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1.
Background: The California mastitis test (CMT) and somatic cell count (SCC) are commonly used for diagnosis of subclinical mastitis in cattle. Acute phase proteins (APPs), as alternative biomarkers of mastitis, may increase in concentration in the absence of macroscopic changes in the milk, or may precede the onset of clinical signs. Objective: The aim of this study was to compare the accuracy of APPs measured in milk and in serum with bacterial culture for the diagnosis of bovine subclinical mastitis. Methods: One hundred and seventy‐five Holstein cows were randomly selected from 7 dairy farms. Quarter milk and serum samples were taken from all cows. Milk samples were analyzed using a CMT and SCC, and for haptoglobin (MHp) and amyloid A (MAA) concentrations, and were also submitted for bacterial culture. Serum samples obtained concurrently were analyzed for haptoglobin (SHp) and amyloid A (SAA). Two‐sample Wilcoxon (Mann–Whitney) test was used to compare SCC, MAA, MHp, SAA, and SHp concentrations between culture‐positive and culture‐negative animals. Receiver‐operating characteristic analysis was used to assess the performance of each test using bacterial culture as the reference method. Results: MAA concentration was the most accurate of the 5 tests, with a sensitivity of 90.6% and specificity of 98.3% at concentrations >16.4 mg/L. MAA and MHp had significantly larger areas under the curve than the respective serum proteins, SAA and SHp. Conclusions: The results suggest that measuring haptoglobin and amyloid A in milk is more accurate than serum analysis for the diagnosis of subclinical mastitis in Holstein cows.  相似文献   

2.
The aim of this study was to investigate total antioxidant (TAC), and oxidant capacity (TOC) and nitric oxide (NO) levels in milk of cows with subclinical mastitis. Brown Swiss and Holstein breed cows were screened with California Mastitis Test (CMT) to determine mammary glands with subclinical mastitis. Moreover, somatic cell counts (SCC) were determined electronically in all milk samples. Mammary quarters were classified as healthy (n = 25) or subclinical mastitis (n = 35) based on CMT scores and somatic cell count (SCC: ?200,000/ml or >200,000/ml) in milk. Nitric oxide, TOC and SCC levels were significantly higher (p < 0.001, p < 0.005 and p < 0.001, respectively) in milk from mammary quarters with subclinical mastitis compared to those from healthy mammary quarters. In conclusion, subclinical mastitis results in higher NO concentrations, TOC and SCC, and NO and TOC were positively correlated with SCC. Moreover, alterations in NO levels and TOC in milk could be used as an alternative diagnostic tool to screen for subclinical mastitis.  相似文献   

3.
The concentrations of haptoglobin (Hp) and serum amyloid A (SAA) and the activity of N-acetyl-β-D-glucosaminidase (NAGase) in milk from 234 cows with spontaneous mastitis caused by different pathogens were measured to assess whether they corresponded with the clinical signs of mastitis and whether there were any differences between pathogens. Ninety-eight of the cows had clinical mastitis and 136 had subclinical mastitis. There were statistically significant positive correlations between the concentrations of SAA and Hp and the activity of NAGase. Significant differences in the concentrations of acute phase proteins and NAGase activity were found in milk from cows with mastitis caused by different pathogens. The highest concentrations of Hp and NAGase were found in cases of mastitis caused by Escherichia coli and Arcanobacterium pyogenes, and the lowest concentrations were from cases of mastitis caused by coagulase-negative staphylococci. Very low SAA concentrations were found in milk from the cases caused by A pyogenes, in contrast to cases caused by other major mastitis pathogens. The median concentration of SAA was over 10 times higher in cases of mastitis caused by E coli than in mastitis caused by other pathogens. There were significant differences in the mean Hp concentration and NAGase activity between clinical and subclinical mastitis. In approximately one-third of the samples, the Hp concentration was below the detection limit, potentially compromising the use of Hp as a mastitis marker.  相似文献   

4.
Mastitis is inflammation of mammary gland affecting all the species of domestic animals. Fragments of the mitochondrial genome released from dying cells are considered surrogate markers of mitochondrial injury. We hypothesized that bovine mastitis would be associated with increased cell free mitochondrial DNA (mtDNA) content in serum and milk. Milk and serum samples were collected from sub-clinical mastitic and normal animals. Mastitis was confirmed by California mastitis test and bacterial isolation. Oxidative stress, nitric oxide and inflammatory cytokines were also estimated. Real time polymerase chain reaction was conducted in serum and milk from sub-clinical mastitic animals and compared with healthy animals targeting the mtDNA genes cytochrome b. Mastitis animals showed higher oxidative stress markers and nitric oxide along with higher level of inflammatory cytokines. Cell free mtDNA was significantly higher in serum and milk of mastitic animals comparing to that of healthy control. The higher cell free relative mtDNA content in mastitis animals indicates injury to the mammary epithelial cells and thereby releasing the mtDNA in the milk and blood. This mtDNA may be a bio-marker of oxidative stress and tissue injury in bovine mastitis.  相似文献   

5.
The purpose of the study was to identify the potential risk factors for subclinical mastitis (SCM) in lactating dairy cows in Bangladesh. A cross-sectional study was carried out on randomly selected 212 smallholder dairy farms of Sadar upazilas of Rangpur, Mymensingh, and Satkhira districts of Bangladesh during January to October 2011. The direct interview using a structured questionnaire and physical examination of the cows were done to collect data on 15 variables. Milk samples collected from study cows were subjected to California Mastitis Test (CMT). The diagnosis of SCM was based on the results of CMT and physical examination of udder and milk. The bivariable followed by multivariable analysis was done using SPSS 17.0. Of the total cows examined, 20.2 % had subclinical mastitis. In bivariable analysis, eight risk factors were identified. However, in the final model of multivariable analysis, four potential risk factors were identified. These were history of previous clinical mastitis (odds ratio (OR) 10.51, p?<?0.001), pendulous type of udder (OR 2.26, p?=?0.008), no grass feeding (OR 1.84, p?=?0.039), and body condition score (BCS) 2.5 or less (OR 7.25, p?=?0.054). Four different factors were significantly associated with the occurrence of subclinical mastitis, which need to be considered in the control of the disease. However, particular emphasis should be given on grass feeding and BCS because these traits can be modified or improved to allow prevention of SCM.  相似文献   

6.
Serum amyloid A proteins (SAA) are very sensitive acute phase proteins, displaying multiple isoforms in plasma and different body fluids. They are currently under investigation as biomarkers of diseases. The aim of the present study was to compare the concentration and isoform expression of SAA in serum and milk of cows with bacteriologically negative milk (control group) and naturally occurring Staphylococcus aureus (S. aureus) subclinical mastitis (subclinical mastitis group). Somatic cell count (SCC) and bacteriological analyses were performed to establish the control and subclinical mastitis group. SAA concentration was evaluated using a commercial ELISA kit, while expression of different isoforms (serum A-SAA and milk M-SAA3 isoforms) was visualized by denaturing isoelectrical focusing and immunoblotting. The SAA concentrations in sera and milk of cows in the subclinical mastitis group were three and 100 times higher than in those from the control group of cows, respectively. Cows in the subclinical mastitis group had more acidic SAA isoforms in serum with the most prominent one at pI 5.5. This isoform was not detected in sera from the control group. Milk samples in the subclinical mastitis group contained abundant highly alkaline M-SAA3 isoforms and most of the serum isoforms, except for that at pI 5.5. In the subclinical mastitis group SAA isoforms with equivalent pI as serum isoforms accounted for 20% of the total SAA concentration in milk. There were significant differences in the concentrations and isoform patterns of SAA in serum and milk between the control and subclinical mastitis groups of cows. Also, we demonstrated that serum SAA isoforms were not transferred to milk proportion to their plasma content.  相似文献   

7.
The major acute phase proteins (APPs) in cattle are haptoglobin (Hp) and serum amyloid A (SAA), and in swine, are Hp, SAA, C-reactive protein (CRP), and Pig major acute phase protein (Pig-MAP). Many methodologic assays are presently available to measure these parameters, which are still being improved to increase their specificity, sensitivity, user-friendliness, and economic availability. In cattle, the main applications are the diagnosis and monitoring of frequent diseases such as mastitis and metritis in dairy cows and respiratory problems in young calves. In pigs, APPs are useful in the control of bacterial and viral infections, and they may be used at the slaughterhouse to monitor subclinical pathologies and improve food safety. The utility of APP in animal production must not be forgotten; optimization of protocols to improve performance, welfare, and nutrition may benefit from the use of APPs. Other sample types besides serum or plasma have potential uses; APP determination in milk is a powerful tool in the control of mastitis, saliva is a non-invasive sample type, and meat juice is easily obtained at the slaughterhouse. Increasing our knowledge of reference intervals and the influence of variables such as age, breed, sex, and the season is important. Finally, worldwide harmonization and standardization of analytical procedures will help to expand the use of APPs.  相似文献   

8.
The development of pyometra in cows depends largely on the state of local immunity of the uterus. The objective of the study was to evaluate the concentration of the following proinflammatory cytokines: tumour necrosis factor (TNF‐α) and interleukin‐6 (IL‐6); anti‐inflammatory cytokine: interleukin‐10 (IL‐10); and acute‐phase proteins (APPs): haptoglobin (Hp) and serum amyloid A (SAA), in serum and uterine washings in cows with pyometra and healthy animals. The study was performed on 20 cows divided into two groups based on the results of cytological and ultrasonographic tests: a pyometra and a healthy group (10 cows per group). Experimental material consisted of blood serum and uterine washings. The levels of the following cytokines, TNF‐α, IL‐6, IL‐10 and APPs – Hp and SAA, in the study material were determined by ELISA. The results showed that the values of TNF‐α, IL‐6, IL‐10 as well as SAA and Hp were significantly higher in serum of cows with pyometra compared to controls (p < 0.001). The uterine washings had significantly higher levels of IL‐6, IL‐10, and Hp in pyometra cows compared to the control (p < 0.001). Our results indicate that it is possible to monitor the course of pyometra in cows based on the evaluation of the concentration of cytokines and Hp in the serum and uterine washings. Simultaneous evaluation of selected indicators of antagonistic interaction can be helpful in determining the current status of local immunity of the uterus. On this basis, it could be possible to properly select an adjunctive therapy in the form of immunomodulating preparations.  相似文献   

9.
This study established the precision and accuracy of a modified latex agglutination turbidimetric immunoassay (LATIA) reagent, and evaluated the ability of the measurement of serum amyloid A (SAA) compared to haptoglobin and α1-acid glycoprotein, which are acute phase proteins (APPs), for diagnosis of clinical mastitis. Concentrations of APPs in cows with mastitis were significantly higher than those in healthy cow. Only the plasma SAA concentration in cows with clinical mastitis (44.90 mg/l; n=15) was significantly higher than that in those with subclinical mastitis (10.70 mg/l; n=16), enabling their diagnosis in contrast to other APPs. Thus, the SAA assay using a LATIA reagent is useful in assessing mastitis severity due to its higher sensitivity and specificity than other APP assays.  相似文献   

10.
Mastitis remains a major cause of economic losses in dairy herds. It is believed, that the enhancement of natural defense mechanisms in mammary gland may be helpful in the treatment of bovine mastitis. Our study was designed to assess the apoptosis of leukocytes isolated from bovine milk during subclinical staphylococcal mastitis. Milk samples were collected from cows naturally infected with one pathogen--Staphylococcus aureus and from animals with mastitis caused by several pathogens, including S. aureus. It has been determined that the rate of apoptosis was lower in mastitic milk, as compared with control samples, although varied significantly between groups. High percentage of apoptotic cells was detected in milk with high counts of pathogenic bacteria. In all groups the rate of apoptosis was dependent on the bacterial load, although various correlations were identified. Thus, it is postulated, that the rate of apoptosis of somatic cells in mastitic milk is related to the etiology of infection and is determined by the bacterial load.  相似文献   

11.
The aim of the present study was to determine the concentration of serum amyloid A (SAA) and the activity of ceruloplasmin (Cp) in milk from cows with subclinical mastitis caused by different pathogens. Eighty-four milk samples from cows with subclinical mastitis and fourteen milk samples from healthy cows were examined. SAA concentration was determined using the commercial ELISA kit (Tridelta Development Ltd., Greystones, Wicklow, Ireland). Cp activity was assessed spectrophotometrically, using the Rice method. The results reveal that the concentration of SAA (with exception of CNS) and activity of Cp in cow milk can be regarded as markers of subclinical mastitis, irrespective of the microorganism inducing the disease. In conclusion, measurement of SAA and Cp in milk samples could be a useful method in diagnosing subclinical mastitis in cows, but the method should be adapted for field use.  相似文献   

12.
Chronic, subclinical intramammary infection depresses fertility. We previously found that 30% of subclinical mastitic cows exhibit delayed ovulation, low circulating estradiol levels, and delayed luteinizing hormone surge. We examined the function of preovulatory follicles of cows experiencing subclinical mastitis or a past event of acute clinical mastitis. Cows were diagnosed for mastitis by somatic cell count and bacteriological examination. All clinical infections were caused by Escherichia coli, and most subclinical infections were caused by Streptococcus dysgalactiae and coagulase-negative staphylococci. On day 6 of the cycle, cows received PGF2α; 42 h later, follicular fluids and granulosa cells or theca cells were aspirated from preovulatory follicles in vivo or following slaughter, respectively. Overall, follicular estradiol and androstenedione concentrations in the subclinical group (n = 28) were 40% lower (P < 0.05) than those in uninfected cows (n = 24) and lower than in past clinical mastitic cows (n = 9). Distribution analysis revealed a clear divergence among subclinical cows: one-third (9/28) exhibited low follicular estradiol; the other two-thirds had normal levels similar to all uninfected (P < 0.01) and most clinical cows (P < 0.08) that had normal follicular estradiol levels. Subclinical normal-estradiol cows had twofold higher (P < 0.05) circulating estradiol concentrations and sevenfold and fourfold higher (P < 0.05) follicular androstenedione levels and estradiol-to-progesterone ratio, respectively, than subclinical low-estradiol cows. Follicular progesterone level was not affected. Reduced expression (P < 0.05) of LHCGR in theca and granulosa cells, CYP11A1 (mRNA and protein) and CYP17A1 in theca cells, and CYP19A1 in granulosa cells may have contributed to the lower follicular steroid production in the subclinical low-estradiol subgroup. StAR and HSD3B1 in theca cells and FSHR in granulosa cells were not affected. Mastitis did not alter follicular growth dynamics, and no carryover effect of past clinical mastitis on follicular function was detected. These data indicate that a considerable proportion (one-third) of subclinical mastitic cows have abnormal follicular steroidogenesis, which can explain the reproductive failure associated with this disease.  相似文献   

13.
The present study was carried to ascertain the association of various risk factors of mastitis in water buffaloes. The milk samples from buffaloes were collected and screened through California Mastitis Test for the presence of mastitis. In the present study, 15.2 % prevalence of subclinical mastitis was recorded both at the government (13.4 %) and private farms (15.5 %). The chi-square analysis showed significantly higher involvement of the right rear and front quarters. The analysis of variance technique showed significant difference in live body weight, milk yield, teat end to floor distance (P?<?0.001), udder depth, teat length, and teat diameter in mastitic and healthy buffaloes. The frequency analysis also revealed significant difference between various groups including lactation stage, teat and/or udder pathology, teat shape, and udder shape (P?<?0.001). The logistic regression analysis revealed significant positive association of mastitis with milk leakage, live body weight, milk yield, parity, calf suckling, pendulous udder, number of attendants at the farm, dirty hind legs, and udder depth.  相似文献   

14.
Udder inflammations, independently of significant losses in milk yield and alteration of its quality, can negatively affect cows' reproduction efficiency. Mastitis causes changes in many active constituents, both in milk and blood. Pathogenic changes in existing constituents and new active chemical compounds, generated during disease, can affect other organs, particularly the reproductive system and its mechanisms. Fertility disorders in mastitic cows are mostly connected with the activity of cytokines (especially TNF-alpha), cortisol, prostaglandin F2-alpha, reactive oxygen species (ROS), and classic inflammatory mediators. The successful treatment of mastitis as well as protection against udder infections should be considered important methods for prophylaxis of fertility disorders in cows. The first Polish trials indicated that injection of supportive drugs (antioxidants or lysozyme dimer or flunixin meglumine) to intramammarily treated cows can increase fertility in cows with mastitis.  相似文献   

15.
The study was undertaken to determine the aetiology and prevalence of mastitis in hand-milked cows (n = 186) in two major Ethiopian dairies. The California Mastitis Test and culturing for bacteria revealed that 21.5% of the cows were clinically infected and 38.2% had subclinical mastitis. Most mastitis pathogens isolated from milk samples testing positive by the California Mastitis Test were Gram-positive cocci. Staphylococci constituted 57% of the isolates, of which the predominant cause of bovine mastitis was Staphylococcus aureus (40.5%). Other mastitis pathogens isolated include streptococci (16.5%), coliforms (9%) and corynebacteria (5%). Retrospective analysis of farm records indicated that mastitis was the second most important cause of culling and accounted for 27% of the cows removed from these two dairies.  相似文献   

16.
Three hundred fifty one (195 local zebu and 156 Holstein x local zebu crosses) lactating cows of smallholder farms in Bahir Dar 'milk shed' were examined from September 2003 to March 2004 to determine mastitis prevalence, isolate pathogens and identify the role of some potential risk factors. Clinical prevalence was determined through examination of abnormalities of milk, udder or cow. California mastitis test (CMT) was used for determination of subclinical mastitis prevalence. Clinical prevalence at cow level was 3.9% in crossbreds and none in local zebu breeds. Subclinical mastitis at cow level based on CMT was high (34.4%) in crossbreds compared to indigenous zebu (17.9%) (p < 0.05). Quarter subclinical prevalence based on CMT was 17.9% and 4.9% in crossbreds and local zebu, respectively. The pathogens isolated from mastitic milk (CMT positive milk) were coagulase negative staphylococci (CNS), S. aureus, Str. agalactiae, Str. dysgalactiae, Str. uberis, Micrococcus species, C. bovis, A. pyogens, B. cereus, and S. intermedius. Among these, the most frequent isolates were CNS (50%), S. aureus (19%), Str. agalactiae (8%) and Str. dysgalactiae (7%). Among potential risk factors considered, stage of lactation, parity and breed were found to affect the occurrence of mastitis significantly (p < 0.05).  相似文献   

17.
New tools are needed to detect chronic sub-clinical mastitis, especially in automatic milking systems. Haptoglobin and serum amyloid A (SAA) are the two most sensitive bovine acute phase proteins, and their concentrations increase in milk from cows with clinical mastitis and in milk from cows with experimentally induced chronic sub-clinical Staphylococcus aureus mastitis. The aim of this study was to further evaluate the potential for haptoglobin and SAA in milk as indicators of chronic sub-clinical mastitis. Quarter milk samples were collected from 41 cows with a mean composite milk somatic cell count (CSCC) above 300,000 cells/mL during at least two months prior to sampling. Quarter milk samples were also taken from eleven cows with a mean CSCC below 80,000 cells/mL during at least two previous months. These samples were analysed for haptoglobin, SAA, adenosine triphosphate (ATP) activity and bacterial growth. The samples were grouped according to their ATP, haptoglobin and SAA status. ATP+ samples had ATP > 2 x 10(-10) mol/mL, Hp+ and SAA+ samples had detectable levels of haptoglobin (> or = 0.3 mg/L) and SAA (> or = 0.9 mg/L), respectively. In udder quarter samples from healthy cows, 42 out of 44 samples belonged to the ATP-Hp-SAA- group. Among cows with chronic sub-clinical mastitis, the ATP+Hp+SAA+ group contained 66 out of 164 samples while 44 samples belonged to the ATP+Hp-SAA- group. Detectable levels of haptoglobin and SAA were found in 92 and 80 samples, respectively. Growth of udder pathogens was detected in 28 samples and Staphylococcus aureus was the most common bacteria. In conclusion, haptoglobin and SAA concentrations below the detection limit were considered as good indicators of healthy udder quarters. A substantial variation in haptoglobin and SAA concentrations in milk was observed in udder quarters with chronic sub-clinical mastitis.  相似文献   

18.
The aim of the present study was to characterise the serum amyloid A (SAA) response to intramammary inoculation of Escherichia coli and to examine the distribution of hepatically and extrahepatically produced SAA isoforms in plasma and milk from cows with mastitis. Milk and plasma SAA concentrations were determined before and after experimental induction of E. coli mastitis in six dairy cows. The milk SAA response was characterised by low or undetectable levels before inoculation, very rapid and large increases in concentration after inoculation, and rapid decline towards baseline levels after resolution of disease. In plasma from cows with experimentally induced E. coli mastitis, four hepatically derived SAA isoforms with apparent isoelectric point (pI) values of 5.8, 6.2, 6.8 and 7.4 were demonstrated by denaturing isoelectric focusing. In milk three highly alkaline isoforms with apparent pI values above 9.3 appeared 12 h post-inoculation. These isoforms were not present in any of the plasma samples, and it therefore seems likely that they were locally produced, tissue-specific isoforms. At 24-36 h post-inoculation one or more acidic isoforms corresponding to those found in plasma appeared in the milk samples. The isoforms demonstrated in plasma from cows with E. coli mastitis were also present in serum obtained from three cows with clinical Streptococcus uberis mastitis. In conclusion, experimentally induced E. coli mastitis is accompanied by a prominent SAA response. The results of the present study indicate that SAA accumulation in mastitic milk is the result of both local synthesis of SAA and of hepatically derived SAA gaining access to the milk due to increased permeability of the blood-milk barrier.  相似文献   

19.
奶牛隐性乳房炎的检测及其治疗方法   总被引：1，自引：1，他引：0  
奶牛乳房炎是困扰奶牛养殖业发展最严重的疾病之一,特别是隐性乳房炎,临床症状不明显,不仅导致奶牛产乳量和乳质量的降低,还可能传染给其他奶牛,导致疾病的扩散.因此,只有对乳汁进行检测,通过其生理、生化等指标变化来判断乳房炎的发生情况.为减少该病的发生、提高乳质量,本文总结了奶牛隐性乳房炎的微生物学检验、细胞学检查、酶学指标...  相似文献   

20.
The effect of naturally acquired bacterial infection of the bovine udder on the activity of 5'-thyroxine monodeiodinase (5'-MD), and on the concentrations of the pro-inflammatory cytokines interleukin (IL)-6 and tumour necrosis factor (TNF)-alpha in milk, from healthy (control) and inflamed quarters, was determined. The diagnostic procedure included history and clinical examination of the udder, macroscopic evaluation of secretions, the Californian Mastitis Test, determination of somatic cell counts and bacteriological examination of milk. It has been found that the milk triiodothyronine (T3) content and the 5'-MD activity from inflamed quarters were decreased when compared with controls. The decrease in the milk T3 from subclinical mastitic quarters was manifested when somatic cell counts were > 10(6) ml(-1). TNF-alpha was on average 2-fold higher in infected milk, and the concentration of IL-6 was unchanged. These results suggest that the decreased T3 content in mammary secretions during naturally occurring mastitis is associated with the severity of inflammation, increased TNF-alpha concentration and impaired enzymatic activity of 5'-MD.  相似文献   

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