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Extracellular vesicles (EVs) have demonstrated unique advantages in serving as nanocarriers for drug delivery, yet the cargo encapsulation efficiency is far from expectation, especially for hydrophilic chemotherapeutic drugs. Besides, the intrinsic heterogeneity of EVs renders it difficult to evaluate drug encapsulation behaviour. Inspired by the active drug loading strategy of liposomal nanomedicines, here we report the development of a method, named "Sonication and Extrusion-assisted Active Loading" (SEAL), for effective and stable drug encapsulation of EVs. Using doxorubicin-loaded milk-derived EVs (Dox-mEVs) as the model system, sonication was applied to temporarily permeabilize the membrane, facilitating the influx of ammonium sulfate solution into the lumen to establish the transmembrane ion gradient essential for active loading. Along with extrusion to downsize large mEVs, homogenize particle size and reshape the nonspherical or multilamellar vesicles, SEAL showed around 10-fold enhancement of drug encapsulation efficiency compared with passive loading. Single-particle analysis by nano-flow cytometry was further employed to reveal the heterogeneous encapsulation behaviour of Dox-mEVs which would otherwise be overlooked by bulk-based approaches. Correlation analysis between doxorubicin auto-fluorescence and the fluorescence of a lipophilic dye DiD suggested that only the lipid-enclosed particles were actively loadable. Meanwhile, immunofluorescence analysis revealed that more than 85% of the casein positive particles was doxorubicin free. These findings further inspired the development of the lipid-probe- and immuno-mediated magnetic isolation techniques to selectively remove the contaminants of non-lipid enclosed particles and casein assemblies, respectively. Finally, the intracellular assessments confirmed the superior performance of SEAL-prepared mEV formulations, and demonstrated the impact of encapsulation heterogeneity on therapeutic outcome. The as-developed cargo-loading approach and nano-flow cytometry-based characterization method will provide an instructive insight in the development of EV-based delivery systems., (© 2021 The Authors. Journal of Extracellular Vesicles published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles.)

The immune response during the onset of coliform mastitis in vaccinated cows was investigated by measuring lactoferrin (LF), interleukin-8 (IL-8), and interleukin-1β (IL-1β) concentrations and somatic cell counts in 28 milk samples at the onset of acute coliform mastitis (ACM) and 73 milk samples at the onset of peracute coliform mastitis (PCM). Vaccinated ACM, unvaccinated ACM, and vaccinated PCM showed significantly higher values for LF and IL-1β levels than unvaccinated PCM (p < .01). The IL-8 concentration was lower in vaccinated PCM than in unvaccinated PCM (p < .05). There was no significant difference in somatic cell counts for each parameter. There were no significant differences in the parameters between vaccinated and unvaccinated ACM cows, or vaccinated ACM and PCM cows. From the above results, it is suggested that mastitis vaccination improved the early immune response, particularly at the onset of PCM, and played a large role in host defense against the initial infection., (© 2021 Japanese Society of Animal Science.)

The aim of this study was to estimate genetic parameters for lactation yields of milk (MY), fat (FY), protein (PY), and somatic cell score (SCS) of New Zealand dairy goats. The analysis used 64,604 lactation records from 23,583 does, kidding between 2004 and 2017, distributed in 21 flocks and representing 915 bucks. Estimates of genetic and residual (co) variances, heritabilities, and repeatabilities were obtained using a multiple-trait repeatability animal model. The model included the fixed effects of contemporary group (does kidding in the same flock and year), age of the doe (in years), and as covariates, kidding day, proportion of Alpine, Nubian, Toggenburg, and "unknown" breeds (Saanen was used as the base breed), and heterosis. Random effects included additive animal genetic and doe permanent environmental effects. Estimates of heritabilities were 0.25 for MY, 0.24 for FY, 0.24 for PY, and 0.21 for SCS. The phenotypic correlations between MY, FY, and PY ranged from 0.90 to 0.96, and the genetic correlations ranged from 0.81 to 0.93. These results indicate lactation yield traits exhibit useful heritable variation and that multiple trait selection for these traits could improve milk revenue produced from successive generations of New Zealand dairy goats., (© 2019 Japanese Society of Animal Science.)

The objectives of this study were to estimate the heritability of mastitis incidence and genetic correlations between the mastitis and the somatic cell score (SCS) statistics, and to compare the practicability between different models. We used test-day records with the mastitis incidence and SCS collected from Holstein cows calving from 1988 to 2015 in Hokkaido, Japan. As indicators of mastitis, the average SCS (avSCS), the standard deviation of SCS (sdSCS), and the maximum SCS (maxSCS) were calculated using test-day records up to the first 305 days in milk within a lactation. We compared a four-trait repeatability animal model (MTRP) with a four-trait multiple-lactation animal model (MTML). The heritability for mastitis was equal to or lower than 0.05 in all the models. Genetic correlations between lactations with MTML within the same trait were positive and close to 1. With MTRP, the estimated genetic correlations of the mastitis incidence with avSCS, sdSCS, and maxSCS were 0.66, 0.79, and 0.82, respectively. A joint evaluation with SCS statistics is expected to give an extra reliability for mastitis because of high and positive genetic correlations among the traits., (© 2019 Japanese Society of Animal Science.)

The current study was conducted to investigate the effect of stocking density (SD) on oxidative stress status and mammary gland permeability in early lactating dairy cows. Thirty-two dairy cows were allocated into 16 blocks, basing on parity, previous milk yield, and body weight, and were then randomly assigned into one of the two treatments as follows: 75% (75SD) and 100% (100SD) SD. The cows were fed with same diet throughout the 8-week experimental period. The milk yield and milk sample were collected on two consecutive days during the 8-week experimental period weekly. Plasma samples were collected on fourth and eighth experimental weeks. Raw, energy-corrected, and 4% fat-corrected milk yield were significantly higher in 75SD-cows than that of 100SD-animals, respectively. The milk somatic cell count was lower in 75SD-cows than that of 100SD-animals. The levels of Na + , Na + /K + , bovine serum albumin and plasmin were lower in 75SD-cows than those of 100SD-cattle, respectively. The 75SD-cows had reduced insulin and insulin/glucagon levels but higher prolactin and growth hormone concentrations, compared with those of 100SD-animals, respectively. In conclusion, compared with low SD-animals, early lactating cows with higher SD had higher oxidative stress status, which further led to a greater mammary gland permeability., (© 2019 Japanese Society of Animal Science.)

The aim of this study was to estimate genetic associations between alternative somatic cell count (SCC) traits and milk yield, composition and udder type traits in Italian Jersey cows. Alternative SCC traits were test-day (TD) somatic cell score (SCS) averaged over early lactation (SCS&#95;150), standard deviation of SCS of the entire lactation (SCS&#95;SD), a binary trait indicating absence or presence of at least one TD SCC >400,000 cells/ml in the lactation (Infection) and the ratio of the number of TD SCC >400,000 cells/ml to total number of TD in the lactation (Severity). Heritabilities of SCC traits, including lactation-mean SCS (SCS&#95;LM), ranged from 0.038 to 0.136. Genetic correlations between SCC traits were moderate to strong, with very few exceptions. Unfavourable genetic associations between milk yield and SCS&#95;SD and Infection indicated that high-producing cows were more susceptible to variation in SCC than low-producing animals. Cows with deep udders, loose attachments, weak ligaments and long teats were more susceptible to an increase of SCC in milk. Overall, results suggest that alternative SCC traits can be exploited to improve cow's resistance to mastitis in Italian Jersey breed., (© 2019 Japanese Society of Animal Science.)

We used daily records from provincial Japanese weather stations and monthly test-day records of milk production to investigate the length of the lags in the responses of cows' milk yield and somatic cell score (SCS) to heat stress (HS). We also investigated the HS thresholds in milk yield and SCS. Data were a total of 17,245,709 test-day records for milk and SCS in Holstein cows that had calved for the first time between 2000 and 2015, along with weather records from 60 weather stations. Temperature-humidity index (THI) values were estimated by using average daily temperature and average daily relative humidity. Adjusted THI values were calculated by using temperature, relative humidity, wind speed, and solar radiation. The model contained herd, calving year, month of test day, age group, days in milk, and THI as a fixed effect. THIs for each day from 14 days before the test day until the test day were used to represent the HS effects. The HS occurring 3 days, and between 8 and 10 days, before the test day had the greatest effect on the milk yield and SCS, respectively. The threshold THI values for the HS effect were about 60-65 for both traits., (© 2019 The Authors. Animal Science Journal published by John Wiley & Sons Australia, Ltd on behalf of Japanese Society of Animal Science.)

The exfoliation rate of mammary epithelial cells (MECs) in milk is affected by physiological, breeding and environmental factors. Little is known about the relationship between the MEC exfoliation into milk and mammary-infected Staphylococcus aureus (S. aureus) load on bovine mastitis caused by S. aureus. The aim of this study was to investigate the relationship between S. aureus load and the proportion of MEC exfoliation in milk using five substantial bovine mastitis models. In 64 randomly extracted milk samples from udders at 3-21 days after S. aureus infusion, there were various samples with different numbers of S. aureus counts and somatic cell counts. No significant correlations were found between the S. aureus counts and somatic cell count (r = 0.338). In contrast, a significant correlation was noted between S. aureus counts and the proportion of cytokeratin-positive cells in the milk from the infused udders (r = 0.734, P < 0.01). In conclusion, the increasing MEC exfoliation rate in milk from mastitis udders caused by S. aureus may contribute to reduced milk yield., (© 2017 Japanese Society of Animal Science.)

Buffalo milk represents an indispensable source of nourishment in many parts of the world and it is the second most consumed milk worldwide. Buffalo milk is actually used for the production of many dairy products such as pasteurized or concentrated milk, butter, yogurt, ice-cream, dehydrated milk products and cheeses. Due to its high nutritional value and the presence of natural bioactive substances, buffalo milk can also provide health benefits to consumers. In Italy, buffalo milk is used mainly for cheese making, mozzarella PDO (Protected Designation of Origin), which is a highly valued dairy product. This 3-year study, carried out between 2011 and 2013, was aimed at evaluating the quality of bulk Italian Mediterranean buffalo milk by monitoring physico-chemical parameters, somatic cell and total bacterial counts. A total of 51 samples of bulk milk were collected from one herd throughout the monitored period. Analysis of variance, carried out to test month, season and year main effects, highlighted remarkable seasonal effects for fat, protein and lactose content, as well as for predicted mozzarella cheese yield, and somatic cell counts. The calculation of simple correlations allowed the identification of positive correlations between estimated cheese yield and fat and protein content., (© 2017 Japanese Society of Animal Science.)

The objective of the present study was to examine the changes in innate immune factors in milk during the estrous cycle in dairy cows. Milk was collected from cows with a normal ovulatory cycle and cows subjected to the OVSYNCH protocol, and somatic cell counts (SCC), lingual antimicrobial peptide (LAP) concentrations and lactoperoxidase (LPO) activity in milk were measured. In cows with a normal ovulatory cycle, there was no significant change in LAP concentrations and SCC during the ovulatory cycle. However, LPO activity at days 0 and 19 (day 0 = day of ovulation) were significantly higher than those at days 10, 12 and 15. In cows subjected to the OVSYNCH protocol, a significant increase in SCC was observed at day 9 (2 days after prostaglandin treatment) compared with that at day 11 (2 days after second administration of gonadotropin). There were no significant changes in LAP concentrations and LPO activity during the OVSYNCH protocol. These results indicate that LPO activity, an innate immune factor, was enhanced in the preovulatory period., (© 2016 Japanese Society of Animal Science.)

We examined the effects of heat stress (HS) on production traits, somatic cell score (SCS) and conception rate at first insemination (CR) in Holsteins in Japan. We used a total of 228 242 records of milk, fat and protein yields, and SCS for the first three lactations, as well as of CR in heifers and in first- and second-lactation cows that had calved for the first time between 2000 and 2012. Records from 47 prefectural weather stations throughout Japan were used to calculate the temperature-humidity index (THI); areas were categorized into three regional groups: no HS (THI < 72), mild HS (72 ≤ THI < 79), and moderate HS (THI ≥ 79). Trait records from the three HS-region groups were treated as three different traits and trivariate animal models were used. The genetic correlations between milk yields from different HS groups were very high (0.91 to 0.99). Summer calving caused the greatest increase in SCS, and in the first and second lactations this increase became greater as THI increased. In cows, CR was affected by the interaction between HS group and insemination month: with summer and early autumn insemination, there was a reduction in CR, and it was much larger in the mild- and moderate-HS groups than in the no-HS group., (© 2016 Japanese Society of Animal Science.)

A survey on ovine dairy farms directly transforming own-produced milk, in the Italian Marche region, was carried out to assess flock and milking practices that may influence milk hygienic-sanitary conditions. A census survey established that 24 dairy farms were located in this region. Bulk milk samples were collected throughout the milking period in each dairy farm in 2013. Analyzed variables were: (i) chemical parameters such as fat, protein and lactose content, dry matter and pH; and (ii) total bacterial (TBC) and somatic cell counts (SCC). Chemical parameter values were in agreement with published data while, geometric mean (GM) log10 SCC was 5.91 and TBC GM was 57 978 colony forming units/mL, in compliance with Eropean Union criteria. A positive correlation was found between SCC and TBC when GMs of all farm data were considered (Spearman's rho = 0.7925; P = 0.0001). Statistical analysis did not show significant correlation between SCC or TBC GM and dairy farm principal characteristics. Although SCC levels detected in the present study should suggest the need to implement mastitis control programs, Marche's dairy sheep flocks revealed a good hygienic condition level. This is an important aspect in implementing safety for end users of the final product., (© 2015 Japanese Society of Animal Science.)

The present study aims to evaluate the suitability of 10 candidate genes, namely GAPDH, ACTB, RPS15A, RPL4, RPS9, RPS23, HMBS, HPRT1, EEF1A1 and UBI as internal control genes (ICG) to normalize the transcriptional data of mammary epithelial cells (MEC) in Indian cows. A total of 52 MEC samples were isolated from milk of Sahiwal cows (major indigenous dairy breed of India) across different stages of lactation: Early (5-15 days), Peak (30-60 days), Mid (100-140 days) and Late (> 240 days). Three different statistical algorithms: geNorm, Normfinder and BestKeeper were used to assess the suitability of these genes. In geNorm analysis, all the genes exhibited expression stability (M) values below 0.5 with EEF1A1 and RPL4 showing the maximum expression stability. Similar to geNorm, Normfinder also identified EEF1A1 and RPL4 as two of the most stable genes. In Bestkeeper algorithm as well, all the 10 genes showed consistent expression levels. The analysis showed that four genes, that is, EEF1A1, RPL4, GAPDH and ACTB exhibited higher coefficient of correlation to the Bestkeeper index, lower coefficient of variance and standard deviation, indicating their superiority to be used as ICG. The present analysis has provided evidence that RPL4, EEF1A1, GAPDH and ACTB could probably act as most suitable genes for normalizing the transcriptional data of milk-derived mammary epithelial cells of Indian cows., (© 2016 Japanese Society of Animal Science.)

The objective of this study was to investigate the milk protein profiles of normal milk and those of milk during the course of subclinical mastitis, caused by natural Streptococcus agalactiae infection. Two-dimensional gel electrophoresis and liquid chromatography mass spectrometry were used to assess protein profiles and to identify the proteins. The results showed that S. agalactiae subclinical mastitis altered the protein profiles of milk. Following Mascot database matching, 11 and 12 protein types were identified in the milk collected from healthy and S. agalactiae subclinical mastitic udders, respectively. The distinct presence of the antibacterial protein cathelicidin-1 was detected in infected milk samples, which in turn was highly correlated to the severity of subclinical mastitis as represented by the milk somatic cell count (r = 0.616), but not the bacterial count. The protein profile of milk reveals changes in the host response to S. agalactiae intramammary infection; cathelicidin-1 could therefore serve as a biomarker for the detection of subclinical mastitis in dairy cows., (© 2015 Japanese Society of Animal Science.)

We estimated the genetic parameters of fat-to-protein ratio (FPR) and the genetic correlations between FPR and milk yield or somatic cell score in the first three lactations in dairy cows. Data included 3,079,517 test-day records of 201,138 Holstein cows in Japan from 2006 to 2011. Genetic parameters were estimated with a multiple-trait random regression model in which the records within and between parities were treated as separate traits. The phenotypic values of FPR increased soon after parturition and peaked at 10 to 20 days in milk, then decreased slowly in mid- and late lactation. Heritability estimates for FPR yielded moderate values. Genetic correlations of FPR among parities were low in early lactation. Genetic correlations between FPR and milk yield were positive and low in early lactation, but only in the first lactation. Genetic correlations between FPR and somatic cell score were positive in early lactation and decreased to become negative in mid- to late lactation. By using these results for genetic evaluation it should be possible to improve energy balance in dairy cows., (© 2015 Japanese Society of Animal Science.)

The spent mushroom substrate of Ganoderma balabacense cultivation (SMSGB) contains a large amount of bioactive substances. However, the potentials of SMSGB for improving milk production in dairy cows have not been thoroughly studied. In this study, the effects of hot water extract (HWE) from spent mushroom substrate after G. balabacense cultivated with JUNCAO, the herbaceous plants long-known to be suitable for cultivating edible and medicinal fungi, on production performance and hematology parameters of dairy cows, were determined. Holstein dairy cows were fed different doses of HWE. After a 60-day administration period with 100 g/day of HWE, milk yield, milk protein and triglyceride (TG) levels increased by 4.02% (P < 0.01), 4.49% (P < 0.05) and 32.65% (P < 0.05), respectively; somatic cell count (SCC) and alanine aminotransferase (ALT) were significantly decreased (P < 0.05). The production performance of dairy cows suggests that HWE with SMSGB treatment is effective in improving milk yield (P < 0.01) and hematology parameters of dairy cows, and may be useful as a functional feed additive., (© 2015 Japanese Society of Animal Science.)

Evidence for the efficacy of antimicrobial treatments in Escherichia coli mastitis is limited. The aim of this double-blinded field trial was to investigate the efficacy of enrofloxacin compared with placebo, with a special focus on survival, in dairy cows with acute clinical mastitis caused by E. coli. Dairy cows (n=116) with acute clinical mastitis were included in the study. A clinical examination was performed and a milk sample from the affected udder quarter was collected for investigation of somatic cell count (SCC) and bacteriology on the first day of treatment (day 0) and at day 3 (clinical examination only), day 22 and day 28. Data regarding culled cows, SCC and daily milk yield were retrieved from monthly milk recording each month until 180 days after treatment. All cows were treated with either enrofloxacin or placebo once a day for three days, starting at day 0. After culturing, 56 cows with confirmed E. coli mastitis remained in the study. Nine (16 per cent) of them died within the first week. Enrofloxacin-treated cows had lower SCC compared with placebo-treated cows at first monthly milk recordings after being treated for mastitis. Treatment with enrofloxacin did not result in a higher probability of survival compared with placebo., (British Veterinary Association.)

Casein (CN) micelles are colloidal aggregates of protein dispersed in milk, the importance of which in the dairy industry is related to functionality and yield in dairy products. The objective of this work was to investigate the correlation of milk CN micelles diameter from Holstein and Zebu crossbreds with milk composition (protein, fat, lactose, total and nonfat solids and milk urea nitrogen), somatic cell count (SCC), age, lactation stage and production. Average casein micelles diameters of milk samples obtained from 200 cows were measured using photon correlation spectroscopy and multiple regression analysis was used to find relationship between variables. CN micelle diameter, SCC and nonfat solids were different between animals with different Holstein crossbreed ratios, which suggests influence of genetic factors, mammary gland health and milk composition. Overall, results indicate the potential use of CN micelle diameter as a tool to select animals to produce milk more suitable to cheese production., (© 2014 Japanese Society of Animal Science.)

The objective was to examine (a) how pregnancy rate on one farm (500 cows) was affected by signs of oestrus and disease stressors and (b) whether pregnancy rate could be maximised by considering cow activity. The signs of oestrus and timings were recorded at artificial insemination (AI), and cow activity was monitored by neck collars. Pregnancy rate tended to be higher in animals that displayed standing oestrus (35 v 26 per cent; P=0.06) but was 10 per cent lower in those cows with an elevated somatic cell count (SCC; >200,000 cells/ml milk) within 0-4 or 4-8 weeks prior to AI (P=0.01 and 0.05, respectively), irrespective of the incidence of clinical mastitis prior to AI. Cow activity data were available for 525 inseminations (from a total of 1299). The mean interval from increased activity to AI in all cows (11 hours 32 minutes; 95 per cent CI 10 hours 40 minutes to 12 hours 24 minutes) was not different for cows that did or did not establish a pregnancy (P=0.90). The pregnancy rate improved to the average of unaffected cows if AI was delayed by about eight hours in animals with an elevated SCC 0-4 weeks prior to AI (P=0.025), indicating that, in cows with prior elevated SCC, AI could be repeated approximately eight hours later to achieve maximum pregnancy rates., (British Veterinary Association.)

The objective of this paper was to evaluate the global milk quality in Tunisian dairy herds. Samples of milk were analyzed for chemical, physical and hygienic parameters. Milk total solids, fat content and density were consistently correlated and one of them can be used as a chemical indicator of milk quality. The somatic cell count value of 689 × 10(3) /mL was higher than the recommended threshold. All milk samples were positive for the major pathogen Staphylococcus aureus. These hygienic parameters were related more closely with chloride content, minerals and electrical conductivity, which allows them to be used as indicators of mammary gland infection. It was concluded that milk producers have at hand rapid and easy tools for assessing the overall quality of milk., (© 2014 Japanese Society of Animal Science.)

Somatic cell counts (SCC) measurements are typically performed using quantitative methods, such as the Breed method (Breed) and the Fossomatic method (FSCC). The DeLaval cell counter (DCC) developed recently is a quantitative somatic cell counter with a low initial cost and superior portability. However, since the DCC was specifically developed for measuring SCC of ≤ 4 × 10(6) cells/mL milk from bulk tanks or individual cows, its reliability for estimating SCC that exceed this concentration has not yet been clarified. This study therefore examined whether it is possible to accurately measure SCC by diluting milk samples with initial SCC of 4 × 10(6) cells/mL, as seen in clinical mastitis milk. We collected milk samples from 99 quarters of 99 Holstein cows with clinical mastitis. These milk samples were diluted 10-fold with saline and thoroughly mixed before performing SCC measurement with the DCC. The correlation coefficients of SCC measured by the FSCC, Breed and DCC methods indicated strong correlations between each pair of methods. The findings showed that DCC can be used to identify bovine clinical mastitis milk and is useful as a quantitative SCC measurement device on farm sites., (© 2013 Japanese Society of Animal Science.)

The present study was undertaken to examine whether potential levels of innate immune factors (lingual antimicrobial peptide (LAP), lactoferrin (LF) and lactoperoxidase (LPO)) in colostrum are associated with subsequent milk somatic cell count (SCC) in dairy cows. Quarter milk samples were collected daily for 1 week postpartum to measure LAP and LF concentrations and LPO activity. SCC in milk was determined weekly for 2 months postpartum and its correlations to concentrations of LAP and LF and LPO activity were examined. Only small variations of all immune factors were found among four udders in each individual cow, whereas there were great differences in these factors among cows. Negative correlation was detected only between LPO activity and mean and maximum SCC, whereas its relationship was not significant. LAP and LF concentrations were significantly correlated positively to mean, maximum and minimum SCC. These results suggest that the great difference in innate immune factors among animals and high LAP and LF concentrations in colostrum may be associated with subsequent high incidence of SCC increase., (© 2013 Japanese Society of Animal Science.)

This study describes the failure of a single jet exit race automatic teat spray (ATS) system resulting in the spread of Staphylococcus aureus infection in a 135-cow dairy herd, which showed an increased herd somatic cell count from 91,000/ml to 554,000/ml over a nine-month period. S aureus was isolated from 34 of 46 high cell count cows. The milking procedures were modified and manual teat spraying was restarted. Bacteriology was used to identify S aureus positive high cell count cows, and first and second lactation cows were treated during lactation. If their cell counts were not reduced, these were then culled. High cell count S aureus cows in lactation three or above were culled. The three-month geometric mean cell count fell to below 150,000/ml within five months. As all replacements were home-bred, S aureus infection must have spread from within the herd itself. All other causes have been eliminated, and this spread is attributed to the failure of the ATS to carry out effective postmilking teat disinfection. The advantages and disadvantages of ATS systems are discussed, especially in relation to robotic or voluntary milking systems.

This study aims to identify single nucleotide polymorphisms (SNPs) and haplotypes in the TLR2 gene, and analyze the association of SNPs or haplotypes and somatic cell scores in 151 Xinjiang Brown cattle and 138 Holsteins to evaluate the role of TLR2 during intramammary infections. TLR2 coding region was amplified by PCR and screened for SNP sequencing. Genotypes and frequencies of SNPs were identified. Finally, the associations of genotypes or haplotypes and somatic cell scores (SCS) were analyzed. The results showed that: (i) 15 SNPs (E+653, E+945, E+978, E+1010, E+1250, E+1688, E+1707, E+1779, E+1782, E+1891, E+1995, E+2025, E+2055, E+2214 and E+2295) were observed and detected from 289 cows; (ii) distribution of the 14 SNPs were significantly different from Xinjiang Brown cattle and Holstein (P<0.001) except for the E+945 (P>0.05); (iii) in 11 SNPs (E+945, E+978, E+1010, E+1688, E+1707, E+1779, E+1782, E+1995, E+2025, E+2055 and E+2214), the SCS of AB genotype was lower than AA (P<0.05) in Xinjiang Brown cattle; and (iv) haplotypes composed of the above-mentioned 11 SNPs were constructed. The SCS of cattle with Hap5 was lower than that of Hap3 (P<0.05). This suggests that Hap5 might play an important role in sub-mastitis resistance in Xinjiang Brown cattle., (© 2011 The Authors. Animal Science Journal © 2011 Japanese Society of Animal Science.)

The effect of treatment with eprinomectin on milk yield, milk composition and somatic cell counts (SCCs) was studied in 105 dairy cows located on seven farms in South Tyrol, Italy. On each farm, half of the animals were treated with eprinomectin and the other half were used as an untreated control group. Three test day records per animal were obtained before treatment (days -117, -75 and -33) and another three test day records were obtained after treatment (days 22, 62 and 131). Test day records comprised milk yield, milk composition, SCC and days in milk. On the day of treatment, blood samples and faecal samples were taken for parasitological analysis. Cows with positive faecal egg counts yielded less milk. A significant effect of eprinomectin on milk yield was observed after treatment and was most pronounced on the second and the third test days after treatment (+1.90 kg [P=0.002] and +2.63 kg [P<0.001], respectively). Furthermore, a significant decrease in SCC was observed on the second test day after treatment.

To determine milk yield and composition, total microbic count (TMC) and somatic cell count (SCC) of browsing goats throughout the first lactation, 100 goats of Sarda breed, equally distributed in four flocks (F1, F2, F3 and F4), were selected. They were exclusively fed pasture and hand-milked once daily. Individual milk samples and daily milk yield were taken from each goat at monthly intervals, from March to July. Milk samples were analyzed for: total protein, fat, lactose, urea, freezing point (FP), pH, TMC and SCC. The data was subjected to analysis of variance and to correlation matrix. On the whole, in all the flocks, milk yield showed the highest production in April and May. Fat content increased (P < 0.01) throughout the lactation. Protein content showed the lowest value (P < 0.01) in June (4.15%). Urea and pH values were fluctuating. FP was lower (P < 0.01) at the start of lactation (-0.562 Hortvet degrees). TMC log10 values were low, considering the hand milking and inadequacy of facilities on the farms. SCC increased (P < 0.01) throughout the lactation and, on the whole, SCC and TMC were not correlated., (© 2010 The Authors; Journal compilation © 2010 Japanese Society of Animal Science.)

A total of 2538 quarter milk samples from 638 lactating dairy cows from 47 farms in the canton of Bern, Switzerland, were investigated for streptococci. A novel, simple and inexpensive laboratory method was used for the differentiation of Streptococcus species, and a risk factor analysis was carried out. The prevalence in the quarter milk samples was 0.2 per cent for Streptococcus agalactiae, 1.3 per cent for Streptococcus uberis, 1.3 per cent for Streptococcus dysgalactiae, 0.1 per cent for Enterococcus species and 2.9 per cent for minor Streptococcus species (designated Streptococcus-Lactococcus-Enterococcus [SLE] group). Based on the somatic cell count (SCC), S uberis and S dysgalactiae were classified as 'major' pathogens and the bacteria in the SLE group as 'minor' pathogens. For S uberis, S dysgalactiae and bacteria in the SLE group, the most significant risk factor was an intramammary infection (IMI) of a neighbouring quarter by the same pathogen. Other significant risk factors for S uberis infection were a positive California Mastitis Test (CMT) result and a SCC of more than 100,000 cells/ml. Significant risk factors for IMI with S dysgalactiae were a positive CMT result, teat injury and palpable abnormalities in the udder. Infection with bacteria in the SLE group was significantly associated with a SCC of more than 100,000 cells/ml, a lactation number of more than 2, the right rear quarter (as the location of infection) and a positive CMT result.

A total of 757 milk samples were taken from 57 farms throughout Wales as part of a mastitis investigation. Problem cows were selected using the most recent milk recording or, if these data were unavailable, the whole herd was assessed using the California mastitis test (CMT) and scored on a scale of 0 to 3. The affected quarter was identified using CMT. The results of the bacteriology were grouped into negative and positive culture. All contaminated samples were excluded. The CMT recording at the time of sampling, the infection status of the cow (new, first, chronic or repeat) as defined by national milk records and the number of quarters infected at the time of sampling were identified for negative and positive samples. Selecting quarters with a higher CMT score increased the likelihood of positive culture. Culturing from a sample with a CMT score of 3 was over three times more likely to yield a positive culture than a score of 1 (odds ratio [OR] 3.74, 95 per cent confidence interval [CI] 1.41 to 9.97) and 1.7 times more likely to yield a positive culture than a score of 2 (OR 1.70, 95 per cent CI 1.16 to 2.50). Culturing from a score of 2 was not statistically different from a score of 1 in terms of the likelihood that a positive culture would be identified (OR 2.20, 95 per cent CI 0.83 to 5.93). There was no statistically significant difference in the proportion of negative samples when comparing chronic, repeat, new and first infections or number of quarters infected.

A survey of dairy farmers assessed whether they were aware of the potential production and economic benefits of adopting and implementing efficient practices to control somatic cell count (SCC), and whether providing them with additional information on projected economic losses on a regular basis might motivate them to implement enhanced control programmes. In-depth interviews revealed that the majority of the dairy farmers perceived cow-specific and herd-specific projected losses due to elevated SCCs, as not very relevant to them. Farmers considered that SCC was already monitored regularly at individual cow level, which provided them with adequate information to support decision-making. The farmers justified their actions with regard to SCC control in terms of their intention to manage the problem, and their belief in whether their efforts would be successful. Actions were rationalised in a specific context comprising the intertwined notions of intentions and efficacy beliefs.

The California mastitis test (CMT) is used on farms to identify subclinical mastitis by an indirect estimation of the somatic cell count (SCC) in milk. Four commercially available detergents were compared with a bespoke cmt fluid for their ability to detect milk samples with a scc above 200,000 cells/ml; differences between the interpretation of the results of the tests by eight operators were also investigated. The sensitivity and specificity of the test were affected by the type of detergent, and by the operators' interpretations. When used by the most sensitive operator, suitably diluted Fairy Liquid performed almost identically to cmt fluid in identifying milk samples with more than 200,000 cells/ml. The average sensitivities achieved by the eight operators for detecting this threshold were 82 per cent for Fairy Liquid and 84 per cent for cmt fluid, and the specificities were 93 and 91 per cent respectively. The other detergents contained less anionic surfactants and were less sensitive but similarly specific.

All the quarters in the cows with high somatic cell counts in 10 herds were treated at drying off with either 600 mg cloxacillin or 600 mg cloxacillin and 4 g of an internal teat sealant containing 65 per cent bismuth subnitrate. The quarters were sampled daily for bacteriological tests for the three days before drying off and twice after calving to establish whether they were infected. Clinical mastitis cases were monitored from drying off until 100 days after calving. The odds of a quarter being bacteriologically negative after calving or developing clinical mastitis in the first 100 days after calving were investigated by multilevel logistic regression. The quarters treated with the internal sealant and cloxacillin were significantly more likely to be bacteriologically negative in the immediate period after calving and were significantly less likely to suffer clinical mastitis during the first 100 days after calving than the quarters treated with cloxacillin alone. There was more variation between cows than between herds in the underlying risk of an infection after calving, but there was more variation between herds than between cows in the underlying risk of clinical mastitis during the 100 days after calving.

Mastitis is a problem in the sheep industry, and its incidence varies widely with how it is recorded, the breed of sheep and the farm. Virtually all the published information about the genetics of mastitis refers to dairy breeds of cattle and sheep, and there is little information for meat sheep breeds. Many dairy breeding programmes worldwide use the somatic cell count (scc) in milk as an indicator of resistance to clinical and subclinical mastitis, but it is difficult to measure in meat sheep breeds. Molecular genetic technologies may therefore be a more practical way to assess susceptibility to mastitis. This paper reviews the genetics of mastitis and considers the opportunities for breeding for resistance to mastitis, with particular reference to sheep. In addition, to investigate the potential economic effects of mastitis in a purebred sheep population, a computer model of flock dynamics was developed. By making a modest set of assumptions about the key farm parameters that influence lowland sheep production, the model showed that breeding for resistance (or other control methods), if it could reduce the risk of contracting mastitis by 10 per cent, would be worth pounds 8.40 per ewe, equivalent annually to pounds 2.7 million for the purebred sector of the Texel breed alone.

An intervention study was carried out on 52 dairy farms in England and Wales to determine whether the implementation of a well-specified mastitis control plan in herds with an incidence of clinical mastitis of more than 35 cases per 100 cows per year would reduce the incidence of clinical mastitis, and also reduce the incidence of increases in the somatic cell counts of individual cows. A clearly defined plan for the diagnosis and control of mastitis was developed by two veterinary specialists from the research literature. The herds were randomly allocated to receive the plan either at the start of the study (intervention herds) or after one year (control herds). Data on mastitis management and the farm environment were collected during farm visits. After one year there was a significant 22 per cent reduction in the proportion of cows affected with clinical mastitis on the intervention farms compared with the control farms. There were also significant reductions of approximately 20 per cent in the incidence of clinical mastitis and in the occurrence of increases in the somatic cell counts of individual cows from below, to above 200,000 cells/ml.

A survey of clinical and subclinical mastitis was carried out on 97 dairy farms in England and Wales, selected at random from members of a national milk recording scheme. The farmers were asked to collect aseptic milk samples from five consecutive cases of clinical mastitis and from five quarters with high somatic cell counts using a defined protocol, and they completed a questionnaire that included information on the cows sampled, the herd and the history of mastitis in the herd. The samples were collected throughout the year. The mean incidence of clinical mastitis was 47 cases per 100 cows per year (estimated from historic farm records) and 71 cases per 100 cows per year (estimated from the samples collected). Streptococcus uberis and Escherichia coli were isolated in pure culture from 23.5 per cent and 19.8 per cent, respectively, of the clinical samples; 26.5 per cent of the clinical samples produced no growth. The most common isolates from the samples with high cell counts were coagulase-negative staphylococci (15 per cent), S uberis (14 per cent) and Corynebacterium species (10 per cent). Staphylococcus aureus and coagulase-positive staphylococci together accounted for 10 per cent of the samples with high somatic cell counts; 39 per cent produced no bacterial growth.

The bacterial causes of subclinical mastitis were determined in samples of milk taken from one half of the udders of 159 goats in three different herds. The mean prevalence of subclinical infection was 33 percent, with prevalences of 26 percent, 39 percent and 42 percent in the three herds. Staphylococcus aureus was isolated from seven (13 percent) of the 53 infected halves, coagulase-negative staphylococci accounted for 47 percent, Corynebacterium species for 31 percent and alpha-haemolytic streptococci for 6 percent of the infected samples. The mean somatic cell count of the uninfected milk samples was 428,000 cells/ml, and 93 percent of uninfected samples had counts less than 1,000,000 cells/ml; the mean cell count of the infected samples was 2,785,000 cells/ml.

A sample of dairy farms in Great Britain with a monthly bulk milk somatic cell count of less than 150,000 cells/ml was enrolled into a 12-month prospective study. At the end of the study, a questionnaire on milking practice and other farm management practices was sent to the 482 farmers who had collected data on the occurrence of mastitis throughout the 12 months. The response rate was 93 per cent. The reported mean incidence of clinical mastitis was 36.9 cases per 100 cow-years. Factors associated with an increase in the incidence of clinical mastitis were cleaning out the straw yard less frequently than every six weeks, more than 5 per cent of cows leaking milk outside the parlour, checking the foremilk, wearing gloves during milking, an average annual milk yield of more than 7000 litres per cow, dipping or spraying teats before milking and keeping milk with a high somatic cell count out of the bulk tank. Factors associated with a decrease in the incidence of clinical mastitis were using a cloth to dry the teats after washing them as part of premilking preparation, using calving boxes for less than 40 per cent of calvings, and using both cubicles and straw yards to house dry cows, as opposed to other housing.

The factors affecting the mortality of cows in Danish dairy herds were investigated by analysing data from 6839 herds. The mean risk of mortality during the first 100 days of lactation was 2.5 per cent. The risk of mortality increased with increasing herd size (odds ratio [OR] 1.05 for an increase in herd size of 50 cows), with the proportion of purchased cows (OR 1.05 for an increase in the proportion of purchased cows of 0.1), and with increases in the somatic cell count (OR 1.16 for an increase in average weighted mean somatic cell count of 100,000 cells/ml). The risk decreased with increasing average milk yield per cow (OR 0.93 for an increase in mean yield per cow-year of 1000 kg). The risk was lower in free-stall barns with deep litter (OR 0.79) than in barns with cubicles (OR 1) or tie-stalls (OR 1.04). Herds with Danish Holstein (OR 1) or Danish Jersey (OR 0.93) cows as the predominant breed had a higher risk of mortality than herds of the Danish red dairy breed (OR 0.67). The risk of mortality was lower in organic herds (OR 1) than in conventional herds (OR 1.17), and lower in herds that grazed pasture during the summer (OR 0.78).

To study the course of clinical mycoplasma mastitis and investigate its potential for persistence, 10(8) colony-forming units (cfu) of an Irish isolate of Mycoplasma bovis was inoculated aseptically into the right fore teat canal of three lactating cows. M bovis rapidly colonised the infected quarters and grew exponentially to more than 10(10) cfu/ml within the first three days, and spread to other quarters of each of the three cows within five to 10 days. After periods of between 24 and 72 hours the infected quarters became distended and sensitive to touch, and their secretions changed from containing visible particles, to a seropurulent exudate, to an aqueous suspension of fine particles which formed a sediment after a sample was collected. M bovis-specific antibody levels increased to varying degrees in all three cows. Subsequently, the concentrations of mycoplasma decreased to less than 10(7) cfu/ml in two of the cows, but remained at more than 10(8) cfu/ml to the end of the lactation of the other cow. Apparently normal milk was secreted by one of the cows within a month of the challenge, and by the other two cows at the start of their next lactation. However, in two of the cows subclinical M bovis infection persisted through the dry periods and into their next lactations.

Cows in a 250-cow Holstein-Friesian herd were allocated at random to be treated with either a homoeopathic nosode or a negative control, both treatments being applied by means of an aerosol spray to the vulval mucous membranes. A total of six treatments were given over a period of three days and milk samples were taken for the determination of somatic cell counts (SCC) on days -3, 3, 7, 9, 14, 21 and 28. Individuals applying the treatments or carrying out the SCC determination were unaware of which animals were receiving which treatment. Owing to the wide natural variations in SCC, the trial had only a 71 per cent possibility of detecting a 30 per cent difference in SCC between the two groups. There were no significant differences between the SCC of the two groups on any sample day, but there were significant variations between the SCC on different days (P=0.003) in both groups.

The accuracy of somatic cell counts in milk samples was investigated in four studies. First, the counts recorded by one milk buyer in one supply over six months ranged from 105,000 to 401,000 cells/ml with no apparent changes in the volume of milk consigned or the level of mastitis in the herd that would explain this wide range. Secondly, the counts in daily samples from one bulk milk supply for 28 days ranged from 84,000 to 282,000 cells/ml, again with no apparent changes in the performance of the herd to explain the wide range. Thirdly, the replicated counts recorded for one sample by three separate laboratories agreed closely; however, when a sample with a high cell count was interspersed then two of the three laboratories reported high cell counts suggestive of 'carry-over' in excess of the 2 per cent 'allowable' Finally, cell count data from three separate laboratories on samples from 21 cows for 33 days revealed problems with the misidentification of samples on the farm in 1 per cent of the samples, and misidentification and mishandling of 1 to 2.6 per cent of the samples in the laboratories. All three laboratories differentiated samples from cows with subclinical and clinical mastitis, but the mean cell count of the uninfected cows varied between the laboratories with one of them recording statistically significantly higher counts over the period.

The inoculation of 2000 colony-forming units of Corynebacterium pseudotuberculosis into one teat canal of each of three cows resulted in severe, chronic, pyogranulomatous mastitis. Within three days the cows had a reduced haematocrit, haemoglobin concentration and red cell count. The anaemia was initially normocytic, normochromic and non-regenerative, and was associated with a brief peak of neutrophilia; a regenerative response became evident two to three weeks later. Clinical signs of mastitis appeared seven to 14 days after the inoculation, with a peak of high fever, more severe anaemia, a second peak of neutrophilia and the complete cessation of milk production from all quarters; extensive and severe pyogranulomatous mastitis developed in the inoculated quarters. No other lesions were detected postmortem, and C pseudotuberculosis was cultured from the affected quarters but not from the supramammary lymph nodes and viscera.

Mastitis was induced experimentally in ewes with Staphylococcus epidermidis, and the concentrations of serum amyloid A (SAA) in milk and serum, and the somatic cell counts and bacteria in the milk were determined for up to 10 weeks in two experiments, each examining five infected and five control ewes. The somatic cell counts peaked eight hours after infection and preceded an increase in SAA in milk. A maximum concentration of 6460 microg/ml SAA was recorded in milk from the infected sheep, compared with a mean concentration of 1.4 microg/ml in the control sheep. The mean peak concentration of SAA in serum (206.8 microg/ml) occurred earlier (one day after infection) than in milk. The serum concentration of SAA in the healthy animals ranged from 0 to 29.4 microg/ml. There was no correlation between the concentrations of SAA in serum and milk.