ویژگی‌یابی مولکولی جهش‌ها در Staphylococcus pasteuri کوآگولاز منفی مقاوم به متی‌سیلین جداشده از نمونه‌های مختلف مواد غذایی

مقالات آماده انتشار

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه زیست‌فناوری، دانشکده علوم، دانشگاه الانبار، رمادی، عراق.

2 دانشکده علوم مهندسی کشاورزی، دانشگاه بغداد، بغداد، عراق.

3 . گروه زیست‌فناوری، دانشکده علوم، دانشگاه الانبار، رمادی، عراق.

10.22103/jab.2026.27378.1944

چکیده

هدف: استافیلوکوک‌ها به‌طور طبیعی در بدن انسان و حیوانات، به‌ویژه در دستگاه‌های گوارش و تنفس، حضور دارند. گونه‌های Staphylococcus از جمله میکروارگانیسم‌های رایج در محیط‌های مرتبط با مواد غذایی هستند و از محصولات متنوعی نظیر گوشت، فرآورده‌های لبنی و غذاهای آماده مصرف جداسازی شده‌اند. هدف این مطالعه، جداسازی و شناسایی Staphylococcus pasteuri از انواع فرآورده‌های غذایی (گوشت و پنیر) و همچنین از دست‌اندرکاران تهیه و عرضه مواد غذایی، و بررسی الگوهای مقاومت آنتی‌بیوتیکی و جهش‌های ژنتیکی آن‌ها با استفاده از داده‌های توالی‌یابی کل ژنوم بود.
مواد و روش‌ها: در مجموع 58 نمونه مواد غذایی شامل گوشت تازه، گوشت پخته و پنیر، به همراه 13 نمونه سواب دست از کارکنان تهیه و عرضه مواد غذایی، از شهر بغداد، عراق، جمع‌آوری شد. برای غربالگری مقاومت به متی‌سیلین از روش انتشار دیسک کربی–باوئر (Kirby–Bauer) استفاده شد و نتایج آن با سیستم خودکار VITEK 2 تأیید گردید. تعدادی از سویه‌های مقاوم برای توالی‌یابی کل ژنوم (Whole-Genome Sequencing; WGS) با استفاده از فناوری Illumina انتخاب شدند تا ژن‌های مقاومت و پلی‌مورفیسم‌های تک‌نوکلئوتیدی (Single-Nucleotide Polymorphisms; SNPs) مورد بررسی قرار گیرند.
نتایج: Staphylococcus pasteuri با موفقیت از نمونه‌های غذایی و نیز از دست‌اندرکاران تهیه مواد غذایی جداسازی شد و نتایج، احتمال انتقال این باکتری از طریق زنجیره غذایی را نشان داد. بررسی‌های فنوتیپی و مولکولی، شیوع بالای ویژگی‌های مقاومت به چند دارو را در جدایه‌ها آشکار ساخت. تحلیل توالی‌یابی کل ژنوم (WGS) وجود مسیرهای متابولیکی مهم، از جمله آنزیم‌های وابسته به NAD⁺ و سامانه انتقال فسفوانول‌پیرووات فسفوترانسفراز (Phosphoenolpyruvate Phosphotransferase System; PTS) را نشان داد. این یافته‌ها بیانگر توان متابولیکی بالا و قابلیت سازگاری گسترده این باکتری با انواع مختلف محیط‌های غذایی است. همچنین، تحلیل ژنومی انعطاف‌پذیری ژنتیکی قابل توجهی را نشان داد که با حضور عناصر ژنتیکی متحرک، تعیین‌کننده‌های مقاومت آنتی‌بیوتیکی و تعداد زیادی پلی‌مورفیسم تک‌نوکلئوتیدی (SNP) همراه بود. از جمله یافته‌های مهم، شناسایی جهش‌های با اثر زیاد در دومین‌های با عملکرد ناشناخته (Domains of Unknown Function; DUFs، به‌ویژه DUF3310 و DUF771 بود که احتمالاً دارای نقش‌های تنظیمی جدید هستند.
نتیجه‌گیری: نتایج این مطالعه نشان داد که Staphylococcus pasteuri دارای ساختار ژنومی بسیار انعطاف‌پذیر و پیچیده‌ای است که به آن امکان بقا، سازگاری با محیط‌های مختلف و انتشار ژن‌های مقاومت آنتی‌بیوتیکی در زنجیره غذایی را می‌دهد. این ویژگی‌ها، S. pasteuri را به یک نگرانی مهم در حوزه سلامت عمومی و ایمنی مواد غذایی تبدیل می‌کند و ضرورت پایش و کنترل آن را بیش از پیش آشکار می‌سازد.

کلیدواژه‌ها

عنوان مقاله [English]

Molecular characterization of mutations in methicillin resistant coagulase negative Staphylococcus pasteuri from various food samples

نویسندگان [English]

  • Zahraa A. Ahmed 1
  • Asmaa S. Ahmaed 2
  • Huda M. Mahmood 3
1 Department of Biotechnology, College of Sciences, University of Anbar, Ramadi, Iraq.
2 College of Agricultural Engineering Sciences, University of Baghdad, Baghdad, Iraq.
3 Department of Biotechnology, College of Sciences, University of Anbar, Ramadi, Iraq
چکیده [English]

Objective
Staphylococci are naturally present in the bodies of animals and humans, particularly in the digestive and respiratory systems. Staphylococcus spp. are widespread microorganisms in food environments, having been isolated from a variety of products, including meat, dairy, and ready-to-eat foods. This aim of the current study was to isolate and identify Staphylococcus pasteuri from various food products (meat and cheese) and food handlers, and to characterize their antibiotic resistance profiles and genetic mutations using the whole-genome data.
Materials and methods
A total of 58 food samples (fresh meat, cooked meat, and cheese) and 13 hand swabs from food handlers were collected in Baghdad, Iraq. The Kirby-Bauer disc diffusion method was used to screen isolates for methicillin resistance. Output of this method was confirmed via the VITEK 2 system. Selected resistant strains were sequenced using Illumina technology to study whole-genome sequencing (WGS) and to analyze resistance genes and single-nucleotide polymorphisms (SNPs).
Results
Staphylococcus pasteuri was successfully isolated from the tested food samples and food handlers. The results highlighted its potential transmission through the food chain. A high prevalence of multi-drug resistance traits was determined by considering phenotypic and molecular characterization. Key metabolic pathways, including NAD+ dependent enzymes and the phosphoenolpyruvate phosphotransferase system (PTS) were revealed by WGS analysis. These results indicated high metabolic versatility and adaptability to different type of food environments. Moreover, significant plasticity was revealed by genomic analysis. These results were characterized by the presence of mobile genetic elements, resistance determinants, and numerous single-nucleotide polymorphisms (SNPs). Notably, high-impact mutations were detected within uncharacterized domains (DUFs), specifically DUF3310 and DUF771, which may possess novel regulatory functions.
Conclusion
The results of the present study showed that S. pasteuri has a highly flexible and comprehensive genomic architecture. This enables it to survive, adapt, and spread antibiotic resistance through the food chain. This is a significant public health concern that must be addressed.

کلیدواژه‌ها [English]

  • food safety
  • methicillin resistance
  • single-nucleotide polymorphisms (SNPs)
  • Staphylococcus pasteuri
  • whole-genome sequencing

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