鸡副鸡Avibacterium paragallinarum选择性培养基的研制。

IF 5.4 2区 医学 Q1 MICROBIOLOGY
Journal of Clinical Microbiology Pub Date : 2025-08-13 Epub Date: 2025-07-18 DOI:10.1128/jcm.00311-25
Mariela E Srednik, Mostafa M S Shelkamy, Amro Hashish, Nubia R De Macedo, Yuko Sato, Mohamed M El-Gazzar, Orhan Sahin, Qijing Zhang
{"title":"鸡副鸡Avibacterium paragallinarum选择性培养基的研制。","authors":"Mariela E Srednik, Mostafa M S Shelkamy, Amro Hashish, Nubia R De Macedo, Yuko Sato, Mohamed M El-Gazzar, Orhan Sahin, Qijing Zhang","doi":"10.1128/jcm.00311-25","DOIUrl":null,"url":null,"abstract":"<p><p>Infectious coryza (IC) caused by <i>Avibacterium paragallinarum</i> (AvP) is an upper respiratory disease in chickens and incurs a significant economic impact on laying hens. Control of IC requires reliable bacterial isolation, but AvP is a fastidious bacterium, and the currently used methods yield poor isolation. To address this need, we developed selective media for the efficient growth and isolation of AvP. Several basal media supplemented with various growth factors were explored, and Mueller Hinton agar (MHA) supplemented with fetal bovine serum (FBS) plus NAD yielded optimal AvP growth, eliminating the need for nurse bacteria. This medium (named MSN) was further supplemented with vancomycin and crystal violet to produce two selective media, named MSNV and MSCV, respectively, to inhibit commensal Gram-positive bacteria that reside in the upper respiratory tract of chickens. MSNV and MSCV were compared with the standard isolation methods used in veterinary diagnostic laboratories for AvP isolation using samples from IC-positive and IC-negative flocks verified by clinical observation and AvP-specific RT-PCR. The two selective media significantly increased the isolation of AvP from clinical cases (59.4% for MSNV and 46.9% for MSCV) compared to the conventional method (37.7%). In addition, MSNV and MSCV successfully isolated the recently discovered non-pathogenic AvP variants, which were difficult to obtain using the standard methods. Notably, both media drastically reduced background bacteria and improved the purity of AvP isolates. These results demonstrate the efficacy and usefulness of the selective media for AvP isolation, which will enhance IC diagnosis, antimicrobial susceptibility testing, and vaccine development.IMPORTANCEInfectious Coryza is an economically important disease in poultry, resulting in poor growth and a significant reduction in egg production. The causative agent (AvP) of the disease is difficult to isolate from clinical samples and often requires a nurse bacteria to provide essential nutrients for optimal growth. In addition, the currently used methods for isolation are inefficient and often result in contamination by residential bacterial species. The selective media (MSNV and MSCV) developed in this study solve these problems by eliminating the need for nurse bacteria and by inhibiting Gram-positive bacteria normally present in the respiratory tract of chickens. The media not only increase isolation efficiency but also improve the purity of the isolates. These advancements will facilitate IC diagnosis and the development of vaccines for effective control of this major poultry pathogen.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0031125"},"PeriodicalIF":5.4000,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345139/pdf/","citationCount":"0","resultStr":"{\"title\":\"Development of selective culture media for efficient isolation of <i>Avibacterium paragallinarum</i> from chickens.\",\"authors\":\"Mariela E Srednik, Mostafa M S Shelkamy, Amro Hashish, Nubia R De Macedo, Yuko Sato, Mohamed M El-Gazzar, Orhan Sahin, Qijing Zhang\",\"doi\":\"10.1128/jcm.00311-25\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Infectious coryza (IC) caused by <i>Avibacterium paragallinarum</i> (AvP) is an upper respiratory disease in chickens and incurs a significant economic impact on laying hens. Control of IC requires reliable bacterial isolation, but AvP is a fastidious bacterium, and the currently used methods yield poor isolation. To address this need, we developed selective media for the efficient growth and isolation of AvP. Several basal media supplemented with various growth factors were explored, and Mueller Hinton agar (MHA) supplemented with fetal bovine serum (FBS) plus NAD yielded optimal AvP growth, eliminating the need for nurse bacteria. This medium (named MSN) was further supplemented with vancomycin and crystal violet to produce two selective media, named MSNV and MSCV, respectively, to inhibit commensal Gram-positive bacteria that reside in the upper respiratory tract of chickens. MSNV and MSCV were compared with the standard isolation methods used in veterinary diagnostic laboratories for AvP isolation using samples from IC-positive and IC-negative flocks verified by clinical observation and AvP-specific RT-PCR. The two selective media significantly increased the isolation of AvP from clinical cases (59.4% for MSNV and 46.9% for MSCV) compared to the conventional method (37.7%). In addition, MSNV and MSCV successfully isolated the recently discovered non-pathogenic AvP variants, which were difficult to obtain using the standard methods. Notably, both media drastically reduced background bacteria and improved the purity of AvP isolates. These results demonstrate the efficacy and usefulness of the selective media for AvP isolation, which will enhance IC diagnosis, antimicrobial susceptibility testing, and vaccine development.IMPORTANCEInfectious Coryza is an economically important disease in poultry, resulting in poor growth and a significant reduction in egg production. The causative agent (AvP) of the disease is difficult to isolate from clinical samples and often requires a nurse bacteria to provide essential nutrients for optimal growth. In addition, the currently used methods for isolation are inefficient and often result in contamination by residential bacterial species. The selective media (MSNV and MSCV) developed in this study solve these problems by eliminating the need for nurse bacteria and by inhibiting Gram-positive bacteria normally present in the respiratory tract of chickens. The media not only increase isolation efficiency but also improve the purity of the isolates. These advancements will facilitate IC diagnosis and the development of vaccines for effective control of this major poultry pathogen.</p>\",\"PeriodicalId\":15511,\"journal\":{\"name\":\"Journal of Clinical Microbiology\",\"volume\":\" \",\"pages\":\"e0031125\"},\"PeriodicalIF\":5.4000,\"publicationDate\":\"2025-08-13\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345139/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Clinical Microbiology\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1128/jcm.00311-25\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2025/7/18 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q1\",\"JCRName\":\"MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Clinical Microbiology","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1128/jcm.00311-25","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/7/18 0:00:00","PubModel":"Epub","JCR":"Q1","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0

摘要

副鸡Avibacterium paragallinarum (AvP)引起的传染性鼻炎(IC)是鸡的一种上呼吸道疾病,对蛋鸡造成重大的经济影响。控制IC需要可靠的细菌分离,但AvP是一种挑剔的细菌,目前使用的方法分离效果很差。为了满足这一需求,我们开发了有效生长和分离AvP的选择性培养基。研究了几种添加了各种生长因子的基础培养基,发现添加了胎牛血清(FBS)和NAD的Mueller Hinton琼脂(MHA)产生了最佳的AvP生长,消除了对护理菌的需求。在此培养基(命名为MSN)中进一步添加万古霉素和结晶紫,得到两种选择性培养基,分别命名为MSNV和MSCV,以抑制鸡上呼吸道共生革兰氏阳性菌。采用经临床观察和AvP特异性RT-PCR验证的ic阳性和ic阴性禽群样本,将MSNV和MSCV与兽医诊断实验室分离AvP的标准方法进行比较。与传统方法(37.7%)相比,这两种选择性培养基显著提高了临床病例AvP的分离率(MSNV为59.4%,MSCV为46.9%)。此外,MSNV和MSCV成功分离出了新发现的标准方法难以获得的非致病性AvP变异体。值得注意的是,这两种培养基都大大减少了背景细菌,提高了AvP分离株的纯度。这些结果证明了选择培养基分离AvP的有效性和实用性,这将加强IC诊断,抗菌药物敏感性试验和疫苗开发。传染性鼻炎在家禽中是一种经济上重要的疾病,导致生长不良和产蛋量显著减少。该疾病的病原体(AvP)很难从临床样本中分离出来,通常需要护士细菌提供最佳生长所需的必需营养素。此外,目前使用的分离方法效率低下,经常导致驻留细菌的污染。本研究开发的选择性培养基(MSNV和MSCV)通过消除对护理菌的需求和抑制通常存在于鸡呼吸道的革兰氏阳性菌来解决这些问题。该培养基不仅提高了分离效率,而且提高了分离物的纯度。这些进展将有助于IC的诊断和有效控制这一主要家禽病原体的疫苗的开发。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Development of selective culture media for efficient isolation of Avibacterium paragallinarum from chickens.

Infectious coryza (IC) caused by Avibacterium paragallinarum (AvP) is an upper respiratory disease in chickens and incurs a significant economic impact on laying hens. Control of IC requires reliable bacterial isolation, but AvP is a fastidious bacterium, and the currently used methods yield poor isolation. To address this need, we developed selective media for the efficient growth and isolation of AvP. Several basal media supplemented with various growth factors were explored, and Mueller Hinton agar (MHA) supplemented with fetal bovine serum (FBS) plus NAD yielded optimal AvP growth, eliminating the need for nurse bacteria. This medium (named MSN) was further supplemented with vancomycin and crystal violet to produce two selective media, named MSNV and MSCV, respectively, to inhibit commensal Gram-positive bacteria that reside in the upper respiratory tract of chickens. MSNV and MSCV were compared with the standard isolation methods used in veterinary diagnostic laboratories for AvP isolation using samples from IC-positive and IC-negative flocks verified by clinical observation and AvP-specific RT-PCR. The two selective media significantly increased the isolation of AvP from clinical cases (59.4% for MSNV and 46.9% for MSCV) compared to the conventional method (37.7%). In addition, MSNV and MSCV successfully isolated the recently discovered non-pathogenic AvP variants, which were difficult to obtain using the standard methods. Notably, both media drastically reduced background bacteria and improved the purity of AvP isolates. These results demonstrate the efficacy and usefulness of the selective media for AvP isolation, which will enhance IC diagnosis, antimicrobial susceptibility testing, and vaccine development.IMPORTANCEInfectious Coryza is an economically important disease in poultry, resulting in poor growth and a significant reduction in egg production. The causative agent (AvP) of the disease is difficult to isolate from clinical samples and often requires a nurse bacteria to provide essential nutrients for optimal growth. In addition, the currently used methods for isolation are inefficient and often result in contamination by residential bacterial species. The selective media (MSNV and MSCV) developed in this study solve these problems by eliminating the need for nurse bacteria and by inhibiting Gram-positive bacteria normally present in the respiratory tract of chickens. The media not only increase isolation efficiency but also improve the purity of the isolates. These advancements will facilitate IC diagnosis and the development of vaccines for effective control of this major poultry pathogen.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
Journal of Clinical Microbiology
Journal of Clinical Microbiology 医学-微生物学
CiteScore
17.10
自引率
4.30%
发文量
347
审稿时长
3 months
期刊介绍: The Journal of Clinical Microbiology® disseminates the latest research concerning the laboratory diagnosis of human and animal infections, along with the laboratory's role in epidemiology and the management of infectious diseases.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:604180095
Book学术官方微信