What Is The Function Of Sealed Collection Bottles And Ethanol Solutions? Maximize Varroa Mite Sampling Accuracy

The function of sealed collection bottles and ethanol solutions is to simultaneously preserve the sample and chemically break the bond between the parasite and the host. In the ethanol wash method, the sealed bottle acts as a secure vessel for vigorous mechanical agitation and transport, ensuring no specimen is lost. The 70 percent ethanol solution serves as a critical detachment agent, effectively reducing the adhesion of Varroa mites to the honey bee's body so they can be dislodged and counted.

Accurate Varroa assessment relies on overcoming the strong physical grip the mite has on the bee without destroying the ability to count them. This method utilizes ethanol's chemical properties to loosen that grip, enabling precise infestation calculations that are vital for identifying mite-resistant bee colonies.

The Mechanism of Detachment

The Chemical Role of Ethanol

The primary function of the ethanol solution—specifically at a 70 percent concentration—is to act as a release agent. It chemically infiltrates the contact point between the mite and the bee, significantly reducing the adhesion that allows the parasite to cling to its host. By wetting the surfaces effectively, the solution prepares the mites for physical separation.

The Necessity of Mechanical Agitation

The sealed collection bottle allows for vigorous shaking without the risk of spillage or sample loss. This mechanical oscillation works in tandem with the chemical action of the ethanol. While the alcohol loosens the mites, the physical force of shaking is what actually dislodges them from the bee's thorax or abdomen.

Ensuring Data Integrity

Biological Preservation

Beyond detachment, the ethanol solution serves as a powerful fixative and preservative. It instantly halts biological degradation, ensuring that the bees and mites do not decay between the time of collection and analysis. This is particularly important if samples must be transported to a laboratory under refrigerated conditions.

Enabling Precise Filtration

Once the mites are detached and suspended in the liquid, the mixture is passed through a filter or mesh screen. The ethanol medium facilitates this separation, allowing the smaller mites to pass through for counting while retaining the larger worker bees. This distinct separation allows for a definitive calculation of the infestation percentage (mites per 100 bees).

Understanding the Trade-offs

Sample Mortality

It is important to recognize that this is a destructive sampling method. The high concentration of ethanol kills the sample bees immediately, meaning they cannot be returned to the colony. This sacrifice is exchanged for the higher accuracy this method offers compared to some non-lethal alternatives.

Chemical Handling Requirements

Using high-concentration ethanol (70-75%) requires adherence to safety protocols regarding flammability and fumes. Furthermore, while some variations suggest 50% ethanol with surfactants, the primary standard for optimal detachment and preservation remains the 70% concentration.

Making the Right Choice for Your Goal

To apply this method effectively, align your procedure with your specific objectives:

If your primary focus is breeding for resistance: Prioritize the 70% ethanol concentration and vigorous shaking to ensure the most accurate mite count possible, as even a small margin of error can affect resistance data.
If your primary focus is remote sampling: Rely on the preservation qualities of the ethanol and the seal of the glass or plastic bottles to transport samples to a lab without spoilage.

The ethanol wash remains the gold standard for accuracy when the precise measurement of infestation levels is more critical than the survival of the sample bees.

Summary Table:

Component	Primary Function	Key Benefit
70% Ethanol Solution	Chemical detachment & preservation	Loosens mite grip and prevents sample decay
Sealed Collection Bottle	Secure vessel for mechanical agitation	Enables vigorous shaking without sample loss
Mechanical Shaking	Physical dislodgement	Forces mites to separate from the honey bee host
Fine Mesh Filtration	Size-based separation	Isolates mites for precise counting per 100 bees

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References

Eliana Bianchi, Silvia B. Lanzavecchia. Phenotypic and genetic characterization of Africanized Apis mellifera colonies with natural tolerance to Varroa destructor and contrasting defensive behavior. DOI: 10.3389/finsc.2023.1175760

This article is also based on technical information from HonestBee Knowledge Base .