Sperm viability, serological, molecular, and modified seminal plasma agglutination tests in the diagnosis of Leptospira in the semen and serum of bovine bulls
Sperm viability, serological, molecular, and modified seminal plasma agglutination tests in the diagnosis of Leptospira in the semen and serum of bovine bulls
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DOI：
10.1007/s42770-021-00562-8
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发表时间：
2021-08-23
影响因子：
2.2
通讯作者：
Ribeiro, Marcio Garcia
中科院分区：
文献类型：
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作者：
Maiolino, Sergio Ricardo;Cortez, Adriana;Ribeiro, Marcio Garcia
本研究对203头自然繁殖的公牛进行了血清显微凝集试验（MAT），这些公牛没有明显的睾丸炎和副生殖腺炎症的迹象。同时，对所有公牛的精液进行精子活力分析和基于16S rRNA基因的PCR。精液样本pcr阳性结果经测序证实。采用改良的精浆凝集（MSPA）试验，用精浆代替MAT中所有公牛的血清。8份（8/203 = 3.9%）公牛精液样本被认为是无活力的（失精症和无精症），与PCR诊断无关。MSPA试验未检测到凝集素滴度。MAT中钩端螺旋体凝集素滴度较高（132/203 = 65%），尤其是Sejroe血清组（Hardjo CTG, 100/203 = 49.3%; Wolffi 74/203 = 36.4%; Guaricura 72/203 = 35.5%; Hardjoprajitno 56/203 = 27.6%）。3份（3/203 = 1.5%）公牛精液标本PCR阳性，但测序结果未得到证实。来自Sejroe血清群的血清型的高频率通常适用于牛，这表明需要在取样农场采取预防/控制病原体的措施。MAT、精子活力和精液中钩端螺旋体分子检测之间的差异突出了诊断公牛钩端螺旋体病的综合方法的必要性。据我们所知，本文首次描述了改良的精浆凝集，以研究通过自然方式繁殖的公牛中生殖道局部产生的抗钩端螺旋体抗体对牛钩端螺旋体病的诊断。
The present study investigated the serum microscopic agglutination test (MAT) among 203 bovine bulls with reproduction by natural means, without apparent signs of orchitis or inflammation of accessory reproductive glands. Simultaneously, the semen of all bulls was subjected to sperm viability analysis and PCR based on the 16S rRNA gene. PCR-positive results of semen samples were confirmed by sequencing. A modified seminal plasma agglutination (MSPA) test, replacing the blood serum of all bulls in the MAT with seminal plasma was performed as well. Eight (8/203 = 3.9%) semen samples from bulls were considered nonviable (necrospermia and azoospermia) without relation to the PCR diagnosis. No agglutinin titers were identified in MSPA test. A high frequency (132/203 = 65%) of leptospiral agglutinin titers was identified in the MAT, particularly for the Sejroe serogroup (Hardjo CTG, 100/203 = 49.3%; Wolffi 74/203 = 36.4%; Guaricura 72/203 = 35.5%; and Hardjoprajitno 56/203 = 27.6%). Three (3/203 = 1.5%) semen samples of bulls were positive in the PCR, but these results were not confirmed by sequencing. The high frequency of serovars from the Sejroe serogroup typically adapted to bovines indicates the need for measures for the prophylaxis/control of the pathogen on the sampled farms. Discrepancies among the MAT, sperm viability, and molecular detection of leptospires in semen highlight the need for a combination of methods to diagnose leptospirosis in bovine bulls. To our knowledge, modified seminal plasma agglutination is described for the first time here to investigate anti-Leptospira antibodies produced locally in the genital tract in the diagnosis of bovine leptospirosis among bulls that reproduce by natural means.