基于荧光定量PCR检测和原位杂交技术分析湖北省蛋鸡滑液支原体的感染情况

doi:10.11843/j.issn.0366-6964.2025.03.038

Abstract

Abstract:

In recent years, the infection rate of Mycoplasma synoviae (MS) in chicken flocks has been increasing. The inventory of laying hens in Hubei Province is relatively large, and all commodity breeds are raised. Currently, the situation of MS infection in laying hens in Hubei Province is not clear. Therefore, this study conducted an investigation into the infection status of MS in chicken flocks of large-scale laying hen farms in Hubei and explored the impact of MS infection on clinical eggshell apex abnormality (EAA). One thousand six hundred and thirty-seven serum samples and tracheal cotton swabs were collected from 82 chicken flocks in 36 large-scale laying hen farms in Hubei Province. The MS load was detected using the established real-time qPCR method, and the serum antibodies of vaccinated and non-vaccinated chicken flocks were detected using ELISA kits. Perform MS loading, pathological sectioning, and MS localization analysis on the oviduct of EAA laying hens. The results showed that in clinical samples, the MS positive rate of tracheal swabs was 9.41%; The positivity rate of serum antibodies was 92.24%, among which the positivity rate of antibodies in non-vaccinated chicken flocks was 89.04%, and the positivity rate of antibodies in 6-month-old chickens was 100%, while the positivity rate of antibodies in vaccinated chicken flocks was 95%; Out of 36 large-scale farms, 26 (72.22%) were infected with MS, and out of 82 chicken flocks, 61 (74.39%) were infected; All 10 breeds of laying hens were infected with MS. MS was detected in the oviduct of EAA laying hens, and the load in the magnum was significantly higher than that in the uterus, isthmus and infundibulum (P < 0.01), the uterus was higher than that in the isthmus and infundibulum (P < 0.05). In the oviduct of EAA laying hens, mucosal epithelial cell degeneration and glandular enlargement occur in the isthmus and uterus, as well as shedding and necrosis of glandular epithelial cells in the magnum. Moreover, MS positive signals were detected in the mucosal epithelium of the uterus and isthmus, as well as in the mucosal epithelium and surrounding tubular glands of the magnum, and among them, the magnum and uterus had the strongest positive signals. The infection of MS in laying hens in Hubei Province is severe, and different breeds are infected. MS mainly colonizes in the uterus and magnum of the laying hens' oviduct, causing pathological damage. This study provides data support for understanding the situation of MS infection in laying hens in Hubei Province and the study of MS induced EAA eggs.

Key words: Mycoplasma synoviae, eggshell apex abnormality, real-time qPCR, in situ hybridization, tissue distribution

CLC Number:

S858.31

WANG Chun, WANG Qing, WU Xiaoqian, HU Ting, CUI Weitao, PEI Jie, SONG Tieping, HU Sishun, ZHANG Wanpo, LI Zili, ZHOU Zutao. Analysis of Mycoplasma synoviae Infection in the Laying Hens in Hubei Province based on Fluorescence Quantitative PCR Detection and in situ Hybridization Technology[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(3): 1396-1407.

Figures/Tables 10

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References 26

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标准浓度/(copies·μL^-1) Standard concentration	批内重复试验 Intra-assay repeatability			批间重复试验 Inter-assay repeatability
标准浓度/(copies·μL^-1) Standard concentration	AVG	SD	CV/%	AVG	SD	CV/%
1.91×10⁶	16.76	0.026	0.16	17.4	0.714	4.10
1.91×10⁵	20.11	0.123	0.61	20.51	0.633	3.09
1.91×10⁴	24.35	0.130	0.53	24.47	0.125	0.51

群体 Flocks	总数 Count	核酸阳性数 Nucleic acid-positive count	阳性率/% Positive rate	抗体阳性数 Antibody-positive count	阳性率/% Positive rate
免疫鸡群	880	94	10.68^a	836	95.00^A
Vaccinated chicken
非免疫鸡群	757	60	7.93^a	674	89.04^B
Non-vaccinated chicken

品种 Breeds	日龄 Days	产蛋率/% Laying rate	破损率/% Damage rate	EAA	核酸阳性率/% Nucleic acid-positive rate	抗体阳性率/% Antibody-positive rate	每克黏液MS载量(×10³拷贝)(${\bar x}$±s) MS load per gram of mucus (×10³ copies) (${\bar x}$±s)
罗曼粉	360	70	12.0	√	54.55	100	3.15±1.29
海兰灰	359	86	6.0	√	30	100	4.79±3.44
罗曼粉	200	83.5	7.8	√	30	100	2.00±0.54
罗曼灰	170	87	1.2		25	100	18.15±29.90
罗曼灰	355	85	10.0	√	25	100	6.39±10.27
罗曼粉	450	82	3.7	√	25	100	5.69±4.74
罗曼粉	491	81	4.5	√	25	100	4.18±2.34
罗曼灰	264	89	3.0		25	100	8.31±4.93
罗曼粉	220	87	0.8		20	100	3.17±1.51
京粉6号	442	83	6.0		20	100	14.95±10.63

部位 Location	Ct值 Ct value	每克黏膜MS载量(×10⁴拷贝) MS load per gram of mucosa (×10⁴ copies)
子宫部Uterus	27.22±0.67^Aa	3.31±1.68^Aa
峡部Isthmus	28.53±0.13^AbC	1.24±0.11^Ab
膨大部magnum	25.64±0.16^Bc	9.26±1.08^Bc
漏斗部Infundibulum	29.24±0.81^bC	0.60±0.14^Ab

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Analysis of Mycoplasma synoviae Infection in the Laying Hens in Hubei Province based on Fluorescence Quantitative PCR Detection and in situ Hybridization Technology

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