The American Journal of Veterinary Sciences and Wildlife Discovery
06
https://www.theamericanjournals.com/index.php/tajvswd
TYPE
Original Research
PAGE NO.
6-13
10.37547/tajvswd/Volume07Issue03-02
OPEN ACCESS
SUBMITED
21 March 2025
ACCEPTED
17 April 2025
PUBLISHED
19 May 2025
VOLUME
Vol.07 Issue03 2025
CITATION
I.B. Butaeva. (2025). Evaluation of the effectiveness of the
hemagglutination inhibition (hi) test in the diagnosis of avian influenza.
The American Journal of Veterinary Sciences and Wildlife Discovery,
7(03), 6
–
13.
https://doi.org/10.37547/tajvswd/Volume07Issue03-02
\
COPYRIGHT
© 2025 Original content from this work may be used under the terms
of the creative commons attributes 4.0 License.
Evaluation of the
effectiveness of the
hemagglutination
inhibition (hi) test in the
diagnosis of avian
influenza
I.B. Butaeva
LLC "Afrosiab Parranda", Samarkand, Uzbekistan
Abstract:
Avian influenza remains one of the most
dangerous viral infections affecting both wild and
domestic birds. Effective diagnostics are essential to
prevent outbreaks and virus dissemination. This article
discusses the use of the hemagglutination inhibition test
(HI test) as a standard serological method for identifying
antibodies against the avian influenza virus. The
methodology for performing the HI test, interpretation
of results, and its significance in the veterinary control
system in the Republic of Uzbekistan are presente.
Keywords:
Avian influenza, HI test, diagnostics,
serology, H5N1 virus, infection control.
Introduction:
Avian influenza is a highly pathogenic viral
disease caused by various subtypes of influenza A virus,
particularly H5 and H7. The disease poses a serious
threat both to poultry farming and human health.
Among
various
diagnostic
methods,
the
hemagglutination inhibition (HI) test holds a prominent
place due to its utility in virus serotyping and
determining antidiv levels in serum from infected or
vaccinated birds.
METHODS
Study subjects: Serum samples were collected from
chickens raised at the poultry farm LLC "Afrosiab
Parranda" (Samarkand region) following vaccination
against avian influenza (strain H5).
Antigens and erythrocytes: Inactivated H5N1 subtype
avian influenza virus was used as the antigen. Chicken
erythrocytes were used at a concentration of 1% in
The American Journal of Veterinary Sciences and Wildlife Discovery
7
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The American Journal of Veterinary Sciences and Wildlife Discovery
Alsever’s solution.
HI test procedure:
•
Serum samples were heat-inactivated at 56°C
for 30 minutes.
•
Serial twofold dilutions of sera were prepared
in 96-well plates.
•
Virus antigen was added, followed by
incubation for 30 minutes at room temperature.
•
Chicken erythrocytes were then added, and
the plates were incubated for an additional 30-45
minutes.
•
Results were assessed based on the presence
or absence of hemagglutination.
RESULTS
Blood samples for HI testing were collected between
21
–
28 days post-vaccination, aligning with the optimal
period for the development of post-vaccination
immunity. The overall immunity rate was found to be
85%, which is a satisfactory level of protection
according to veterinary norms.
According to the vaccine manufacturer's instructions,
immunity duration post-vaccination is up to 6 months,
provided that vaccine administration protocols are
strictly followed and no immunosuppressive factors
are present.
The analysis included two age categories: young birds
(up to 6 months old) and adults. A total of 120 serum
samples were examined, of which 70 were from adult
chickens and 50 from young birds. Positive antidiv
detection in adults was observed in 64 cases (91.4%),
whereas among young birds, antibodies were detected
in 38 cases (76%). Thus, it can be concluded that adult
chickens developed reliable immunity, while younger
birds demonstrated a lower immunity level, indicating
the need for additional monitoring and potential
revaccination.
Among the 120 samples studied, the average antidiv
titer was 1:64, confirming the vaccine’s effectiveness.
DISCUSSION
The HI test has proven to be a reliable serological
diagnostic method for avian influenza, demonstrating
high sensitivity, ease of execution, and accessibility.
However, the method requires stringent adherence to
protocol conditions, particularly concerning antigen
and erythrocyte quality. Given the limited laboratory
resources available in Uzbekistan, the HI test remains
the primary method for post-vaccination immunity
monitoring.
Furthermore, the HI test complies with international
WOAH (OIE) standards and is widely used in serological
monitoring for export purposes.
CONCLUSION
The hemagglutination inhibition test is a reliable
diagnostic tool for avian influenza. At the poultry farm
LLC "Afrosiab Parranda," the method showed high
informativeness and reproducibility. The obtained
results can be utilized to adjust vaccination protocols
and enhance biosecurity measures.
Conducting the HI test allowed for the assessment of
post-vaccination immunity against avian influenza virus.
An overall immunity rate of 85% confirmed the
effectiveness of the vaccination program. Nevertheless,
younger birds showed a slightly reduced seropositivity
rate (76%), indicating a necessity for ongoing
monitoring and, if needed, adjustment of the
vaccination scheme.
REFERENCES
Capua I., Alexander D.J. Avian influenza and human
health. Acta Tropica, 2002.
WHO Manual on Animal Influenza Diagnosis and
Surveillance, 2002.
Дадабаев Ш.Х. и др. Методические рекомендации
по диагностике гриппа птиц. Ташкент, 2021.
Office International des Epizooties (WOAH). Manual of
Diagnostic Tests and Vaccines for Terrestrial Animals,
2021.
