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1.
A cross-over study was conducted to investigate the effect of intramammarily infused lipopolysaccharide (LPS) on the acute phase reaction in early (EL) and in late (LL) lactation. Nine cows received intramammary injections of 100 microg of Escherichia coli 0111:B4 LPS during EL and LL. The severity of each cows systemic and local signs and change in milk appearance were recorded and scored throughout the experiment. Systemic and local signs were found to be more serious in EL cows. Tumor necrosis factor alpha (TNF alpha) was detected in milk but not in serum. Serum amyloid A (SAA) concentrations increased both in serum and in milk. The milk TNF alpha concentrations peaked at 8 h post-challenge (PC). SAA concentrations started to increase at 8 h PC, and peak concentrations were seen at 32 and 48 h PC in milk and serum, respectively. The milk TNF alpha and SAA seemed to be correlated, being on average higher in EL. Serum SAA concentration was not correlated with milk TNF alpha or SAA, nor with the severity of local or systemic signs, but was correlated with changes in milk appearance.  相似文献   

2.
The concentrations of the two acute phase proteins, serum amyloid A and haptoglobin, in serum and milk were compared in 10 cows with clinical mastitis, 11 cows with extramammary inflammatory conditions and 10 clinically healthy control cows. The concentrations of both acute phase proteins were higher in the serum and milk of the cows with mastitis than in the cows in the other two groups. Four of the cows with extramammary inflammatory conditions had serum amyloid A concentrations in serum above 100 microg/ml, but negligible concentrations in milk, indicating that a pathogen must be present in the mammary gland for serum amyloid A to accumulate in milk. The acute phase protein concentrations in milk increased significantly with increasing somatic cell count, suggesting that they may be indicators of the severity of an infection.  相似文献   

3.
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   

4.
The aim of the study was to evaluate the concentrations of amyloid A in serum (SAA) and in milk (MAA) of cows with mastitis caused by Streptococcus agalactiae, Streptococcus dysgalactiae and Streptococcus uberis and healthy cows. The blood and milk samples were obtained from Holstein-Friesian cows with clinical signs of mastitis from two tie-stall housing systems herds in the Lublin region in Poland. A total of 80 milk and serum samples from 30 cows with mastitis and 10 healthy cows were selected for study. In the quarter milk samples from cows with mastitis Streptococcus strains were isolated: Strep. agalactiae (7 cows), Strep. dysgalactiae (9 cows) and Strep. uberis (14 cows). The present study indicates that amyloid A concentration was significantly higher in milk of cows with mastitis compared to control cows (1134.25 ng/mL and 324.50 ng/mL, P < 0.001). The highest concentration of amyloid A was found in milk of cows with mastitis caused by Strep. agalactiae and Strep. uberis whereas lowest in the milk of cows with mastitis caused by Strep. dysgalactiae (3882.50 ng/mL, 2587.75 ng/mL and 812.00 ng/mL, respectively). No statistically significant difference in amyloid A concentration in serum was revealed between all unhealthy cows and control group (2140.00 ng/mL and 2510.00 ng/mL, P > 0.05). There was also no statistically significant difference between the level of amyloid A in serum and in milk of cows with mastitis caused by Strep. agalactiae and Strep. uberis. Whereas, in the case of Strep. dysgalactiae, like in the group of healthy cows, the level of amyloid A was significantly higher in serum compared to this in milk (2100 ng/mL and 812.00 ng/mL, P < 0.001; 2510.00 ng/mL and 324.50 ng/mL, P < 0.001; respectively).  相似文献   

5.
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   

6.
Acute phase proteins in serum and milk from dairy cows with clinical mastitis   总被引：17，自引：0，他引：17  
The serum concentrations of haptoglobin, serum amyloid A and alpha1 acid glycoprotein were determined in serum collected from healthy dairy cows and cows with clinical mastitis, graded as mild (clots in milk) or moderate (clots in milk and visible signs of inflammation in the mammary gland/s) to assess their relative diagnostic value in detecting the disease. The concentrations of haptoglobin and serum amyloid A were also measured in milk collected from infected and uninfected quarters. The concentrations of haptoglobin and serum amyloid A were higher in the serum and milk from the cows with mild or moderate mastitis. The diagnostic value of haptoglobin in differentiating between healthy animals and those with mastitis gave sensitivities and specificities of 82 per cent and 94 per cent respectively with serum and 86 per cent and 100 per cent with milk. The diagnostic value of serum amyloid A in differentiating between healthy animals and those with mastitis gave sensitivities and specificities of 83 per cent and 90 per cent with serum and 93 per cent and 100 per cent with milk. The diagnostic value of serum alpha1 acid glycoprotein in differentiating between healthy animals and those with mastitis gave sensitivities and specificities of 62 per cent and 91 per cent.  相似文献   

7.
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   

8.
An experimental infection model was developed to study host response to intramammary infection in cows caused by Staphylococcus chromogenes. CNS intramammary infections have become very common in modern dairy herds, and they can remain persistent in the mammary gland. More information would be needed about the pathophysiology of CNS mastitis, and an experimental mastitis model is a means for this research. Six primiparous Holstein-Friesian cows were challenged with S. chromogenes 4 weeks after calving. One udder quarter of each cow was inoculated with 2.1 x 10(6)cfu of S. chromogenes. All cows became infected and clinical signs were mild. Milk production of the challenged quarter decreased on average by 16.3% during 7 days post-challenge. Cows eliminated bacteria in a few days, except for one cow which developed persistent mastitis. Milk indicators of inflammation, SCC and N-acetyl-beta-D-glucosaminidase (NAGase) returned to normal within a week. Milk NAGase activity increased moderately, which reflects minor tissue damage in the udder. Concentrations of serum amyloid A (SAA) and milk amyloid A (MAA) were both elevated at 12h PC. MAA was affected by the milking times, and was at its highest before the morning milking. In our experimental model, systemic acute phase protein response with SAA occurred as an on-off type reaction. In conclusion, this experimental model could be used to study host response in CNS mastitis caused by the main CNS species and also for comparison of the host response in a mild intramammary infection and in more severe mastitis models.  相似文献   

9.

Background

Acute phase proteins haptoglobin (Hp), serum amyloid A (SAA) and lipopolysaccharide binding protein (LBP) have suggested to be suitable inflammatory markers for bovine mastitis. The aim of the study was to investigate acute phase markers along with clinical parameters in two consecutive intramammary challenges with Escherichia coli and to evaluate the possible carry-over effect when same animals are used in an experimental model.

Methods

Mastitis was induced with a dose of 1500 cfu of E. coli in one quarter of six cows and inoculation repeated in another quarter after an interval of 14 days. Concentrations of acute phase proteins haptoglobin (Hp), serum amyloid A (SAA) and lipopolysaccharide binding protein (LBP) were determined in serum and milk.

Results

In both challenges all cows became infected and developed clinical mastitis within 12 hours of inoculation. Clinical disease and acute phase response was generally milder in the second challenge. Concentrations of SAA in milk started to increase 12 hours after inoculation and peaked at 60 hours after the first challenge and at 44 hours after the second challenge. Concentrations of SAA in serum increased more slowly and peaked at the same times as in milk; concentrations in serum were about one third of those in milk. Hp started to increase in milk similarly and peaked at 36–44 hours. In serum, the concentration of Hp peaked at 60–68 hours and was twice as high as in milk. LBP concentrations in milk and serum started to increase after 12 hours and peaked at 36 hours, being higher in milk. The concentrations of acute phase proteins in serum and milk in the E. coli infection model were much higher than those recorded in experiments using Gram-positive pathogens, indicating the severe inflammation induced by E. coli.

Conclusion

Acute phase proteins would be useful parameters as mastitis indicators and to assess the severity of mastitis. If repeated experimental intramammary induction of the same animals with E. coli is used in cross-over studies, the interval between challenges should be longer than 2 weeks, due to the carry-over effect from the first infection.  相似文献   

10.
Serum amyloid A (SAA) and haptoglobin (Hp) levels were determined in 25 cows suffering from amyloidosis. SAA levels in cows with amyloidosis ranged between < 0.3 and 225.8 microg/ml, with a median level of 105.1 microg/ml, and Hp levels ranged between < 20 and 1860 microg/ml, with a median level of 950 microg/ml. These levels were significantly higher than the levels observed in healthy cows (SAA levels ranged from < 0.3 to 13.5 microg/ml, with median of 1.4 microg/ml, and Hp levels were undetectable in all cases), but were not significantly different from the levels observed in control cows with chronic inflammation. There was a significant correlation between SAA and Hp levels in cows with chronic inflammation , but not in cows with amyloidosis. It was concluded that the serum SAA levels in cows with amyloidosis might be changed by some factor other than inflammation.  相似文献   

11.
BACKGROUND: Haptoglobin and serum amyloid A are major acute phase proteins in cattle. Dairy cattle often develop pathologic conditions in the peripartum period; acute phase proteins may be useful in their diagnosis. OBJECTIVES: The purpose of this study was to compare the accuracy of serum haptoglobin (Hp) and serum amyloid A (SAA) concentrations with clinical health status for diagnosing disease during the peripartum period in dairy cattle. METHODS: Dairy cows from 4 herds were evaluated every 15 days over a 6-month period. Health status was determined by thorough clinical examination. Haptoglobin and SAA concentrations were measured in serum using validated methods and the results were classified as positive or negative based on defined cutoff points. Disease prevalence, sensitivity, and specificity were compared using clinical examination as the gold standard. RESULTS: A total of 1896 samples from 158 cows were analyzed. Significant increases in mean Hp and SAA concentrations were observed in the week following parturition in both primiparous and multiparous cows, although high interindividual variability was observed. Both Hp and SAA had low sensitivity but higher specificity in determining disease status compared with clinical examination. Increased concentrations of Hp and SAA were found in <10% of samples from clinically healthy cows, except in the week after parturition. CONCLUSIONS: Haptoglobin and serum amyloid A should be used with caution as markers of inflammation in the week after calving. Poor sensitivity in other postpartum periods could be related to the higher incidence of chronic (vs acute) inflammation. Haptoglobin may be appropriate for routine screening, but further work needs to be done to assess its value as an indicator of herd health.  相似文献   

12.
The concentrations of serum amyloid A (SAA) in 4 cows given Escherichia coli endotoxin as an acute-phase stimulant were quantitatively evaluated by use of an indirect micro-ELISA method and compared with other clinical hematologic values. Serum amyloid A concentration changed minimally after intradermal infection of endotoxin. The concentration of SAA was increased 5 hours after IV injection of endotoxin, with maximal concentration after 17 to 20 hours. The increase in SAA concentration coincided with decreasing serum Zn and Fe concentrations; however, with decreasing serum Zn and Fe concentrations; however, Zn and Fe concentrations appeared to be restored when SAA concentration was still maximal. It was concluded that the SAA response of cattle is comparable with that of other species and can be used for monitoring the activity of clinical inflammation and tissue injury.  相似文献   

13.
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   

14.
The objective of this study was to determine the effect of evaporative cooling and dietary supplemental Zn source on blood metabolites, insulin and mineral concentrations, and milk mineral concentrations following intramammary lipopolysaccharide (LPS) infusion. Seventy-two multiparous Holstein cows were assigned to one of four treatments with a 2 × 2 factorial arrangement. Treatments included two environments: with or without evaporative cooling using fans and misters over the freestall and feedbunk, and two dietary sources of supplemental Zn: 75 mg/kg of dry matter (DM) supplied by Zn hydroxychloride (inorganic Zn; IOZ) or Zn hydroxychloride (35 mg of Zn/kg of DM) + Zn–Met complex (ZMC; 40 mg of Zn/kg of DM). A subset of cows (n = 16; 263 ± 63 d in milk) was infused with 10 μg of LPS or a saline control in the left or right rear quarters on day 34 of the environmental treatment. Individual milk samples collected from LPS-infused quarters at −4, 0, 6, 12, 24, 48, 72, 96, and 144 h relative to infusion were analyzed for minerals. Blood samples were collected at the same time with an additional sample collected at 3 h post-infusion to analyze glucose, nonesterified fatty acids (NEFA), insulin, and minerals. Cooling by time interactions (P ≤ 0.07) were observed for plasma glucose, NEFA, and serum insulin. Compared with cooled cows, non-cooled cows had lower concentrations of plasma glucose except at 3 h following intramammary LPS infusion, greater serum insulin at 3 and 12 h, and lower plasma NEFA at 24 and 48 h after infusion. Relative to cooled cows, non-cooled cows tended (P = 0.07) to have lower serum K concentration and had lower (P < 0.01) serum Zn 6 h following infusion (cooling by time interaction: P < 0.01). Relative to ZMC cows, IOZ cows had greater (P ≤ 0.09) concentrations of plasma Se, skim milk Na and Se, and skim milk Na to K ratio. Regardless of treatment, intramammary LPS infusion reduced (P < 0.01) serum or plasma concentrations of Ca, Mg, Zn, Fe, and Se, but increased (P < 0.01) their concentration in skim milk. In conclusion, deprivation of cooling resulted in more rapid and prolonged insulin release and influenced the systemic and mammary mineral metabolism during mammary inflammation induced by LPS of lactating dairy cows. Dietary supplementation of Zn–Met complex reduced blood and milk Se concentrations compared with cows fed Zn from an inorganic source.  相似文献   

15.
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.  相似文献   

16.
OBJECTIVE: To determine the anti-inflammatory effects of glycyrrhizin (GL) in lactating cows with mastitis attributable to naturally occurring infection with coagulase-negative staphylococci (CNS). ANIMALS: 12 lactating Holstein cows with mastitis attributable to infection with CNS and 2 healthy cows without mastitis. PROCEDURE: Clinical signs, number of bacteria in milk, somatic cell count (SCC) in milk, concentrations of alpha-lactalbumin and lactoferrin in milk, and concentration of histamine in milk were investigated before and after intramammary infusion of GL (6 cows) or antimicrobials (6 cows). Glands of 2 healthy cows were infused with staphylococcal enterotoxin; milk leukocytes were then harvested and incubated with various doses of GL. RESULTS: In cows infected with CNS that had a low bacterial concentration in milk, infusion of GL alone resulted in significant improvements in swelling, firmness of glands, and number of clots in milk, and it decreased the SCC, but not significantly. Percentage of neutrophils decreased significantly (to < 30%) by 2 days after infusion. Use of lactoferrin as a marker of inflammation in mammary glands revealed a decrease in concentrations, whereas use of alpha-lactalbumin as a marker of recovery for mammary glands revealed significant increases in concentrations in the GL-infused group. Accompanying these anti-inflammatory effects, a decrease in the concentration of histamine in milk was observed in the GL-infused group. Glycyrrhizin decreased histamine production by milk leukocytes in a concentration-dependent manner. CONCLUSIONS AND CLINICAL RELEVANCE: Infusion of GL may regulate intramammary inflammation through modulation of inflammatory mediators such as histamine.  相似文献   

17.
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18.
Indicators of inflammation in the diagnosis of mastitis   总被引：18，自引：0，他引：18  
Mastitis affects the quality of milk and is a potential health risk for the other cows. In a well managed dairy herd, in addition to clinical mastitis, subclinical mastitis should be efficiently detected. Bacteriological sampling is not feasible as a routine test to identify subclinical mastitis, and indirect tests of mastitis are more suitable for selecting cows with intramammary infections for subsequent bacteriological sampling. Mastitis affects the composition of milk, and the degree of changes depends on the infecting agent and the inflammatory response. Indicators of inflammation in the milk which can be determined using rapid, reliable and easy routine techniques, can be used for the early detection of mastitis. The measuring of the somatic cell count in milk is the standard method, but the analysis technique is problematic for routine use in herds. The most promising parameters for monitoring subclinical mastitis are milk N-acetyl-beta-D-glucosaminidase activity, lactose, and electrical conductivity along with some other indicators such as optical and milk flow measurements, preferably with an inter-quarter evaluation included in the test. Acute phase proteins, haptoglobin and serum amyloid A, are also potential candidates for mastitis monitoring. New mastitis detection systems which can be adapted into on-line use are urgently needed, since dairy units are growing bigger and automatic milking systems are being taken into use.  相似文献   

19.
Acute mastitis was induced in lactating cows by intramammary challenge with 10 g of Escherichia coli lipopolysaccharide. The cows were monitored clinically prior to and for 96 hours after challenge. Milk production, complete blood counts, serum enzyme activities and milk indicators of inflammation were evaluated.Endotoxin challenge was in 7 groups of 3 cows each. Within the groups, cows were randomly assigned to 3 intravenous treatments: saline controls, steroid (one dose of dexamethasone at 0.44 mg/kg) and non-steroidal agent (two doses of flunixin meglumine at 1.1 mg/kg, 8 h apart).Anti-inflammatory therapy reduced rectal and mammary gland surface temperatures. Milk production was significantly reduced (p<0.05) in cows treated with dexamethasone. Although dexamethasone treatment produced significant increases (p<0.05) in blood leukocytes and segmented neutrophils, milk somatic cell concentrations were not significantly altered. Flunixin meglumine did not alter milk production or blood or milk leukocytes.  相似文献   

20.
ABSTRACT: Coagulase-negative staphylococci (CNS) are in several countries the most common bacteria isolated in subclinical mastitis. To investigate the innate immune response of cows to infections with two common mastitis-causing CNS species, Staphylococcus epidermidis and Staphylococcus simulans, experimental intramammary infection was induced in eight cows using a crossover design. The milk somatic cell count (SCC), N-acetyl-β-D-glucosaminidase (NAGase) activity, milk amyloid A (MAA), serum amyloid A (SAA) and proinflammatory cytokines interleukin (IL)-1β, IL-8, and tumor necrosis factor α (TNF-α) were determined at several time points before and after challenge. All cows became infected and showed mild to moderate clinical signs of mastitis. The spontaneous elimination rate of the 16 infections was 31.3%, with no difference between species. Infections triggered a local cytokine response in the experimental udder quarters, but cytokines were not detected in the uninfected control quarters or in systemic circulation. The innate local immune response for S. simulans was slightly stronger, with significantly higher concentrations of IL-1β and IL-8. The IL-8 response could be divided into early, delayed, or combined types of response. The CNS species or persistency of infection was not associated with the type of IL-8 response. No significant differences were seen between spontaneously eliminated or persistent infections.  相似文献   

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