بررسی مولکولی مقاومت به سموم پایرتروئیدی درلاروهای مگس خانگی (Musca Domestica) در مرغداری‌های تخمگذار شمالغرب ایران

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه پاتوبیولوژی، دانشکده دامپزشکی، دانشگاه ارومیه، ارومیه، ایران

2 گروه پاتوبیولوژی ،دانشکده دامپزشکی ،دانشگاه ارومیه، ارومیه،ایران

3 گروه پاتوبیولوژی، دانشکده دامپزشکی ،دانشگاه ارومیه،ارومیه، ایران

4 گروه پاتوبیولوژی، دانشکده دامپزشکی، دانشگاه ارومیه،ارومیه،ایران

5 گروه بهداشت و مواد غذایی،دانشکده دامپزشکی، دانشگاه ارومیه، ارومیه، ایران

چکیده

مطالعه حاضر با هدف بررسی حساسیت مگس خانگی به حشره­کش­های پایرتروئیدی با استفاده از آنالیز مولکولی انجام شد. مگس­ها از مرغداری­های چهار شهر مختلف آذربایجان غربی جمع­آوری شدند و برای ارزیابی مقاومت آنها به پایروتروئیدها با استفاده از سنجش زیستی مقاومت به حشره­کش­ها به آزمایشگاه منتقل شدند. DNA ژنومی از مگس­های حساس و مقاوم استخراج شد. در این مطالعه برای تکثیر الل اختصاصی مگس خانگی، از تکنیک واکنش زنجیره­ای پلیمراز(PCR)  استفاده شد که وجود تغییرات آلل ژنتیک در ژن کانال سدیم نوع  پارا (para) را برای شناسایی جهش از نوع هموزیگوت با حساسیت کامل(sus/sus) به نوع هموزیگوت جهش یافته با مقاومت کامل(kdr/kdr) یا نوع هتروزیگوت جهش یافته(kdr/sus)  نشان میدهد. در این مطالعه، این ژنوتیپ­ها با استفاده از پرایمرهای خاص برای شناسایی این تغییرات ژنتیکی مورد هدف قرار گرفتند. نتایج حاصل از سنجش زیستی مقاومت بالای 50% را در همه­ی جمعیت­های مورد مطالعه نشان داد که بالاترین میزان مقاومت از مگس­های خانگی شهرستان ارومیه  با فراوانی (86 درصد) گزارش شد.  از آزمایش واکنش زنجیره­ای پلیمراز برای شناسایی  kdr همو یا هتروزیگوت در مگس خانگی به منظور تعیین فراوانی kdr استفاده شد وآلل L1014 F در تمام جمعیت مورد آزمایش یافت شد. فراوانی kdr در جمعیت­های مورد مطالعه بالا و از 37 درصد تا 81 درصد متغیر بود. بیشترین درصد (71 درصد) ژنوتیپ هموزیگوت (kdr/kdr) در ارومیه و پس از آن تکاب (61 درصد) و کمترین درصد در سردشت  (30 درصد) رخ داد. مگس­ها با ژنوتیپ kdr/kdr در تمام جمعیت‌های آزمایش شده شناسایی شدند.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Molecular analysis of pyrethroid resistance in the larvae of house fly (Musca Domestica) collected from Poultry farms in northwestern Iran

نویسندگان [English]

  • Maryam Keyghobadi 1
  • mousa tavassoli 2
  • Bijan Esmaeilnejad 3
  • Farnaz Malekifard 4
  • Rahim Molaie 5
1 Department of Pathobiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran.
2 Department of Pathobiology, Faculty of Veterinary Medicine, Urmia University, urmia.Iran
3 Department of Pathobiology, Faculty of Veterinary Medicine, Urmia University, urmia.Iran
4 Department of Pathobiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran.
5 Department of Health and Food, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran.
چکیده [English]

The present study was aimed to assess the house fly susceptibility to pyrethroid insecticides using molecular analysis. Flies were collected from poultry farms in four different cities of West Azerbaijan and were transferred to the laboratory to evaluate their resistance to pyrethroids using insecticide resistance bioassay. Genomic DNA was extracted from susceptible and resistant flies. Using a technique called polymerase chain reaction (PCR) amplification of specific allele (PASA), PCR was employed to reveal the presence of allelegenetic variations in the para-type sodium channel (para) gene to recognize knockdown resistance (kdr) mutation from the homozygous type of complete susceptibility (sus/sus) to the mutated-homozygous type of complete resistance (kdr/kdr) or to the mutated-heterozygous type (kdr/sus). In this study, these genotypes were targeted using specific primers to identify these genetic variations. The results of bioassay showed more than 50% resistance in all studied populations , and the highest level of resistance was reported from houseflies in Urmia city (86%). A polymerase chain reaction test for identifying kdr homo- or heterozygote house flies was used to determine the frequency of kdr. The L1014 F allele was found in all populations tested. The frequency of KDR in the studied populations was high and varied from 37% to 81%.The maximum percentage (71%) of homozygous genotype (kdr/kdr) occurred at Urmia, followed by Takab (61%), and the minimum percentage occurred at Sardasht (30%). Flies with kdr/kdr genotype were identified in all tested populations.

کلیدواژه‌ها [English]

  • house fly
  • insecticide
  • pyrethroid
  • resistance

Extended Abstract

Introduction

The Diptera order is divided into three sub-orders, Cyclorapha, Brachycera and Nematocera. The housefly is in the Cyclorapha sub-order and the Musidae family. The house fly, Musca domestica, is a common pest that is necessary to control due to its role as a disease vector in humans, livestock, and poultry. Pyrethroid insecticides combine high insecticidal activity with low mammalian toxicity and lack of environmental persistence, thus promoting their widespread use to control many pests that are important to agriculture and to human health. However, their intensive use has led to development Resistance in many insect species, which often leads to Control failures. The present study was aimed to assess the house fly susceptibility to pyrethroid insecticides using molecular analysis.

 

Materials and Methods

In this study, sampling was done from four different geographical areas of West Azarbaijan province in 2023 (2023) and in the summer season and were transferred to Urmia University parasitology laboratory for bioassay and genotype determination. The resistance of collected housefly larvae was evaluated using Whatman papers impregnated with insecticide. Genomic DNA of the larvae that survived after the effect of the insecticide, as well as the larvae that died, was extracted using a DNA extraction kit (MBST, Tehran, Iran) according to the manufacturer's instructions. After extracting the DNA of house fly larvae, the PCR kit of Cinagen Company (Cinagen, Tehran) was used to perform PCR. For this purpose, Two outer allele-specific primers (kdr1,kdr4) and two inner allele-specific primers(kdr2,kdr3) were designed to detect the Kdr mutation (L1014 F; CTT to TTT) in house flies. Eventually, PCR products were electrophoresed using 1.5% agarose gel.

 

Results

The results of bioassay showed more than 50% resistance in all studied populations , and the highest level of resistance was reported from houseflies in Urmia city (86%) And the lowest level of resistance reported was related to Takab city(65%). The frequency of the L1014 F kdr mutation in four housefly populations that showed different levels of pyrethroid resistance was determined using molecular analysis. The L1014 F allele was found in all tested populations. The frequency of Kdr in the studied populations was high and varied from 37% (Sardasht) to 81%(Urmia). In addition, houseflies with the kdr/kdr genotype were identified in all tested populations. The highest number of houseflies with kdr/kdr genotype was reported from Urmia(71%), followed by Takab (61%). While the lowest number was related to Sardasht (30%).

 

Conclusion

 The results obtained from this study indicate that resistance has been developed in houseflies of West Azerbaijan province to different groups of pyrethroid insecticides, which are the most common insecticides used for pest control. When an insect population shows resistance to an insecticide, control programs using this insecticide are no longer effective. Based on this, the list of pyrethroid insecticides used to control house flies in the province should be re-evaluated to reduce the frequency of kdr resistance allele in the population of flies.

مراجع

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علوم دامی ایران
دوره 55، شماره 3
مهر 1403
صفحه 477-492

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  • تاریخ دریافت: 16 مرداد 1402
  • تاریخ بازنگری: 26 دی 1402
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