MXPA01006893A - Concurrent in ovo injection and detection method and apparatus - Google Patents

Abstract

A method of injecting a plurality of bird eggs comprises:(a) orienting a plurality of avian eggs in a predetermined position;(b) forming an opening in the shell of each of the eggs;(c) extending an elongate delivery device through each of the openings and into the eggs, each of the delivery devices comprising a detector and an injection needle, with the injection needle having a lumen formed therein;(d) detecting with the detector information from the interior of each of the plurality of eggs;and (e) injecting a substance into each of the plurality of eggs through the lumen of said injection needle. The detected information can be used for a variety of purposes, including adjusting the depth of penetration of the injection needle to more precisely control the location of the injection, identifying the gender of the eggs for subsequent sorting of the eggs, distinguishing viable from non-viable eggs, etc. When used for controlling depth penetration, the method positions the needle tip for any purpose, including withdrawing biological material as well as injecting substances. Apparatus, particularly high-speed apparatus, for carrying out the method is also disclosed.

Description

INOVO INJECTION CONNECTION AND METHOD AND DETECTION APPARATUS Related requests This application claims the benefit of the provisional application for E.U.A. No 60 / 114,933 filed on January 6, 1999.

FIELD OF THE INVENTION The present invention concerns methods and apparatus for the injection of eggs, particularly eggs of live embryonic birds.

BACKGROUND OF THE INVENTION The injection of substances into bird eggs are used for many reasons, including decreasing post-hatching mortality rates, increasing the potential growth ratio or the eventual size of the resulting chicken, and even influencing the determination of the embryo's genus. Similarly, viruses have been injected into live eggs to produce viruses for use in vaccines. Examples of substances that have been introduced into embryonic poultry eggs via in ovo injection include live vaccine cultures, antibiotics, vitamins and competitive exclusion medium (e.g., a living replicating organism). Specific examples of the treatment substances are described in the patent of E.U.A. No. 4,458,630 to Sharma et al, and in the U.S. patent. No. 5,028,421 to Fredericksen et al. See also the patent of E.U.A. No. 4,458,630 to Sharma et al., U.S. Patent. No. 4,681, 063 of Hebrank, and patent of E.U.A. No. 5,158,038 to Sheeks et al. When using the injection in ovo, the location of the injection will vary depending on the desired result and the injectable to be used. The patent of E.U.A. No. 4,458,630 to Sharma describes injection within the region defined by either the amnion or the yolk sac. The published PCT application W093 / 15185 describes the injection of substances into the air chamber of embryonic eggs; PCT application W093 / 14629 describes the injection into the muscle tissue of the embryo contained within the egg. The patent of E.U.A. No. 5,136,979 to Paul describes a modular injection system for poultry embryos. The system comprises a horizontally oriented platform tool with an opening therethrough, an injector that generally rests vertically in the opening in the tool platform with a lower portion of the injector suspended below the platform tool and an upper portion of the "injector resting on the platform tool." Means are included for raising and lowering the platform tool and the injector together with it so that when the platform is lowered and the lower portion of the injector at rest hits an egg to be injected , the injector at rest stops while the platform tool proceeds downwards until the injector separates from the platform tool and is free to move in a translational direction independent of the platform tool. This is coupled back to the injector and takes it up and away from the egg This apparatus, described in Paul et al., Has been used in commercial high-speed injection of eggs of live birds, and has gained wide acceptance in the industry. The success of this device has made possible the in ovo injection of highly sophisticated and expensive materials, such as vaccines. The size and location of the internal compartments and bird egg structures are, however, inherently variable, and some eggs are occasionally injected into non-optimal sites. The increase of the precise injection is desirable to minimize misdirected injections, in order to avoid the waste of injectables and avoid ineffective injection. In order to achieve an increase in the precision of the injection, it is necessary to obtain useful, real-time information of the interior of the egg during the injection process, which has not been achieved by the methods and automated injection devices in ovo above.

BRIEF DESCRIPTION OF THE INVENTION A first aspect of the present invention is a method for injecting a plurality of bird eggs. The method comprises: (a) orienting a plurality of bird eggs in a predetermined position; (b) forming an opening in the shell of each of the eggs; (c) extending an elongate delivery device through each of the openings and into the eggs, each of the delivery devices comprising a detector and an injection needle, with the injection needle having a lumen formed therein; (d) detecting with the detector information from the inside of each of the plurality of eggs; and (e) injecting a substance into each of the plurality of eggs through the lumen of said injection needle. The injection can be to any suitable location, including the amnion, allantois, embryo, yolk sac, albumen, etc. The detected information can be used for a variety of purposes, including but not limited to adjusting the penetration depth of the injection needle for more precise control of the injection location, stopping the movement of the injection needle to control the depth of the injection, identify the genus of the eggs for subsequent classification of the eggs, and distinguish viable from non-viable eggs so that the non-viable eggs do not need to be injected and / or can be subsequently separated from the viable eggs.

In addition, the stage of development of the embryo can be determined. For example, the air chamber and the allantois become larger as the embryo is older, with the air chamber having different electrical properties than other compartments of the egg. Therefore, the stage of embryonic development can be determined by measuring properties such as the size of the air chamber with an electrical probe. The method may further comprise the steps of: (f) withdrawing the delivery device from each of the eggs and then (g) repeating steps (a) through (e) with a second plurality of eggs to provide a means fast, high speed to automatically inject and concurrently detect information from a large number of eggs. A second aspect of the present invention is a method for precisely locating the tip of a needle within a plurality of bird eggs to inject a substance therein or withdrawing a sample from a plurality of eggs. The method comprises (a) orienting a plurality of bird eggs in a predetermined position; (b) forming an opening on the shell of each of the eggs; (c) extending an elongated delivery device through each of the openings and within the "eggs," each of the delivery devices comprises a detector and a needle, with the needle having a portion of the tip and a lumen formed inside, said lumen extending through the tip portion; (d) detecting the depth detector information from within each of said plurality of eggs, and then (e) independently controlling the depth of penetration of each of the needles based on the corresponding information of the depth of the needle, wherein a substance can be injected or biological material is removed from a particular position of the tip portion of the needle with each of the plurality of eggs through the lumen of said needle A third aspect of the present invention is an apparatus for injecting a plurality of bird eggs with a substance, m While useful information is also detected from inside the injected egg. The apparatus includes an aligned assembly for orientation of a plurality of eggs of birds to be injected. A plurality of injectors are associated with the aligned assembly, while the injectors are configured to inject each of a plurality of bird eggs in a predetermined location. A detector is operatively associated with each of the injection needles to detect information from within each of the plurality of eggs. A fourth aspect of the invention is an apparatus for locating the tip of a needle to inject a substance into or withdraw biological material from a particular location in a plurality of bird eggs. The apparatus comprises an aligned assembly for orientation of a plurality of eggs of birds to be injected. A plurality of. injectors are associated with the aligned assembly and are configured to inject each of a plurality of bird eggs at a predetermined location, each of the injectors includes a needle having a lumen through which the substance is injected or the material is removed. A location detector is connected to and operatively associated with each of the injection needles to detect information on the location of the needle from the interior of said egg. A controller is operatively associated with each of the location detectors to independently control the depth of penetration of each of the needles. The foregoing and other objects and aspects of the present invention are explained in detail in the drawings herein and in the attached specifications below.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a schematic diagram of an apparatus for determining the penetration of the needle into the fluids of an egg and providing a signal to stop the movement of the needle inwardly.

Note that the conductive bearing rises to the egg just before the injection sequence starts. Figure 2 is a side view of an apparatus with multiple injection heads useful for carrying out the present invention.

Figure 3 is a perspective view of an apparatus with multiple injection heads of Figure 2. Figure 4 is a schematic diagram of a current sensor, rectifier, average circuit and threshold circuit to carry out the apparatus of the figure 1. Figure 5 is a schematic diagram of a pneumatic steering control valve that raises or lowers an injection needle and that can hold the injection needle in any intermediate position. Note that the movement is slow enough to allow an accurate stop signal (faster movement can be handled by anticipating the degree of travel before sending the stop signal). Figure 6A illustrates an optical detector according to an embodiment of the present invention. Figure 6B is a side elevational view of the optical detector of Figure 6A. Figure 6C illustrates an exemplary light intensity curve indicating the distance to a membrane. Figure 7 is a schematic diagram of a pressure sensor that is configured to deliver a small amount of fluid into the medium surrounding a needle, according to one embodiment of the present invention. Figure 8 is a schematic diagram of a pressure sensor that is configured to deliver a small amount of fluid into the medium surrounding a needle, according to another embodiment of the present invention.

DETAILED DESCRIPTION OF THE INVENTION The invention will now be described more fully hereinafter with reference to the accompanying drawings, in which preferred embodiments of the invention are shown. As noted below, the invention is particularly adapted to the injection of, or removal of, material from, a plurality of eggs in a high-speed apparatus, such as that described in the US Pat.

E.U.A. No. 5,136,979 to Paul et al., The description of which is incorporated herein by reference herein in its entirety. The invention is practiced with eggs, particularly bird or poultry eggs, and more particularly poultry eggs, such as chicken, turkeys, ducks, geese, quail, pheasant, or ostrich eggs. Eggs can be viable eggs; this is eggs that contain a live bird embryo. Viable eggs can be at any stage of embryonic development, including both early embryonic development and late embryonic development. In addition, non-viable eggs may be injected by the present invention for any suitable purpose, such as growth of vaccine cultures therein.

Any substance can be injected by the present injection, including but not limited to vaccines, hormones, growth promoting agents, probiotic cultures such as competitive exclusion media, antibiotics, heterologous nucleic acids including gene transfer vectors, markers such as dyes, etc. . The substances can be injected alone, or in combination. The substance can be injected into any suitable compartment of the egg, including intraperitoneally, intramuscularly, or subcutaneously within the embryo, within the yolk sac or yolk, within the liver or lungs of the embryo, inside the air chamber, the allantoic sac, the amniotic fluid, albumen, blastoderm, etc. Similarly, biological materials or samples can be removed from any of these compartments or materials, when the invention is used to remove material from the egg. The detection step time will depend on the particular purpose of the method or characteristic to be detected, and the nature of the material to be injected or the sample to be removed. In general, the detection step can be carried out before, after, or concurrently with the injection step. Electrical sensors, optical sensors, chemical sensors, temperature sensors, acoustic sensors, pressure sensors (particularly suitable for detecting the embryonic muscle), or any other device to detect a physical or chemical parameter can serve as detection or detector means for carrying out the present invention. The detector or sensor may be connected to the lateral external wall of the injection needle, or in the case of an electrical detector, and wherein the needle is formed from a conductive metallic material rather than an insulating or polymeric material, the The side of the needle itself can serve as the detector, with suitable circuits connected to it. The detector may be one or two (or more) electrodes carried by a non-conductive needle, or carried in an insulating portion of a driving needle. It will be appreciated that, for the purpose of sensing the depth or location within the anatomy of the egg, for the purposes of distinguishing normal eggs from abnormal eggs (and therefore, eggs to be rejected), and for a variety of other purposes , numerous different physical or chemical parameters can be evaluated or detected, while providing a useful indication of whether or not the egg should be injected, or a useful indication that a particular depth or position has been reached. As used herein, "location" within the egg refers to the various anatomical compartments within an egg (such as the air chamber, yolk sac, allantois, amniotic fluid, albumin, bird embryo, blastoderm), or various tissues of the same bird embryo (for example, muscle). The sensor may be located at the top of the needle, or at any predetermined position along the side wall thereof, and / or with a space apart from the tip of the needle.

The biosensors can be used to carry out the present invention. Numerous biosensors are known. See for example, patent of E.U.A. Nos. 5,804,453, 5,770,369, 5,496,701, 4,682,895, 5,646,039, ,587,128, and 4,786,396 (the description of which is incorporated herein by reference). When an electrical detector is used, it may be desirable to provide a second electrode in operative association with a first electrode. When two electrodes are used, both can be connected to the injection needle, or one can be connected to the injection needle and the other inserted separately through the same opening into the egg shell. In a preferred embodiment, the second electrode is in contact with the outside of the egg. An electrical signal can pass through the two electrodes, and the presence or absence of conduction between the two electrodes can be detected. When the second electrode is simply contacted to the outside of the egg, the signal is preferably an alternating current signal so that the second electrode is capacitively coupled to the contents of the interior of the egg. Preferably each egg (or a platform containing a plurality of eggs) is placed on top of a conductive material prior to detection using an electrical detector (see, for example, US Patent No. 5,591, 482 with respect to the conductive polyurethane foam.) When an electrical detector is used to detect the location of a compartment filled with fluid such as the allantois, the electrical detector detects the entrance of the probe into the fluid compartment and thus serves as a detector depth (the term "depth detector" here encompasses a "position detector".) In one embodiment of the present invention, the mobility of the detector and / or the associated injection needle stops as the detector and / or the needle In this way, an injection needle can stop just before the allantois penetrate, to prevent the penetration of the amnion. It falls into the creation of a complete circuit, and the measure of the totality or current flow in the circuit. For example, the circuit may comprise the conduction in the metallic needle, the conduction from the tip of the needle inside the salted allantoic fluid, a capacitative coupling between the allantois and the amnion through a thin membrane that separates the allantois and the amnion, a capacitive coupling from the amnion through the shell to a conductive pad on the outer membrane, and then back through a circuit that captures the flow of an alternating current (AC) signal through the entire circuit . Figure 1 shows this circuit and the current sensor, and the scheme of the electrical sensor system is shown in Figure 4, both are discussed in detail below. In one embodiment, an air cylinder slowly presses the detector and the associated injection needle into the egg. The entry into the fluid can be detected by monitoring the conductivity. The entire circuit path is detected for example when the tip of the needle enters the amnion or allantois. When the entry is detected within the desired compartment, the air is exhausted on both sides of the air cylinder so that movement stops. Movement stops due to friction in the piston and seals the air cylinder. Preferably, both sides of the air cylinder are depleted to prevent back pressure from directing the needle back out of the egg compartment. A magnetic brake can be used instead of, or as a supplement to, the brake action of the friction cylinder. When the detector is used to sense a compartment of the desired egg for injection, and the injection needle is not aligned in the same vertical position with the detector, a delay between the sensor of the associated compartment and the detection of movement of the needle of the Injection can be used to allow the injection needle to enter the sensor compartment. An optical sensor can comprise an optical fiber, and may be connected to a portion of the outer wall of the injection needle. A light source can be provided through a second optical fiber inserted concomitantly with the needle into the egg, or an external source of illumination can be directed into the egg. The light conduction or transmission properties can be used to determine egg viability, with different light transmission properties or thermal properties that distinguish between live and dead eggs. Light can also detect a color marker for a physiological measurement, a measure of disease, or a measure of gender.

With reference to Figures 6A and 6B, an optical detector 100 is schematically illustrated according to one embodiment of the present invention. Two thin optical fibers (about 0.0025 mm to 0.010 mm in diameter) 101, 102 are linked to the sides of an injection needle 29. The optical fibers 101, 102 are oriented such that the light is emitted or received in the direction of the tip 20a of the needle 29. A light source 103 sends visible or infrared light within a fiber 101 (the emitting fiber) to illuminate the area of an egg in front of the tip of the needle 29a. The second fiber 102 (the optical fiber) collects the light from the area of the egg in front of the tip of the needle 29a. The collected light is directed by an optical fiber 102 to a photodetector 104. The photodetector 104 provides an electrical signal proportional to the intensity of the received light. In open air, the electrical signal is low since only a small portion of the light from the emitting fiber 101 is dissipated to the side of the needle 29 and is received by the optical fiber 102. According to the tip of the needle 29 and the fibers 101, 102 approach the membrane within an egg, progressively more light from the emitting fiber 101 is reflected back to the optical fiber 102. Just before the fibers 101, 102 contacted a membrane, the The level of the signal drops abruptly since the light from the emitting fiber 101 is no longer reflected to the optical fiber 102. As the ends of the fibers 101, 102 penetrate through a membrane, the high levels will change again depending on from entry into clear amnion, rotten opaque egg, opaque muscle, or yellow yolk.

In addition, the configuration of the optical detector is illustrated in Figures 6A and 6B which can be used as a penetration signal to a membrane or to act as a measure for the distance from a needle to a membrane. Figure 6C illustrates an exemplary light intensity curve 105 indicating the distance to a membrane. The intensity of the light is plotted along the Y axis 106 and the distance from the tip of a needle to a membrane is plotted along the X axis 107. As illustrated, the light intensity increases as A needle approaches a membrane and then rapidly decreases just before penetration through the membrane. A chemical sensor can be provided in any of a variety of ways known to those skilled in the biosensing art. For example, a chemical sensor may be provided through the BBL® liposome technology available from Becton Dickinson Microbiology Systems, Cockeysville, Maryland USA, or as described in the US patent. Nos. 4,703,017 and 4,743,560, the description of which is incorporated by reference herein in its entirety. The results of said test, when the components are mounted on the injection needle, can be determined by reading with an optical fiber as discussed above. Another chemical test can be carried out by electrochemical detection. Such sensors can be used, for example, to determine the genus of the embryo within the egg, and to detect potential microbiological infection within the egg.

The chemical sensor can be a pH sensor mounted on the injection needle, with a pH measurement used to detect potential microbial contamination, distinguish live from dead eggs, etc. Ion-specific electrons for detecting various anionic or cationic species can also be used, as discussed further below. The ion and pH waves feel the movement between the compartments inside an egg because of the differences in the chemistry of the biological fluids present in the various areas and compartments of an egg. A temperature sensor can be used to distinguish live from dead eggs based on the temperature of the eggs, or for the classification of the genus of the eggs. An acoustic sensor can be used as a passive or active sensor (ie, coupled with an acoustic signal source such as a transducer making contact with the outer portion of the egg) to determine depth, to distinguish viable eggs from non-viable ones, etc. A location or depth sensor can be implemented by any of a variety of techniques. The electrical contact with the membrane of the air chamber can be used to relatively control the penetration of the needle into the preselected compartments of the egg, for example at a predetermined depth below the membrane of the air chamber, to ensure an additional injection precise inside, of the allantois, within the embryo, within the amnion or amniotic fluid, etc. alternatively, the depth can be detected with a pressure sensor to evaluate pressure changes during the transition of the needle from a compartment to a compartment within the egg (e.g., air chamber to allantois, fluid compartment to muscle tissue; etc.). A suitable method to detect the location of the sensor measures the pressure exerted on the sensor by the middle of the egg surrounding the sensor. For example, the pressure retained to emit a gas or liquid into the medium surrounding an outlet opening located in the sensor can be measured using either the injection needle or a tube containing gas or filled with fluid. The pressure discharge requires increments as the exit opening moves from a compartment filled with gas (eg, air chamber), to a compartment filled with liquid, and increases again as the exit opening moves to from a compartment filled with liquid to a solid tissue (such as embryonic muscle tissue). Changes in pressure can be measured by a pressure measuring device located outside the egg. The pressure needed to force a small amount of fluid from the tip of a needle can indicate the type of material that surrounds the end of the needle. The pressure needed to dispense fluid into muscle is greater than the pressure needed to dispense fluid within amnion or allantois or air. A pressure sensor 110 configured to deliver a small amount of fluid (such as distilled water) 111 into the medium surrounding the needle 29, according to one embodiment of the present invention, is illustrated schematically in Figure 7. As shown in FIG. illustrated, a pressure transducer 112 is connected to the inlet of the needle 29b and a fixed volume of fluid is dispensed by a positive displacement pump 114. A fluid pulse determines whether the needle 29 is in the muscle just prior to dispensing the vaccine 115 via pump 116. A pressure sensor 120 configured to dispense a small amount of fluid into the medium surrounding the needle 29, according to another embodiment of the present invention, is schematically illustrated in Figure 8. A small volume of fluid (for example, five to ten microliters) is recycled into a needle 29 by a cyclic positive displacement pump 122. The pump is preferably a solenoid or platform. Handling device by cams pressing on flexible tubes 124. Alternatively, a cyclic displacement piston can be used. The tube 124 is flexible and is also filled by pumping the fluid back into the needle 29 from fluid (or air) surrounding the tip of the needle 29a. Using either the illustrated modalities of Figures 7 or 8, while a needle is in the amnion, the fluid moves in and out of the tip of the needle with little pressure. However, when the needle is in muscle, the fluid does not flow as easily in and out of the needle and more pressure will be needed. This increase in pressure identifies the entry into muscle.

A light sensor can be used in conjunction with an external light source or a light source carried by the needle to distinguish whether the needle is in a compartment filled with air such as the air chamber, a compartment filled with liquid such such as the amnion or the allantois, or a compartment of solid tissue such as muscle or an internal organ of the same embryo. The sensor can be a diagnostic sensor for the detection of a bacterial contamination or other microbiological contamination of the eggs, such as contamination by Escherichia coli, Salmonella, or Listeria monocytogenes of the eggs. The diagnostic sensor can be implemented by any suitable means, typically a chemical sensor or biosensor. The detection of a contaminated egg can be used to trigger a signal for the subsequent selection of contaminated eggs from uncontaminated eggs. A plurality of sensors may be associated with the needle. For example. , when it is desired to detect microbial contamination of the egg or when it is desired to select the genus of the egg, it may be beneficial to provide two different or different types of data to provide a more accurate indication of the desired condition. For example, a combination of a pH sensor and a temperature sensor can be combined, or either combined with an optical sensor (with the optical sensor used, for example, to simply detect the clarity of fluids such as albumin. ).

The detection step can be carried out by removing a biological sample from the egg within a processing system in which subsequent analyzes are carried out, for example, a liquid sample can be removed and analyzed to obtain the desired information of the same in the same way as the analytical systems available to process small liquid samples (for example, in which the samples are separated by air spaces in the liquid processing line). In such cases it is necessary to provide a way to identify the egg from which each biological sample is removed, for example, by providing hardware, software or combinations of hardware and software to count the eggs and the relative position of each egg, in association with the time of sampling and storage of that information for a short or long period of time until it is used appropriately (for example, to reject a particular egg, or to provide a large database of information about the quality or other parameters of the injected eggs). Information on the composition of the allantoic fluid, detected with ionic sensors, can be used to indicate the true age of the embryo, estimate the time required for hatching, evaluate the operation of the incubation system, etc. It will be appreciated that the present invention can provide a way to record and store large amounts of information about the eggs to be injected. For example, population data can be obtained so that they can be used for quality control programs, or to modify previous treatments of eggs, or to modify selective cross-linking programs. In such cases, the identity of the injected egg may be in association with a particular batch of eggs, rather than with its identity as a particular individual within that batch of eggs. A preferred embodiment of the present invention for injecting multiple eggs concurrently is illustrated in Figures 1-4. As shown in Figures 2-3, the apparatus 10 includes a platform 15 for carrying eggs, a stationary base 16, and a plurality of conventional injection administration devices, or heads 25 with fluid delivery means such as lumens or needle (s) located on it according to known techniques. The platform 15 maintains a plurality of eggs 20 in a substantially upward position. The platform 15 is configured to provide external access to predetermined areas of the eggs 20. Each egg is held by the platform 15 so that the respective end thereof is in a proper alignment relative to one corresponding to the injection devices 25 such as the Injection device 25 advances towards base 16 of the apparatus. As used herein, a "lumen" is an internal cavity or open space of a tube that can be provided by a syringe or needle. A lumen for the administration of a treatment substance can be inside a needle, or between a needle and another external guide or sleeve. Multiple lumens can be formed inside the singular needles, with the exit ports located over different locations of the needle. Each of the plurality of injection device 25 have opposite initial and secondary ends 26,27. The device 25 has an initial extended position and a secondary retracted position, as is known in the art. After the extension of the injection device 25, the end 26 is configured to contact and to stay against predetermined areas of the outer shell. When not injecting, the injection device 25 is retracted to remain at a predetermined distance above the eggs and the stationary base 16. Alternatively, the base 16 can slide longitudinally to move from the position of the eggs in a suitable position relative to the injection of the administration apparatus or injection administration needle 29 (see figure 5). As shown in figure 1, the second end 27 of the injection management device includes primary and secondary input ports 28a, 28b which are configured to receive tubes respectively from chambers of treatment substances. The treatment substances can be administered into the needle along with separate administration routes, such as the lumen of an internal needle, and the space between the internal needle and a guide punch. In a preferred embodiment, only a single substance is injected together with the singular route, and a sanitized fluid (such as chloride solution) is carried in the external lumen, according to known techniques.

As shown in Figure 1, the in ovo injection head for administering compounds within an egg comprises a body member 40 having an opposite edge 41 and bottom end portion 43 and an elongated longitudinal opening formed therein, and a device of administration located in said opening. The device includes a member for locating an egg, or a member for adjusting an egg, on a final portion 26, which is slidably connected to the body member and includes a spring 42 for both the fixation pad that holds the egg in place during the down stroke of the injection head. An external guide is preferably provided to pierce the egg shell, and a needle then extends downwardly of the outer guide and into the desired compartments of the egg, all in accordance with known techniques. The pneumatic lines 51, 52 are provided for advancing or withdrawing the injection needle into the egg in the usual manner (see Figures 1 and 5), by providing a baseline 53 that is in electrical contact with the injection needle, and an electrical line 54, isolated from the environment, which is brought into electrical contact with an egg by assembling the plunger 55, the electrical contact of an injection needle to the egg can be detected. In addition, the position of the injection needle within several compartments of the egg can be detected by changes in conductivity, resistance, capacitance, etc. In the preferred embodiment, an alternating power current supplement 60 (specifically, a voltage peak of 6.100 kilohertz to a sine wave peak) is employed, so that capacitive coupling of the current through the egg is achieved. via line 54. In the preferred embodiment of FIG. 1 depth information is detected through the resistance of the current sensor 61 by the current threshold monitor 62 to provide a stop signal of the tool 63. The monitor Current threshold 62, which is illustrated in great detail in Figure 4, comprises a portion of an amplified current signal 71, a rectifier signal and an average current signal of portion 72, and a threshold detection portion. 73, all of which can be assembled from resistors 74, operational amplifiers 75, capacitors 76, diodes 77, etc., according to known techniques, and power is provided at an additional cost of energy (not shown). The stop signal of the tool can be delivered by a pneumatic directional control valve 80 as illustrated in FIG. 5. The pressure supply together with the supply of the line 81 can be directed between the three positions of the needle downwards 82, at standstill 83, and upward needle 84, by rotating the pressure through lines 51, 52, or through exhaust lines 85, 86. The foregoing is illustrative of the present invention, and is not considered as limiting the same The invention is defined by the following claims with equivalents of the claims to be included herein.

Claims (38)

NOVELTY OF THE INVENTION CLAIMS

1. - A method for injecting a plurality of bird eggs, comprising: a) orienting a plurality of bird eggs in a predetermined position; b) forming an opening in the shell of each of said eggs; c) extending an elongated delivery device through each of said openings and inside the egg, each of said administration devices comprises a detector and an injection needle, with said injection needle having a lumen formed inside: d) detecting with said detector information from the inside of each of said plurality of eggs; and e) injecting a substance into each of said plurality of eggs through the lumen of said injection needle.

2. A method according to claim 1, further characterized in that said detection step is carried out before said injection step.

3. A method according to claim 1, further characterized in that said step is carried out concurrently with said injection step.

4. A method according to claim 1, further characterized in that said detection step is carried out after said injection step.

5. - A method according to claim 1, further characterized in that said detector is an electrical sensor.

6. A method according to claim 5, further characterized in that it comprises the step of contacting a secondary electric sensor to the shell of said egg.

7. A method according to claim 6, further characterized in that said second electrical sensor is capacitively coupled to the content of said egg.

8. A method according to claim 1, further characterized in that said detector is an optical sensor.

9. A method according to claim 1, further characterized in that said detector is a chemical sensor.

10. A method according to claim 1, further characterized in that said detector is a temperature sensor.

11. A method according to claim 1, further characterized in that said detector is an acoustic sensor.

12. A method according to claim 1, further characterized in that said detector is a pressure sensor.

13. A method according to claim 1, further characterized in that said detection step comprises: the removal of a biological sample from said egg; identify the egg from which said biological sample was removed; obtain information from said withdrawn biological sample; and then storing said information in association with the identification of the egg from said biological sample that was removed.

14. A method according to claim 1, further characterized by comprising the steps of: identifying the egg from which said information was obtained, and then storing said information in association with the identity of the egg from which said information it was obtained.

15. A method according to claim 1, further characterized in that it comprises the steps of: f) removing said elongated delivery device from each of said eggs; and then g) repeating steps (a) through (e) with a second plurality of eggs.

16. A method according to claim 1, further characterized in that said bird eggs are live embryonic bird eggs.

17. A method for locating the tip of a needle to inject a substance into or withdrawing a sample from a particular location within each of a plurality of bird eggs, comprising: a) orienting a plurality of eggs from bird in a predetermined position; b) forming an opening in the shell of each of said eggs: c) extending an elongate delivery device through said openings and into the eggs, each of said administration devices comprising a depth detector and a needle, with said needle having a frontal portion and a lumen formed therein, said lumen extending through said frontal portion; d) detecting the depth information of the needle from the inside of each of said plurality of eggs with said depth detector; and then e) independently controlling the depth of penetration of each of said needles based on said corresponding information of the depth of the needle; wherein a substance can be injected or a biological material can be withdrawn from a particular position of said portion of the tip of the needle into each of said plurality of eggs through the lumen of said needle.

18. A method according to claim 17, further characterized in that said portion of the tip of the needle is located in the allantois.

19. A method according to claim 17, further characterized in that said portion of the needle tip is located in the amnion.

20. A method according to claim 17, further characterized in that said portion of the tip of the needle is located in the embryonic muscle.

21. A method according to claim 17, further characterized in that said portion of the tip of the needle is located in the air chamber.

22. - A method according to claim 17, further characterized in that said portion of the tip of the needle is located in the vitelino sac.

23. A method according to claim 17, further characterized in that said portion of the tip of the needle is located in the blastoderm.

24. An injection apparatus for injecting a plurality of bird eggs with a substance, said apparatus comprising: an assembled alignment for orienting a plurality of eggs of birds to be injected; a plurality of injectors associated with said alignment assembled and configured to inject each of said plurality of bird eggs at a predetermined location, each of said injectors including an injection needle having a lumen through which said substance is injected; and a detector operatively associated with each of said injection needles that detect information from inside said egg.

25. An injection device according to claim 24, further characterized in that said detector comprises a depth detector.

26.- An apparatus according to claim 25, further characterized in that it comprises the control modes operatively associated with said detector that controls the depth of penetration of said injection needle.

27. - An apparatus according to claim 24, further characterized in that said detector comprises an electrical sensor.

28. An apparatus according to claim 24, further characterized in that said detector comprises an optical sensor.

29. Apparatus according to claim 24, further characterized in that said detector comprises a chemical sensor 30. An apparatus according to claim 24, further characterized in that said detector comprises a temperature sensor. with claim 24, further characterized in that said detector comprises an acoustic sensor. 32. An apparatus according to claim 24, further characterized in that said detector comprises a pressure sensor. 33. An apparatus according to claim 24, further characterized in that said detector comprises: sampling means for removing a biological sample from said egg; means of identification to identify the egg from which said biological sample was removed; analytical means operatively associated with said sampling means for obtaining information from said biological sample; and storage means operatively associated with said means of analysis and said identification means for storing said information in association with the identification of the egg from which said biological sample was removed. 34.- An apparatus for locating the tip of a needle for injecting a substance into or removing a biological material from a particular location in a plurality of bird eggs with a substance, said apparatus comprising: an aligned assembly for orientation of a plurality of bird eggs to be injected; a plurality of injectors associated with said assembly aligned and configured to inject each of said plurality of water eggs at a predetermined location, each of said injectors including a needle having a lumen through which said substance is injected or said material it is removed; a depth detector connected to and operatively associated with each of said injection needles to detect the depth information of the needle from the interior of said egg; and control means operatively associated with said depth detector to independently control the depth of penetration of each of said needles. 35.- An apparatus according to claim 34, wherein said depth detector comprises an electrical sensor connected to said needle. 36.- An apparatus according to claim 35, further characterized in that said depth detector further comprises a second electrical sensor configured for capacitive coupling to the outside of each of said eggs. 37.- An apparatus according to claim 34, further characterized in that said depth detector comprises a pressure sensor. 38.- An apparatus according to claim 34, further characterized in that said depth detector comprises an acoustic sensor.