急性期蛋白及其在动物应激过程中的作用

检测急性期蛋白( acute phase proteins,APPs)水平可作为估计动物机体对疾病先天性免疫反应的一种方法.本文主要对APPs的结构和功能,及其在动物应激中的调节机制等方面进行综述,并指出APPs经进一步研究,有望成为评价动物健康的一项新技术.     

Acute phase proteins in cattle and swine: A review

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.     

Acute phase proteins in dogs and cats: current knowledge and future perspectives

The acute phase response is a nonspecific inflammatory reaction of the host that occurs shortly after any tissue injury. The response includes changes in the concentration of plasma proteins called acute phase proteins (APPs), some of which decrease in concentration (negative APPs), such as albumin or transferrin, and others of which increase in concentration (positive APPs), such as C-reactive protein, serum amyloid A, haptoglobin, alpha-1-acid glycoprotein, and ceruloplasmin. Most positive APPs are glycoproteins synthesized mainly by hepatocytes upon stimulation by proinflammatory cytokines and released into the bloodstream. The acute phase response and clinical application of monitoring APPs in dogs and cats are reviewed in this article, including biochemical characteristics, assays developed for each individual APP, and preanalytic and analytic factors influencing APP results that should be taken into account for proper and adequate clinical interpretation. In addition, the diagnostic use of APPs and their possible application in monitoring treatment, which can be considered one of the most interesting and promising practical applications of these proteins, will be discussed. Finally, challenges and future developments of APPs in dogs and cats will be considered, because it is expected that new and cheaper automated assays for determination of the main APPs in small animals will contribute to a wider use of these proteins as biomarkers of infection and inflammatory lesions.     

Meat juice: An alternative matrix for assessing animal health by measuring acute phase proteins. Correlations of pig-MAP and haptoglobin concentrations in pig meat juice and plasma

Quantification of acute phase proteins (APPs) in blood can be used for monitoring animal health and welfare on farms, and could be also of interest for the detection of diseased animals during the meat inspection process. However serum or plasma is not always available for end-point analysis at slaughter. Meat juice might provide an adequate, alternative matrix that can be easily obtained for post-mortem analysis at abattoirs. The concentrations of pig Major Acute phase Protein (pig-MAP) and haptoglobin, two of the main APPs in pigs, were determined in approximately 300 paired samples of plasma and meat juice from the diaphragm (pars costalis), obtained after freezing and thawing the muscle. APPs concentrations in meat juice were closely correlated to those in plasma (r = 0.695 for haptoglobin, r = 0.858 for pig-MAP, p < 0.001). These results open new possibilities for the assessment of animal health in pig production, with implications for food safety and meat quality.     

Pharmacological treatments and risks for the food chain

Veterinarians play a pivotal role in public health control, in particular in the management of risks deriving from pharmacological treatments of food-producing animals. Veterinary medicinal products can represent a risk for animal health and welfare (side effects, decreased efficacy), for farmers and practitioners administering the drug, for consumers of food of animal origin (presence of residues, occurrence of antibiotic resistance) and for the environment. According to pending European guidelines, risk management starts from marketing authorisation that must be based on risk evaluation and can be denied when the risk/benefit ratio is not favourable considering the advantages for animal health and welfare and for safety of consumers. Veterinarians can prevent and control risks by using correct pharmacological criteria to choose and administer medicinal products and undertaking risk-based inspection of residues of drugs in food of animal origin. Moreover, a major tool for veterinarians to prevent and control drug-borne risk is “pharmacovigilance”. Risks for the environment are usually assessed during the pre-marketing approval process, however veterinarians, as risk managers, should educate farmers about correct drug handling and disposal, and periodically verify that suggested measures are applied.     

Pharmacological treatments and risks for the food chain

Veterinarians play a pivotal role in public health control, in particular in the management of risks deriving from pharmacological treatments of food-producing animals. Veterinary medicinal products can represent a risk for animal health and welfare (side effects, decreased efficacy), for farmers and practitioners administering the drug, for consumers of food of animal origin (presence of residues, occurrence of antibiotic resistance) and for the environment. According to pending European guidelines, risk management starts from marketing authorisation that must be based on risk evaluation and can be denied when the risk/benefit ratio is not favourable considering the advantages for animal health and welfare and for safety of consumers. Veterinarians can prevent and control risks by using correct pharmacological criteria to choose and administer medicinal products and undertaking risk-based inspection of residues of drugs in food of animal origin. Moreover, a major tool for veterinarians to prevent and control drug-borne risk is "pharmacovigilance". Risks for the environment are usually assessed during the pre-marketing approval process, however veterinarians, as risk managers, should educate farmers about correct drug handling and disposal, and periodically verify that suggested measures are applied.     

Acute phase protein response in goats

Acute phase proteins (APPs) are important diagnostic indicators of inflammatory disturbances in animals. The objectives of the current study were to validate analytical methods for measuring haptoglobin (Hp), serum amyloid A (SAA), acid soluble glycoprotein (ASG), fibrinogen, and albumin concentrations in goats and to evaluate their response to an inflammatory stimulus in this species. Intra- and interassay coefficients of variation (CVs) were in the range 0.07-9.31% and 1.83-12.68%, respectively, for all APPs and showed good precision. All assays determined APPs in a linear manner at different sample dilutions with high correlation coefficients with the exception of fibrinogen, which was measured by the heat precipitation method. Subcutaneous injection of turpentine oil induced an increase in Hp, SAA, ASG, and fibrinogen serum concentrations and a decrease in albumin concentration.     

Acute phase proteins as indicators of calf herd health

The potential for using acute phase proteins (APPs) in the assessment of herd health was studied by examining the levels of serum haptoglobin, serum amyloid A (SAA) and plasma fibrinogen in relation to clinical findings and leukocyte counts in calves. Two groups of calves from conventional dairy farms were studied. The animals were examined 10 times during the first six weeks after introduction into a new environment. Haptoglobin, SAA and fibrinogen were analysed and weight gain, disease symptoms and treatments were recorded. Analysis of antibodies against viral infections was performed. An acute phase reaction (APR) score was established at each sampling by combining the APP results and total leukocyte counts. The health status differed between the two groups, although no manipulation of health had been performed, except that the group with a higher incidence of disease had a concurrent experimental infection with lungworm as part of another study. In the group with a higher incidence of disease, the mean weight gain was significantly lower, and the number of sampling days with elevated serum concentrations of APPs, and the mean maximum concentrations of haptoglobin and fibrinogen were significantly higher compared to the healthier group. The APR score was significantly higher at days 4 and 8 of the study in the group with a higher incidence of disease. The results indicate that measurement of APPs could be a useful tool for evaluation of health in calf herds.     

Porcine acute phase protein concentrations in different diseases in field conditions

Five acute phase proteins (APPs) [C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin (Hp), pig-MAP and albumin] were measured in pigs with naturally occurring infections by porcine reproductive and respiratory syndrome virus (PRRSV), Aujeszky's disease virus (ADV), porcine circovirus type 2 (PCV2) and Mycoplasma hyopneumoniae, and in animals with tail and ear bites, arthritis and other acute inflammatory processes. Healthy specific pathogen-free (SPF) pigs were used as controls. In PRRSV-infected pigs, all APPs with the exception of pig-MAP exhibited significant changes compared with controls. In animals affected with ADV only Hp presented changes of statistical significance, whereas pigs with PCV2 showed marked modifications in all APPs tested. Animals affected with Mycoplasmosis showed concentrations of all positive APPs significantly above levels obtained in SPF pigs, though albumin concentrations did not differ from controls. Finally, all APPs studied showed substantial changes in pigs with acute inflammation. The results indicated that an acute phase response was developed in the different diseases studied, this response being higher in animals with clinical signs and concurrent bacterial processes. Haptoglobin would be the APP that better reflects pathological states; however, to get more complete and valuable information it might be advisable to perform APPs profiles including another APP, such as CRP or SAA.     

欧盟禁用饲料药物添加剂的历史和法规

欧盟考虑到抗生素耐药性的发展和抗生素耐药基因可能从动物到人微生物的转移,决定从2006年1月1日起禁止抗生素作为生长促进剂,甚至还建议禁止在动物饲料中全面使用所有的抗菌剂。收集和整理欧盟饲料药物添加剂的禁用清单和法规,介绍欧盟禁用饲料药物添加剂的历史及其对动物健康的影响,以期为中国饲料药物添加剂的使用和管理提供借鉴和参考。     

Changes in haptoglobin, C-reactive protein and pig-MAP during a housing period following long distance transport in swine

The aim of this study was to investigate the effects of a housing period following long distance transport on haptoglobin (Hp), C-reactive protein (CRP) and pig major acute phase protein (pig-MAP) in swine. After transportation, 80 gilts were allotted to group A, B, C, or D. Blood samples were collected on arrival and 28 days later; additional samples were collected from Group C on day 14, and from Group D on days 3, 5 and 14. Acute phase proteins (APPs) in Group A were significantly lower on day 28 than on day 1; the opposite occurred in Group B because of a tail biting episode. In Group C, values remained elevated on day 14 and showed a reduction on day 28; in Group D elevated levels detected on day 14 were preceded by a decrease from days 1 to 5. The results indicate that stressors associated with transportation and new accommodation can cause an increase in APPs that could be useful indicators of welfare during transport and routine management.     

Analytical validation of commercially available methods for acute phase proteins quantification in pigs

The aim of this study was to validate commercially available methods for porcine haptoglobin (Hp), C-reactive protein (CRP), serum amyloid A (SAA) and major acute phase protein (Pig-MAP) determinations. Intra and inter assay coefficients of variation (CVs) were lower than 20% in all cases with exception of inter assay CVs for CRP and Pig-MAP assays with samples of low acute phase proteins concentration, and for SAA assay at any acute phase proteins concentration. All methods showed good linearity and detection limits were low enough to detect APPs levels in healthy animals. Hp and SAA were very affected by haemolysis. Lipaemia influenced mainly on SAA determination. Over 15-fold increase was observed in CRP and SAA concentrations after artificially induced inflammation by a single subcutaneous dose of turpentine, whereas Hp and Pig-MAP increased less than 5-fold.     

Diagnostic utility of measuring serum amyloid A with a latex agglutination turbidimetric immunoassay in bovine mastitis: Comparison with haptoglobin and alpha 1 acid glycoprotein

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.     

Evaluation of goose serum amyloid a acute phase response by enzyme-linked immunosorbent assay

Serum amyloid A (SAA) is of interest as the circulating precursor of amyloid A protein, the fibrillar component of AA (secondary) amyloid deposits, and also as an extremely sensitive and rapid major acute phase protein. Serum concentrations of acute phase proteins (APPs) provide valuable information about the diagnosis and prognosis of various diseases, and thus the relevance of APPs for monitoring the health status of domestic animals is widely accepted. More importantly, the measurement of SAA concentration assists in assessing the prognosis in secondary amyloidosis, which is a common disease of geese, affecting an increasing number of animals. In the present study we introduce a highly sensitive goose-specific ELISA method for measuring SAA concentration in goose serum or plasma samples. Samples were taken from geese of the Landes Grey and Hungarian White breeds, which were stimulated for an acute phase reaction by administration of a commercially available fowl cholera vaccine containing inactivated Pasteurella multocida. Strong and characteristically rapid acute phase responses were measured in both breeds, peaking at approximately 24 h after inoculation. The maximum SAA concentration was 1200 microg/ml. At 72 h postinoculation, the concentrations returned to pre-inoculation values. There was significantly (p = 0.004) less intense response in the control groups; however, a very mild increase of SAA levels was detected due to the stress inevitably caused by the sampling procedure.     

Acute phase proteins,proinflammatory cytokines and oxidative stress biomarkers in sheep,goats and she-camels with Coxiella burnetii infection-induced abortion

Acute phase proteins (APPs) and oxidative stress are helpful markers in diagnosis of several infectious diseases. APPs, proinflammatory cytokines and oxidative stress markers were evaluated for their role in the diagnosis of naturally acquired Coxiella burnetii (Q fever) associated with abortion in sheep, goats and she-camels. Blood, aborted materials and vaginal swabs were collected from mixed herds in the Eastern Province of Saudi Arabia. Antioxidant biomarkers showed significant decline in cases of abortion compared to control animals at delivery time. The correlation between disease status and all parameters ranged from moderate to high. The APPs, cytokines and the oxidative stress marker malondialdehyde (MDA) displayed a high degree of distinction between aborted sheep and goat and normal delivered animals (AUC > 0.90). However, only MDA showed a high degree of differentiation (AUC > 0.90) between aborted she-camels and normal delivered controls. In conclusion, results from our study allow us to recommend using APPs, cytokines and oxidative stress markers as an additional tool for diagnosis of naturally occurring C. burnetii infection in sheep, goats and she-camels. However, it does not replace standard procedures for detection of C. burnetii.     

Acute phase proteins in cats: Diagnostic and prognostic role,future directions,and analytical challenges

While clinical studies on acute phase proteins (APPs) have significantly increased in the last decade, and most commercial labs are now offering major APPs in their biochemical profiles, APP testing has not been widely adopted by veterinary clinical pathologists and veterinarians. Measurement of APP concentration is a useful marker for detecting the presence or absence of inflammation in cats with various diseases. APPs can also be reliably measured in different biological fluids (eg, effusions and urine) to improve their diagnostic utility. Measurement of APPs can be extremely beneficial in cats with feline infectious peritonitis (FIP) to discriminate between FIP and non-FIP cats with similar clinical presentations. Additional benefits come from multiple and sequential measurements of APPs, particularly in the assessment of therapeutic efficacy. APPs are more sensitive than WBC counts for early detection of inflammation and to demonstrate an early remission or recurrence of the diseases. Given the potential utility of APPs, more studies are warranted, with a particular focus on the applications of APPs to guide the length of antimicrobial therapies, as suggested by the antimicrobial stewardship policy. New inflammatory markers have been discovered in human medicine, with a higher specificity for distinguishing between septic versus nonseptic inflammatory diseases. It is desirable that these new markers be investigated in veterinary medicine, to further test the power of APPs in diagnostic setting.     

Temporal changes in serum concentrations of acute phase proteins in newborn dairy calves

Age-dependent changes in blood concentrations of four bovine acute phase proteins (APPs), serum amyloid A (SAA), lipopolysaccharide binding protein (LBP), haptoglobin (Hp) and alpha(1)-acid glycoprotein (AGP), were examined using two groups of newborn dairy calves. APP concentrations were monitored for either 3 weeks (Group A, n=13) or 2 months (Group B, n=13) after birth. Blood was collected at day 0-1 (Group A only), day 3 and then once or twice weekly until the end of the study. The possible transfer of colostral SAA as a source of SAA in the offspring was investigated by determining SAA isoforms in the serum of calves and in colostrum of their dams. Serum concentrations of all four APPs were high after birth, and concentrations showed a gradual decrease during the first 3 weeks of life. The lowest concentrations were at 21 days of age, after which concentrations stabilized. The calves synthesized adult SAA isotypes, and circulating SAA was not derived from colostrum. The results indicated that post-partum elevation of APPs is associated with the birth process and/or factors in colostrum and not necessarily with disease-related processes. This stresses the importance of considering a calf's age when interpreting APP concentrations in serum.     

Analysis of the acute phase responses of Serum Amyloid A, Haptoglobin and Type 1 Interferon in cattle experimentally infected with foot-and-mouth disease virus serotype O

ABSTRACT: A series of challenge experiments were performed in order to investigate the acute phase responses to foot-and-mouth disease virus (FMDV) infection in cattle and possible implications for the development of persistently infected "carriers". The host response to infection was investigated through measurements of the concentrations of the acute phase proteins (APPs) serum amyloid A (SAA) and haptoglobin (HP), as well as the bioactivity of type 1 interferon (IFN) in serum of infected animals. Results were based on measurements from a total of 36 infected animals of which 24 were kept for observational periods exceeding 28 days in order to determine the carrier-status of individual animals. The systemic host response to FMDV in infected animals was evaluated in comparison to similar measurements in sera from 6 mock-inoculated control animals.There was a significant increase in serum concentrations of both APPs and type 1 IFN in infected animals coinciding with the onset of viremia and clinical disease. The measured parameters declined to baseline levels within 21 days after inoculation, indicating that there was no systemically measurable inflammatory reaction related to the carrier state of FMD. There was a statistically significant difference in the HP response between carriers and non-carriers with a lower response in the animals that subsequently developed into FMDV carriers. It was concluded that the induction of SAA, HP and type 1 IFN in serum can be used as markers of acute infection by FMDV in cattle.     

Antimicrobial activity of red-tailed phascogale (Phascogale calura) serum

Antimicrobial substances in serum include circulating complement proteins and acute phase proteins (APPs). We identified gene sequences for APPs, haptoglobin (Hp), C-reactive protein (CRP) and serum amyloid A (SAA) in marsupial genomes. Hp and SAA levels were measured in red-tailed phascogale (Phascogale calura) sera using commercially available assays. Hp levels were higher in males than females, while SAA levels suggest the phascogales used in this study were healthy. Serum was co-cultured with four bacterial species. Bacterial growth was inhibited after incubation at 37 °C, however effectiveness differed with bacteria and incubation time. The least amount of bacterial growth was noticed after introduction to K. pneumoniae, and most when introduced to P. aeruginosa. Despite marsupials not having mature immune tissues at birth, and unable to mount specific immune responses, this study suggests other immune strategies, such as APPs in serum likely aid marsupials in their defence against pathogens.     

Strategies for reduced antibiotic usage in dairy cattle farms

The need for antibiotic treatments in dairy cattle farms can be reduced by a combined intervention scheme based on: (1) timely clinical inspections, (2) the assessment of animal-based welfare parameters, and (3) the use of predictive laboratory tests. These can provide greater insight into environmental adaptation of dairy cows and define animals at risk of contracting disease. In the long-term, an improved disease control justifies the adoption of such a combined strategy. Many antibiotic treatments for chronic disease cases are often not justified with a cost/benefit analysis, because the repeated drug administration does not give rise to the expected outcome in terms of animal health. In particular, compared with untreated cases, antibiotics may not lead to greater cure rates for some forms of mastitis. Lastly, a substantial reduction of antibiotic usage in dairy farms can be achieved through the proper use of immunomodulators, aimed at increasing immunocompetence and disease resistance of cows.