Identification of subpopulations of bovine mammary-gland phagocytes and evaluation of sensitivity and specificity of morphologic and functional indicators of bovine mastitis

The number and function of bovine mammary-gland phagocytes were assessed in 8 lactating cows, each tested at least twice within an 8-mo period (total number of observations, 20). Macrophages and polymorphonuclear (PMN) cells were evaluated by conventional cytology, flow cytometry, fluorescent microscopy, and somatic-cell count (SCC). Phagocytosis was evaluated from the uptake of fluorescent beads and expressed as median fluorescence intensity (MFI). Two major subpopulations of phagocytes, of low or high MFI (LFI or HFI), were observed, and there were up to 4 sub-subpopulations within the HFI subpopulation of both macrophages and PMN cells. Fluorescent microscopy identified phagocytes containing up to 4 beads per cell. Cows showing ≤ 72.3% phagocytes by cytology were regarded as non-mastitic (11 observations), and those showing ≥ 80.7% phagocytes were considered to be mastitic (8 observations). Phagocyte MFI was negatively associated with mastitis; that is, the higher the MFI, the lower the SCC. The percentage of HFI PMN cells was the only indicator of mastitis with 100% sensitivity and specificity. Thus, bovine mammary-gland phagocytes consist of several subpopulations of different phagocytic ability, whose assessment more adequately predicts bovine mastitis than do morphologic indicators.     

Normal somatic cell count and subclinical mastitis in Murrah buffaloes

This study was conducted to investigate the normal somatic cell count (SCC) and to define subclinical mastitis in Murrah buffaloes. Data were collected from 60 clinically normal buffaloes stationed at five farms of Chitwan Nepal and Buffalo Research Center, Hissar, India. Somatic cell count was measured using the Newman-Lampert staining technique. The upper limit of SCC was determined >or=200 000/ml of milk based on the mean +/- 2SD of a total SCC. Abnormal data of the SCC was repeatedly removed, which lie beyond the values of more than mean + 2SD until all the data come to lie within (mean + 2SD). Averages of SCC of right front and right hind quarters were significantly higher than left front and left hind quarters. Nearly 94% of California mastitis test (CMT) negative quarters were having somatic cells >or=200 000/ml. The mean SCC of CMT positive quarter was significantly higher (P < 0.01) than CMT negative quarters. Subclinical mastitis was diagnosed on the basis of samples with SCCs >or=200 000/ml with positive bacterial cultures. Subclinical mastitis was found in 21.7% buffaloes and 8% of the quarter foremilk samples. Neutrophil counts were significantly higher in subclinical mastitis milk.     

Incidence and types of clinical mastitis in dairy herds with high and low somatic cell counts

Eighteen dairy herds were studied, 12 with a 12-month Dairy Herd Improvement Association herd mean somatic cell count (SCC) less than or equal to 150,000 cells/ml (low SCC) and 6 with a 12-month mean SCC greater than 700,000 cells/ml (high SCC). At the outset of the study, quarter samples for bacteriologic culture were collected (in duplicate) from all quarters of all lactating cows (whole herd culture). Subsequently, quarter milk samples for culture from all cows with clinical mastitis were collected for a period of 6 months. In the herds with low SCC, results of whole herd culture revealed low prevalence of intramammary infection attributable to all major pathogens (less than 4% of all quarters). Prevalence of infection with Streptococcus agalactiae (22.2% of all quarters) and Staphylococcus aureus (6.6% of all quarters) was significantly (P less than 0.05) higher in the herds with high SCC. Mean incidence of clinical mastitis in the herds with low SCC was 4.23 infections/100 cows/month (range, 0.42 to 10.25 infections). In the herds with high SCC, mean incidence was 2.91 infections/100 cows/month (range, 1.33 to 3.92 infections). In the herds with low SCC, infection type, as mean percentage of total clinically infected quarters sampled for culture/herd, was 0.0%, 2.2%, 12.3%, 43.5%, and 28.6% for Str agalactiae, S aureus, streptococci other than Str agalactiae, coliforms, and organisms not isolated, respectively. Respective percentages for the herds with high SCC were 41.5%, 18.3%, 12.6%, 8.0%, and 8.8%. During the study period (from April through January), incidence of clinical mastitis and clinical mastitis caused by coliform bacteria were highest in July and August for herds with low SCC.(ABSTRACT TRUNCATED AT 250 WORDS)     

A method to estimate the somatic cell count of milk from a mastitic quarter using composite somatic cell count.

The purpose of the study was to estimate the extent to which one quarter could be inflamed relative to the other three quarters of a cow, given that knowledge only of the composite milk somatic cell count (SCC) was available. An algebraic relationship, which incorporated the parameters of composite milk SCC, production loss associated with composite milk SSC, SCC of milk from an inflamed quarter, and SCC from three non-inflamed quarters, was used to derive hypothetical estimates of the SCC in one inflamed quarter. A simple case was considered in which one quarter was mastitic, the SCC in milk of the noninflamed quarters was equal, and there was no production compensation by the noninflamed quarters. Previously published estimates were used for production loss associated with composite milk SCC. For moderate composite milk SCC (300,000-500,000 cells/mL) and low-to-moderate SCC in milk of noninflamed quarters (100,000 cells/mL), the SCC of milk from one inflamed quarter was predicted to be very high, ranging from about 1.26 million (log10 SCC = 6.1) to 6.3 million (log10 SCC = 6.8) cells/mL, and compatible with signs of clinical mastitis. These results suggest that in order to screen cows with clinical mastitis in only one quarter, composite milk SCC should be considerably lower than values presented previously by other investigators.     

Development of a Staphylococcus aureus vaccine against mastitis in dairy cows. I. Challenge trials

A vaccine composed of three field isolates of Staphylococcus aureus (S. aureus) derived from cases of mastitis in cows was developed. The vaccine was administered to nine uninfected cows while 10 other cows were used as controls. All cows were challenged with a highly virulent S. aureus strain administered into two quarters of each cow. Quarters were tested for clinical signs, secretion of S. aureus, and somatic cell count (SCC). No systemic effects were observed in any of the cows, vaccinated or control. Vaccinated cows had 70% protection from infection compared with fewer than 10% in the controls. Moreover, all quarters challenged in the vaccinated cows, regardless of whether they were successfully infected or not with S. aureus, exhibited very mild inflammatory reactions, identified by their low SCCs (<100,000).     

Prevalence of subclinical mastitis and isolated udder pathogens in dairy cows in Southern Vietnam

Dairy production is not traditional in Vietnam. The farmers have little practical knowledge and udder health control is generally lacking. In order to give the farmers appropriate advice, knowledge about the distribution of udder pathogens is crucial. The aim of the study was to investigate the prevalence of sub-clinical mastitis and to identify udder pathogens isolated from smallholder dairy herds in Southern Vietnam. Twenty farms with a herd somatic cell count (SCC) ranging from low (≤400?×?10³?cells/mL) to high (>400?×?10³?cells/mL) were randomly selected. Milk samples were collected from 458 quarters of 115 clinically healthy cows. SCC was analyzed on farm by a portable cell counter. Bacteriological samples were taken using Mastistrip^© cassettes and sent to Sweden for examination. For all herds the mean herd SCC was 632?×?10³/mL milk. The prevalence of subclinical mastitis at quarter SCC basis was 63.2 % and at cow basis 88.6 %. Only 40 % of all cows were bacteriologically negative in all quarters. Streptococcus agalactiae was the most commonly found bacteria species, isolated from 96 of the 458 quarter samples, in 13 of the 20 farms. The results indicate pronounced subclinical mastitis problems among the dairy cows in this region mainly due to infections with S. agalactiae. The high prevalence of this highly contagious pathogen is probably attributable to the generally poor milking hygiene and low awareness of proper measures to prevent occurrence and spread of udder infections. A strict, targeted action program for the herds in this area is required in order to lower the prevalence of subclinical mastitis.     

Quarter milk somatic cell count in infected dairy cows: a meta-analysis

The aim of this paper was to evaluate the effects associated with intramammary infection (IMI) by a bacterium or a group of bacteria (Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, coliforms, Staphylococci other than S. aureus, and Corynebacterium bovis) on the somatic cell count (SCC) in quarter milk of dairy cows. Papers selected for analysis had to provide SCC values associated with the natural infection in quarters by different bacteria. Sampling for measurement of SCC and determination of the infection had to be done on the same day. Only papers published in English or in French after 1971 were considered. Twenty-one papers fulfilled the selection criteria. The animals sampled, the measurement techniques for SCC and the bacteriological identification, as well as the definition of the infection, all differed widely among the selected studies. The meta-analysis method was used to estimate both the mean SCC (arithmetic and geometric) value and the average increase on SCC of each type of infection. The geometric mean SCC in bacteriologically negative quarters was 68 000 c/mL. In case of IMI, the retained SCC was 357 000, 857 000, 547 000, 1 024 000, 1 151 000, 138 000 and 105 000 c/mL in quarters infected by Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, coliforms, staphylococci other than S. aureus and Corynebacterium bovis, respectively. The variation factors that could influence these SCC values and the bacteriological results are discussed.     

Efficacy of intramuscular treatment of beef cows with oxytetracycline to reduce mastitis and to increase calf growth

Spring-calving multiparous Angus x Hereford cows were used to determine the efficacy of intramuscular treatment with oxytetracycline to reduce the incidence of mastitis-causing bacteria, decrease milk somatic cell counts (SCC), and increase calf growth. During 2 yr, milk samples were collected from each quarter from a total of 319 cows at 8 to 14 d after calving and at weaning, to determine the presence of bacteria and SCC. A California mastitis test (CMT) was performed on milk from each quarter of each cow at the initial sample collection. Cows with a CMT score of 1, 2, or 3 in at least one quarter, were randomly assigned to receive either an intramuscular injection of oxytetracycline (n = 63) or the control vehicle (n = 60), and cows with a CMT score of 0 or trace in all four quarters were not treated (n = 196). Calf weights were determined at birth, early lactation, and weaning. The number of somatic cells in milk and the percentage of quarters that were infected increased as CMT score increased (P < 0.01). The presence of mastitis-causing bacteria at calving increased (P < 0.05) the incidence of infection at weaning. The presence of mastitis-causing bacteria at weaning was associated with increased SCC for quarters and average SCC for cows (P < 0.01). Average SCC per cow at weaning increased (P < 0.05) as the number of infected quarters per cow increased. Treatment did not alter (P > 0.10) the percentage of cows or quarters infected with mastitis-causing bacteria or SCC of cows or quarters at weaning. Average SCC per cow was negatively correlated (P < 0.05) with calf weights at early lactation, but not with weaning weights of calves. Treatment did not influence (P > 0.10) calf weights at early lactation or at weaning. Cows with one or more dry quarters after calving had calves that weighed less at early lactation and weaning than cows with four functional quarters (P < 0.01). Intramuscular oxytetracycline treatment of beef cows that had CMT scores of 1 or greater after calving did not reduce intramammary infection rates or increase calf weights at weaning.     

The use of Markov chain Monte Carlo for analysis of correlated binary data: patterns of somatic cells in milk and the risk of clinical mastitis in dairy cows

Two analytical approaches were used to investigate the relationship between somatic cell concentrations in monthly quarter milk samples and subsequent, naturally occurring clinical mastitis in three dairy herds. Firstly, cows with clinical mastitis were selected and a conventional matched analysis was used to compare affected and unaffected quarters of the same cow. The second analysis included all cows, and in order to overcome potential bias associated with the correlation structure, a hierarchical Bayesian generalised linear mixed model was specified. A Markov chain Monte Carlo (MCMC) approach, that is Gibbs sampling, was used to estimate parameters.

The results of both the matched analysis and the hierarchical modelling suggested that quarters with a somatic cell count (SCC) in the range 41,000–100,000 cells/ml had a lower risk of clinical mastitis during the next month than quarters <41,000 cell/ml. Quarters with an SCC >200,000 cells/ml were at the greatest risk of clinical mastitis in the next month. There was a reduced risk of clinical mastitis between 1 and 2 months later in quarters with an SCC of 81,000–150,000 cells/ml compared with quarters below this level. The hierarchical modelling analysis identified a further reduced risk of clinical mastitis between 2 and 3 months later in quarters with an SCC 61,000–150,000 cells/ml, compared to other quarters.

We conclude that low concentrations of somatic cells in milk are associated with increased risk of clinical mastitis, and that high concentrations are indicative of pre-existing immunological mobilisation against infection. The variation in risk between quarters of affected cows suggests that local quarter immunological events, rather than solely whole cow factors, have an important influence on the risk of clinical mastitis. MCMC proved a useful tool for estimating parameters in a hierarchical Bernoulli model. Model construction and an approach to assessing goodness of model fit are described.     

Quarter Milking for Improved Detection of Increased SCC

The aim of the present study was to investigate whether milk composition and milk yield are changed in relation to a moderate increase in milk somatic cell count (SCC) in separate udder quarters. During a period of 13 weeks, 4158 bulk quarter milk samples from 68 cows were collected and analysed for milk SCC and milk composition. The sampling was done twice weekly. The cows were in different stages of lactation and in different lactation numbers. For calculations, three groups of cows were formed according to their SCC value. Group 1 cows, where all quarters had an SCC <100,000 cells/ml at all sampling occasions, were considered to be non-affected. Group 2 cows had one udder quarter with an increased SCC >100,000 cells/ml and 1.5-fold higher than the opposite quarter at one sampling occasion. For group 3 cows, the increase in SCC remained for several consecutive sampling occasions. Data from group 1 cows revealed that front and rear quarters were similar when compared with each other. For group 3 cows, the lactose content in milk decreased significantly, simultaneously with the increase in SCC and remained decreased for two sampling occasions after the initial increase in SCC. It was concluded that deviations in lactose content within front and rear quarters, respectively, may be a useful tool for detection of moderately increased SCC in separate udder quarters.     

Milk leucocyte population patterns in bovine udder infection of different aetiology

This study compared the different leucocyte populations in milk from udders infected with different mastitic pathogens and in different stages of infection. Milk samples were collected from quarters free of intramammary infection, acutely infected with Escherichia coli or Staphylococcus aureus and chronically infected with S. aureus, coagulase-negative staphylococci (CNS) or Streptococcus dysgalactiae. Udder bacteriological status was confirmed after three consecutive bacteriological examinations from weekly quarter milk samples. At the time of the trial, milk samples were tested for somatic cell count (SCC) and differential cell count by both light microscopy (LM) and flow cytometry. Monoclonal antibody (mAb) CD11a/CD18 was used in order to differentiate between leucocytes and epithelial cells when tested by flow cytometry. Udder quarters free of intramammary infection had a mean SCC lower than 107 x 10(3) cells/ml in which the epithelial cells were the main cell type followed by polymorphonuclear cells (PMNs), while macrophages and lymphocytes had a lower concentration. Only 56% of the cells were labelled with the mAb anti-CD11a/CD18. In either acute E. coli- or S. aureus-infected quarters, SCC were significantly higher (P < 0.0001) than in samples from the time of inoculation, with over 90% of the cells labelled with the mAb anti-CD11a/CD18. The main cell type was neutrophils. In chronically infected cows, differences in SCC and in leucocyte patterns were found between infecting pathogens as well as between quarters harbouring the same pathogen. In all the chronically infected quarters, SCC was significantly higher (P < 0.05) than in uninfected ones. The distribution of the leucocyte patterns in the quarters infected with S. dysgalactiae did not differ from that in quarters with acute infection with both E. coli and S. aureus. In the cows chronically infected with S. aureus or CNS, the proportion of PMN was higher but not significantly different from quarters free of intramammary infection, while epithelial cells were significantly lower (P < 0.05). The T lymphocytes bearing CD4+ or CD8+ were significantly higher in quarters chronically infected with S. aureus than in quarters free of intramammary infection and in quarters acutely infected with either E. coli or S. aureus. In all samples B cells were negligible.     

The contribution of mammary infections by coagulase-negative staphylococci to the herd bulk milk somatic cell count

Quarter foremilk samples were taken at 2–3 weekly intervals for several years in a experimental herd comprising about 45 cows. The samples were submitted to bacteriological analysis and somatic cell counting. The most prevalent quarter infections from 1982 to 1988 were by coagulase-negative staphylococci (15–20% of all the quarters sampled). Most of these (75.6%) persisted until drying-off Dry cow therapy eliminated 86.5% of these infections. Comparison of udder quarters within cows, involving 775 samples from pairs of non-infected quarters and quarters infected by coagulase-negative staphylococci, yielded geometric means of somatic cell counts of 210 000 and 420 000 cells/ml, respectively. The correlation (r=0.87) between the herd bulk milk somatic cell count (SCC) and its estimation from the quarter milk somatic cell count performed on the same day allowed us to evaluate the contribution of the different categories of quarters, according to their infection status, to the herd bulk milk SCC. Quarters infected by a major pathogen (8.5% of samples) gave rise to 46.6% of the total number of cells, while quarters infected by coagulase-negative staphylococci (17.8% of samples) gave rise to 18.1%. Although coagulase-negative staphylococci represented only a secondary source of somatic cells as compared to major pathogens, they were not a negligible source considering the threshold of 300 000 somatic cells advocated for herd milk of good quality.     

Reliability in somatic cell count measurement of clinical mastitis milk using DeLaval cell counter

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 × 10⁶ 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 × 10⁶ 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.     

Immune cell differentiation in mammary gland tissues and milk of cows chronically infected with Staphylococcus aureus

This study identifies and compares the distribution of mononuclear cells in the mammary gland tissues and milk of healthy and chronically infected with Staphylococcus aureus cows. Somatic cell counts (SCCs) during the 3 months before the study were > 1 x 10(6) cell/ml in the infected quarters and < 1 x 10(5) cell/ml in the infection-free quarters. Immediately after slaughter, samples from the tissues above the gland cistern and supra-mammary lymph node were collected. No histological differences were found between the supra-mammary lymph nodes of the healthy and infected udders, and both appeared normal. In the milk of the healthy infection-free mammary glands, SCC was < 50,000 cells/ml) while epithelial cells were the predominant type. The percentage of CD18+ was low than 45%, of which over three-quarters were polymorphonuclear (PMN), and less than one- quarter were mononuclear cells. The later comprised CD4+ or CD8+ T-lymphocytes, macrophages (Mo) but not B-cells. In the tissues, there were few CD18+ leukocytes, and most of the cells were T-lymphocytes. The number of B-lymphocytes bearing CD21+ was similar to that of CD8+ and were localized in the connective tissue as clusters of 2-5 cells, mainly in areas with no alveoli, or as single cell having a dendritic like form. The number of Mos was negligible. In the milk of the infected glands, SCC exceeded 700,000 cells/ml, of which > 95% were CD18+ positive. The distribution of the leukocytes had two patterns: one presented (> 80%) of PMN cells and a small number of mononuclear cells; the second had less than 50% PMN and many mononuclear cells. The CD8+ cells in these infected sections were observed throughout the mammary epithelial cells (MEc) around the alveoli and in the alveolar lumen (AL). The numbers and the location of CD21+ B-lymphocytes were similar to those in the infection-free mammary glands. The number of CD5+ positive cells was lower than T and B- cells combined and were located throughout the mammary epithelial cells, around the alveoli and within the connective tissue. Mo numbers were high in most of those infected quarters, and were localized around the connective tissue and within the AL.     

Association of endometritis and ovarian follicular cyst with mastitis in dairy cows

The occurrence of multiple metabolic and inflammatory diseases in dairy cows is higher during the periparturient period, which may be triggered by bacterial components, but not a viable bacterium. This study aimed to determine the association of endometritis and ovarian follicular cyst (OFC) with mastitis in dairy cows. Ninety-eight Holstein dairy cows were clinically examined for endometritis and OFC approximately 30–50 days after calving. Blood and milk samples were collected for the determination of milk somatic cell count (SCC); milk interleukin-1β (IL-1β), tumor necrosis factor-α (TNFα), and interleukin-8 (IL-8) concentrations; and plasma haptoglobin (Hp) and lipopolysaccharide-binding protein (LBP) concentrations. Of the 98 dairy cows included in this study, 12 were diagnosed with endometritis and 37 cows were identified as OFC-positive, whereas the remaining 49 cows were healthy (without endometritis or OFC). The average and maximum SCCs and plasma Hp and LBP concentrations were not significantly different between the healthy cows and those with endometritis or OFC. However, when the maximum SCC was classified as <300, 300–1,000, or >1,000 × 10³ cells/ml, the percentage of cows with the maximum SCC <300 × 10³ cells/ml was significantly lower in the endometritis and OFC-positive groups than in the healthy group. These results suggested that cows with endometritis and OFC during the postpartum period exhibit high SCC, indicating that some bacterial components can be transferred between organs.     

Benchmarks for evaluation and comparison of udder health status using monthly individual somatic cell count

The objectives of this study were to propose benchmarks for the interpretation of herd udder health using monthly individual somatic cell counts (SCC) from dairy herds in Quebec, Canada and to evaluate the association of risk factors with intramammary infection (IMI) dynamics relative to these benchmarks. The mean and percentiles of indices related to udder infection status [e.g., proportion of healthy or chronically infected cows, cows cured and new IMI (NIMI) rate] during lactation and over the dry period were calculated using a threshold of ≥ 200 000 cells/mL at test day. Mean NIMI proportion and proportion of cows cured during lactation were 0.11 and 0.27. Benchmarks of 0.70 and 0.03 for healthy and chronically infected cows over the dry period were proposed. Season and herd mean SCC were risk factors influencing IMI dynamics during lactation and over the dry period.     

Prevalence and herd-level risk factors for intramammary infection with coagulase-negative staphylococci in Dutch dairy herds

In this study, the prevalence of intramammary infection (IMI) with coagulase-negative staphylococci (CNS) in The Netherlands was estimated on 49 randomly selected herds with at least 40 lactating cows. In total, 4220 quarter milk samples were collected. The prevalence of CNS IMI in The Netherlands was estimated at 10.8% at quarter level and 34.4% at cow level, making it the most frequently isolated group of pathogens. Fourteen species of CNS were identified; the most frequently isolated species was Staphylococcus chromogenes (30.3%) followed by Staphylococcus epidermidis (12.9%) and Staphylococcus capitis (11.0%). Prevalence of CNS IMI was higher in heifers compared to older cows. Geometric mean quarter SCC of CNS-positive quarters was 109,000 cells/ml, which was approximately twice as high as culture-negative quarters. Quarters infected with S. chromogenes, S. capitis and Staphylococcus xylosus had a higher SCC (P<0.05) than culture-negative quarters, while quarters that were culture-positive for S. epidermidis and Staphylococcus hyicus tended to have a higher SCC than culture-negative quarters. An increased prevalence of CNS IMI was associated with the herd-level variables source of drinking water not being tap water, housing of dry cows in one group instead of multiple groups, measurement of cow SCC every month, udder health monitoring by the veterinarian, pasturing during outdoor season, percentage of stalls contaminated with milk, and BMSCC>250,000 cells/ml. Although a causal relation between these factors and prevalence of CNS is not proven and for some factors not even likely, knowledge of the associations found may be helpful when approaching CNS problems on dairy farms.     

松针多糖对小鼠腹腔巨噬细胞免疫调节作用的研究

本试验旨在研究松针多糖对正常状态及脂多糖(LPS)刺激状态下小鼠腹腔巨噬细胞的免疫调节作用。试验采用不同浓度的松针多糖作用于正常的和经LPS刺激的小鼠腹腔巨噬细胞,设空白对照组(加入100μL RPMI-1640培养基)、阳性对照组(加入100μL终浓度为5μg/m L的LPS)、松针多糖组(分别加入100μL 25、50、100、200μg/m L的松针多糖)和LPS+松针多糖组(加入与松针多糖组相同浓度的松针多糖和终浓度为5μg/m L的LPS,液体终体积为200μL)。噻唑蓝(MTT)比色法检测细胞活力,中性红吞噬试验检测巨噬细胞吞噬能力,Griess法检测一氧化氮(NO)的分泌量,酶联免疫吸附测定(ELISA)法检测巨噬细胞培养上清液中白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)的分泌量。结果表明:1)对空白对照组相比,各松针多糖组巨噬细胞相对增殖率均显著或极显著提高(P0.05或P0.01),50、100和200μg/m L松针多糖组巨噬细胞中性红吞噬率显著或极显著提高(P0.05或P0.01),各LPS+松针多糖组巨噬细胞中性红吞噬率均极显著提高(P0.01),200μg/m L松针多糖组巨噬细胞NO分泌量极显著提高(P0.01),50、100和200μg/m L松针多糖组巨噬细胞TNF-α和IL-1β分泌量显著或极显著提高(P0.05或P0.01),50、100和200μg/m L松针多糖组巨噬细胞IL-10分泌量极显著降低(P0.01)。2)与阳性对照组相比,各松针多糖组巨噬细胞相对增殖率均极显著降低(P0.01),100和200μg/m L LPS+松针多糖组巨噬细胞中性红吞噬率极显著升高(P0.01),50、100和200μg/m L LPS+松针多糖组巨噬细胞NO分泌量极显著提高(P0.01),各LPS+松针多糖组巨噬细胞TNF-α分泌量显著或极显著提高(P0.05或P0.01),50、100和200μg/m L LPS+松针多糖组巨噬细胞IL-1β分泌量显著或极显著提高(P0.05或P0.01),100和200μg/m L LPS+松针多糖组巨噬细胞IL-10分泌量极显著降低(P0.01)。由此可见,松针多糖通过发挥其促炎作用调节巨噬细胞的免疫功能,进而增强机体抗疾病的能力。     

The association between quarter somatic-cell counts and clinical mastitis in three British dairy herds

The association between quarter somatic-cell counts (QSCCs) of milk and the risk of clinical mastitis (CM) was investigated in a 1-year study on three dairy herds in Somerset, UK. The three herds had 95-130 milking cows and an annual mean bulk milk somatic-cell count (BMSCC) of <150 x 10(3)cells/ml. The farms were visited every 4-6 weeks at morning milking when quarter-milk samples were collected. The farmers recorded all cases of CM and were trained to collect sterile milk samples from affected quarters, before treatment for bacteriology.The three herds had CM incidence rates of 25.4, 55.2, and 67.6 quarter-cases per 100 cow-years. Escherichia coli and Streptococcus uberis were cultured from approximately 50% of cases. QSCC was categorised and the risk of CM occurring in the month after the QSCC was examined using multilevel models to account for the correlated nature of the dependent data. Three models were developed: one for all cases of CM, one for those caused by coliforms and one for those caused by S. uberis. When all cases of CM were considered, quarters with somatic-cell count (SCC) 21-100 x 10(3)cells/ml had reduced odds (OR=0.60, P=0.06) and quarters with SCC >200 x 10(3)cells/ml has over three time the odds (OR=3.7, P<0.01) of CM compared with QSCC 1-20 x 10(3)cells/ml. When only coliform CM were investigated, quarters with SCC 6-200 x 10(3)cells/ml had reduced odds of coliform CM (OR=0.47, P=0.04) compared with QSCC 1-5 x 10(3)cells/ml, and SCC >200 x 10(3)cells/ml were not significantly different from the baseline. Finally, when S. uberis CM were investigated, quarters with SCC >200 x 10(3)cells/ml had more than three times the odds of S. uberis CM compared with QSCC 1-20 x 10(3)cells/ml (OR=3.73, P<0.01). QSCC <21 x 10(3) and >200 x 10(3)cells/ml are associated with increased odds of CM in the following 4-6 weeks; this association may be pathogen specific.     

Effects of antibiotics on phagocyte recruitment, function, and morphology in the bovine mammary gland during the early nonlactating period

The effects of 2 antibiotic preparations administered intramammarily on phagocyte recruitment, function, and morphology were evaluated at the beginning of the nonlactating period. Twelve cows with no clinical or microbiologic evidence of mastitis were assigned to 1 of 2 treatment groups. At the end of lactation, 1 of the antibiotic preparations was infused in a fore- and hind quarter of each cow; the remaining quarters were untreated controls. One group was given benzathine cephapirin; the second group was given sodium novobiocin. Secretion samples were collected from 1 treated and 1 control quarter at 16 hours, and from the remaining 2 quarters at 64 hours after treatment. Total and differential somatic cell counts were determined, and morphology of mammary polymorphonuclear neutrophils (PMN) and macrophages was observed by transmission electron microscopy. In vitro ingestion and killing of Staphylococcus aureus by mammary PMN and macrophages were assessed by fluorescent microscopy, using acridine orange stain. Cells resident in a fixed volume of secretion were incubated with a known concentration of S aureus. Total cell and PMN concentrations were higher in treated than in control quarters. Neutrophils were the predominant cell type in both treated and control quarters over the sampling period. As measured in this study, in vitro ingestion and killing of S aureus by individual PMN from treated quarters was reduced. Antibiotic treatment also increased the proportion of morphologically abnormal phagocytes. There were significant correlations among PMN ingestion, killing, and morphology. However, increased PMN concentrations tended to compensate for the reduced phagocytic function of individual cells. Therefore, efficacy of antibiotic treatment of nonlactating cows may depend, at least in part, on increased PMN concentration, which may tend to compensate for reduced phagocytic function. Compared with PMN, macrophages appeared to have only a minor role in phagocytosis of bacteria.