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To date, no study has examined mosquito resistance after pre-exposure to PBO and subsequent enzymatic activity when exposed to PBO-synergized insecticides. We used Culex quinquefasciatus Say (Diptera: Culicidae), an important vector of arboviruses and lymphatic filariasis, as a model to examine the insecticide resistance mechanisms of mosquitoes to PBO-synergized pyrethroid using modified World Health Organization tube bioassays and biochemical analysis of metabolic enzyme expressions pre- and post-PBO exposure. Mosquito eggs and larvae were collected from three cities in Orange County in July 2020 and reared in insectary, and F0 adults were used in this study. A JHB susceptible strain was used as a control. Mosquito mortalities and metabolic enzyme expressions were examined in mosquitoes with\/without pre-exposure to different PBO concentrations and exposure durations. Except for malathion, wild strain Cx quinquefasciatus mosquitoes were resistant to all insecticides tested, including PBO-synergized pyrethroids (mortality range 3.7 \u00b1 4.7% to 66.7 \u00b1 7.7%). Wild strain mosquitoes had elevated levels of carboxylesterase (COE, 3.8-fold) and monooxygenase (P450, 2.1-fold) but not glutathione S-transferase (GST) compared to susceptible mosquitoes. When wild strain mosquitoes were pre-exposed to 4% PBO, the 50% lethal concentration of deltamethrin was reduced from 0.22% to 0.10%, compared to 0.02% for a susceptible strain. The knockdown resistance gene mutation (L1014F) rate was 62% in wild strain mosquitoes. PBO pre-exposure suppressed P450 enzyme expression levels by 25~34% and GST by 11%, but had no impact on COE enzyme expression. Even with an optimal PBO concentration (7%) and exposure duration (3h), wild strain mosquitoes had significantly higher P450 enzyme expression levels after PBO exposure compared to the susceptible laboratory strain. These results further demonstrate other studies that PBO alone may not be enough to control highly pyrethroid-resistant mosquitoes due to multiple resistance mechanisms. Mosquito resistance to PBO-synergized insecticide should be closely monitored through a routine resistance management program for effective control of mosquitoes and the pathogens they transmit.<\/jats:p>","DOI":"10.1093\/jme\/tjab231","type":"journal-article","created":{"date-parts":[[2022,1,20]],"date-time":"2022-01-20T09:01:40Z","timestamp":1642669300000},"page":"638-647","source":"Crossref","is-referenced-by-count":22,"title":["Emerging Mosquito Resistance to Piperonyl Butoxide-Synergized Pyrethroid Insecticide and Its Mechanism"],"prefix":"10.1093","volume":"59","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9283-5520","authenticated-orcid":false,"given":"Guofa","family":"Zhou","sequence":"first","affiliation":[{"name":"Program in Public Health, University of California, Irvine, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yiji","family":"Li","sequence":"additional","affiliation":[{"name":"Program in Public Health, University of California, Irvine, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brook","family":"Jeang","sequence":"additional","affiliation":[{"name":"Program in Public Health, University of California, Irvine, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1232-3125","authenticated-orcid":false,"given":"Xiaoming","family":"Wang","sequence":"additional","affiliation":[{"name":"Program in Public Health, University of California, Irvine, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert F","family":"Cummings","sequence":"additional","affiliation":[{"name":"Orange County Mosquito and Vector Control District, Garden Grove, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daibin","family":"Zhong","sequence":"additional","affiliation":[{"name":"Program in Public Health, University of California, Irvine, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guiyun","family":"Yan","sequence":"additional","affiliation":[{"name":"Program in Public Health, University of California, Irvine, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2022,1,20]]},"reference":[{"key":"2022031611050692400_CIT0001","first-page":"346","article-title":"Monitoring of insecticide resistance of Culex pipiens (Diptera: Culicidae) colonies-collected from California","volume":"3","author":"Ahmed","year":"2012","journal-title":"Int. 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