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分析猪口腔液中的生物标志物:采集策略和猪龄的影响。

Analysing biomarkers in oral fluid from pigs: influence of collection strategy and age of the pig.

作者信息

Ornelas Mario Andre S, López-Martínez María José, Franco-Martínez Lorena, Cerón José J, Ortín-Bustillo Alba, Rubio Camila Peres, Manzanilla Edgar Garcia

机构信息

Pig Development Department, Teagasc Grassland Research and Innovation Centre, Moorepark, Fermoy, Co. Cork, Ireland.

School of Veterinary Medicine, University College Dublin, Belfield, Dublin, Ireland.

出版信息

Porcine Health Manag. 2023 Aug 30;9(1):39. doi: 10.1186/s40813-023-00333-x.

Abstract

BACKGROUND AND OBJECTIVES

Oral fluid (OF) is an easy-to-collect, inexpensive, fast and non-invasive sample to characterize health and welfare status of the pig. However, further standardisation of the collection methods is needed in order to use it regularly in veterinary practice. Cotton ropes are routinely used to collect OF for pathogen detection but they may not be optimal for biomarker analysis due to sample contamination. This study compared two methods (cotton ropes and sponges) to collect porcine OF for biomarker analysis. A panel of 11 biomarkers of stress, inflammation, sepsis, immunity, redox status and general homeostasis was studied.

MATERIALS AND METHODS

Eighteen farrow-to-finish pig farms were included in the study. In each farm, three (for sponges) or four pens of pigs (for ropes) were sampled at four age categories: the week after weaning (5 weeks), before (11-12 weeks) and after (12-13 weeks) moving to finisher facility and the week before slaughter (22-25 weeks). In total, 288 OF samples were collected with cotton ropes and 216 with sponges and analysed for the biomarkers: cortisol, alpha-amylase, oxytocin (stress), haptoglobin (inflammation), procalcitonin (sepsis), adenosine deaminase, immunoglobulin G (immune system), ferric reducing antioxidant power (redox status), and creatine kinase, lactate dehydrogenase and total protein (general homeostasis). Samples were also scored visually for dirtiness using a score from 1 (clean) to 5 (very dirty).

RESULTS

Rope-collected OF had higher levels of dirtiness (3.7 ± 0.04) compared to sponge-collected OF (2.7 ± 0.15) and had higher values than sponges for cortisol, procalcitonin, oxytocin, haptoglobin, total protein, lactate dehydrogenase and ferric reducing antioxidant power. All biomarkers decreased in value with age. Immunoglobulin G did not perform well for any of the two collection methods.

DISCUSSION AND CONCLUSION

The results showed a clear effect of age on the biomarkers in OF collected with both, sponges or ropes. Sponges provided a cleaner sample than cotton ropes for biomarker analysis. Both methods are easy to apply under the commercial conditions in pig farms although sponges may take more time in early weaner stages. From a practical point of view, sampling with sponges achieved the best combination of reduced sampling time and low contamination.

摘要

背景与目的

口腔液(OF)是一种易于采集、成本低廉、快速且非侵入性的样本,可用于表征猪的健康和福利状况。然而,为了在兽医实践中常规使用,需要进一步规范采集方法。棉绳常用于采集口腔液以进行病原体检测,但由于样本污染,它们可能并非生物标志物分析的最佳选择。本研究比较了两种采集猪口腔液以进行生物标志物分析的方法(棉绳和海绵)。研究了一组11种与应激、炎症、败血症、免疫、氧化还原状态和一般内环境稳定相关的生物标志物。

材料与方法

本研究纳入了18个从仔猪到育肥猪的猪场。在每个猪场,在四个年龄阶段对三栏(用于海绵采集)或四栏猪(用于棉绳采集)进行采样:断奶后一周(5周龄)、转入育肥舍前(11 - 12周龄)和后(12 - 13周龄)以及屠宰前一周(22 - 25周龄)。总共用棉绳采集了288份口腔液样本,用海绵采集了216份,并对以下生物标志物进行分析:皮质醇、α - 淀粉酶、催产素(应激)、触珠蛋白(炎症)、降钙素原(败血症)、腺苷脱氨酶、免疫球蛋白G(免疫系统)、铁还原抗氧化能力(氧化还原状态)以及肌酸激酶、乳酸脱氢酶和总蛋白(一般内环境稳定)。还使用从1(清洁)到5(非常脏)的评分对样本的脏污程度进行了视觉评分。

结果

与海绵采集的口腔液(2.7±0.15)相比,棉绳采集的口腔液脏污程度更高(3.7±0.04),并且在皮质醇、降钙素原、催产素、触珠蛋白、总蛋白(TP)、乳酸脱氢酶(LDH)和铁还原抗氧化能力方面的值高于海绵采集的样本。所有生物标志物的值均随年龄增长而降低。免疫球蛋白G在两种采集方法中表现均不佳。

讨论与结论

结果表明年龄对用海绵或棉绳采集的口腔液中的生物标志物有明显影响。对于生物标志物分析,海绵提供的样本比棉绳更清洁。尽管在早期断奶仔猪阶段使用海绵可能需要更多时间,但两种方法在猪场的商业条件下都易于应用。从实际角度来看,使用海绵采样在减少采样时间和低污染方面实现了最佳组合。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c24/10466680/48b6bd2773cd/40813_2023_333_Fig1_HTML.jpg

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