Automated in ovo injection apparatus

Animal husbandry – Avian egg treatment or production

Reexamination Certificate

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Reexamination Certificate

active

06286455

ABSTRACT:

FIELD OF THE INVENTION
The present invention relates to treatment of avian embryos and, more particularly, relates to in ovo injection devices and methods for delivering various substances to live embryonated eggs.
BACKGROUND OF THE INVENTION
Injections of various substances into avian eggs have been employed to decrease post-hatch mortality rates, increase the potential growth rates or eventual size of the resulting chicken, and even to influence the gender determination of the embryo. Similarly, injections of antigens into live eggs have been employed to incubate various substances used in vaccines which have human or animal medicinal or diagnostic applications.
Examples of substances which have been proposed as viable treatment (or harvestable vaccine material) alternatives for delivery via in ovo injection of avian embryos include live culture vaccines, antibiotics, vitamins, and even competitive exclusion media (a live replicating organism). Specific examples of treatment substances are described in U.S. Pat. No. 4,458,630 to Sharma et al, and U.S. Pat. No. 5,028,421 to Fredericksen et al.
Conventionally, the physical injection has been typically targeted at preferred positions within the egg in order to administer the substance into specific developing regions of the embryo. As understood by those of skill in the art, as the incubation period progresses towards maturity (i.e., hatching), the embryo and its membranes, e.g., the air cell, the allantois, and yolk sac, correspondingly change in both volume and position within the egg shell. Additionally, the quantitative volume of the enclosed fluids vary as well; for example, the density of the allantois (fluid, solid) varies as a function of time over the incubation period.
Thus, selection of both the site and time of treatment can impact the effectiveness of the injected substance as well as the mortality rate of the treated embryos. See e.g., U.S. Pat. No. 4,458,630 to Sharma et al., U.S. Pat. No. 4,681,063 to Hebrank, and U.S. Pat. No. 5,158,038 to Sheeks et al.
SUMMARY OF THE INVENTION
The present invention, recognizes that there is a need to introduce multiple substances into a live egg with a minimum of trauma thereto, including substances which are effective treatment alternatives when separately injected but become biologically noxious when combined. Thus, a first object of the present invention is to provide a multi-site in ovo injection device for delivering a variety of treatment substances to avian embryos while minimizing the risk of injury thereto.
Additionally, the present invention recognizes that there is a need to withdraw multiple samples from a live egg with a minimum of trauma thereto, including withdrawing two samples from different compartments of, or locations in, the egg at the same time. Thus, a further object of the present invention is to provide a multi-site in ovo sampling device for withdrawing a variety of samples from avian embryos while minimizing the risk of injury thereto.
It is another object of the present invention to introduce, without mixing, biologically incompatible products in ovo to embryos.
It is a further object of the present invention to separately introduce without mixing at least two different treatment materials into different locations in the egg, through either a single or two separate delivery paths.
It is another object of the present invention to introduce at least two different treatment substances which are separately delivered by one or more of time and spatial separation into an opening in the egg shell.
These and other objects, advantages, and features are provided by a multi-site or multi-dosage injection or withdrawal methods and apparatus disclosed herein. The methods and apparatus of the invention deliver at least two different substances into predetermined areas within the egg, or withdraw samples from at least two different predetermined locations in the egg.
In particular, a first aspect of the present invention is a multi-injection method for treating avian embryos in ovo. In the method, an avian egg is oriented into a predetermined position and a small first opening is introduced into the shell of the avian egg. A delivery device which has either a single or a plurality of lumens therein is extended through the first opening and into the egg a predetermined depth. Predetermined dosages of a first substance and a second substance are separately released into the egg and the delivery device is retracted from the egg, thereby treating the avian embryo. Advantageously, a plurality of lumens can include separate needles that separately deliver the first and second substances to spatially separate areas, or different compartments, of the egg. In one embodiment, a first needle extends longitudinally a greater distance in the egg than a second needle; alternatively, one or more of the needles can include a side port to dispense the substance transversely spatially separated from the other substance. Alternatively, the needles can be adapted to withdraw a sample of material from the egg.
Another aspect of the present invention includes a multi-injection method for treating avian embryos in ovo that first orients an avian egg into a predetermined position and then introduces a small first opening into the shell of an avian egg. Additionally, a small second opening is introduced into the shell of the avian egg, the second opening being spaced apart from the first opening. Respective ones of the first and second delivery devices are extended through corresponding first and second openings and into the egg a predetermined depth. A predetermined dosage of a first substance and a second substance is released from respective ones of the first and second delivery devices into the egg. The delivery devices are retracted from the egg, thereby treating the avian embryo. Alternatively, the delivery devices can be adapted as sampling devices, to withdraw a sample of material from the egg.
Yet another aspect of the present invention includes a multi-injection method for treating avian embryos in ovo which orients an avian egg into a predetermined position and introduces a small first opening into the shell of an avian egg. A delivery device is extended through the first opening and into the egg a predetermined depth. Predetermined dosages of a first substance and a second substance are released into the egg and the delivery device is retracted from the egg, thereby treating the avian embryo. Advantageously, this method temporally combines the different substances to minimize degradation of the substances attributed to reactance therebetween. Thus, preferably, this method allows the first and second substances to be stored in separate chambers and temporally combines or mixes the first and second substances, either with an active mixing chamber, or by introducing them into a common delivery path, prior to delivery into the egg.
An additional aspect of the present invention is directed towards an automated in ovo injection apparatus. The apparatus comprises a fixture for holding a plurality of eggs in a substantially upright and aligned position. The fixture is configured to provide external access to predetermined areas of the eggs. The apparatus also includes a plurality of injection delivery devices configured to contact predetermined areas of the egg; at least one of the injection devices corresponds to each egg in the fixture. Each of the delivery devices comprises a first and second lumen that is adapted to be received into the egg. The apparatus further comprises a first treatment substance container for holding a first treatment substance. The first container is in fluid communication with each of the plurality of injection delivery devices. The apparatus also includes a second treatment substance container for holding a second treatment substance. The second container is in fluid communication with each of the plurality of injection devices. The first container and each of the plurality of injection devices defines a first fluid pathway therebetween. Similarly, the se

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