Local and Systemic Inflammation in Finnish Dairy Cows with Digital Dermatitis.

Digital dermatitis is a disease of the digital skin and causes lameness and welfare problems in dairy cattle. This study assessed the local and systemic inflammatory responses of cows with different digital dermatitis lesions and compared macroscopical and histological findings. Cow feet (n = 104) were evaluated macroscopically and skin biopsies histologically. Serum samples were analyzed for acute phase proteins (serum amyloid A and haptoglobin) and pro-inflammatory cytokines (interleukin-1 beta, interleukin-6, and tumor necrosis factor-alpha). Cows with macroscopically graded active lesions (p = 0.028) and non-active lesions (p = 0.008) had higher interleukin-1 beta levels in their serum compared to healthy cows. Interleukin-1 beta serum concentrations were also higher (p = 0.042) when comparing lesions with necrosis to lesions without necrosis. There was no difference when other cytokine or acute phase protein concentrations in healthy cows were compared to those in cows with different digital dermatitis lesions. A novel histopathological grading was developed based on the chronicity of the lesions and presence of necrosis and ulceration. The presence and number of spirochetes were graded separately. In the most severe chronic lesions, there was marked epidermal hyperplasia and hyperkeratosis with necrosis, deep ulceration, and suppurative inflammation. Spirochetes were found only in samples from necrotic lesions. This study established that digital dermatitis activates proinflammatory cytokines. However, this did not initiate the release of acute phase proteins from the liver. A histopathological grading that takes into account the age and severity of the lesions and presence of spirochetes was developed to better understand the progression of the disease. It is proposed that necrosis of the skin is a result of ischemic necrosis following reduced blood flow in the dermal papillae due to pressure and shear stress caused by thickened epidermis, and that the spirochetes are secondary invaders following tissue necrosis.

Effect of Farm Management Practices on Morbidity and Antibiotic Usage on Calf Rearing Farms.

Antimicrobial resistance has been recognized as one of the top health threats to human society. Abundant use of antibiotics in both humans and animals has led to ever-increasing antibiotic resistance in bacteria. In food production, decreasing morbidity in beef herds would be an effective way to reduce the use of antibiotics. The objective of this retrospective observational study was to determine overall morbidity on calf rearing farms and to identify associated risk factors. Data were collected by questionnaire, meat companies' databases and the national cattle register for 28,228 calves transported to 87 calf rearing farms. All medications given to these calves were retrospectively followed for 180 days from calf arrival to the farm. In total, 34,532 parenteral antibiotic medications were administered to the 28,228 study calves (122.3%), and 17,180 calves (60.9%) were medicated with antibiotics at least once during the follow-up. Higher numbers of calves transported to the same farm and larger age variation in calves in the same arrival batch were both associated with increased morbidity. In contrast, higher arrival age of individual calves was associated with decreased morbidity. Our study identifies several factors to consider in decreasing morbidity and antibiotic usage on calf rearing farms.

Genomic Analysis of Staphylococcus aureus Isolates Associated With Peracute Non-gangrenous or Gangrenous Mastitis and Comparison With Other Mastitis-Associated Staphylococcus aureus Isolates.

Staphylococcus aureus is a highly prevalent cause of mastitis in dairy herds worldwide, capable of causing outcomes that vary from subclinical to peracute gangrenous mastitis. We performed a comparative genomic analysis between 14 isolates of S. aureus, originating from peracute bovine mastitis with very severe signs (9 gangrenous, 5 non-gangrenous) and six isolates originating from subclinical or clinical mastitis with mild to moderate signs, to find differences that could be associated with the clinical outcome of mastitis. Of the 296 virulence factors studied, 219 were detected in all isolates. No difference in the presence of virulence genes was detected between the peracute and control groups. None of the virulence factors were significantly associated with only a single study group. Most of the variation in virulence gene profiles existed between the clonal complexes. Our isolates belonged to five clonal complexes (CC97, CC133, CC151, CC479, and CC522), of which CC522 has previously been detected only in isolates originating from caprine and ovine mastitis, but not from bovine mastitis. For statistical analysis, we sorted the CCs into two groups. The group of CCs including CC133, CC479, and CC522 was associated with gangrenous mastitis, in contrast to the group of CCs including CC97 and CC151. The presence of virulence genes does not explain the clinical outcome of mastitis, but may be affected by allelic variation, and especially different regulation and thus expression in the virulence genes.

Effect of farm and animal-level factors on youngstock mortality and growth on calf rearing farms.

Both calf mortality and daily weight gain have a crucial impact on profitability of calf rearing farms. In addition, high calf mortality rates represent an animal welfare problem. Mortality rates on calf rearing farms have been reported in several studies in different countries, but scant data regarding daily weight gain of the calves are available. The objectives of this observational retrospective study were to determine the average mortality and daily weight gain of calves and youngstock on Finnish calf rearing farms and to identify factors associated with these production parameters. National cattle register and national herd health register databases together with meat companies' databases were used to collect weight, age, breed, medication, and origin farm data for 28 228 calves transported to the 87 calf rearing farms between 1 January and 1 October 2016. A telephone questionnaire was completed by selected farms to collect management and farm-specific data. Calves were retrospectively followed for maximum 180 days since arrival to the farm. Average arrival age of the calves were 24 days (SD 14). Average calf mortality on Finnish calf rearing farms was 4.5 %. Mortality was 5.3 % on fattening farms buying milk calves, 4.6 % on specialized calf rearing farms, and 2.5 % on fattening farms buying weaned calves. Size of the calf rearing farms varied, being smallest on fattening farms for weaned calves and largest on specialized calf rearing farms. Average daily gain of the study calves was 1.074 kg/day (SD 0.166). Multilevel mixed effects logistic and linear regression models, where herd and calf batch were used as random effects, were generated to study calf level mortality and daily gain, respectively. Activities preventing transmission of pathogens between arrival batches and different age groups of animals, including application of the all in/all out principle and proper washing and disinfection of compartments for milk feeding calves between arrival batches, were associated with lower mortality and increased daily gain. In addition, higher arrival age was associated with lower mortality during the rearing period and relatively higher arrival age of the calf, compared to other calf in a same arrival batch, was associated with higher daily gain. By contrast, increased number of individual medications during the rearing period was associated with both increased mortality and decreased daily gain. There was no significant difference in mortality between farm types. Current study highlights several factors that can be affected in future to further develop the beef production chain.

Prevalence of digital dermatitis using mirror scoring in Finnish freestall dairy herds.

Digital dermatitis (DD) is a severe bacterial hoof disease found worldwide. The disease can be classified into 5 different stages, denoted as M1 to M4 and M4.1, by clinical examination. The main objective of this study was to estimate prevalence of DD lesions in Finnish freestall dairy cattle population through hind feet inspection of standing cows with a mirror. Another aim was to estimate the sensitivity and specificity of mirror scoring on standing cows in a pen or in a milking parlor without washing the feet. Three veterinarians visited 81 randomly selected herds across the country. During the herd visits, hind feet of standing cows (n = 7,010) were scored with a mirror without washing the feet, either when the cows were standing in a pen (n = 4,992) or in the milking parlor (n = 2018). In total, 128 cows (111 from pen and 17 from milking parlor) including 256 feet were chosen with cross-sectional sampling and scored in a trimming chute. Animal-level sensitivity for scoring M2 lesions with a mirror was 55% and specificity was 97%; for all active DD lesions (M1, M2, or M4.1), sensitivity was 36% and specificity was 96%. Sensitivity for scoring any DD lesions was 90% and specificity was 82%. The bias-corrected sensitivity and specificity for scoring any DD lesions were 79% and 92%, respectively. The bias-corrected sensitivity and specificity for scoring M2 DD lesions were 10% and 100%. We found M2 lesions in 12.1% of the study herds, and true herd-level prevalence was the same. Altogether, 33.3% (true prevalence 28.4%) of the herds had either M1, M2, or M4.1 DD lesions. However, only 0.7% (true prevalence 5.4%) of cows in total had active M2 lesions. The within-herd prevalence of M2 lesions (in herds where at least 1 cow had a M2 lesion) was 5.7% and varied between 0.4% and 18.8%. Herds with active DD lesions also had more any DD lesions than herds without active DD lesions. The herd-level prevalence was higher than previously thought, with only 1 herd without any DD lesions. However, the animal-level prevalence of active DD lesions was relatively low. Farmers and veterinarians need to be informed of the disease and possible control measures. Because of the low within-herd prevalence, the control of the disease might be easier than in countries where DD is widespread. Further studies are needed to identify factors associated with DD prevalence in Finnish dairy herds.

Mycoplasma bovis infection in dairy herds-Risk factors and effect of control measures.

As Mycoplasma bovis spreads to new countries and becomes increasingly recognized as a disease with major welfare and economic effects, control measures on dairy farms are needed. To minimize the risk of infection spread to naive herds, all possible risk factors for M. bovis infection should be identified and controlled. Mycoplasma bovis was first diagnosed in dairy cattle in Finland in 2012, and by January 2020, 86 Finnish dairy farms (<1.5%) supporting M. bovis infections were identified. We evaluated risk factors for M. bovis infection using a questionnaire provided to 40 infected and 30 control dairy farms. Control measures were advised for 19 of the infected dairy farms during visits by a veterinarian. The course of the infection on those farms was followed by analyzing calf nasal swabs with PCR for presence of M. bovis 4 times at 6-mo intervals. Control measures included culling of M. bovis mastitic cows, isolation of new calves from older animals after initial M. bovis mastitic cows had been culled, prevention of nose-to-nose contact with infected animals, early detection of mastitis cases using M. bovis PCR, and hygiene measures mainly related to milking, calf pens, feeding buckets, and teats. Farms implemented the control measures related to the isolation of calves or avoidance of nose-to-nose contact in various ways, according to farm structures and financial circumstances.In our study, the control measures recommended to the dairy farms appeared effective, such that 13 of 19 farms reached a low risk level during at least 3 consecutive negative samplings from calves, with no M. bovis mastitis detected subsequently. Among risk factors, insemination with an M. bovis-positive bull indicated a trend of increasing the odds of M. bovis infection on the farm in a multivariable logistic model. In contrast, higher herd average milk yield had an association with lower odds for M. bovis infection. Occurrence of other infectious diseases affecting several animals on the dairy farm in the previous 6 mo before M. bovis infection were more frequent on M. bovis-infected farms.

Field trial to evaluate the effect of an intranasal respiratory vaccine protocol on bovine respiratory disease incidence and growth in a commercial calf rearing unit.

BACKGROUND: Bovine respiratory disease (BRD) continues to be great challenge in calf rearing units. The urgent need to decrease the use of antibiotics and increase animal welfare in beef production has forced us to introduce new preventive methods. Vaccinations could contribute to the solution, but the high incidence of BRD already at an early age has made it difficult to introduce suitable vaccination programs. Challenge studies have shown promising results in 3-14 day old calves vaccinated with intranasal BRD vaccine, but very few field trials are available to assess the efficacy of the intranasal vaccines in field conditions. We evaluated the effect of one dose of commercial intranasal vaccination on calf mortality, daily gain, and treatment incidence for BRD in one calf rearing unit. In total, 497 calves (mean age 19 days) were included in our study, 247 of which were vaccinated at the time of arrival to the unit and 250 served as negative controls (unvaccinated). Vaccinated and unvaccinated calves were situated in separate compartments until weaning. Daily gain, treatment incidence, and mortality were recorded until the calves were transported to the finishing unit, which averaged 154.5 days from arrival. RESULTS: Average daily gain over the complete study period was 1151.9 g/day (SD 137.9) for the vaccinated calves and 1139.5 g/day (SD 135.9) for the unvaccinated calves. Intranasal vaccination combined with older arrival age (17 days or older) resulted in a higher daily gain (47.8 g/day) compared with unvaccinated calves (coef. 0.0478, p = 0.003). This association was not recorded in calves that were younger than 17 days upon arrival. Intranasal vaccination was not significantly associated either with mortality (OR 0.976, p = 0.968) or treatment incidence for BRD (OR 1.341, p = 0.120). In total, six vaccinated calves (2.43%) and six unvaccinated calves (2.40%) died during the study period. CONCLUSIONS: Vaccinating arriving calves with intranasal vaccine in the calf rearing unit did not decrease the mortality or treatment incidence for BRD, but it significantly improved the weight gain in calves transported to the unit at the age of 17 days or older.

Acute phase response and clinical manifestation in outbreaks of interdigital phlegmon in dairy herds.

Several Finnish dairy herds have suffered from outbreaks of interdigital phlegmon (IP). In these new types of outbreaks, morbidity was high and clinical signs severe, resulting in substantial economic losses for affected farms. In our study, we visited 18 free stall dairy herds experiencing an outbreak of IP and 3 control herds without a similar outbreak. From a total of 203 sampled cows, 60 suffered from acute stage IP. We demonstrated that acute phase response of bovine IP was evident and therefore an appropriate analgesic should be administered in the treatment of affected animals. The response was most apparent in herds with high morbidity in IP and with a bacterial infection comprising Fusobacterium necrophorum and Dichelobacter nodosus, indicating that combination of these two bacterial species affect the severity of the disease.

An atypical Bacillus anthracis infection in a bull-A potential occupational health hazard.

Bacillus anthracis infecting cattle is usually identified based on the typical symptom: sudden death. Bacillus anthracis causing atypical symptoms may remain undiagnosed and represent a potential occupational health hazard for, that is veterinarians and producers, butchers and tanners. In the year 2004, one case of sudden death in a dairy farm in southern Finland was diagnosed as bovine anthrax. Four years later 2008, an atypical case of anthrax was diagnosed in the same holding. The bull was taken to the Production Animal Hospital of the Faculty of Veterinary Medicine, University of Helsinki because of fever, loss of appetite and a symmetrically swollen scrotal sac. Penicillin treatment cured the fever but not the swollen scrotum. Before the intended therapeutic castration, a punctuate consisting of 10 ml fluid collected into a syringe from the scrotal sac was cultivated on blood agar at 37°C. After 24 hr, an almost pure culture of a completely non-hemolytic Bacillus cereus-like bacteria was obtained. The strain was identified as B. anthracis using Ba-specific primers by the Finnish Food Safety Authority (RUOKAVIRASTO). After the diagnosis, the bull was euthanized and destroyed, the personnel were treated with prophylactic antibiotics and the clinic was disinfected. In this particular case, treatment with water, Virkon S and lime seemed to be effective to eliminate endospores and vegetative cells since no relapses of anthrax have occurred in 10 years. This case is the last reported anthrax case in Finland.

Bovine milk microbiome: a more complex issue than expected.

The aim of this study was to analyze bacterial profiles of bovine mastitic milk samples and samples from healthy quarters using Next Generation Sequencing of amplicons from 16S rRNA genes and to compare results with microbiological results by PCR assays of the same samples. A total of 49 samples were collected from one single dairy herd during the same day. The samples were divided in two sample sets, which were used in this study. The DNA extraction as well as the library preparation and sequencing of these two sets were performed separately, and results of the two datasets were then compared. The vast majority of genera detected appeared with low read numbers and/or in only a few samples. Results of PCR and microbiome analyses of samples infected with major pathogens Staphylococcus aureus or Streptococcus uberis were consistent as these genera also covered the majority of reads detected in the microbiome analysis. Analysis of alpha diversity revealed a much higher species richness in set 1 than in set 2. The dominating bacterial genera with the highest read numbers clearly differed between datasets, especially in PCR negative samples and samples positive for minor pathogens. In addition to this, linear discriminant analysis (LDA) was conducted between the two sets to identify significantly different genera/family level microbes. The genus Methylobacterium was much more common in set 2 compared to set 1, and genus Streptococcus more common in set 1. Our results indicate amplification of contaminating bacteria in excess in samples with no or minor amounts of pathogen DNA in dataset 2. There is a need for critical assessment of results of milk microbiome analyses.

Bacterial species associated with interdigital phlegmon outbreaks in Finnish dairy herds.

BACKGROUND: Severe outbreaks of bovine interdigital phlegmon (IP) have occurred recently in several free stall dairy herds in Finland. We studied the aetiology of IP in such herds, and the association of bacterial species with the various stages of IP and herds of various morbidity of IP. Nineteen free stall dairy herds with IP outbreaks and three control herds were visited and bacteriological samples collected from cows suffering from IP (n = 106), other hoof diseases (n = 58), and control cows (n = 64). The herds were divided into high morbidity (morbidity ≥50%) and moderate morbidity groups (9-33%) based on morbidity during the first two months of the outbreak. RESULTS: F. necrophorum subspecies necrophorum was clearly associated with IP in general, and T. pyogenes was associated with the healing stage of IP. Six other major hoof pathogens were detected; Dichelobacter nodosus, Porphyromonas levii, Prevotella melaninogenica, Treponema spp. and Trueperella pyogenes. Most of the samples of acute IP (66.7%) harboured both F. necrophorum and D. nodosus. We found differences between moderate morbidity and high morbidity herds. D. nodosus was more common in IP lesion in high than in moderate morbidity herds. CONCLUSIONS: Our result confirms that F. necrophorum subspecies necrophorum is the main pathogen in IP, but also T. pyogenes is associated with the healing stage of IP. Our results suggest that D. nodosus may play a role in the severity of the outbreak of IP, but further research is needed to establish other bacteriological factors behind these severe outbreaks.

Elimination of experimentally induced bovine intramammary infection assessed by multiplex real-time PCR and bacterial culture.

Diagnosis of bovine intramammary infection (IMI) has traditionally been based on bacterial culture, but currently IMI can also be detected with DNA based methods, such as multiplex real-time PCR. The aim of this study was to describe the elimination of bacteria in experimentally induced IMI on the quarter level, using conventional bacterial culture (BC) and multiplex real-time PCR. Two coagulase-negative staphylococcal species, Staphylococcus epidermidis and Staphylococcus simulans, were experimentally inoculated into 14 healthy quarters of 8 dairy cows during 4 consecutive study periods. Intramammary infections were followed with 20 milk samplings per each quarter. Milk somatic cell count was monitored to evaluate the inflammation process in the quarters. Four quarters cured spontaneously during the study period according to the culture. The PCR detected staphylococcal DNA from these quarters for several days after they were defined as cured in BC. Agreement between BC and PCR results varied from substantial to almost perfect agreement for the first 36 h postchallenge, decreasing to moderate levels toward the end of the sampling period. Based on this study, we recommend collecting possible follow-up samples to assess the bacteriological cure from IMI not until 2 to 3 wk after the onset of mastitis or after the quarter milk somatic cell count has normalized when PCR is used.

Semen as a source of Mycoplasma bovis mastitis in dairy herds.

Mycoplasma bovis infections are responsible for substantial economic losses in the cattle industry, have significant welfare effects and increase antibiotic use. The pathogen is often introduced into naive herds through healthy carrier animals. In countries with a low prevalence of M. bovis, transmission from less common sources can be better explored as the pathogen has limited circulation compared to high prevalence populations. In this study, we describe how M. bovis was introduced into two closed and adequately biosecure dairy herds through the use of contaminated semen during artificial insemination (AI), leading to mastitis outbreak in both herds. Epidemiological analysis did not reveal an infection source other than semen. In both farms the primary clinical cases were M. bovis mastitis in cows inseminated with the semen of the same bull four weeks before the onset of the disease. One semen straw derived from the semen tank on the farm and other semen lots of this bull were positive for M. bovis. In contrast, semen samples were negative from other bulls that had been used for insemination in previous or later oestrus to those cows with M. bovis mastitis. Furthermore, cgMLST of M. bovis isolates supported the epidemiological results. To our knowledge this is the first study describing the introduction of M. bovis infection into a naive dairy herd via processed semen. The antibiotics used in semen extenders should be re-evaluated in order to provide farms with M. bovis-free semen or tested M. bovis-free semen should be available.

A comparison of unheated loose housing with stables on the respiratory health of weaned-foals in cold winter conditions: an observational field-study.

BACKGROUND: Newly weaned horses in Finland are often moved to unheated loose housing systems in which the weanlings have free access to a paddock and a shelter. This practice is considered to be good for the development of young horses. The daily temperatures can stay below - 20 °C in Finland for several consecutive weeks during the winter season. However, the effect of unheated housing in a cold climatic environment on the respiratory health of weanlings under field conditions has not been studied before. This investigation was an observational field-study comprising 60 weanlings among 11 different voluntary participant rearing farms in Finland. Weanlings were either kept in unheated loose housing systems (n = 36) or in stables (n = 24) and were clinically examined on two separate occasions 58 days apart in cold winter conditions. RESULTS: The odds of clinical respiratory disease were lower in the older foals (loge days); OR = 0.009, P = 0.044). The plasma fibrinogen concentration was higher when the available space (m2/weanling) in the sleeping hall was smaller (P = 0.014) and it was lower when the sleeping hall was not insulated (P = 0.010). The plasma fibrinogen concentrations at the second examination were lower with a body condition score above 3 (P = 0.070). Standardbreds kept in loose housing systems had a lower body condition score than Finnhorses or Standardbreds kept in stables at both examinations (P = 0.026 and P = 0.007, respectively). Haemoglobin level was lower in weanlings in loose housing systems compared to their counterparts at the first examination (P = 0.037). Finnhorses had higher white blood cell count than Standardbreds at first (P = 0.002) and at the second examination (P = 0.001). CONCLUSIONS: Keeping weanling horses in cold loose housing systems does not seem to increase the occurrence of respiratory disease, but special attention should be focused on ventilation, air quality and feeding-practices. Our field study data suggest it might be advantageous to keep Standardbred foals born late in the season in a stable over the Finnish winter.

Survey of interdigital phlegmon outbreaks and their risk factors in free stall dairy herds in Finland.

BACKGROUND: Severe outbreaks of interdigital phlegmon (IP) associated with a high morbidity and major economic losses have occurred in Finland in the past decade. A survey was performed to indicate the current occurrence of infectious hoof diseases and to identify herd level risk factors predisposing to an outbreak of IP. RESULTS: Responses to a questionnaire revealed that an outbreak of IP defined as morbidity ≥5% within the 1st month of the outbreak, had occurred in 18.0% of the respondent study farms. Risk factors for an outbreak included animal transport between herds, i.e. either animal purchase or contract heifer rearing, enlargement or renovation of the barn, and if the fields of the farm had been organically cultivated. Having any kind of mechanical ventilation in comparison to natural ventilation seemed to lower the risk of IP. Additionally, the farms that had experienced an outbreak of IP often had other infectious hoof diseases. However, it was unclear which disease appeared first. CONCLUSIONS: More attention is needed before and during enlargement or renovation of the barn and substantial planning is crucial for every part of the enlargement process in dairy farms.

Prevalence of subclinical mastitis in Finnish dairy cows: changes during recent decades and impact of cow and herd factors.

BACKGROUND: The dairy industry has undergone substantial structural changes as intensive farming has developed during recent decades. Mastitis continues to be the most common production disease of dairy cows. Nationwide surveys of mastitis prevalence are useful in monitoring udder health of dairy herds and to study the impact of structural changes on the dairy industry. This survey on bovine subclinical mastitis was the first based on cow composite milk somatic cell count (SCC) data from the Finnish national health monitoring and milk recording database. A cow with composite milk SCC ≥200,000 cells/ml in at least one of the four test milkings during the year was considered to have subclinical mastitis and a cow with composite milk SCC ≥200,000 cells/ml in three or in all four test milkings during the year to have chronic subclinical mastitis. The aim of the study was to determine the prevalence of subclinical mastitis and chronic subclinical mastitis in Finland in 1991, 2001 and 2010 and to investigate cow and herd factors associated with elevated SCC. RESULTS: Prevalence of subclinical mastitis in Finland decreased over recent decades from 22.3% (1991) and 20.1% (2001) to 19.0% (2010). Prevalence of chronic subclinical mastitis was 20.4% in 1991, 15.5% in 2001 and 16.1% in 2010. The most significant cow and herd factors associated with subclinical mastitis or high milk SCC were increasing parity, Holstein breed, free-stalls with an automatic milking system and organic production. Milk SCC were highest from July to September. Main factors associated with chronic mastitis were increasing parity and Holstein breed. CONCLUSIONS: Prevalence of subclinical mastitis in Finland decreased over recent decades, the greatest change taking place during the first decade of the study. Prevalence of chronic subclinical mastitis significantly decreased from 1991. The most significant factors associated with both types of mastitis were increasing parity and Holstein breed, and for subclinical mastitis also free-stalls with automatic milking. National surveys on mastitis prevalence should be carried out at regular intervals to monitor udder health of dairy cows and to study the impact of the ongoing structural changes in the dairy industry to enable interventions related to udder health to be made when needed.

Health and growth of Finnish beef calves and the relation to acute phase response.

Healthy, thriving calves are essential for beef calf production. We studied the health status and factors associated with the growth of beef calves in six cow-calf herds during the first month of the calves' lives and at weaning age (200 days). The six herds were visited three times, when calves were approximately 3 days, 16 days and 30 days of age. On each visit calves (n=37) were clinically examined, weighed or measured, blood samples were collected, faecal samples obtained and deep nasopharyngeal swabs were taken. Each blood sample was analysed for acute phase proteins (haptoglobin, serum amyloid-A, fibrinogen), total proteins and albumin, the faecal sample for intestinal tract pathogens (rotavirus, bovine coronavirus, enterotoxigenic Escherichia coli and Salmonella, oocysts of Eimeria coccidia and Cryptosporidium, and nematode eggs), and the nasopharyngeal swab for respiratory tract pathogens (bovine coronavirus (BCV), respiratory syncytial virus (RSV), bacteria and mycoplasma). Clinical diagnosis of respiratory tract disease, diarrhoea or umbilical disease was set at 15.0% for all the three consecutive examinations combined (n=107), but only few pathogens were detected from the samples. The increased levels of acute phase proteins were neither associated with any of the diseases nor with the pathogens. Random intercept linear models were used to explore factors affecting early (3-30 days) and long-term (3-200 days) growth, showing that calves with elevated serum amyloid-A concentrations at the age of 16 days had lower long-term growth. Increased albumin concentration at 30 days of age and higher parity of the dam increased early-term growth. The lack of association between a disease and the acute phase protein may stem from the low disease prevalence in the beef calves examined. The measurement of acute phase proteins of a young calf can help identify animals with possible future growth deficiencies, although the mechanisms through which the association between acute phase proteins and growth has yet to be explained.

N-acetyl -ß-D-glucosaminidase activity in cow milk as an indicator of mastitis.

Activity of lysosomal N-acetyl-ß-d-glucosaminidase (NAGase) in milk has been used as an indicator of bovine mastitis. We studied NAGase activity of 808 milk samples from healthy quarters and quarters of cows with spontaneous subclinical and clinical mastitis. Associations between milk NAGase activity and milk somatic cell count (SCC), mastitis causing pathogen, quarter, parity, days in milk (DIM) and season were studied. In addition, the performance of NAGase activity in detecting clinical and subclinical mastitis and distinguishing infections caused by minor and major bacteria was investigated. Our results indicate that NAGase activity can be used to detect both subclinical and clinical mastitis with a high level of accuracy (0·85 and 0·99). Incomplete correlation between NAGase activity and SCC suggests that a substantial proportion of NAGase activity comes from damaged epithelial cells of the udder in addition to somatic cells. We therefore recommend determination of NAGase activity from quarter foremilk after at least six hours from the last milking using the method described. Samples should be frozen before analysis. NAGase activity should be interpreted according to DIM, at least during the first month of lactation. Based on the results of the present study, a reference value for normal milk NAGase activity of 0·1-1·04 pmoles 4-MU/min/µl for cows with ≥30 DIM (196 samples) could be proposed. We consider milk NAGase activity to be an accurate indicator of subclinical and clinical mastitis.

The effect of sampling technique on PCR-based bacteriological results of bovine milk samples.

The aim of the study was to evaluate the effect of sampling technique on the microbiological results of bovine milk samples using multiplex real-time PCR. Comparison was made between a technique where the milk sample was taken directly from the udder cistern of the udder quarter using a needle and vacuum tube and conventional sampling. The effect of different cycle threshold (Ct) cutoff limits on the results was also tested to estimate the amount of amplified DNA in the samples. A total of 113 quarters from 53 cows were tested pairwise using both techniques, and each sample was studied with real-time PCR. Sampling from the udder cistern reduced the number of species per sample compared with conventional sampling. In conventional samples, the number of positive Staphylococcus spp. results was over twice that of samples taken with the needle technique, indicating that most of the Staphylococcus spp. originated from the teat or environmental sources. The Ct values also showed that Staphylococcus spp. were present in most samples only in low numbers. Routine use of multiplex real-time PCR in mastitis diagnostics could benefit from critical evaluation of positive Staphylococcus spp. results with Ct values between 34.0 and 37.0. Our results emphasize the importance of a careful aseptic milk sampling technique and a microbiologically positive result for a milk sample should not be automatically interpreted as an intramammary infection or mastitis.

The costs of interdigital phlegmon in four loose-housed Finnish dairy herds.

BACKGROUND: The aim of the study was to provide detailed herd level cost information about an outbreak of interdigital phlegmon (IP), which has been an emerging problem with enlarged loose house barns in Finland in recent years. During enlargement, the farmer's financial situation is sensitive because of the large investments to the farm business and unexpected costs can risk the farm's survival. RESULTS: The University of Helsinki research herd and three commercial herds having outbreaks of IP in 2012 or 2013 were visited to collect detailed information about the costs and economic impact of the outbreaks. The majority of the costs came from the discarded milk due to the antibiotic treatments. In Finland IP is usually treated with parental benzylpenicillin for 5 days which result in discarded milk for a total of 11 days. Third generation cephalosporins, widely used in other countries, have no milk withdrawal time. However, the use of these antibiotics is not recommended in Finland since these antimicrobials are critically important for human health. Herd-level costs varied between 4560 and 28,386 € depending on the herd size, the frequency of the infected cows, the antibiotics used and other costs involved. The average cost per infected cow was 489 €. CONCLUSIONS: The outbreaks of IP cause severe economic losses to dairy farms and the costs are lower if cows are treated with antibiotics with no withdrawal time. However, other costs, such as involuntary culling, reduced production and fertility also produce substantial costs to the farms. Early detection of sick animals, rapid treatment and control measures to limit the outbreak of IP can lower the costs. Because of the high costs farms should concentrate on preventing the disease.