This invention relates to a container having multiple chambers for containing biological liquids, more particularly, biological liquids used in the artificial insemination of animals.
Artificial insemination (AI) and embryo transfer are widely used techniques for delivering biological liquids containing semen or embryos into the reproductive tract of a female animal. Such techniques are routinely used in livestock breeding and introducing new genetic lines into an animal herd, especially dairy cattle and swine (pigs).
Artificial insemination techniques for livestock typically utilize liquid extended semen. In practice, fresh semen collected from a boar is combined with a semen extender, an aqueous solution used to dilute the ejaculate to obtain a large number of doses that can be used for inseminating multiple sows, e.g., up to 200 sows.
The extender functions to provide membrane stabilization in cool storage temperatures, an energy source for sperm metabolism, pH buffering, ions for membrane and cell balance, and antibiotics to prevent microbial growth. The proper extension of semen is critical to the success of the insemination procedure, and it is vital that extended semen doses possess acceptable characteristics to be capable of impregnating a gilt or sow which is inseminated at the appropriate time. In the U.S., artificial insemination protocols for swine recommend carrying out inseminations with a dose of 70 to 90 ml (cc) containing 3 to 3.5 billion spermatozoa per dose. European protocols typically use about 1.5 billion sperm per dose.
Extending the semen needs to be done while preserving the functional characteristics of the sperm cells (spermatozoa). Seminal plasma within the ejaculate supplies the sperm cells with nutrients for a limited period. To preserve the sperm cells for an extended period, their metabolic activity is reduced by diluting the cells in an appropriate liquid extender medium and lowering the temperature. To perform its function, the extender should supply the nutrients needed for the metabolic maintenance of the sperm cell (glucose), afford protection against cold shock (BSA), control the pH (bicarbonate, TRIS, HEPES) and osmotic pressure (NaCl, KCl) of the medium, and inhibit microbial growth (antibiotics).
In current semen extension processing, the fresh semen is diluted with a semen extender to the desired dosage concentration and the dose amount (e.g., 80 cc) is dispensed into and sealed within a semen container as a ready-to-use semen dose. In a livestock insemination procedure, the container holding the semen is attached to a semi-rigid tube or catheter which has been inserted into the female animal (sow/gilt) to be inseminated. Muscular contractions then aid in drawing the seminal fluid into the body of the sow or gilt.
Excessive or abrupt dilution of semen or the exposure of spermatozoa to a solution with a relatively high concentration of solutes (hypertonic solution) can lead to what is known as a ‘dilution effect’ or osmotic shock and a permanent loss of motility, metabolic activity and fertilizing capacity. Generally, when sperm cells are exposed to a hypertonic solution (e.g., an extender solution), water is initially drawn out of the cells into the surrounding medium resulting in dehydration and shrinkage of the cells, followed by an influx of water causing the cells to swell to balance the internal and external osmotic pressures. If the semen extender is abruptly added (or the semen is excessively diluted), cell shrinkage and swelling can occur rapidly and excessively resulting in physical damage to the cells with a loss of sperm motility and function. For this reason, the semen should be diluted slowly or by step-wise addition of the extender to allow for a gradual osmotic adjustment between intra- and extracellular fluids to minimize osmotic shock. However, this is both tedious and time consuming.
Other problems in extending the semen concern the protracted exposure of the sperm cells to certain solutes, which can be detrimental causing toxic injuries to the cells. In addition, the amount and composition of semen extender and number of sperm cells for currently practiced insemination dose amounts gives rise to high production costs and an economic impact on a breeding operation.
Containers for holding semen are known. For example, U.S. Pat. No. 8,025,855 (Kuhlow) discloses a semen container having a hollow body with an open, sealable end and a nozzle at the opposing end fitted with a removable tip. In another example, U.S. Pat. No. 6,551,819 (Simmet) discloses a container for semen and other biological liquids that includes a nozzle for delivery of the liquid with means for bending of the nozzle such that the container can be positioned in a vertical alignment while attached to a horizontally aligned catheter without kinking of the container or catheter which can hinder delivery of the biological liquid from the container to the animal. The vertical positioning of the container facilitates gravity flow, which assists the transfer of the biological liquid from the container into the catheter and the sow/gilt being bred.
Known sperm containers are structured to contain an insemination dose of 80 to 100 ml containing 3 to 3.5 billion sperm cells, which requires careful processing to avoid osmotic shock during the dilution process. In addition, with current containers, sperm cells are continuously exposed to the solutes of the extender solution without the ability to modify the solution during the storage period.
It would be desirable to provide a container for an insemination dose that overcomes the foregoing problems.
The present invention provides a multi-chambered container for containing two different liquids and allowing for the flow of one liquid into the other liquid.
The containers of this invention advantageously may be used to store and transport semen for the artificial insemination of animals, particularly swine and other livestock. In such applications, the containers of the invention are structured for containing an aliquot of extended semen in one compartment, and an aliquot of semen extender or other fluid in a separate but connected compartment.
In embodiments, the biological liquid within one of the compartments comprises extended semen, and the biological liquid within the other compartment comprises a fluid without sperm cells. In embodiments, the construction of the containers allows for the containment of an aliquot of about 15 to 35 ml of extended semen having a concentration of spermatozoa as low as 500 million sperm cells in one compartment, and an aliquot of about 30 to 50 ml of semen extender or other biological liquid (without sperm cells) in the other compartment. A conduit section interconnects the two compartments. In embodiments, the conduit section can be completely closed (sealed) (e.g., with a plug) to contain the biological liquid separate from the extended semen, with the plug being optionally semi-permeable to allow passage of nutrients into the extended semen fraction. In other embodiments the conduit section can be partially closed to provide separate containment of the biological liquid from the extended semen while providing a channel for a minimal but continuous flow of the biological liquid into the extended semen solution, or by an exchange of liquid and/or nutrients through diffusion and osmotic pressure equilibration. In yet other embodiments, the conduit section can be constructed with an inner diameter that provides an air pocket that separates the two fluids by surface tension.
Through the use of the containers of the invention, the number of sperm cells and volume of the inseminate can be reduced by six- to seven-fold compared to current standard amounts without lowering the fertility result or litter size, which allows for a larger number of insemination doses that can be obtained from a single ejaculate. The containers also eliminate the need for multi-step dilution of the semen. In addition, embodiments of the container structure provide for continuous feeding of nutrients to the extended sperm fraction to maintain viability of the sperm cells, e.g., through the use of a porous plug or by providing a channel around the plug within the conduit section.
In embodiments, the container comprises a first compartment connected by a conduit section to a second compartment, with the first compartment having a first end being sealable and a second end connected to one end of the conduit section, the second compartment having a first end connected to an opposing end of the conduit section and a second end comprising a nozzle, and the conduit section having a channel extending therethrough and structured to contain a fluid within the first compartment substantially separate from a fluid within the second compartment.
In embodiments, the container has a hollow body of a molded plastic material and a first compartment connected by a conduit section to a second compartment. The first compartment has a first end that is sealable and a second end that is integrally molded to one end of the conduit section. The first end of the second compartment is integrally molded to the opposing end of the conduit section and the second end has an integrally molded nozzle having an inner diameter and a closed end. The conduit section has a channel that extends therethrough, which, in embodiments, is fully closed (sealed) to contain a fluid within the first compartment, or partially open to maintain a passageway for the fluid to pass into the second compartment, or fully open but with an inner diameter that maintains an air pocket between the two fluids. In embodiments, the second compartment is structured to contain extended semen, and the first compartment is structured to contain a biological fluid that does not contain sperm cells.
In one embodiment, the container is structured with an unattached and movable spherical mass (sphere) situated within the body, which is initially located within the first compartment, and can be sequentially positioned within the conduit section and then within the second compartment. In embodiments, the first compartment is structured with one or more ridges or protuberances on the inner surface proximal to the conduit section, to block entry of the spherical mass into the conduit section. The conduit section is sized to receive the spherical mass therein to seal the conduit section or, in other embodiments, to maintain a channel between the spherical mass and the wall of the conduct section to allow passage of liquid therethrough. The spherical mass can be porous to allow passage of fluid therethrough. In embodiments, the second compartment is structured with one or more ridges or protuberances on the inner surface proximal to the nozzle, to block entry of the spherical mass into the nozzle.
In another embodiment, the conduit section of the container is structured with opposing arcuate sidewalls with at least one of said sidewalls configured such that an applied force against one of the sidewalls inverts and positions the sidewall into a nesting relationship within the opposing sidewall while maintaining a channel for passage of liquid from the first compartment into the second compartment.
In yet another embodiment, the conduit section of the container is structured with opposing sidewalls with an adhesive element on the inner surface of one or both of the sidewalls. Upon the application of force onto the outer surface of the sidewalls, the adhesive element(s) seals the conduit section or, in other embodiments, a channel can be maintained for passage of liquid therethrough.
In a further embodiment, a rod-shaped plug is inserted through the conduit section to fully or partially seal the channel, and is secured in the end tab or flange of the nozzle of the container. To open the channel of the conduit section, the end tab or flange can be twisted off and removed with the attached rod-shaped plug.
In another embodiment, the conduit section can be structured with a channel having a small inside diameter (i.d.) that allows for insertion therethrough of a fluid delivery nozzle to deliver fluid into the second compartment, and maintains an air pocket to separate the fluids in the first and second compartments without the need for a plug or other closure element.
In yet another embodiment, the container comprises two sheets of plastic sealed together to define the first and second compartments and the conduit section of the container. The container can be formed by heat sealing plastic sheets together to define the container body, i.e., the compartments, conduit section and nozzle.
In use, the second compartment can be first filled (partially or completely) with an aliquot of a biological fluid (e.g., extended semen, etc.) and, in embodiments, the channel through the conduit can be closed or semi-closed, the first compartment filled with an aliquot of a second biological fluid (e.g., semen extender without spermatozoa), and the open end of the container sealed. For delivery of the biological fluids from the container, the closed end of the nozzle of the container can be opened, the nozzle inserted into a catheter and the container positioned in a vertical orientation. The fluid within the second compartment can be dispensed through the nozzle into the catheter, and pressure then applied to the fluid within the first compartment to force the fluid through the channel into the second compartment and through the nozzle into the catheter. In embodiments, pressure applied to the fluid in the first compartment can force a plug that may be present out of the channel of the conduit section and into the second compartment.
In embodiments in which the container is structured with a rod-shaped plug inserted through the channel of the conduit section and attached to a severable end tab or flange, the end tab or flange can be twisted off to open the nozzle end of the second compartment and withdrawn to remove the interconnected rod-shaped plug from the channel of the conduit. In embodiments in which the channel of the conduit section is open (without a plug or other sealing element), pressure applied to the fluid in the first compartment can force the fluid through the open channel into the second compartment and out through the nozzle.
In embodiments using a container having an open channel through the conduit section without a plug or other closure element, the first and second compartments can be separately or simultaneously filled with the first and second biological fluids. The compartments can be filled, for example, by delivering the fluid (e.g., extended semen) into the second compartment through a first delivery nozzle inserted through the channel of the conduit section, and simultaneously delivering fluid (e.g., extended semen) into the first compartment through a second delivery nozzle. As the delivery nozzles are removed, the first delivery nozzle is withdrawn through the channel of the conduit section and an air pocket remains within the channel to maintain a separation of the fluid (e.g., extended semen) within the second compartment from the fluid (e.g., semen extender) in the first compartment without the use of a plug within the channel.
In embodiments of the use of the multi-compartment containers in an artificial insemination procedure of an animal, the second compartment can be filled with an aliquot of extended semen, the channel of the conduit can be closed or semi-closed where applicable (e.g., in use of a container structured with a plug), and the first compartment can be filled with an aliquot of semen extender or other biological fluid to provide the necessary physiological volume for insemination. In an insemination procedure, the closed end of the nozzle of the container can be opened and an insemination catheter that has been inserted into a female animal to be inseminated, seated into the inner diameter of the nozzle. The container can then positioned in a vertical alignment to the horizontally aligned catheter. Muscular contractions aid in drawing the fluids from the container into the animal to be inseminated.
In embodiments, the fluid (e.g., semen extender) contained in the first compartment can be formulated as an activation medium (“AM” fluid) to stimulate sperm cell activity and motility and the extended semen contained in the second compartment can be formulated in a preservation medium (“PM” fluid) to maintain the sperm cells at a low metabolism and enhance toleration of stressors during storage and transport.
In embodiments in which the container is structured with a spherical mass to partially close the conduit section, with the nozzle of the container attached to the insemination catheter, the spherical mass can be dislodged from the conduit section into the second compartment by squeezing the sides of the first compartment, which results in the discharge of the biological liquid (e.g., semen extender) from the first compartment through the channel of the conduit section into the second compartment and through the nozzle into the insemination catheter. The aliquots of extended semen and biological fluid (with sperm cells) are drawn into the catheter and the body of the sow or gilt by muscular contractions of the animal.
In embodiments in which the conduit section of the container is structured with opposing arcuate sidewalls that are positioned in a nested relationship to partially close the channel through the conduit section, the sidewalls can be forced out of their nested relation to discharge the biological liquid from the first compartment through the channel of the conduit section into the second compartment and through the nozzle into the insemination catheter.
In other embodiments in which the conduit section of the container is structured with opposing sidewalls releasably attached by an adhesive element to fully or partially close the channel through the conduit section, the sidewalls are forced apart to discharge the biological liquid from the first compartment through the channel of the conduit section into the second compartment and through the nozzle into the insemination catheter.
In another embodiment of a container configured for simultaneous filling of the first and second compartments, the conduit section of the container has a channel with an inner diameter (i.d.) effective to maintain an air pocket therein between fluids within the first and second compartments during filling, and the nozzle at the second end of the second compartment comprises an opening that is sized for receiving a tube (e.g., straw) therethrough for delivery of fluid into the second compartment. In embodiments in which the nozzle terminates in an end having a severable tab extending therefrom, an opening for receiving the tube therethrough can extend through the tab to the second compartment. While filling the second compartment with fluid through the tube, fluid can be simultaneously delivered into the first compartment through a cannula or other filling device. After filling, the opening to the first compartment can be sealed, and the first end of the tube can be advanced through the second compartment and into the channel of the conduit section, which is sized for receiving the tube (or straw) therethrough, to fully or partially close the channel, and the second end of the tube (positioned within the opening in the nozzle or through the severable tab) can be sealed. In use, the nozzle of the second compartment can be opened and a catheter connected to an animal inserted to deliver the fluid from the container as described previously.
Embodiments of the invention relate to a multi-compartment container for holding biological liquids.
The term “biological liquid(s)” refers to a biological fluid or liquid medium or biological cells in a liquid or fluid medium. Examples of biological liquids include semen and seminal fluids that may include sperm cells and seminal plasma, extended semen, cultured tissue such as biological cells useful for the purposes of breeding and reproduction such as oocytes, embryos or combinations thereof, and culture media, semen extenders, diluents, stimulating solutions (e.g., caffeine, etc.), and antimicrobial agents, pH buffering agents, fillers, solvents, dispersants and other additives, and combinations thereof.
A first embodiment of a multi-compartment container 10 according to the invention is described with reference to
As shown in
The first compartment 16 has a first end 22, a second end 24 and a body 26. The first end 22 is adapted and configured for receiving a liquid into the body 26 of the container 10, and is also adapted for forming a fluid tight seal at a sealing section 28 near the end of the container. The second end 24 is integrally molded to one end 30a of the conduit section 20 having a channel 31 therethrough A spherical mass 32, which is unattached and freely movable, is initially situated within the body 26 of the first compartment 16. The spherical mass can be composed of a solid or resiliently compressible material, for example, of a biocompatible silicone, wax, gel, glass, metal, plastic, thermoplastic elastomer (TPE such as chips or pellets of thermoplastic polyurethane (TPU), among other materials, and can be porous or permeable to allow the flow of fluid therethrough. Another useful spherical mass can be composed of an absorbent, water-insoluble polymer that swells or gels in aqueous fluid such as water but does not dissolve in the fluids, for example, polyacrylamides, starch graft copolymers and starch-based polymers, such as Soil Moist™ starch polymers (JRM Chemical Inc., Cleveland, Ohio) and polymers such as those described in U.S. Pat. No. 8,017,553 (Doane), and polymeric materials such as H-100 or H-600 polymers (JRM Chemical Inc.). In use, granules of an absorbent polymer can be reconstituted in water to provide a gel-like mass that can be delivered into the first compartment. The spherical mass can be heat sterilized prior to insertion into the container.
Referring to
In another embodiment illustrated in
As depicted in
Referring to
In embodiments, the nozzle (second end) 40 includes bendable elements 54, for example, one or more accordion folds, corrugations, or a combination thereof, which permits the nozzle 40 to be bent without forming kinks in either the nozzle or the body of the container 10. “Corrugations” refer to a rounded protrusion extending either inward or outward, preferably outward, from the nozzle surface, for example, as shown in
In use during an artificial insemination procedure, with the container 10 and nozzle 40 in a horizontal alignment, an insemination catheter 56 which has been inserted into the animal's body, is inserted into the channel 50 and seated in the first end 44 of the nozzle 40 (with reference to
The nozzle 40 should have sufficient rigidity to allow insertion of the insemination catheter 56 into the first end 44 of the nozzle 40. In embodiments, the body 42 of the second compartment 18 proximal to the nozzle (second end) 40 can be reinforced, for example, by increasing the body wall thickness and/or by reinforcement ribs 58 as shown in
In embodiments, the end 46 of the nozzle 10 is sealed by a severable section 60 such as a tab or flange extending therefrom, which can be manually twisted off along a score line 62 to open the end 46 without the use of tools. This embodiment allows the user to avoid cutting the container nozzle with a knife, thereby decreasing the occurrence of microbial contamination or cross contamination of the semen held within the container.
The walls of the container body are typically made from a non-spermicidal material, preferably a flexible plastic material. Examples of suitable plastics include polyolefins (such as polyethylene, polypropylene etc.), polyvinylchloride, nylons, polyfluorocarbons, thermoplastic polyurethanes, polystyrene, elastomers, cellulosic resins, acrylic resins and silicones, preferably polyolefins and elastomers. The containers of the invention can be produced by conventional molding techniques such as injection molding, blow molding, extrusion molding, thermoforming and other processing techniques that are well known to those skilled in the art of plastics processing.
During an artificial insemination procedure, the pumping action of muscular contractions within the animal and the drainage of the biological liquid from the container 10 are believed to exert a partial vacuum within the container. Preferably, the walls are thin enough such that the container body will collapse upon the application of a partial vacuum within the lumen of the container. If the walls are too rigid, the partial vacuum can persist and hinder the flow of the liquid from the container. The collapse of the flexible walls of the container relieves the partial vacuum and facilitates a rapid flow and near complete drainage of the biological liquid from the container. The rigidity of the walls can be varied by the wall thickness according to the application. An example of a tube having a suitable wall thickness for the desired wall collapse is the ULTRAFLEX™ boar semen tube available from Minitube of America, Inc., Verona, Wis. USA.
The body wall thickness near the first end 22 of the first compartment 16 to provide a more stable opening for filling the container 10 and a more stable material for sealing the container. The body wall thickness of the first and second compartments 16, 18 can also be increased at the junction with the conduit section 20 for added stability.
As illustrated in
The container 10 can be provided to a stud farm with the first end 22 of the first compartment 16 open, and the opposing end 46 of the nozzle 40 sealed, e.g., by a severable tab or flange 60. The container can then be filled with extended semen in one compartment 18 and a semen extender or other biological fluid in the other compartment 16, and sealed for storage and later use.
Referring now to
The second compartment 18 can also be filled by manually pouring the liquid into the first compartment 16 and allowing the liquid to flow through the conduit section 20 into the second compartment 18. The protuberances 34 function to block the spherical mass 32 from sealing the opening 36 to the conduit section 20.
Upon filling the second compartment 18, the filling device 70 can then be withdrawn from the container 10, as depicted in
Referring to
In the embodiment illustrated in
As illustrated in
In embodiments, the second compartment 18 is filled with a volume of about 15 to 20 mls of extended semen 68 containing at least about 500 million sperm cells up to about 875 million, or up to about 750 million sperm cells or more, and in some embodiments up to 3 billion sperm cells. The first compartment 16 can be filled with a volume of about 30 mls up to about 50 mls of a compatible semen extender 76 to provide a total combined volume (with the extended semen) of at least about 50 mls up to about 70 mls to ensure adequate transport of spermatozoa for insemination of the sow/gilt. Where the extended semen 68 is at the lower concentration of 500 million sperm cells per 15 to 20 mls, one or more additives and/or nutrients can be included which will function to reduce or prevent dilution shock and maintain semen quality.
As shown in
The sealed container 10 can be used immediately or stored according to protocol (temperature, etc.). During the storage period, the second biological liquid 76 can flow or diffuse through the channel 74 into the biological liquid 68 (e.g., extended semen) within the second compartment 18. Clearance of the spherical mass 32 from the sidewalls of the conduit section 20 provides a channel 74 that allows a desired exchange of fluids between the first and second compartments through equilibration of osmotic pressure differences between the two fluids. The channel 74 can be, for example, about 0.25 mm to 1 mm in width. The channel 74 feature of the containers of the invention can provide for a continuous inflow of fresh fluid containing, for example, fresh semen extender, a pH buffering agent, a stimulant such as caffeine, etc., into the extended semen 68 to maintain the viability of the spermatozoa during the storage period.
In use during an artificial insemination procedure, the container 10 is held in a horizontal alignment, the removable tab 60 is twisted off along the perforations 62, and an artificial insemination (AI) catheter 56 (which has been inserted into the female animal's body) is inserted into the channel 50 and seated in the first end 44 of the nozzle 40, as illustrated in
As depicted in
Removal of the spherical mass 32 from the conduit section dispenses the second biological liquid 76 from the first compartment 18 into the second compartment 18 and then into the insemination catheter 56 to flush the remaining extended semen 68 from the second compartment 18 and to dilute the extended semen 68 to a total volume of about 50 mls to about 65 or 70 mls for delivery into the animal. As illustrated in
The collapse of the flexible walls of the container 10 relieves the partial vacuum within the container and enables a rapid flow and near complete drainage of the biological liquids 68, 76 from the container. Tearing along the cut or score line 80 to open the sealed section 28 during the insemination procedure releases the vacuum formed inside of the container 10.
A second embodiment of a multi-compartment container 10″ according to the invention is described with reference to
As shown in
In the present embodiment, the conduit section 20″ is structured with opposing arcuate (curved) sidewalls 84a″, 84b″. As illustrated in
As shown in
To allow the biological liquid 76″ to freely flow out of the first compartment 18″, a force (arrow “d” in
As in the first embodiment, the collapse of the walls of the container 10″ and tearing the scoring 80″ to open the sealed section 28″, as shown in
A third embodiment of a multi-compartment container 10′ according to the invention is described with reference to
The inner surface 12′″ of one or both of the sidewalls 88′″ of the conduit section 20′″ (shown as both sidewalls) is structured with one or more adhesive elements 90′″. As depicted in
In embodiments, the adhesive element 90′″ is made from a non-spermicidal, biocompatible and water-resistant material that will adhere together or to the sidewall 88″ to form a temporary and releasable bond, for example, a wax, glue, gum, pressure-sensitive adhesive, or other highly viscous material, as known and used in the art, that will releasably adhere together by applying pressure. The adhesive material can be applied, for example, by introducing a delivery nozzle into the conduit section 20′″ after forming the container 10′″ and depositing a small amount of material onto the inner surface 12′″ of the conduit section.
In use during an artificial insemination procedure, the sidewalls 88′″ can be manually pulled apart to fully open the conduit section 20′″ to allow the liquid 76′″ to flow out of the first compartment 16′″.
In another embodiment of a container illustrated in
In use, the second compartment 18 can be filled by injecting biological fluid 68 (e.g., extended semen) through a fluid delivery nozzle inserted through the channel 31 (similar to the illustration in
In other embodiments, as depicted in
Referring now to
Perforations 104 can be formed through the plastic sheets 96a, 96b as a line between adjacent containers 10a, 10b to allow ready separation of the containers. Index holes 106 can also be provided at or near the edges 100a, 100b of the sheets to facilitate the placement of the sheet of containers on a conveyance mechanism, for example, in conjunction with filling the containers or other processing.
As illustrated in
In use, the containers 10a, 10b can be separated along the perforations 104, the sealed flaps 102a, 102b opened to expose the opening to the nozzle 40, and an AI catheter (not shown) inserted into the nozzle.
The channel 31 of the conduit section 20 is configured with a small inside diameter (i.d.) that maintains air therein to separate the fluids in the first and second compartments, and allows for insertion therethrough of a rod-like tube 92 for fluid delivery and to fully or partially seal the channel 31. The severable section 60 includes an opening or channel 94 which is sized to receive the rod-like tube 92 therethrough to fully or partially seal the opening 94 but allow the tube 92 to be advanced into the second compartment 18 and through the channel 31 of the conduit section 20.
The rod-like tube 92 is a hollow tube or straw with first and second open ends 96a,b, to allow passage of fluid 68 therethrough for delivery into the chamber of the second compartment 18. In embodiments, the rod-like tube 92 is constructed of a rigid to semi-rigid plastic to facilitate being advanced through the opening 94 in the severable section (e.g. tab) 60 and inserted through the channel 31 of the conduit section 20. In embodiments, the rod-like tube 92 can be cut from a length of tubing after placement in the container (as shown in
To fill the container 10, the first end 96a of the rod-like tube 92 is inserted through the opening 94 of the severable section (e.g., tab) 60 and positioned within the chamber of the second compartment 18. The rod-like tube 92 is connected to a source 98 for fluid 68, for example, by means of a tubing 100. A filling device 70 such as a cannula, delivery nozzle or an automatic filler, connected to a source 102 for fluid 76, can be introduced into the open end 22 of the container 10 for filling the first compartment 16.
The first and second compartments 16, 18 can then be filled with fluid 68, 76, respectively. During filling, air being displaced from the second compartment 18 can bubble up through the fluid 76 in the first compartment 16. During filling, air remains within the channel 31 to maintain separation of the fluids 68, 76. Upon filling the first and second compartments 16, 18, the flow of fluids 68, 76 is terminated and the rod-like tube 92 is advanced (arrow “g”) further into the chamber of the second compartment 18 and through the channel 31, as depicted in
Referring now to
The invention provides multi-compartment containers for low dose insemination that maintain lower dose volumes with high motility and cell viability in one compartment and a necessary physiological volume for insemination in a second compartment. To optimize results and ensure successful fertilization with low dose volumes, an extender is utilized that prevents dilution shock and allows storage in a lower concentration of sperm cells per ml.
The semen can be packaged in a dual-stage tube according to the invention, which is optimized for low dose semen delivery. The low volume dose is combined with additional transport medium to facilitate peristaltic movement of the uterus to transport the semen cells to the site of fertilization. The dual-stage containers of the invention conveniently package a low volume of extended semen in a preservation medium (“PM fluid”) and a separate volume of an activation medium (“AM fluid”) in a single tube/container.
The dual-stage low dose semen tubes of the invention store the necessary volume of media in one convenient easy to use package, and are designed to optimize low dose IUI. For example, an about 20-ml reservoir compartment can be used for low dose/low concentrated semen volumes down to about 500 million sperm cells/dose, and a 50-ml reservoir compartment for sperm cell activation and reconstitution medium. The two-compartment containers maintain a uniform temperature of both packaged extenders. A removable or dislodgeable sealing element separates the low dose semen volume from the activation and reconstitution medium until insemination.
A sow can be inseminated utilizing the SafeBlue Clear glide IUI insemination catheter in combination with a dual-stage container of the invention, for gentle insertion and deep intrauterine semen delivery. The inner catheter's smoothly rounded tip is designed to easily glide through the cervix. With fewer sperm cells per dose, the semen is advantageously delivered beyond the cervix via intra-uterine insemination in a hygienic manner.
A two-stage extender system, AndroPRO IUI™ PM/AM (available commercially from Minitube of America, Verona, Wis.), can be used to protect and preserve sperm cells from the negative effects of low dose concentrations during storage and then to activate sperm cells for optimal fertilization just prior to insemination. The first stage, PM, is formulated with an additive to preserve sperm cell concentrations as low as 500 million per dose. The second stage, AM, is formulated to provide two functions: activation of the sperm cells from storage and to provide an adequate volume of medium to support the peristaltic motion of the uterus, which transport the semen to the site of fertilization.
In applications, the AndroPRO IUI™ PM extender is combined with the sperm cells to maintain and stabilize sperm cell motility at low dose concentrations. To successfully reach the oviduct during IUI, the low volume, low concentration dose is combined with a larger volume of medium, such as the AndroPRO IUI™ AM extender, to add the necessary volume to produce the physiological conditions needed to transport sperm cells to the site of fertilization. An extender such as the AndroPRO IUI™ AM extender is formulated with additives to boost sperm cell activity and stimulate an increase in progressive motility, e.g., an increase in progressive motility at low dose concentrations stored up to five days.
A low concentration of sperm cells in a high volume dose can adversely affect semen shelf life. The PM extender, such as the AndroPRO IUI™ PM extender, is formulated to maintain the sperm cells at low metabolism (at “rest”) and can include an additive such as the Cell Shield Plus (CSP) extender additive (available commercially from Minitube of America, Verona, Wis.) to enhance the effect of the extender to protect low dose sperm motility and viability from the effects of dilution, and increase the ability of sperm cells to tolerate stressors associated with semen preservation and transport such as temperature fluctuations (e.g., between 10° C. to 25° C.), low dose concentrations, cold storage and oxidative stressors.
It is specifically intended that the present invention not be limited to the embodiments and illustrations contained herein, but include modified forms of those embodiments including portions of the embodiments and combinations of elements of different embodiments as come within the scope of the following claims.
Number | Date | Country | |
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61656465 | Jun 2012 | US |