| China is a country with severe rodent problems. Serious ecological problems and huge economic losses are caused by rodents yearly, while rodents transmitting diseases also poses a threat to human health. Eutamias sibiricus commonly known as five eyebrow, chipmunk, etc, is a kind of agriculture and forestry rodents, but it has certain economic value due to its beautiful fur, because it’s cute and easy to tame, it also has ornamental value, as well as edible and can be used as medicine. In view of the above characteristics, how to control and utilize the Eutamias sibiricus reasonably, has become a pressing problem.The traditional methods of rodents control, it now seems that there are many drawbacks. Thus, to find an effective, specific, non-toxic, and non-polluting method controlling rodents is of great significance. Using rodents pheromone to develop sex-attractant just has these advantages, open up a new way for Eutamias sibiricus prevention. Currently, using insect pheromone for pest control is relatively mature and widely used, in comparison rodent pheromone is more complex, the research on Eutamias sibiricus pheromone is still blank.Therefore, this study under laboratory conditions, with wild Eutamias sibiricus as the experimental subjects, using headspace-solid phase micro-extraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC/MS) technology, behavioral test, ect, tested the attraction effect of heterosexual urine, and analyzed the volatile components in urine of Eutamias sibiricus, furtherly verified the attraction effect of the different composition of urine, aimed at preliminarily screening the sex pheromone components, which provided a theoretical basis for developing sex-attractant, biological controlling rodents. Main results and conclusions were listed below:1. Using GC/MS to analyze the composition of Eutamias sibiricus urine,35 of them were putatively identified, and mainly contained 7 kinds of esters,6 kinds of alcohols,6 kinds of ketones,4 kinds of aldehydes,4 kinds of nitrogen compounds,3 kinds of phenols,2 kinds of sulfur compounds,2 kinds of olefins, and 1 kind of quinones substances. Among them, the levels of phenols, esters and alcohols were relatively higher; sulfur compounds and olefins were relatively lower. Among them, 2,5-Dimethylpyrazine,4-Methylphenol,4-Ethylphenol showed significant sexual difference. Thus, these compounds might be pheromone in Eutamias sibiricus, and played some roles in sex attractiveness. The content of phenols, nitrogen compounds floated relatively larger, the content of most of the rest of the compounds was less variable. The results showed that all the compounds detected were shared by Eutamias sibiricus, regardless of the reproductive status and gender, but the levels of each one was different. Thus, the information in Eutamias sibiricus urine was encoded by the content of volatile compounds.2. Comparing the compounds detected in Eutamias sibiricus urine, the results showed when at the same reproductive status, in the urine of estrous male and female Eutamias sibiricus,2,5-Dimethylpyrazine,4-Methylphenol and 4-Ethylphenol showed significant sexual difference. Among them,2,5-Dimethylpyrazine,4-Ethylphenol in male urine was significantly higher than female,4-Methylphenol in male urine was significantly lower than female, the rest of the compounds did not show significant gender differences; however, the compounds in the urine of non-estrous male and female Eutamias sibiricus, without significant gender differences.Comparing the compounds identified in Eutamias sibiricus urine of the same sex, the content of 2-Heptanone,2,5-Dimethylpyrazine,4-Methylphenol,4-Ethylphenol, Pseudoionone in the estrous and non-estrous male Eutamias sibiricus urine showed significant difference, among them,2-Heptanone,2,5-Dimethylpyrazine, Pseudoionone in estrous male urine was significantly higher than non-estrous,4-Methylphenol, 4-Ethylphenol was opposite; in addition, the content of 2-Heptanone,4-Ethylphenol, Pseudoionone in the estrous and non-estrous female Eutamias sibiricus urine were significantly different, among them,2-Heptanone, Pseudoionone in estrous female urine was significantly higher than the non-estrous,4-Ethylphenol was opposite. Thus, these five compounds might be potential pheromone in Eutamias sibiricus, and played some roles in sex attractiveness.3. Studying urine odor preference by Eutamias sibiricus for estrous and non-estrous heterosexual individual, the results showed that regardless of the gender the subjects preferred heterosexual urine odors to distilled water (ck), the attraction effect of estrus urine was more obvious than non-estrus urine.4. Studying behavioral responses to heterosexual urine odors by Eutamias sibiricus in a Y-maze, the results showed when compared with the blank control (ck), the frequency and duration of exploratory behavior, territorial behavior and aggressive behavior of the subjects to heterosexual urine had significant difference, while smoothing behavior and resting behavior indicated no differences. Furtherly, compared the behavioral response of Eutamias sibiricus to estrous with non-estrous urine, the results showed regardless of the gender, the subjects spent much more time exploring estrous urine than non-estrous urine, while there was no significant difference in frequency; but there was no significant difference of the other four behavior between the two reproductive status. This suggested that under heterosexual urine odors, especially estrous urine odors, the exploratory behavior of Eutamias sibiricus associated with sex-attractant was more frequent.5. Studying urine odor preference by Eutamias sibiricus for five potential pheromone ingredients, the results showed although Pseudoionone in the estrous male and female Eutamias sibiricus urine was richer than non-estrous, the compound didn’t obviously attract the subjects in this study; but 2-Heptanone (200 ppm), 2,5-Dimethylpyrazine(800 ppm & 400 ppm) could attract the males,2-Heptanone(800 ppm),4-Methylphenol(400 ppm & 200 ppm),4-Ethylphenol(800 ppm) could attract the females. Among them,2-Heptanone showed two-way sexual attraction, the rest of the three components showed one-way sexual attraction. Nevertheless, the attraction effect of these odor stimulus was still not obvious than heterosexual urine odors by Eutamias sibiricus. This suggested that a single component might not achieve the effect of enduringly and effectively attracting the opposite sex, thus, it might be necessary multi-components combine through rigorous proportion. |
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