Mosquito and egg trapping solution and preparation method thereof

Abstract

The invention relates to a mosquito and egg trapping solution, and solves the problems of high monitoring cost and low monitoring efficiency of aedes albopictus in the prior art. The mosquito and egg trapping solution is prepared by fermenting 3%-10% by mass of bamboo leaf powder and 90%-97% by mass of water in a natural environment at the temperature of 20-24 DEG C for 24-96 hours. The preparation method comprises the following steps: step A, collecting bamboo leaves and drying; step B, grinding the dried bamboo leaves into powder; and step C, uniformly mixing 3%-10% by mass of bamboo leaf powder and 90%-97% by mass of water, and fermenting in a natural environment at the temperature of 20-24 DEG C for 24-96 hours. The Aedes albopictus monitoring device is simple in structure, easy in material taking and simple to manufacture, especially the Aedes albopictus can achieve the spawning effect of dozens of times of clear water within one day, the Aedes albopictus monitoring efficiency is greatly improved, meanwhile, dengue fever can be timely and accurately pre-warned and forecasted, the cost is low, and monitoring is convenient and efficient. In addition, the mosquito killer can be daily used for luring and killing mosquitoes.

Description

Technical Field

[0001] This invention relates to the field of disease prevention and control monitoring technology, specifically to a mosquito attractant and egg-attracting solution. Background Technology

[0002] Dengue fever is a vector-borne infectious disease that is a key area for prevention and control in subtropical regions. In recent years, the dengue fever situation in southern tropical areas has been severe. The main vector for dengue fever is the Aedes albopictus mosquito, which transmits the virus to humans through the bite of female mosquitoes carrying the dengue virus. Therefore, mosquito prevention and control are the main measures for dengue fever prevention and control. How to conduct effective vector surveillance is a key factor in controlling the epidemic and preventing its spread. Strengthening the surveillance of Aedes albopictus mosquitoes is an important means of effectively monitoring and providing early warning of the occurrence and spread of dengue fever.

[0003] The Dengue Fever Emergency Response Plan stipulates that in areas where dengue fever outbreaks have occurred or have been prevalent, mosquito vector density surveys and mosquito virus-carrying status monitoring should be conducted year-round to assess the potential severity of dengue fever outbreaks and analyze the epidemic situation. Numerous reports and preliminary field trials suggest that when using ovitraps for dengue vector monitoring, they should be removed after four days, and the captured adult mosquitoes and eggs should be examined and collected. Early warnings should be issued promptly based on mosquito vector density and dengue fever epidemic trends. During dengue fever outbreaks, the ovitrap method can serve as a safe, effective, and convenient evaluation method for emergency mosquito control, especially advantageous in evaluating adult mosquito control effectiveness and detecting virus-carrying in captured adult mosquitoes.

[0004] The national standard (GB / T 23797-2009) for vector density monitoring (mosquitoes) mentions that the current method for monitoring Aedes albopictus eggs is the ovipositor method. The procedure is as follows: fill the ovipositor with 20 mL of tap water that has been left overnight or clear water from the natural environment, place two ovipositors at a distance of ≥10 m between them, and leave them for 4 consecutive days. Check the retrieved ovipositors and collect Aedes albopictus eggs, and calculate the ovipositor positivity rate and ovipositor density.

[0005] Currently, the national standard for mosquito monitoring has adopted the ovipositor method as a routine method for trapping adult mosquitoes and assessing their density. However, this method is time-consuming (requiring 4-5 days), costly, and not very sensitive, making it unsuitable for rapid assessment of Aedes albopictus mosquitoes (especially during dengue fever outbreaks). Improving monitoring efficiency to provide accurate and timely dengue fever early warnings and forecasts has become an urgent problem to be solved. Summary of the Invention

[0006] The purpose of this invention is to solve the problems of high cost and low efficiency in existing monitoring of Aedes albopictus mosquitoes.

[0007] The technical solution adopted to solve the technical problem proposed in this invention is as follows: The mosquito and egg attractant solution of this invention is made by fermenting bamboo leaf powder (3%-10% by weight) and water (90%-97% by weight) in a natural environment at a temperature of 20-24 degrees Celsius for 24 to 96 hours.

[0008] A method for preparing a mosquito attractant and egg-attracting solution, characterized in that the method includes the following steps: Step A: Collect bamboo leaves and dry them; Step B: Grind the dried bamboo leaves into powder; Step C: Mix 3%–10% bamboo leaf powder with 90%–97% water until homogeneous, and ferment in a natural environment at 20–24 degrees Celsius for 24 to 96 hours.

[0009] The technical solutions that further define the present invention include: The bamboo leaf powder mentioned is powder of Conghua Maozhu bamboo, green-skinned bamboo, Buddha's belly bamboo, or golden-green bamboo.

[0010] The mosquito and egg attractant solution is made by fermenting 3%-5% Conghua bamboo powder and 95%-97% water in a natural environment at a temperature of 20-24 degrees Celsius for 48 hours.

[0011] The mosquito and egg attractant solution is made by fermenting 10% Conghua bamboo powder and 90% water in a natural environment at a temperature of 20-24 degrees Celsius for 24 to 72 hours.

[0012] The mosquito and egg attractant solution is made by fermenting 3% green bamboo powder and 97% water in a natural environment at a temperature of 20-24 degrees Celsius for 48 hours.

[0013] The mosquito and egg attractant solution is made by fermenting 3%-5% of Buddha's belly bamboo powder and 95%-97% of water in a natural environment at a temperature of 20-24 degrees Celsius for 72 hours.

[0014] The mosquito and egg attractant solution is made by fermenting 10% by weight of golden bamboo powder and 90% by weight of water in a natural environment at a temperature of 20-24 degrees Celsius for 24 hours.

[0015] The powder is 50-200 mesh.

[0016] The beneficial effects of this invention through the above technical solution are as follows: The mosquito and egg-attracting solution of this invention is made by fermenting 3%-10% bamboo leaf powder and 90%-97% water in a natural environment at a temperature of 20-24 degrees Celsius for 24 to 96 hours. The materials are readily available, and the preparation is simple. In particular, the bamboo from Conghua achieves an egg-laying effect several times greater than that of clean water within one day, greatly improving the monitoring efficiency of Aedes albopictus. Simultaneously, it enables timely and accurate early warning and forecasting of dengue fever. It is low-cost, convenient, and efficient in monitoring. Furthermore, it can be used daily for attracting and killing mosquitoes. Attached Figure Description

[0017] Figure 1 Bar graph showing mosquito egg count with bamboo species as a variable factor; Figure 2 A bar graph showing the number of mosquito eggs with time as a variable factor; Figure 3 A bar graph showing the number of mosquito eggs with solution concentration as a variable factor; Figure 4 Multiplex plots showing the number of mosquito eggs under different experimental conditions; Figure 5 Three-dimensional bar chart of mosquito egg count under different experimental conditions. Detailed Implementation

[0018] The structure of the present invention will be further described below with reference to the accompanying drawings.

[0019] Reference Figures 1 to 5 The mosquito and egg attractant solution of the present invention is made by fermenting 3%-10% bamboo leaf powder and 90%-97% water in a natural environment at a temperature of 20-24 degrees Celsius for 24 to 96 hours.

[0020] The method for preparing the mosquito attractant and egg-attracting solution of the present invention includes the following steps: Step A: Collect bamboo leaves and dry them; Step B: Grind the dried bamboo leaves into powder; in practice, a pulverizer can be used to grind the dried bamboo leaves into powder of 50-200 mesh.

[0021] Step C: Mix 3%–10% bamboo leaf powder with 90%–97% water until homogeneous, and ferment in a natural environment at 20–24 degrees Celsius for 24 to 96 hours.

[0022] In this embodiment, the bamboo leaf powder is Conghua Maozhu powder, Qingpizhu powder, Foduzhu, or Huangjinjianbizhu.

[0023] The following are experiments on the oviductivity of four bamboo leaf extracts with different fermentation times and concentrations: The experimental methods are as follows: (1) Take bamboo leaf extracts with different concentrations and fermentation times: Take the same amount of bamboo leaf extracts with concentrations of 3%, 5% and 10% and fermentation times of 96 hours, 72 hours, 48 ​​hours and 24 hours respectively into different triangular beakers, and label them. Take three beakers of each solution.

[0024] (2) Prepare three cages, hereinafter referred to as cage 1, cage 2 and cage 3. Place one cup of each of the above solutions in each cage, and add another cup of dechlorinated overnight water in the same cage for the test.

[0025] (3) Place a certain number of Aedes albopictus mosquitoes in the cage during their egg-laying period and allow them to lay eggs for 24-48 hours. Then collect and count the number of eggs laid by Aedes albopictus in different water bodies.

[0026] The following are experimental data on the oviduction ability of four different bamboo leaf powder extracts at different fermentation times and concentrations.

[0027] The preferred mosquito and egg-attracting solution is prepared by fermenting 3% (by weight) green bamboo powder and 97% water in a natural environment at 20-24 degrees Celsius for 48 hours. The experimental data in the table above show that a 3% concentration of fermented green bamboo powder solution can achieve a relatively good mosquito and egg-attracting effect after 48 hours of fermentation.

[0028]

[0029] Preferred: The mosquito attractant and egg-attracting solution is prepared by fermenting 3%-5% by weight of *Phyllostachys edulis* powder and 95%-97% by weight of water in a natural environment at a temperature of 20-24 degrees Celsius for 48 hours. Alternatively, the mosquito attractant and egg-attracting solution is prepared by fermenting 10% by weight of *Phyllostachys edulis* powder and 90% by weight of water in a natural environment at a temperature of 20-24 degrees Celsius for 24 to 72 hours.

[0030] The experimental data in the table above show that fermentation with 3% and 5% concentrations of Conghua bamboo powder yielded good mosquito and egg-attracting effects after 24, 72, and 96 hours of fermentation. In particular, the 3% concentration of Conghua bamboo powder fermented for 24 hours achieved several times the mosquito and egg-attracting effect of fermented water from other bamboo leaf powders, and dozens of times the effect of plain water. Furthermore, the 3% concentration of Conghua bamboo powder fermented for 96 hours showed the most significant mosquito and egg-attracting effect, greatly improving the monitoring efficiency of Aedes albopictus mosquitoes. This enables timely and accurate dengue fever early warning and forecasting. Additionally, it can be used daily for mosquito attraction and killing, is low-cost, convenient, and effective.

[0031]

[0032] Preferably, the mosquito and egg-attracting solution is prepared by fermenting 3%–5% (by weight) of *Bambusa textilis* powder and 95%–97% (by weight) of water in a natural environment at a temperature of 20–24 degrees Celsius for 72 hours. As shown in the table above, fermented *Bambusa textilis* powder at concentrations of 3% and 5% after 72 hours of fermentation yields relatively good mosquito and egg-attracting effects.

[0033]

[0034] Preferably, the mosquito and egg attractant solution is made by fermenting 10% by weight of golden bamboo powder and 90% by weight of water in a natural environment at a temperature of 20-24 degrees Celsius for 24 hours.

[0035] The above table shows the fermented water of golden bamboo powder. Fermented water of Buddha's Belly bamboo powder at a concentration of 10% after 24 hours of fermentation can achieve a relatively good mosquito and egg attraction effect.

[0036] The following is an analysis report of experimental data on the oviduction ability of four types of bamboo leaf extracts with different fermentation times and concentrations. This research report aims to screen and verify the optimal formulation of bamboo leaf extract that can significantly attract female mosquitoes to lay eggs, through controlled experiments and statistical analysis. The effects of bamboo leaf species, fermentation time, and solution concentration on mosquito egg production were systematically investigated. A general linear model (GLM) was used for analysis, and to ensure the absolute reliability of statistical inferences, the HC3 robust standard error optimized for heteroscedastic data was employed for calculation. The core findings are as follows: 1. The model was highly significant overall (p<0.001, R² = 0.976), indicating that the difference in egg production was almost entirely explained by the experimental design factors.

[0037] 2. A significant third-order interaction effect exists (p<0.001), indicating that the optimal effect depends on a specific combination of bamboo leaf species, time, and concentration, rather than a simple superposition of individual factors.

[0038] 3. The main effects and interaction effects at all levels of all factors were extremely significant (all p < 0.001).

[0039] 4. The optimal global combination was: Conghua bamboo extract, with a fermentation time of 96 hours and a concentration of 3%, whose predicted egg production (average 162.667 eggs) was significantly higher than all other combinations (p<0.001). The conclusions of this study are robust, providing solid data support and statistical evidence for the patent application of this highly effective mosquito and egg attractant formulation.

[0040] 1. Introduction Based on previous observations and literature review, this study proposes the hypothesis that the extract of certain types of bamboo leaves, after fermentation under specific conditions, can produce chemical substances that are highly effective at attracting female mosquitoes to lay eggs.

[0041] This study aims to quantitatively analyze the effects of three key variables—bamboo leaf species (including four types of bamboo and a water control), fermentation time (four levels), and solution concentration (three levels)—on the number of eggs laid by female mosquitoes through rigorous experimental design, and to determine the optimal formula combination using advanced statistical models.

[0042] 2. Materials and Methods 2.1 Experimental Materials The bamboo leaves used in the experiment (Conghua Maozhu, Foduzhu, Huangjinjianbizhu, and Qingpizhu) were collected from Ayiliu Village in Conghua District, Qide Road in Baiyun District, and Hengzhigang Community in Yuexiu District. The water used was overnight dechlorinated water as a control. After removing impurities and soil, the dried bamboo leaves were initially cut into small pieces and then ground into a fine, easily soluble powder using a stirrer. The powder was then dissolved in water and allowed to ferment. Four different bamboo leaf powders were weighed into Erlenmeyer flasks at concentrations of 3%, 5%, and 10% using an electronic balance and fermented under the same temperature and humidity. The fermentation times were 24 hours, 48 ​​hours, 72 hours, and 96 hours, respectively. Each flask was labeled, and the concentration of the bamboo leaf extract and the start and end times of fermentation were recorded for later use.

[0043] 2.2 Experimental Design This study employed a completely randomized experimental design with five factors (bamboo material) × four levels (time) × three levels (concentration). Each experimental unit (i.e., each combination) was repeated three times. For all experimental groups, one cup each of bamboo leaf extract and dechlorinated overnight water were placed in the same cage. A certain number of gestating Aedes albopictus mosquitoes were placed in the cages and allowed to lay eggs for 24-48 hours. The number of eggs laid by Aedes albopictus in different water bodies was collected and statistically analyzed to determine significant differences.

[0044] 2.3 Data Collection: The dependent variable was the number of mosquito eggs. The total number of mosquito eggs produced in each experimental unit was collected and counted.

[0045] 2.4 Statistical Analysis All data analyses were performed in IBM SPSS Statistics 27.

[0046] Statistical model: Univariate analysis was performed using a general linear model (GLM). The dependent variable was "mosquito egg count", and the fixed factors were "bamboo leaf species", "time", and "concentration".

[0047] Heteroscedasticity handling: The Levin homogeneity of variance test indicates the presence of heteroscedasticity in the data (test based on the mean, based on the p-cut mean, p < 0.05; based on the median, based on the median with adjusted degrees of freedom, p > 0.9). Therefore, significance tests for all parameter estimates were calculated based on the HC3 heteroscedasticity robust standard error to ensure the reliability of all p-values ​​and confidence intervals.

[0048] Post-hoc testing: For significant main effects, Tukey HSD method was used for post-hoc multiple comparisons.

[0049] Optimal combination determination: The global optimal combination is determined by calculating and comparing the estimated marginal means (EMMs) of the third-order interaction terms of "bamboo leaf type × time × concentration".

[0050] 3. Results 3.1 Descriptive Analysis Descriptive statistical analysis was performed on the mosquito egg count data collected from 180 valid experimental units in this study, and the results are shown in the table below.

[0051] Table 1: Descriptive statistics of mosquito egg count under different experimental conditions (mean ± standard deviation)

[0052] Note: The mean represents the average number of eggs laid, and the standard deviation reflects the degree of fluctuation in the data within the group.

[0053] As shown in the table above, there are significant differences in the number of eggs laid by female mosquitoes under different experimental conditions.

[0054] (1) From the overall trend, the average number of eggs laid for all 60 observations was 2383, and the standard deviation was 299, indicating that there is a certain degree of variability in the data.

[0055] (2) From the perspective of factor level: Bamboo species: The experimental groups using bamboo materials generally had a higher average number of eggs laid than the water control group (10.75 eggs). Among them, Conghua bamboo showed the highest average egg-laying level (98.34 eggs) among all bamboo species.

[0056] Fermentation time: The average number of eggs laid fluctuated as the fermentation time increased. The group with 72 hours of fermentation had the highest average number of eggs laid (45.07), while the group with 48 hours of fermentation had the lowest average number of eggs laid (36.47).

[0057] Solution concentration: The average number of eggs laid decreased with increasing concentration. The 3% concentration group had the highest average number of eggs laid (49.15). See Figures 1 to 4 Bar graph showing the number of mosquito eggs under different experimental conditions.

[0058] (3) From the perspective of combined effects, preliminary observations revealed that the *Imperata cylindrica* from Conghua, under a specific combination of 72 hours of fermentation and a concentration of 3%, achieved the highest average number of eggs laid (163) among all combinations, and its within-group standard deviation (16) was relatively small, indicating good data stability under these conditions. In stark contrast, the water control group, under the conditions of the longest fermentation time and the highest concentration, had the lowest average number of eggs laid (2). See Figure 5 Three-dimensional bar chart of mosquito egg count under different experimental conditions.

[0059] 3.2 Overall Model Performance Evaluation Between-subjects effects tests (see Table 1) showed that the calibrated model had a very high explanatory power for mosquito egg count (R² = [0.984], adjusted R² = [0.976]), and the model as a whole was extremely significant (F = 38.670, p < 0.001). More importantly, all main effects (bamboo leaf species, time, concentration) and all order of interaction effects (including second- and third-order interactions) were extremely significant (all p < 0.001), indicating that the three factors must be considered as a whole to determine their optimal combination.

[0060] Table 1: Inter-subject effects test (based on Type III sum of squares and HC3 robust standard error) Table 1-1 Heteroscedasticity test a, b, c; Table 1-2 Between-subjects effect test

[0061] 3.3 Determination of the optimal spawning induction combination An analysis of the estimated marginal means of all 60 combinations of "bamboo leaf species × time × concentration" (see Table 2) revealed that the optimal combination for oviposition attraction was: Conghua bamboo, fermented for 96 hours at a concentration of 3%, which predicted a maximum of 162.667 mosquito eggs. The lower limit of the 95% confidence interval (155.958, 169.375) for this value is still much higher than the upper limit of the confidence interval (146.042) for the second-best combination ([Conghua bamboo], [96 hours], [5%]), demonstrating its superiority and high statistical robustness.

[0062] Table 2: Estimated Marginal Mean (Bamboo Leaf Species * Time * Concentration) - Optimal and Suboptimal Combinations

[0063] 3.4 Main Effects and Post-hoc Comparison of Bamboo Leaf Species Factors Post-hoc multiple comparisons (Tukey HSD) showed (see Table 3) that the attraction effect of "Conghua Maozhu" as a whole was significantly better than that of the water control group and all other bamboo species (all comparisons p<0.001).

[0064] Table 3: Post-hoc multiple comparisons of mosquito oviposition (bamboo leaf factor - Tukey HSD, Bonfrenney)

[0065] 4. Discussion This study irrefutably demonstrates through advanced statistical models that the Conghua bamboo-96-hour-3% formulation is the globally optimal formula for attracting female mosquitoes to lay eggs. Its predicted egg production is far superior to other combinations, and this advantage has extremely high statistical confidence (p<0.001, CI non-overlapping).

[0066] The significant third-order interaction indicates that the optimal effect depends on the precise matching of bamboo leaf species, time, and concentration, which constitutes a narrow "parameter window." This scientifically explains why similar highly effective formulations have not been widely reported before, as their effects are not a simple superposition of single factors, but rather the result of synergistic effects of multiple factors, demonstrating the novelty and inventiveness (non-obviousness) of this invention.

[0067] Furthermore, a large number of comparisons in the parameter estimates were not significant (7 / 60), which precisely proves that some experimental formulations had mediocre effects and little difference from each other, thus further highlighting the outstanding performance of the optimal combination.

[0068] In summary, this optimal formula not only significantly increases mosquito egg production, providing an efficient tool for laboratory mosquito breeding, but also demonstrates enormous application potential and market value in mosquito population monitoring and green control based on the "attract-kill" method.

[0069] 5. Conclusion 1. The type of bamboo leaves, fermentation time, and solution concentration all had a highly significant impact on the oviposition rate of female mosquitoes, and there was a significant interaction between them.

[0070] 2. Among all experimental combinations, the combination of extract from Conghua bamboo, fermented for 96 hours and prepared at a concentration of 3%, showed the most significant effect in attracting female mosquitoes to lay eggs, and was undoubtedly the optimal solution globally.

[0071] This experiment, through rigorous experimental design, quantitatively analyzed the effects of three key variables—common bamboo leaf species (including four types of bamboo leaf materials and a control with water), fermentation time (four fermentation times), and solution concentration (three solution concentrations)—on the number of eggs laid by female mosquitoes. It also analyzed the attraction effect of chemical substances produced by specific types of bamboo leaf extracts fermented under specific conditions on Aedes albopictus mosquitoes. The experiment showed that fermented water containing bamboo leaf powder at specific concentrations could produce an egg-laying effect several times greater than that of water. In particular, fermented water containing Conghua bamboo powder achieved an egg-laying effect dozens of times greater than that of water within one day. Furthermore, fermented water containing 3% Conghua bamboo powder after 96 hours of fermentation showed the most significant mosquito and egg-laying effect, greatly improving the monitoring efficiency of Aedes albopictus and enabling timely and accurate dengue fever early warning. In addition, the materials are readily available, low-cost, and fast-acting, and can be used routinely for mosquito attraction and control.

[0072] While specific embodiments of the present invention have been described in detail, this should not be construed as limiting the scope of protection of the invention. Various modifications and variations that can be made by those skilled in the art without inventive effort within the scope described in the claims still fall within the scope of protection of the present invention.

Claims

1. A mosquito attractant and egg-attracting solution, characterized in that: The mosquito and egg attractant solution is made by fermenting 3%-10% bamboo leaf powder and 90%-97% water in a natural environment at a temperature of 20-24 degrees Celsius for 24 to 96 hours.

2. The mosquito attractant and egg-attracting solution as described in claim 1, characterized in that: The bamboo leaf powder mentioned is powder of Conghua Maozhu bamboo, green-skinned bamboo, Buddha's belly bamboo, or golden-green bamboo.

3. The mosquito attractant and egg-attracting solution as described in claim 2, characterized in that: The mosquito and egg attractant solution is made by fermenting 3%-5% Conghua bamboo powder and 95%-97% water in a natural environment at a temperature of 20-24 degrees Celsius for 24 to 96 hours.

4. The mosquito attractant and egg-attracting solution as described in claim 2, characterized in that: The mosquito and egg attractant solution is made by fermenting 10% Conghua bamboo powder and 90% water in a natural environment at a temperature of 20-24 degrees Celsius for 24 to 72 hours.

5. The mosquito attractant and egg-attracting solution as described in claim 2, characterized in that: The mosquito and egg attractant solution is made by fermenting 3% green bamboo powder and 97% water in a natural environment at a temperature of 20-24 degrees Celsius for 48 hours.

6. The mosquito attractant and egg-attracting solution as described in claim 2, characterized in that: The mosquito and egg attractant solution is made by fermenting 3%-5% of Buddha's belly bamboo powder and 95%-97% of water in a natural environment at a temperature of 20-24 degrees Celsius for 72 hours.

7. The mosquito attractant and egg-attracting solution as described in claim 2, characterized in that: The mosquito and egg attractant solution is made by fermenting 10% by weight of golden bamboo powder and 90% by weight of water in a natural environment at a temperature of 20-24 degrees Celsius for 24 hours.

8. A method for preparing a mosquito-attracting and egg-attracting solution as described in any one of claims 1-7, characterized in that: The manufacturing method includes the following steps: Step A: Collect bamboo leaves and dry them; Step B: Grind the dried bamboo leaves into powder; Step C: Mix 3%–10% bamboo leaf powder with 90%–97% water until homogeneous, and ferment in a natural environment at 20–24 degrees Celsius for 24 to 96 hours.

9. The method for preparing the mosquito-attracting and egg-attracting solution as described in claim 8, characterized in that: The powder is 50-200 mesh.

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