition. Bulk tank milk samples were categorized according to their SCC, as well as polymorphonuclear leucocyte count (PMNC), to study relationships between SCC, PMNC and various raw milk quality traits, i.e. contents of total protein, whey protein, casein, fat and lactose, casein number, proteolysis and rheological properties. The proportion of PMN, obtained by direct microscopy, was significantly higher in samples with high SCC compared with low SCC samples. SCC and PMNC were strongly correlated, yielding a correlation coefficient of 0.85. High SCC samples had lower lactose and casein contents, lower casein number and more proteolysis than low SCC samples. Samples with high PMNC had a lower casein number than low PMNC samples. Samples with high and low SCC or PMNC did not differ in respect to rheological properties. Our results do not indicate that PMNC is a better biomarker than SCC for raw bulk tank milk quality, as previously proposed.

Incidence of mastitis and bacterial findings at clinical mastitis in Swedish primiparous cows- influence of breed and stage of lactation.

Mastitis is a common disease also among primiparous dairy cows. Identification of the extent and type of problem is important to initiate correct control measures. In Sweden, unique national production and disease databases are available. The main aim of the study was to investigate the occurrence of mastitis, measured by the annual incidence of veterinary-treated clinical mastitis (VTCM) and geometric mean of monthly milk somatic cell count (SCC) recordings in Swedish primiparous cows in relation to older cows during 2002-2006 with emphasis on breed differences. Other aims were to study differences between primiparous and older cows in the distribution of bacterial findings at clinical mastitis, and the occurrence of VTCM and bacterial findings in relation to stage of lactation using data from a Swedish field study performed 2002-2003. Descriptive statistics and univariable analyses were used in the investigations. During 2002-2006 approximately 10% of Swedish primiparous cows experienced VTCM each year, while the geometric mean SCC of Swedish primiparous cows was approximately 65,000 ml(-1). Both parameters were lower than in older cows. Primiparous cows of the Swedish Red (SR) breed had better udder health than cows of the Swedish Holstein (SH) breed. The overall distribution of udder pathogens was similar in primiparous and older cows. In primiparous cows, most VTCM occurred during the first week after calving, and Staphylococcus aureus and Streptococcus dysgalactiae were the most common udder pathogens during this period. Better control measures directed at these infections are warranted around calving to reduce the risk of mastitis in primiparous cows.

The xMAP technique can be used for detection of the inflammatory cytokines IL-1beta, IL-6 and TNF-alpha in bovine samples.

Infectious diseases can cause large health problems in cattle. The infections cause an acute inflammatory response, mediated by pro-inflammatory cytokines such as IL-1beta, IL-6 and TNF-alpha. By mapping the pattern of cytokines during inflammations, valuable information about the course of an infection is gained. The aim of the present study was to evaluate a particle-based flow cytometric method, the xMAP technique, using ovine/bovine reagents, for quantification of IL-1beta, IL-6 and TNF-alpha, for application in studies on ruminant infectious diseases with emphasis on bovine milk and plasma samples. Singleplex, duplex and triplex xMAP assays were evaluated, and limits of detection (LODs) as well as intra- and inter-assay variabilities were determined for each assay. Cross-reactivity between reagents in multiplex assays was also tested. In addition, presence of cytokines in milk and plasma samples from healthy and mastitic cows was studied. The LODs were significantly lower for singleplex xMAP assays than for duplex and triplex assays. In singleplex assays, the LODs were 0.08, 0.2 and 0.5 ng/ml, for IL-1beta, IL-6 and TNF-alpha, respectively. Corresponding LODs in triplex assays were 2.0, 6.5 and 3.5 ng/ml. Data indicate that the linear ranges of the multiplex assays were narrower than in singleplex assays. The intra-assay coefficients of variation were < or =10.7% for singleplex assays, while they ranged from 6.2 to 23.2% in the triplex assay. The inter-assay variance ranged from 5.1 to 35.8% in singleplex assays, and from 8.8 to 78.4% in triplex assays. Cross-reactivity between reagents was not observed, and all three cytokines were detected in bovine milk and plasma samples collected from cows with clinical mastitis. In conclusion, our results show that the xMAP technique can be used for quantification of IL-1beta, IL-6 and TNF-alpha in bovine samples, and that further work is required to optimize the multiplex assays.

Haptoglobin and serum amyloid A in relation to the somatic cell count in quarter, cow composite and bulk tank milk samples.

Milk somatic cell count (SCC) is the gold standard in diagnosis of subclinical mastitis, and is also an important parameter in quality programmes of dairy cooperatives. As routine SCC analysis is usually restricted to central laboratories, much effort has been invested in the search for alternative biomarkers of mastitis and milk quality, including the presence in the milk of the acute phase proteins (APP), haptoglobin (Hp) and serum amyloid A (SAA). The aim of this study was to investigate relationships between Hp, SAA and SCC in quarter, cow composite, and bulk tank milk samples. Cows (n=165), without any clinical signs of disease or abnormalities in the milk or udder, from three different dairy farms, were used. Cow composite milk samples from all cows delivering milk at the sampling occasion were taken once in each herd. In one of the farms, representative quarter milk samples (n=103) from 26 cows were also collected. In addition, bulk tank milk samples from 96 dairy farms were included in the study. Samples were analysed for Hp, SAA and SCC, and relationships between the parameters were evaluated at quarter, cow and tank milk levels using Chi-square analysis. Milk samples were categorized according to their SCC, and the presence, or no presence, of SAA and Hp, based on the detection limits of the screening methods (0.3 mg/l and 1.0 mg/l for SAA and Hp, respectively). Hp and SAA were found in milk at quarter, cow composite and bulk tank levels. A large proportion (53%) of the animals had detectable milk concentrations of APP, and SAA was detected more frequently, and at higher concentrations than Hp, regardless of sample type. SAA was detected in as many as 82% of the bulk tank milk samples. Significant relationships were found between Hp, SAA and SCC at quarter and cow composite milk levels, but only between SAA and SCC at bulk tank milk level. Detectable levels of APP were more common at high SCC.

Acute phase proteins as indicators of calf herd health.

The potential for using acute phase proteins (APPs) in the assessment of herd health was studied by examining the levels of serum haptoglobin, serum amyloid A (SAA) and plasma fibrinogen in relation to clinical findings and leukocyte counts in calves. Two groups of calves from conventional dairy farms were studied. The animals were examined 10 times during the first six weeks after introduction into a new environment. Haptoglobin, SAA and fibrinogen were analysed and weight gain, disease symptoms and treatments were recorded. Analysis of antibodies against viral infections was performed. An acute phase reaction (APR) score was established at each sampling by combining the APP results and total leukocyte counts. The health status differed between the two groups, although no manipulation of health had been performed, except that the group with a higher incidence of disease had a concurrent experimental infection with lungworm as part of another study. In the group with a higher incidence of disease, the mean weight gain was significantly lower, and the number of sampling days with elevated serum concentrations of APPs, and the mean maximum concentrations of haptoglobin and fibrinogen were significantly higher compared to the healthier group. The APR score was significantly higher at days 4 and 8 of the study in the group with a higher incidence of disease. The results indicate that measurement of APPs could be a useful tool for evaluation of health in calf herds.

Alpha-tocopherol concentration and stereoisomer composition in plasma and milk from dairy cows fed natural or synthetic vitamin E around calving.

The aim of this study was to compare the effects of supplementing dairy cows with 1000 IU/day of all-rac-alpha-tocopheryl acetate (SynAc), RRR-alpha-tocopheryl acetate (NatAc), or RRR-alpha-tocopherol (NatAlc), from approximately 3 weeks before estimated calving until 2 weeks after calving, on the concentration of alpha-tocopherol and its stereoisomers (RRR-, RSS-, RRS-, RSR- and the four 2S-forms of alpha-tocopherol) in blood and milk. An unsupplemented group was included as control. Blood samples were collected at 3, 2 and 1 weeks before estimated calving, at calving, and 3, 7 and 14 days after calving, while milk samples were taken twice within 24 h after calving and at 7 and 14 days in milk. Overall, time and treatment had significant effects on plasma alpha-tocopherol with higher concentrations in NatAc than in the other groups. In addition, SynAc had higher concentrations than Control, and NatAlc tended to be higher than Control. The lowest plasma concentrations were observed at calving and 3 days after calving. Independent of treatment, the concentration was higher in colostrum than in milk day 7 and 14 after calving. Analyses of the stereoisomer distribution in plasma and milk showed that, irrespective of dietary treatment, RRR-alpha-tocopherol was the most predominant form, constituting more than 86%, whereas the remaining part of alpha-tocopherol was made up by the three synthetic 2R isomers, while the 2S isomers only contributed less than 1% of the total alpha-tocopherol. In control cows and cows supplemented with natural vitamin E, the proportion of RRR-alpha-tocopherol in plasma and milk constituted more than 98% of the total alpha-tocopherol. In conclusion, the results indicate that daily oral supplementation of dairy cows with RRR-alpha-tocopheryl acetate gives the highest blood concentrations of alpha-tocopherol in the periparturient period. Analyses of the distribution of the individual stereoisomers of alpha-tocopherol further indicate that the bioavailability of RRR-alpha-tocopherol relative to synthetic stereoisomers in cattle is considerably higher than officially accepted until now.

Biosensor assay for determination of haptoglobin in bovine milk.

Despite more than 30 years of research into mastitis diagnostics, there are few alternatives to the somatic cell count (SCC) in practical use for identification of cows with subclinical mastitis. Mastitis is not only an animal welfare problem, but also affects the yield, composition and technological properties of milk. Hence, dairy cooperatives give farmers a premium quality payment to encourage low SCC although there is no clear scientific data defining the level of SCC in bulk tank milk that is associated with additional benefits in terms of milk quality. Recent research on alternative markers for inflammatory reactions in the lactating cow, e.g. in mastitis, includes investigations of the acute phase protein, haptoglobin (Hp). So far, the content of Hp in milk has mainly been studied in relation to mastitis diagnostics, with little attention given to its importance for milk composition and technological properties. At present, Hp in milk is measured using ELISA, but this technique is not suitable for routine large-scale analysis. In recent years, optical biosensor technology has been used for automated and rapid quantitative analysis of different components in milk, but so far not for analysis of acute phase proteins. The aim of the present study was to develop a rapid and sensitive biosensor method to determine Hp in milk. An affinity sensor assay based on the interaction between Hp and haemoglobin was developed using surface plasmon resonance (SPR) biosensor technology. The assay was used to analyse Hp in composite milk samples from cows without any clinical signs of mastitis and quarter milk samples with a weak to strong reaction in the California Mastitis Test (CMT). A commercial ELISA for determination of Hp in milk was used for comparison. The limit of detection (LOD) of the biosensor assay was determined as 1.1 mg/l. Within-assay and between-day variations were determined both with bulk tank milk spiked with human Hp and with composite milk samples containing bovine Hp. Coefficients of variation varied between 3.6 and 8.6% at concentrations between 4.0 and 12 mg/l, respectively. Agreement between the results obtained by the biosensor assay and the ELISA was satisfactory; however, the results obtained by the biosensor were generally lower than the results obtained by the ELISA. Possible explanations for this observation are discussed.