Subclinical mastitis (SCM) is a major health problem of dairy animals in India and across the globe. An identification of potential risk factors of SCM can help for efficient udder health management in dairy animals. In this study, apparently healthy cows (HF crossbred: n = 45; Deoni: n = 43) were screened for SCM during different seasons through milk somatic cell count (SCC: reference test using 200 × 103 cells/ml as cut off value), California mastitis test (CMT) and differential electrical conductivity (DEC) test at an organized research farm. SCM positive milk samples (n = 34) were inoculated in selective media for Coliform sp., Streptococcus sp. and Staphylococcus sp. and DNA was isolated (n = 10) for species confirmation by 16s rRNA method. Both bivariate and multivariate models were used for risk assessment. We found the cumulative prevalence of 31 and 65% SCM in Deoni and crossbred cows, respectively. Screening of 328 crossbred cows under field conditions revealed point prevalence of 55% SCM. Multivariate analysis revealed stage of lactation (SOL), milk yield in previous lactation and test day milk yield in Deoni cows, as well as parity and mastitis treatment history in current lactation in HF crossbred cows as risk factors. SOL was a significant factor under field conditions. Receiver operated characteristic curve analysis revealed better accuracy of CMT than DEC. We found more mixed infections due to Staphylococcus sp. and Streptococcus sp. in culture, while 16s rRNA based molecular method revealed lesser-known pathogens associated with SCM. It is concluded that SCM prevalence rate is higher in crossbred than indigenous cows and these breeds have different risk factors for SCM. HF crossbred cows had similar SCM prevalence rate under different farming conditions, where CMT can be used for SCM diagnosis with excellent accuracy. The 16s rRNA method is useful for specific identification of lesser known and emerging mastitis pathogens.

This research aims to test the hypothesis that neural therapy (NT) on the auricular branch of the vagus nerve (ABVN) in dairy cows diagnosed with subclinical mastitis (SCM) results in a reduction of the somatic cell count. Therapeutic options for SCM are mostly based on use of antibiotics and often lead to unsatisfactory results. An alternative therapy targeting the anti-inflammatory properties of the vagus nerve showing good efficacy, economic viability and without major side effects would be of considerable interest. Auriculotherapy (AT) was performed using three repeated infiltrations of 8.0 mg (0.4 ml) procaine hydrochloride (2%) at the location of the ear tag associated with the auricular acupuncture point (AAP) of the udder. Some 85 clinically healthy cows from nine dairy farms were sampled for evaluation of quarter somatic cell count (QSCC) on four days (d0, d2, d4 and d6). Quarters with a QSCC > 100 000 cells/ml on d0 were included in the analysis. Over the study period, a total of 784 quarters were analysed, 385 control (CON) quarters from 40 cows and 399 treated (TRE) quarters from 45 cows. Results showed that AT of the ABNV resulted in a significant reduction of the QSCC after three treatments. The effect was independent of bacteriological culture results of the quarter milk samples. The bacteriological cure rate, however, was not influenced by AT. To our knowledge this is the first report of AT reducing QSCC in dairy cows with SCM. Before AT can be regarded as an alternative therapeutic approach, further research should focus on possible long-term effects of AT on the reduction of SCC, any bacterial elimination and the neural pathways of AT in dairy cows with SCM.

Variations in the levels of acute phase proteins and lactoferrin in serum and milk for diagnosis of subclinical mastitis in dairy cows are described in this research paper. Milking animals from two organized dairy farms in Kerala, India, were screened by California Mastitis Test (CMT), Electrical Conductivity test (EC) and Somatic Cell Count (SCC) test to identify animals affected with sub clinical mastitis (SCM). The concentrations of acute phase proteins (APP) Haptoglobin (Hp), C- reactive protein (CRP), Albumin, Lactoferrin (Lf) and α- 1 acid glycoprotein (AGP) in milk and Hp, Albumin, Serum Amyloid A (SAA) and CRP in the serum of 40 normal cows and 40 cows affected with sub clinical mastitis were assessed. Solid phase ELISA was employed for assessment of all parameters except the albumin levels, for which spectrophotometry was used. The values of Hp in milk; and SAA, AGP and Lf in serum, were significantly elevated in the group with sub clinical mastitis. Such variations were found to be independent of the specific bacterial organism causing the disease. These results show that significant variations exist in the levels of acute phase proteins Hp, AGP and Lf in milk, and SAA in serum of animals affected with subclinical bovine mastitis that are not affected by specific bacterial etiology.

Dairy goat farming is an important sector of the agricultural industry in Greece, with an annual total milk production exceeding 450 000 l and accounting for over 25% of all goat milk produced in the European Union; this milk is used mainly for cheese production. Despite the importance of goat milk for the agricultural sector in Greece, no systematic countrywide investigations in the bulk-tank milk of goats in Greece have been reported. Objectives were to investigate somatic cell counts (SCC) and total bacterial counts (TBC) in raw bulk-tank milk of goat herds in Greece, study factors influencing SCC and TBC therein and evaluate their possible associations with milk content. Throughout Greece, 119 dairy goat herds were visited for milk sampling for somatic cell counting, microbiological examination and composition measurement. Geometric mean SCC and TBC were 0.838 × 106 cells ml−1 and 581 × 103 cfu ml−1, respectively. Multivariable analyses revealed annual frequency of check-ups of milking system and total milk quantity per goat (among 53 variables) to be significant for increased SCC; no factor emerged (among 58 variables) to be significant for increased TBC. Negative correlation of SCC with total protein was found; mean total protein content in the bulk-tank milk in herds with SCC >0.75 × 106 cells ml−1 was 5.1% lower and in herds with SCC >1.5 × 106 cells ml−1, it was 7.8% lower.

Biofilm formation is a central feature to guarantee staphylococcal persistence in hosts and is associated with several diseases that are difficult to treat. In this research paper, biofilm formation and antimicrobial susceptibility were investigated in staphylococcal strains belonging to several species. These strains were isolated from the milk of cows with subclinical mastitis and most of them were coagulase-negative, with the prevalence of Staphylococcus chromogenes. High genetic diversity was observed among the strains by pulsed field gel electrophoresis. Antimicrobial resistance was assessed by disk diffusion and more than 50% of the strains were resistant to ampicillin and penicillin G, with multi-resistance profiles (13.6%) also being observed. Most strains (65.9%) formed biofilms when cultivated in BHI supplemented with 1% glucose. Most strains (72.7%) carried the intercellular adhesion gene (icaA), while less than half (36.3%) carried the biofilm-associated protein gene (bap). Concentrations of up to 10xMIC of erythromycin and tetracycline were not sufficient to suppress cell viability in preformed biofilms. Our results revealed that a genetically diverse group of biofilm-forming Staphylococcus species can be involved in subclinical mastitis. Since high antimicrobial concentrations cannot eradicate biofilm cells in vitro, their use in dairy animals may be ineffective in controlling infections, while supporting selection of resistant microorganisms. These data reinforce the need for alternative therapies aiming at disrupting biofilms for effective disease control.

The objectives of this work were (a) to determine the presence of streptococci in samples from small ruminant dairy farms (bulk-tank milk and, where possible, teatcup swabs), (b) to investigate the potential adverse effects of streptococci on milk quality and (c) to investigate the importance of some husbandry factors for the isolation of streptococci. Bulk-tank milk samples and teatcups swab samples were examined bacteriologically for the presence of streptococci. Somatic cell counting and milk composition measurements were also performed. The husbandry factors present in each farm were assessed for potential associations with the isolation of streptococci. Streptococci were isolated from milk samples from 31.4% of sheep and 17.4% of goat farms and from 4.8% of sheep and 5.9% of goat teatcups. Streptococci were isolated more frequently from the upper part than the lower part of teatcups: 5.0% vs. 1.9%. Most isolates (57.9%) were identified as Streptococcus uberis. Most isolates (68.4%) were slime-producing; slime-production was more frequent among isolates from teatcups (83.3%) than from bulk-tank milk (55.0%). Somatic cell counts and milk composition did not differ between farms in which streptococci were or were not isolated. Machine-milking was associated with the isolation of streptococci from bulk-tank milk samples. The initial stage of the milking period (first two months) was found to be associated with the isolation of streptococci from milking machine teatcups in sheep farms only.

We evaluated the effects of chronic subclinical mastitis (CSM) caused by different types of pathogens on milk yield and milk components at the cow level. A total of 388 Holstein cows had milk yield measured and were milk sampled three times at intervals of two weeks for determination of SCC and milk composition, and microbiological culture was performed. Cows were considered healthy if all three samples of SCC were ≤200 000 cells/ml and were culture-negative at the third milk sampling. Cows with one result of SCC > 200 000 cells/ml were considered to suffer non-chronic subclinical mastitis whereas cows with at least 2 out of 3 results of SCC > 200 000 cells/ml had CSM. These latter cows were further sorted according to culture results into chronic negative-culture or chronic positive-culture. This resulted in four udder health statuses: healthy, non-chronic, chronicNC or chronicPC. The milk and components yields were evaluated according to the udder health status and by pathogen using a linear mixed effects model. A total of 134 out of 388 cows (34.5%) were chronicPC, 57 cows (14.7%) were chronicNC, 78 cows (20.1%) were non-chronic and 119 cows (30.7%) were considered healthy, which resulted in a grand total of 1164 cow records included in the statistical model. The healthy cows produced more milk than each of the other groups (+2.1 to +5.7 kg/cow/day) and produced higher milk component yields than the chronicPC cows. The healthy cows produced more milk than cows with chronicPC caused by minor (+5.2 kg/cow/day) and major pathogens (+7.1 kg/cow/day) and losses varied from 5.8 to 11.8 kg/cow/day depending on the pathogen causing chronicPC mastitis. Chronic positive-culture cows had a reduction of at least 24.5% of milk yield and 22.4% of total solids yield.

Mastitis is a costly disease and in many areas of the world, these costs have been quantified to support farmers in their decision making with regard to prevention of mastitis. Although for subsaharan circumstances estimates have been made for the costs of subclinical mastitis (SCM), farm-specific cost estimations comprising both clinical mastitis (CM) and SCM are lacking. In this paper, we quantified failure costs of both CM and SCM on 150 Ethiopian market-oriented dairy farms keeping Holstein Friesian × Zebu breed cows. Data about CM were collected by face-to-face interviews and the prevalence of SCM was estimated for each farm using the California mastitis test. All other relevant information needed to calculate the failure costs, such as the consequences of mastitis and price levels, was collected during the farm visits, except for the parameter for milk production losses due to SCM, which was based on literature estimates and subjected to sensitivity analyses. The average total failure costs of mastitis was estimated to be 4 765 Ethiopian Birr (ETB) (1 ETB = 0.0449 USD) per farm per year of which SCM contributed 54% of the costs. The average total failure costs per lactating cow per farm per year were 1 961 ETB, with a large variation between farms (range 0 to 35 084 ETB). This large variation in failure costs between farms was mainly driven by variation in incidence of CM and prevalence of SCM. Milk production losses made the largest contribution (80%), whereas culling contributed 13% to 17% to the total failure costs. In our estimates, costs of veterinary services, drugs, discarded milk and labour made a minor contribution to the total failure costs of mastitis. Relative to the income of dairy farmers in North Western Ethiopia; the total failure costs of mastitis are high. In general, Ethiopian farmers are aware of the negative consequences of CM, but creating awareness of the high costs of SCM and showing large variation between farmers may be instrumental in motivating farmers to also take preventive measures for SCM.

This research communication reports the evaluation of cathelicidin in dairy goat milk for its relationship with the somatic cell count (SCC) and microbial culture results. Considering the limited performances of SCC for mastitis monitoring in goats, there is interest in evaluating alternative diagnostic tools. Cathelicidin is an antimicrobial protein involved in innate immunity of the mammary gland. In this work, half-udder milk was sampled bimonthly from a herd of 37 Alpine goats along an entire lactation and tested with the cathelicidin ELISA together with SCC and bacterial culture. Cathelicidin and SCC showed a strong correlation (r = 0.72; n = 360 milk samples). This was highest in mid-lactation (r = 0.83) and lowest in late lactation (r = 0.61), and was higher in primiparous (0.80, n = 130) than in multiparous goats (0.71, n = 230). Both markers increased with stage of lactation, but cathelicidin increased significantly less than SCC. In addition, peak level in late lactation was lower for cathelicidin (5.05-fold increase) than for SCC (7.64-fold increase). Twenty-one (5.8%) samples were positive to bacteriological culture, 20 for coagulase-negative staphylococci and one for Streptococcus spp.; 18 of them were positive to the cathelicidin ELISA (85.71% sensitivity). Sensitivity of SCC >500 000 and of SCC >1 000 000 cells/ml was lower (71.43 and 23.81%, respectively). Therefore, the high correlation of cathelicidin with SCC during the entire lactation, along with its lower increase in late lactation and good sensitivity in detecting intramammary infection (IMI), indicate a potential for monitoring subclinical mastitis in dairy goats. However, based on this preliminary assessment, specificity should be improved (40.41% for cathelicidin vs. 54.57 and 67.85% for SCC >500 000 and >1 000 000 cells/ml, respectively). Therefore, the application of cathelicidin for detecting goat IMI will require further investigation and optimization, especially concerning the definition of diagnostic thresholds.

The possible association between teat morphometric traits and subclinical mastitis (SCM) in dairy buffaloes was studied. Teat morphometric parameters, i.e. teat shape (bottle, conical, cylindrical, and others), teat-end shape (flat, round, and pointed), teat length (TL), teat diameter (TD), and teat-end to floor distance were measured before milking, but after proper milk let-down, in clinically healthy buffaloes (47 Murrah and 34 Nili-Ravi breeds). Subclinical mastitis was defined on the basis of bacteriology and somatic cell count (SCC) of quarter foremilk samples. A high proportion of cylindrical teats (40%) and pointed teat-ends (64·4%) was observed. Hind teats were longer and thicker than fore teats (P < 0·05). A significant breed effect was found with respect to teat shape, length and diameter (P < 0·05). Teats were mostly cylindrical (43·3 vs. 35·4%) and conical (34·2 vs. 30·8%) shaped, smaller (mean 8·2 vs. 9·5 cm) and thinner (mean 3·3 vs. 3·6 cm) in the Murrah breed compared with the Nili-Ravi breed. Teats that had ‘other’ shapes and were longer, wider, and placed closer to the floor were more associated with SCM (P < 0·05). Mean SCC was significantly higher (P < 0·05) in Nili-Ravi buffaloes, teat shapes classified as ‘others’, and quarters with SCM. Teat morphometric traits seem to be associated with indicators of udder health in buffaloes, thus, their inclusion in breeding programmes for selection against undesirable dairy type traits may be of value in reducing susceptibility to intramammary infections in Indian buffaloes.

The objective of the research described in this Research Communication was to describe potential associations of subclinical mastitis with sheep breeds in Greece. A countrywide survey (2198 ewes in 111 farms) was performed. Prevalence of subclinical mastitis was 0·260. Results did not indicate any difference in the prevalence of subclinical mastitis between farms with pure-bred and farms with cross-bred animals, nor any difference in prevalence between farms with Greek pure-bred animals and farms with imported pure-bred animals. Results indicated that prevalence of subclinical mastitis was smaller in farms with Assaf-breed (0·100) and higher in farms with Frisarta-breed (0·625) (P < 0·02). Prevalence of mastitis was smaller in farms with Greek traditional indigenous breeds (0·221) (P = 0·007). In a model that included sheep breed and management system in farm, breed emerged as a significant factor for prevalence of subclinical mastitis (P = 0·003).

A total of 90 cows from three commercial farms were used to evaluate the relationship between subclinical mastitis and clinical mastitis and thermal nociceptive threshold. Milk strips from all udder quarters were tested for clinical mastitis with visual inspection of milk and udder alterations and for subclinical mastitis using California Mastitis Test. Milk yield was recorded, milk was sampled and further analyzed for somatic cells count (SCC). Cows were considered healthy when SCC<200 000 cells/ml and no visual alterations in milk and/or udder, with mild subclinical mastitis when SCC>200 000 cells/ml and no visual alterations in milk and/or udder, with moderate subclinical mastitis when SCC>500 000 cells/ml and no visual alterations in milk and/or udder and with clinical mastitis when visual alterations in milk and/or udder were detected. Nociceptive threshold was evaluated with the thermal threshold meter apparatus applied to the rear legs. Thermal threshold (TT) decreased when we compared healthy cows with cows presenting clinical mastitis and tended to decrease when we compare healthy cows with those with moderate subclinical mastitis. TT was lower at the ipsilateral rear leg compared with the contralateral leg to the infected mammary gland. TT linearly decreases as log10SCC increased and it showed sharp decrease as log10SCC exceed the value of 6.4. Increase in one unit of log10SCC increased the odds of low thermal threshold (lower than 55.8°C). Subclinical mastitis might be a welfare issue as it tended to decrease nociceptive thermal threshold.

Subclinical mastitis (SM) is one of the most important diseases affecting dairy ewes worldwide, with negative impact on the animal health, farm income and public health. Animals with SM often remain untreated because the disease may not be revealed. Increase in somatic cell count (SCC) and positive bacteriology for mastitis pathogens in milk samples are indicative of SM but the evidence of only one of these alterations must suggest an uncertain SM (UM). UM is defined when positive bacteriological examination (Latent-SM) or SCC>500 000 cells/ml (non-specific-SM) are detected in milk. Nevertheless, SCC and bacteriological examination are expensive, time consuming and are not yet in use at the farm level in dairy ewes. Recently, a sensitive acute phase protein, amyloid A, displaying multiple isoforms in plasma and different body fluids including mammary secretion (milk amyloid A-MAA), has been investigated as a marker of mastitis in cows and, in a few studies, in sheep. The aim of this trial was to compare the concentration of MAA of single udder-halves in ewes with healthy udder-halves (HU-control group) and naturally occurring subclinical mastitis, both confirmed (SM group) and uncertain (UM groups: Latent-SM and non-specific-SM), for monitoring udder health. The reliability of a specific ELISA kit for the measurement of MAA was also tested. During a 3-month trial period, 153 udder halves were assigned to the experimental groups based on their health status: 25 with SM, 40 with UM (11 with latent-SM and 29 with non-specific-SM) and 88 HU. SCC and bacteriological analysis were performed to establish the control and subclinical mastitis groups. MAA concentrations in milk samples were measured using a specific commercially milk ELISA kit. The data were submitted to statistical analysis. Significant (P<0·05) differences among the groups SM, non-specific-SM and HU were detected with the SM having the highest level and HU the lowest. MAA concentration is affected by the udder health status and is a useful indicator of subclinical mastitis and increased SCC in sheep.

Somatic cell count (SCC) is generally regarded as an indicator of udder health. A cut-off value of 100×103 cells/ml is currently used in Germany to differentiate between normal and abnormal secretion of quarters. In addition to SCC, differential cell counts (DCC) can be applied for a more detailed analysis of the udder health status. The aim of this study was to differentiate somatic cells in foremilk samples of udder quarters classified as normal secreting by SCC <100×103 cells/ml. Twenty cows were selected and 72 normal secreting udder quarters were compared with a control group of six diseased quarters (SCC >100×103 cells/ml). In two severely diseased quarters of the control group (SCC of 967×103 cells/ml and 1824×103 cells/ml) Escherichia coli and Staphylococcus aureus were detected. DCC patterns of milk samples (n=25) with very low SCC values of ⩽6·25×103 cells/ml revealed high lymphocyte proportions of up to 92%. Milk cell populations in samples (n=41) with SCC values of (>6·25 to ⩽25)×103 cells/ml were also dominated by lymphocytes (mean value 47%), whereas DCC patterns of milk from udder quarters (n=6) with SCC values (>25 to ⩽100)×103 cells/ml changed. While in samples (n=3) with SCC values of (27–33)×103 cells/ml macrophages were predominant (35–40%), three milk samples with (43–45)×103 cells/ml indicated already inflammatory reactions based on the predominance of polymorphonuclear leucocytes (PMN) (54–63%). In milk samples of diseased quarters PMN were categorically found as dominant cell population with proportions of ⩾65%. Macrophages were the second predominant cell population in almost all samples tested in relationship to lymphocytes and PMN. To our knowledge, this is the first study evaluating cell populations in low SCC milk in detail. Udder quarters classified as normal secreting by SCC <100×103 cells/ml revealed already inflammatory processes based on DCC.

Subclinical mastitis (SM) is a major concern in the dairy industry because it causes economic losses and can lead to clinical mastitis. The mechanisms of the onset and progression of SM are not fully understood, and a new procedure for the detection and appropriate prediction of SM leading to clinical mastitis is necessary for dairy cow management. Inflammatory cytokines such as interleukin (IL)-6 are known to be present in inflamed mammary glands at the onset of mastitis, and IL-6 concentrations correlate with the levels of inflammation. In this study, the detection of IL-6 was examined for the evaluation for the future prediction of SM in 77 quarter milk samples from 20 cows. IL-6 concentrations in quarter milk were measured by sandwich ELISA, and the data were compared with milk somatic cell count (SCC) levels to diagnose SM. Average IL-6 concentration was significantly higher in SM quarter milk (207·0±441·6 pg/ml) than in healthy control quarter milk (12·6±33·4 pg/ml, P<0·01). Results of the cross-tabulation table show that SM prediction accuracy based on IL-6 concentration was almost equal or superior to SM prediction accuracy based on SCC. The detection of IL-6 in milk indicated SM earlier than did the detection of elevated SCC. Thus, the detection of IL-6 in milk could be a future prediction marker for SM.

The objective was to investigate the changes occurring in the activities of the enzymes lactate dehydrogenase (LDH), alkaline phosphatase (ALP) and aspartate aminotransferase (AST) in sheep and goat milk as a result of subclinical intramammary infections (IMI) and to evaluate the use of these enzymes for the diagnosis of subclinical IMI in dairy sheep and goats. A total of 206 samples of sheep milk and 162 samples of goat milk, obtained from equal udder halves, were used in the study. For each species they were divided into two groups: a no-infection group and a subclinical infection group. Activities of LDH, ALP and AST were significantly higher in the subclinical infection group than in the no-infection group (P<0·05) in both sheep (LDH: 350·42±11·25 v. 120·91±4·41; ALP: 2773·43±105·18 v. 2189±94·24; AST: 29·57±0·74 v. 17·32±0·46) and goats (LDH: 354·07±13·33 v. 103·79±3·75; ALP: 311·13±25·74 v. 137·24±19·62; AST: 27·59±6·42 v. 15·87±0·45). The activity of LDH was identified as indicator for subclinical IMI in both sheep and goats. The optimum cut-off values for LDH activity, offering the highest diagnostic sensitivity (DSn) and diagnostic specificity (DSp), determined by receiver operating characteristic (ROC) analysis, were at 197 U/l, 185 U/l and 197 U/l for sheep, goats and both species, respectively. DSn for sheep, goats and both species at these cut-off values was 92·8%, 98·2% and 94·0%, whereas DSp was 95·4%, 96·3% and 96·3%, respectively. It was concluded that the determination of LDH activity in milk serum is a sensitive and reliable method for the detection of subclinical IMI in dairy sheep and goats.

Mastitis is the most frequent and costly disease in dairy herds, as it negatively affects yield and milk quality. The presence of clinical mastitis is quite easy to asses, whereas the diagnosis of the subclinical form can be more difficult and requires laboratory assays. Somatic cell count (SCC) is widely used as a rapid and low-cost indicator of mastitis, even if is not useful in discriminating between the clinical and subclinical form. As amyloid A has been investigated as a marker of mastitis, the aim of this study was to assess the potential value of measuring amyloid A in serum and milk and the correlation with SCC in the diagnosis of subclinical mastitis. The reliability of two different ELISA kits for the measurement of amyloid A in milk was also tested. During a 1-month trial period, 21 cows were assigned to three experimental groups according to their health status: 6 cows with clinical mastitis (CM), 10 cows with subclinical mastitis (SM) and 5 healthy cows (HE). Amyloid A was measured both in serum (SAA) and in quarter milk samples (mAA) with a serum ELISA kit, and in quarter milk samples (MAA) with a milk ELISA kit. SCC, total microbial count (TMC) and bacterial examination of the milk were also carried out. After a log transformation, the data were submitted to ANOVA and linear regression. TMC was significantly higher in cows with clinical mastitis, while no differences were observed between the other two experimental groups. SCC and MAA levels were significantly different among the three groups. mAA concentrations were similar between cows with subclinical and clinical mastitis, and SAA was not affected by mastitis. A significant correlation between SCC and MAA or mAA was detected, while no correlation was recorded between SAA and mAA. A close relationship between MAA and mAA was noticeable even at low concentrations, suggesting MAA as a potential physiological marker of subclinical mastitis.

The main objective was to determine the prevalence of intramammary infections (IMI) in dairy cows in Flanders, Belgium. Data were obtained from quarter milk samples of dairy herds subjected to a mandatory yearly screening of all lactating cows. A total of 178 668 quarter milk samples were collected at 1087 cross-sectional dairy herd screenings performed in three consecutive years. Of the dairy cows, 40% had at least one culture-positive quarter. More than 50% of all IMI were caused by non-aureus staphylococci. Streptococcus agalactiae is almost eradicated in Flanders, whereas Staphylococcus aureus was isolated from 18% of the culture-positive quarters. In addition, the distribution of mastitis pathogens in quarter milk samples from selected dairy cows with an elevated somatic cell count (SCC) is described. From 6390 cows with a geometric mean composite SCC ⩾250 000 cells/ml, nearly 65% had at least one culture-positive quarter. The majority of the IMI were caused by non-aureus staphylococci (41·1%), whereas Staph. aureus and aesculin-positive cocci were found in respectively 25% and 18% of the culture-positive milk samples. We conclude that more efforts are needed in the prevention and control of subclinical mastitis in Flanders. Non-aureus staphylococci are the predominant cause of IMI, warranting more research regarding the epidemiology and pathogenicity of those species.

Chronic subclinical mastitis is usually not treated during the lactation. However, some veterinarians regard treatment of some types of subclinical mastitis to be effective. The goal of this research was to develop a stochastic Monte Carlo simulation model to support decisions around treatment of chronic subclinical mastitis caused by Streptococcus uberis. Factors in the model included the probability of cure after treatment, probability of the cow becoming clinically diseased, transmission of infection to other cows, and physiological effects of the infection. Using basic input parameters for Dutch circumstances, the average economic costs per cow of an untreated chronic subclinical mastitis case caused by Str. uberis in a single quarter from day of diagnosis onwards was €109. With treatment, the average costs were higher (€120). Thus, for the average cow, treatment was not efficient economically. However, the risk of high costs was much higher when cows with chronic subclinical mastitis were not treated. A sensitivity analysis showed that profitability of treatment of chronic subclinical Str. uberis mastitis depended on farm-specific factors (such as economic value of discarded milk) and cow-specific factors (such as day of diagnosis, duration of infection, amount of transmission to other cows and cure rate). Therefore, herd level protocols are not sufficient and decision support should be cow specific. Given the importance of cow-specific factors, information from the current model could be applied to automatic decision support systems.

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.