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Immunology and Infection

Intranasal Immunization and Milk Collection in Studies of Maternal Immunization in New Zealand White Rabbits (Oryctolagus cuniculus)

Published: July 31st, 2021



1Division of Laboratory Animal Resources, Duke University Medical Center, 2Department of Pathology, Duke University Medical Center, 3Department of Immunology, Duke University Medical Center, 4Duke Human Vaccine Institute, Duke University Medical Center

This article describes and demonstrates the administration of intranasal vaccines and the collection of milk from lactating rabbits (Oryctolagus cuniculus) as a means to assess mucosal immunity in a translationally appropriate model of maternal immunization.

Due to similarities in placentation and antibody transfer with humans, rabbits are an excellent model of maternal immunization. Additional advantages of this research model are the ease of breeding and sample collection, relatively short gestation period, and large litter sizes. Commonly assessed routes of immunization include subcutaneous, intramuscular, intranasal, and intradermal. Nonterminal sample collection for the chronological detection of the immunologic responses to these immunizations include the collection of blood, from both dams and kits, and milk from the lactating does. In this article, we will demonstrate techniques our lab has utilized in studies of maternal immunization in New Zealand White rabbits (Oryctolagus cuniculus), including intranasal immunization and milk collection.

Studies of maternal immunization and antibody transfer are invaluable for numerous reasons, as this is the initial route of immunity transfer and subsequent protection from pathogens and diseases in newborns and infants. Maternal immunization has the potential to positively impact both maternal and infant/child health at the global level by reducing morbidity and mortality associated with certain pathogens during this vulnerable period1. The main goal of this strategy is to increase the levels of specific maternal antibodies throughout pregnancy. These antibodies can then be transferred to the newborn and infant at levels sufficient enough to p....

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All procedures were approved and performed in accordance with the Duke University IACUC policies.

NOTE: Materials needed are provided in the Table of Materials.

1. Rabbit Sedation and Anesthesia

  1. Sedate the female rabbit (sexually mature; approximately 5-30 months old) by administering acepromazine intramuscularly (IM) at a dose of 1 mg/kg. Depending on the size of the animal, use a 1 or 3 mL syringe with a 25 G needle. Epaxial muscles ar.......

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An overview of a typical maternal intranasal immunization study design is depicted in Figure 1, incorporating the immunizations, breeding, kindling, lactation, and antibody transfer. Though not illustrated, blood should be collected prior to the initial immunization for baseline measurements and throughout the remainder of the study at regular intervals. Blood is easily obtained via the central ear artery with mild sedation and a topical analgesic agent (e.g., lidocaine 2.5% and prilocaine 2.......

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Although not described in the above protocol, successful breeding of the rabbits is necessary for this maternal model and to allow for milk collection. Rabbits are easily bred by live cover in a research setting. It is recommended that does be transferred to the buck's cage for breeding, as does can be territorial and aggressive if kept in their own cage with the buck. If females are non-receptive after 15 minutes (as indicated by running away biting, or vocalizing), the doe should be placed back into her own cage. T.......

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The authors would like to acknowledge the Division of Laboratory Animal Resources at Duke University and their husbandry team for their assistance and great care provided to the animals. Additionally, the authors would like to recognize the PhotoPath team within the Department of Pathology for their assistance with the audio and video portions of the manuscript. This work was supported by discretionary research funds from the Staats laboratory.


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Name Company Catalog Number Comments
Intranasal Immunization
Anesthesia Machine/Vaporizer Vet Equip 901807
Hypodermic Needle (25 g) Terumo 07-806-7584
Isoflurane (250 mL Bottle) Patterson Veterinary 07-893-1389 2-4%
Luer Lock Syringe (1 mL) Air-Tite 07-892-7410
Mucosal Vaccine N/A N/A Experimental Vaccine
Nose Cone McCulloch Medical 07891-1097
Pipette Tips VWR 53503-290
Pipettor VWR 89079-962
PromAce (Acepromazine maleate) Boehringer Ingelheim 07-893-5734 1mg/kg IM
Puralube Sterile Ophthalmic Ointment Dechra 07-888-2572
Milk Collection
Alcohol Prep 2-ply Covidien 07-839-8871
Anesthesia Machine/Vaporizer Vet Equip 901807
Hypodermic Needles (25 g) Terumo 07-806-7584
Isoflurane (250 mL Bottle) Patterson Veterinary 07-893-1389 2-4%
Luer Lock Syringe (1 mL) Air-Tite 07-892-7410
Non-Woven Sponge (4x4) Covidien 07-891-5815
Nose Cone McCulloch Medical 07891-1097
PromAce (Acepromazine Maleate) Boehringer Ingelheim 07-893-5734 1mg/kg IM
Puralube Sterile Ophthalmic Ointment Dechra 07-888-2572
Sterile Conical Vial (15 mL) Falcon 14-959-49B

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