<?xml version="1.0"?>
<ownershipDocument>

    <schemaVersion>X0508</schemaVersion>

    <documentType>4</documentType>

    <periodOfReport>2025-03-10</periodOfReport>

    <issuer>
        <issuerCik>0000803578</issuerCik>
        <issuerName>FIREFLY NEUROSCIENCE, INC.</issuerName>
        <issuerTradingSymbol>AIFF</issuerTradingSymbol>
    </issuer>

    <reportingOwner>
        <reportingOwnerId>
            <rptOwnerCik>0002037643</rptOwnerCik>
            <rptOwnerName>Vnook Stella</rptOwnerName>
        </reportingOwnerId>
        <reportingOwnerAddress>
            <rptOwnerStreet1>C/O FIREFLY NEUROSCIENCE, INC.</rptOwnerStreet1>
            <rptOwnerStreet2>1100 MILITARY ROAD</rptOwnerStreet2>
            <rptOwnerCity>KENMORE</rptOwnerCity>
            <rptOwnerState>NY</rptOwnerState>
            <rptOwnerZipCode>14217</rptOwnerZipCode>
            <rptOwnerStateDescription></rptOwnerStateDescription>
        </reportingOwnerAddress>
        <reportingOwnerRelationship>
            <isDirector>1</isDirector>
        </reportingOwnerRelationship>
    </reportingOwner>

    <aff10b5One>0</aff10b5One>

    <derivativeTable>
        <derivativeTransaction>
            <securityTitle>
                <value>Deferred Stock Units</value>
            </securityTitle>
            <conversionOrExercisePrice>
                <footnoteId id="F1"/>
            </conversionOrExercisePrice>
            <transactionDate>
                <value>2025-03-10</value>
            </transactionDate>
            <transactionCoding>
                <transactionFormType>4</transactionFormType>
                <transactionCode>A</transactionCode>
                <equitySwapInvolved>0</equitySwapInvolved>
            </transactionCoding>
            <transactionAmounts>
                <transactionShares>
                    <value>16667</value>
                </transactionShares>
                <transactionPricePerShare>
                    <value>0</value>
                </transactionPricePerShare>
                <transactionAcquiredDisposedCode>
                    <value>A</value>
                </transactionAcquiredDisposedCode>
            </transactionAmounts>
            <exerciseDate>
                <footnoteId id="F1"/>
            </exerciseDate>
            <expirationDate>
                <footnoteId id="F1"/>
            </expirationDate>
            <underlyingSecurity>
                <underlyingSecurityTitle>
                    <value>Common Stock</value>
                </underlyingSecurityTitle>
                <underlyingSecurityShares>
                    <value>16667</value>
                </underlyingSecurityShares>
            </underlyingSecurity>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>16667</value>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </derivativeTransaction>
    </derivativeTable>

    <footnotes>
        <footnote id="F1">On March 10, 2025, the Reporting Person was granted 16,667 DSUs under the Issuer's 2024 Long-Term Incentive Plan. A total of 8,333 DSUs vested immediately on the date of grant. Another 4,167 DSUs will vest on June 10, 2025, and an additional 4,167 DSUs will vest on September 10, 2025. Each DSU represents a contingent right to receive one share of common stock of the Issuer.</footnote>
    </footnotes>

    <ownerSignature>
        <signatureName>/s/ Stella Vnook</signatureName>
        <signatureDate>2025-03-12</signatureDate>
    </ownerSignature>
</ownershipDocument>
