بررسی حضور کلی‌فرم‌ها و ایشریشیا کولای مقاوم به آنتی بیوتیک در ساحل دریای خزر - استان گیلان

نوع مقاله : مقاله کامل علمی پژوهشی

نویسندگان

1 گروه علوم خاک، دانشکده کشاورزی، دانشگاه گیلان، رشت، ایران.

2 گروه علوم خاک، دانشکده کشاورزی، دانشگاه گیلان

چکیده

سابقه و هدف: در سال‌های گذشته بسیاری از نواحی ساحلی در معرض خطرات ناشی از آلاینده‌ها قرار گرفته‌اند. حضور باکتری‌های بیماری‌زا در سواحل تفریحی مشکلاتی را در این نواحی ایجاد کرده است. مقاوم شدن این باکتری‌ها در برابر آنتی‌بیوتیک‌ها شرایط را بد‌تر نیز کرده است. بنابراین، این پژوهش با هدف بررسی حضور کلی‌فرم‌ها و باکتری ایشریشیا کولای (E. coli) مقاوم به آنتی‌بیوتیک به عنوان باکتری شاخص در خط ساحلی دریای خزر در استان گیلان انجام شد.
مواد و روش‌ها: نُه منطقه ساحلی پربازدید در استان گیلان شامل رودسر، چمخاله، امین آباد، جفرود، ساحل قو (انزلی)، رضوان‌شهر، گیسوم، ساحل زمرد (حَویق) و آستارا برای مطالعه انتخاب شدند. نمونه‌برداری در سه تکرار از عمق 20-0 سانتی‌متر شن ساحلی انجام شد. تعداد کلی‌فرم‌ها در نمونه‌ها در محیط ائوزین-متیلن بلو شمارش شد و pH و EC نمونه‌ها نیز در سوسپانسیون دو به یک شن به آب مقطر اندازه‌گیری شد. سپس باکتری E. coli از کلی‌فرم‌ها انتخاب شد و مقاومت آن به عنوان باکتری شاخص در برابر آنتی‌بیوتیک‌ها به روش پخشیدگی دیسک روی محیط مولر-هینتون آگار بررسی شد. آنتی‌بیوتیک‌های وسیع الطیف شامل سفالکسین، جنتامایسین، داکسی‌سایکلین، سفتریاکسون، سیپروفلوکساسین و تری‌متوپریم تست شدند. داده‌های pH، EC و تعداد کلی‌فرم‌ها به‌صورت طرح کاملاً تصادفی و داده‌های شاخص ZOI به‌صورت فاکتوریل در قالب طرح کاملاً تصادفی آنالیز شدند و مقایسه میانگین‌ها به روش توکی (05/0>p) انجام شد.
یافته‌ها: در همه نقاط نمونه‌برداری شده pH قلیایی بود و بیشترین میزان آن در شن ساحل جفرود (4/8) به دست آمد. بیشترین میزان EC در شن ساحل امین‌آباد (dS m-1 6/3) به دست آمد و تفاوت آن با دیگر نقاط معنی‌دار بود (05/0>p). بیش‌ترین تعداد کلی‌فرم‌ها در ساحل زمرد در حویق (Log CFU g-1 8/5) به دست آمد. برهم‌کنش نوع منطقه نمونه‌برداری در آنتی‌بیوتیک بر قطر ZOI باکتری E. coli نشان داد که در سه منطقه ساحلی آستارا، گیسوم و رضوانشهر باکتری E. coli به هیچ‌کدام از آنتی‌بیوتیک‌های بررسی شده مقاوم نبود. در شش نقطه ساحلی دیگر در همه آنها به جز ساحل قو (انزلی) باکتری E. coli به سفالکسین مقاوم بود. در جفرود باکتری E. coli تنها به سفالکسین مقاوم بود. درحالی‌که در رودسر، چمخاله (سواحل شرق گیلان) ساحل قو و ساحل زمرد باکتری E. coli به سفتریاکسون نیز مقاومت نشان داد.
نتیجه‌گیری: در کل، سواحل بخش غربی نسبت به شرق و مرکز گیلان pH و EC کمتر و تعداد کلی‌فرم‌های بیشتری داشتند. در شن ساحلی همه مناطق بررسی شده تعداد کلی‌فرم‌ها بیشتر از حد استاندارد مناطق تفریحی بود. در شن سواحل شرق گیلان یعنی سواحل چمخاله و رودسر، باکتری E. coli به سفالکسین و سیپروفلوکساسین مقاومت نشان داد. بنابراین، مدیریت آلودگی‌های میکروبی به ویژه آنهایی که در برابر آنتی بیوتیک‌ها مقاوم هستند، در سواحل شرق گیلان نیاز به توجه بیشتری دارد.

کلیدواژه‌ها


عنوان مقاله [English]

Investigation of coliforms and antibiotic resistant Escherichia coli occurrence in the Caspian Sea coast - Guilan province

نویسندگان [English]

  • Mohammad Bagher Farhangi 1
  • Nasrin Ghorbanzadeh 2
1 Soil Science Department, Faculty of agricultural Science, University of Guilan, Rasht, Iran.
2 Soil Science Department, Faculty of agricultural Science, University of Guilan, Rasht, Iran.
چکیده [English]

Background and Objective: In recent years, many coastal areas have been exposed to the pollutant threats. The occurrence of pathogenic bacteria in recreational beaches has made difficulties in these areas. Resistance of these bacteria to antibiotics has also worsened the situation. Therefore, this study aimed to investigate the occurrence of coliforms and antibiotic resistant Escherichia coli, as an indicator bacterium, in the swash zone of Caspian Sea, Guilan province.
Materials and Methods: Nine popular coastal areas in Guilan province including Rudsar, Chamkhaleh, Aminabad, Jafroud, Sahele-Ghou (Anzali), Rezvanshahr, Gisum, Sahele-Zomorrod (Haviq), and Astara were selected for this study. The sand beaches were sampled in three replications from 0-20 cm depth in the swash zone. The coliform number was counted on the EMB medium. pH and EC were also measured in 2:1 sand:distilled water suspension. Then, E. coli as an indicator bacteria was isolated and its resistance to antibiotics was determined by disk diffusion method on Mueller-Hinton agar medium. Broad spectrum antibiotics including cephalexin, gentamicin, doxycycline, ceftriaxone, ciprofloxacin and trimethoprim were tested. Data for pH, EC and coliform number were analyzed in a completely randomized design and for the zone of inhibition (ZOI) test a factorial arrangement experiment in a completely randomized design was adopted. Also, mean comparisons were done by Tukey test (p<0.05).
Results: In all sampled areas, the pH was alkaline and its highest value was obtained in the Jafroud beach sand. The highest EC value was obtained in Aminabad beach sand which was significantly different from EC values in other sampled sites (p<0.05). The highest number of coliforms was obtained in Sahele-Zomorrod sand beach (5.8 Log CFU g-1). The effect of sampling site and antibiotic interactions on the ZOI of E. coli showed that in three sampling sites including Astara, Gisum and Rezvanshahr the isolated E. coli was resistant to none of the tested antibiotics. In the other six sites except Sahele-Ghou (Anzali), it showed resistance to cephalexin. The isolated E. coli showed resistance only to cephalexin in Jafroud. While it also showed resistance to ceftriaxone in Rudsar, Chamkhaleh (eastern beaches), Sahele-Ghou, and Sahele-Zomorrod.
Conclusion: Overall, the beach sand of the western coastal areas had a lower pH and EC values and higher total coliform number compared to the eastern and central coastal areas. In all studied places, the coliform number was above the standard level for recreational sites. In eastern coastal areas; Chamkhaleh and Roudsar, isolated E. coli showed resistance to both cephalexin and ceftriaxone antibiotics. Thus, the management of microbial contamination especially those that are resistant to antibiotics needs more attention in the eastern coastal areas in Guilan province.

کلیدواژه‌ها [English]

  • Ceftriaxone
  • Cephalexin
  • Chamkhaleh
  • Roudsar
  • ZOI
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