The present disclosure relates generally to methods for improving traits or fitness in cattle, pigs, or aquatic organisms via gene editing. In particular, the methods comprise modifying genes in embryonic stem cells of livestock (e.g., cattle, pigs, or fish) for the purpose of improving a trait (e.g., promoting pathogen resistance, fertility, lactation, and native traits that support more rapid growth or feed efficiency). The methods particularly include CRISPR-CAS9 editing of porcine and bovine embryonic stem cells. Also disclosed herein are methods of producing and cultivating porcine embryonic stem cells under feeder free conditions.
The present teachings provide for a method of breeding livestock in vitro. Provided are steps to create embryonic stem cells from a plurality of blastocysts, genotype the embryonic stem cells to select the best embryos for mating to create offspring, and induce the embryonic stem cells into primordial germ cell-like cells (PGCLCs). Male PGCLCs are further induced into spermatogonial stem cell-like cells, and then spermatid-like cells. Female PGCLCs are induced into oocytes, which are then matured. The resulting gametes can then be mixed with each other or with opposite sex gametes from animals with desirable genetics to create the next generation of embryos, which can then be run through the process again. This method of in vitro breeding can be used to increase the speed of genetic progress in livestock.
A method and related apparatus for confirming whether a kill laser successfully destroys an undesired population of cells includes introducing fluorescent dye into cells, exciting the cells with a detection laser or a light emitting diode to cause the cell to fluoresce for a first time, measuring the amount of fluorescence in the cells with a detector capable of emitting a detection pulse, classifying the cells via embedded processing as undesired or desired cells based on the amount of fluorescence, firing a kill beam with a kill laser at any undesired cells, measuring the amount of fluorescence in the cells a second time to determine whether a fluorescent event was generated from the kill beam striking the cells, and providing feedback to an operator of the kill laser as to whether any fluorescent events were generated from the kill beam striking the cells.
Modular flow cytometry systems and methods for processing samples are described herein. The systems include automated or semi-automated modules that are replaceable and removable. A sample pathway module may be removed and placed in a microfluidic device cleaning module for cleaning, and then reinstalled or stored for later use. The systems further include optical modules, electronics modules, and mixing and collection modules. The optical module includes a photo-damaging assembly and detection laser assembly that may be on the same side relative to a plane or surface of a flow cytometry device and opposite of a detection assembly. The laser beam may have a beam waist that is wider in a direction perpendicular to a flow direction than in the flow direction. The mixing and collection module can automatically mix a sample being collected in a sample tube and switch to another sample tube when the other tube is full.
Disclosed are gene edited ungulate animals and methods of editing genes that control various ungulate traits. Such methods include CRISPR, zinc fingers, or TALENS. Exemplary traits for editing include polled, sterility or fertility, milk production, growth (which increases meat production), fat production, conception rates, stillborn rates, calving ease, or content of produced milk such as the amount of protein or the amount of fat. Other traits include backfat thickness, intramuscular fat, ultrasound loin muscle area, loin muscle area and intramuscular fat content, chest circumference, withers height, body length, hip height, rump length, and heart girth. Other exemplary native traits include, but are not limited to, high altitude adaptation and response to hypoxia, cold acclimation, body size and stature, resistance to disease and bacterial infection, reproduction, milk yield and components, and feed efficiency.
What is provided is a microweir structure or a set of one or more microweir structures disposed in a microfluidic flow channel, such as a channel in one or more substrate layers of a microfluidic chip. The microweir structure or set of microweir structures orient particles suspended in a sample fluid stream in the microfluidic flow channel, or focus a sample stream which is in laminar flow with a buffer fluid stream in the microfluidic flow channel. A set of one or more lower pressure zones or vortices downstream of a microweir structure contribute to the focusing and orientation effects of the microweir structures.
A method of choosing which undesired cell to destroy in a multi-cell fluorescent event includes detecting fluorescence of cells, converting photons detected in the fluorescence into an analog voltage output signal, and identifying at least two discernable peaks associated with the cells. By looking solely at properties measured within the multi-cell fluorescent event, a decision of which cell to target for elimination can be made. Using this method with large population sizes can result in an effective sex skewed product. The sex skewed product can, for example, be formed from bull semen which is then later used to inseminate cows which results in an increased likelihood of giving birth to female cattle.
Disclosed are methods of increasing production of sex selected semen for elite animals without a loss of plant efficiency. Assignment of elite animal ejaculate to a number of instruments so that a set number of cells is applied to each instrument is disclosed. Also disclosed is allowing the instrument to run until the ejaculate sample is exhausted, the instrument reports a dead percent of 25, or the sample has been processing for eight hours. Methods of pairing elite bulls with less valuable animals to preserve plant efficiency are also disclosed.
Provided herein are methods of producing heterozygous edited animals by injecting zygotes with low amounts of ribonucleoprotein (RNP) complex. The data cited herein show that more heterozygous animals are obtained from zygotes injected with 100,000 – 450,000, or approximately 111,250 copies of an RNP complex. The methods disclosed herein are particularly useful for editing of bovine or porcine zygotes. Heterozygous animals are preferred when homozygous embryos fail to develop, or when homozygous animals are sterile, therefore making it difficult to breed the edit into a line of animals.
The present teachings provide for methods of separating live and dead spermatozoa cells from non-human mammalian sex-selected semen. Sex-selected semen is first treated with proteinase K and TCEP, then applied to a density gradient media for separation. Separation is typically accomplished through centrifugation. Commercially available density gradient media include PERCOLL® and BOVIPURE®. Separation of live and dead cells in sex selected semen allows for precise quantification of sex skew using ddPCR™.
Disclosed are methods of creating embryonic stem cells through nuclear transfer or embryo complementation. In general, methods of complementation include contacting embryonic stem cells with a host embryo. Host embryos can be wild-type embryos, tetraploid embryos, parthenogenetic embryos, trophoblastic vesicles, or embryos that have been gene edited to prevent them from making germ cells. Trophoblastic vesicles are typically made by ablating the inner cell mass of an embryo via heat shock, chemical treatment, laser ablation, or microdissection of the ICM. The embryonic stem cells can then be injected into the host embryo or fused with the host embryo by placing them in contact in culture. Once fused and matured, the resulting mosaic embryo is then implanted into a surrogate mother.
05 - Pharmaceutical, veterinary and sanitary products
29 - Meat, dairy products, prepared or preserved foods
31 - Agricultural products; live animals
44 - Medical, veterinary, hygienic and cosmetic services; agriculture, horticulture and forestry services
Goods & Services
(1) Animal semen
(2) Meat
(3) Animal embryos; cattle; live cattle
(4) Dairy cows; (1) Animal husbandry; artificial insemination services; artificial insemination services for animals; cattle breeding services; information, advice and consultancy services in the field of artificial insemination services for animals
(2) Dairy cow breeding services; Consultancy and advisory services in the field of animal breeding and animal reproductive management;
14.
Methods of sperm cell sensing utilizing a semiconductor detector and cytometer apparatus
A cytometer includes an avalanche photodiode, a switching power supply, a filter, and voltage adjustment circuitry. The switching power supply includes a feedback loop. The filter is electrically connected between the switching power supply and the avalanche photodiode. The voltage adjustment circuitry adjusts a voltage on the feedback loop based at least in part on a voltage measured between the filter and the avalanche photodiode.
H01L 31/107 - Devices sensitive to infrared, visible or ultraviolet radiation characterised by only one potential barrier or surface barrier the potential barrier working in avalanche mode, e.g. avalanche photodiode
G01N 15/14 - Optical investigation techniques, e.g. flow cytometry
G01N 15/1404 - Handling flow, e.g. hydrodynamic focusing
An integrated system and method for preparing sperm cells to improve their survivability during cryopreservation are described herein. The system features a vessel, a controlled dispenser, and a dispense tube. The dispense tube has a first end fluidly connected to the dispenser and a second end disposed inside the vessel. The second end can be submerged in the sperm cell fluid. The controlled dispenser may be a syringe pump that includes a syringe for containing a cryoprotectant and a pushing mechanism for displacing the syringe. The syringe pump is configured to discharge the cryoprotectant through the dispense tube and into the vessel, thereby dispensing the cryoprotectant into the sperm cell fluid. Mixing of the fluids is achieved using a shaker table agitation.
Provided are genetically edited animals, particularly dairy animals such as cows or heifers, that lactate without having first been pregnant. Also provided are methods and reagents for creating genetically modified animals. Specifically provided are genetically edited dairy animals that have a histidine to arginine substitution in the prolactin receptor gene.
05 - Pharmaceutical, veterinary and sanitary products
44 - Medical, veterinary, hygienic and cosmetic services; agriculture, horticulture and forestry services
Goods & Services
Live animals; Animal embryos Animal semen; animal healthcare products, namely, teat dip and bactericidal or germicidal preparations for domestic animals Artificial insemination services; consultancy and advisory services in the field of animal breeding and reproductive management
18.
SYSTEMS, METHODS, AND APPARATUS FOR A MICROFLUIDIC CHIP HAVING A MICROCHANNEL DESIGN WHICH ASYMMETRICALLY FOCUSES PARTICLES
What is provided is an asymmetrically focusing microfluidic chip and a method of asymmetrically focusing and processing particles using an asymmetrically focusing microfluidic chip. The microfluidic chip comprises a microfluidic channel structure with a sheath fluid channel structure originating upstream at a sheath fluid inlet, the sheath fluid channel structure comprising a first side channel structure and a second side channel structure each having a plurality of tines. A sheath fluid flows through the sheath fluid channel structure into a flow channel at an intersection with a sample fluid flowing from a sample fluid channel. The sample fluid is asymmetrically focused in the intersection. The sample fluid may be analyzed and processed before flowing out of the channel structure via one or more outlets.
01 - Chemical and biological materials for industrial, scientific and agricultural use
29 - Meat, dairy products, prepared or preserved foods
31 - Agricultural products; live animals
44 - Medical, veterinary, hygienic and cosmetic services; agriculture, horticulture and forestry services
Goods & Services
Animal semen. Meat. Live animals, namely cattle and dairy cows; animal embryos. Artificial insemination services; animal breeding; consultancy and advisory services in the field of animal breeding and reproductive management.
05 - Pharmaceutical, veterinary and sanitary products
29 - Meat, dairy products, prepared or preserved foods
31 - Agricultural products; live animals
44 - Medical, veterinary, hygienic and cosmetic services; agriculture, horticulture and forestry services
Goods & Services
animal semen meat live animals, namely, cattle and dairy cows; animal embryos artificial insemination services; animal breeding; consultancy and advisory services in the field of animal breeding and reproductive management
A method and related apparatus for confirming whether a kill laser successfully destroys an undesired population of cells includes introducing fluorescent dye into cells, exciting the cells with a detection laser or a light emitting diode to cause the cell to fluoresce for a first time, measuring the amount of fluorescence in the cells with a detector capable of emitting a detection pulse, classifying the cells via embedded processing as undesired or desired cells based on the amount of fluorescence, firing a kill beam with a kill laser at any undesired cells, measuring the amount of fluorescence in the cells a second time to determine whether a fluorescent event was generated from the kill beam striking the cells, and providing feedback to an operator of the kill laser as to whether any fluorescent events were generated from the kill beam striking the cells.
The present disclosure relates generally to potassium channel blocker compounds and compositions thereof; and methods for improving reproductive cell viability and quality, during and/or after one or more of staining, freezing, thawing, cell sample enrichment, packaging, or in vitro fertilization using the potassium channel blocker compounds.
A61K 31/14 - Quaternary ammonium compounds, e.g. edrophonium, choline
A61P 15/08 - Drugs for genital or sexual disordersContraceptives for gonadal disorders or for enhancing fertility, e.g. inducers of ovulation or of spermatogenesis
e.g.in vitroin vitro fertilization using the fertilization enhancing compounds. These compounds are especially useful for preserving sperm cell viability during a sex selection or sexing process.
A method of inseminating an animal including flowing a stream of a population of sperm cells through a channel, differentiating the sperm cells into two subpopulations of X-chromosome containing sperm cells and Y-chromosome containing sperm cells, selecting a desired subpopulation, ablating an undesired subpopulation, and collecting both the subpopulations of sperm cells including the desired subpopulation and the ablated undesired subpopulation together, wherein the collected population of sperm cells is used to fertilize an egg.
Microfluidic devices and methods for focusing components in a fluid sample are described herein. The microfluidic devices feature a microfluidic chip having a micro-channel having a constricting portion that narrows in width, and a flow focusing region downstream of the micro-channel. The flow focusing region includes a positively sloping bottom surface that reduces a height of the flow focusing region and sidewalls that taper to reduce a width of the flow focusing region, thereby geometrically constricting the flow focusing region. The devices and methods can be utilized in sex-sorting of sperm cells to improve performance and increase eligibility.
e.g.,e.ge.g., promoting pathogen resistance, fertility, lactation, and native traits that support more rapid growth or feed efficiency). The methods particularly include CRISPR-CAS9 editing of porcine and bovine embryonic stem cells. Also disclosed herein are methods of producing and cultivating porcine embryonic stem cells under feeder free conditions.
in vitro in vitro produced embryos. Cell-free DNA can be obtained from the culture medium of blastocysts, and then added to a LAMP or PCR amplification reaction to detect the presence or absence of a certain DNA sequence. Colorimetric or fluorescent indicators such as metallochromatic dyes, intercalating dyes, or pH indicator dyes can be added to the reaction so that amplification can be tracked without additional steps. Provided are methods of sex determination and detection of a specific gene edit, as well adverse or desirable phenotypes. Methods of genotyping using cell-free DNA are also provided.
09 - Scientific and electric apparatus and instruments
42 - Scientific, technological and industrial services, research and design
44 - Medical, veterinary, hygienic and cosmetic services; agriculture, horticulture and forestry services
Goods & Services
Computer hardware for tracking livestock location and health; Computer software for tracking livestock location and health; Computer software applications for tracking livestock location and health; Downloadable mobile application for tracking livestock location and health. Software as a service (SAAS) services for use in the tracking, analysis, and reporting of livestock location and health. Consulting services related to livestock management, performance, and health.
31.
COMPUTATIONAL TECHNIQUES FOR IDENTIFYING UNOBSERVED HEREDITARY INFORMATION BASED ON ANALYSIS OF LIMITED DATA
Systems and methods for organism genotyping and using genomic data for genotype imputation are disclosed. The system can maintain first genetic sequence information of a sire and second genetic sequence information for a dam, the first and second genetic sequence information indicating one or more single nucleotide polymorphisms (SNPs) of interest. The system can sequence, based on a skim sequencing technique, a sample of genetic information of a progeny of the sire and the dam. The system can identify informative variants based on the first genetic sequence information and the second genetic sequence information. The system can identify informative reads in the sequence of the progeny based on the informative variants. The system can construct a genotype of the progeny based on the informative reads and the one or more SNPs of interest in the first and second genetic sequence information.
A cytometer includes an avalanche photodiode, a switching power supply, a filter, and voltage adjustment circuitry. The switching power supply includes a feedback loop. The filter is electrically connected between the switching power supply and the avalanche photodiode. The voltage adjustment circuitry adjusts a voltage on the feedback loop based at least in part on a voltage measured between the filter and the avalanche photodiode.
G01N 21/00 - Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
H01L 31/107 - Devices sensitive to infrared, visible or ultraviolet radiation characterised by only one potential barrier or surface barrier the potential barrier working in avalanche mode, e.g. avalanche photodiode
H01L 31/02 - SEMICONDUCTOR DEVICES NOT COVERED BY CLASS - Details thereof - Details
H01L 31/024 - Arrangements for cooling, heating, ventilating or temperature compensation
09 - Scientific and electric apparatus and instruments
42 - Scientific, technological and industrial services, research and design
44 - Medical, veterinary, hygienic and cosmetic services; agriculture, horticulture and forestry services
Goods & Services
Computer hardware for tracking livestock location and health; Computer software for tracking livestock location and health; Computer software applications for tracking livestock location and health; Downloadable mobile application for tracking livestock location and health. Software as a service (SAAS) services for use in the tracking, analysis, and reporting of livestock location and health. Consulting services related to livestock management, performance, and health.
A method of choosing which undesired cell to destroy in a multi-cell fluorescent event includes detecting fluorescence of cells, converting photons detected in the fluorescence into an analog voltage output signal, and identifying at least two discernable peaks associated with the cells. By looking solely at properties measured within the multi-cell fluorescent event, a decision of which cell to target for elimination can be made. Using this method with large population sizes can result in an effective sex skewed product. The sex skewed product can, for example, be formed from bull semen which is then later used to inseminate cows which results in an increased likelihood of giving birth to female cattle.
A microfluidic chip orients and isolates components in a sample fluid mixture by two step focusing, where sheath fluids compress the sample fluid mixture in a sample input channel in one direction, such that the sample fluid mixture becomes a narrower stream bounded by the sheath fluids, and by having the sheath fluids compress the sample fluid mixture in a second direction further downstream, such that the components are compressed and oriented in a selected direction to pass through an interrogation chamber in single file formation for identification and separation by various methods. The isolation mechanism utilizes external, stacked piezoelectric actuator assemblies disposed on a microfluidic chip holder, or piezoelectric actuator assemblies on-chip, so that the actuator assemblies are triggered by an electronic signal to actuate jet chambers on either side of the sample input channel, to jet selected components in the sample input channel into one of the output channels.
The present disclosure relates generally to methods of improving reproductive cell sample quality. The methods include improving the quality of mammalian semen cell samples by discriminating between live and dead sperm cells in a semen sample obtained from livestock, such as bovine or porcine animals. The methods include improving reproductive cell sample quality based on the exposure or availability of an acrosome associated molecule by contacting the sample with a binding agent that binds to the acrosome associated molecule to form a mixture of the semen and the binding agent and separating sperm cells bound to the binding agent from sperm cells not bound to the binding agent.
A method of inseminating an animal including flowing a stream of a population of sperm cells through a channel, differentiating the sperm cells into two subpopulations of X-chromosome containing sperm cells and Y-chromosome containing sperm cells, selecting a desired subpopulation, ablating an undesired subpopulation, and collecting both the subpopulations of sperm cells including the desired subpopulation and the ablated undesired subpopulation together, wherein the collected population of sperm cells is used to fertilize an egg.
The present teachings provide for a method of breeding livestock in vitro. Provided are steps to create embryonic stem cells from a plurality of blastocysts, genotype the embryonic stem cells to select the best embryos for mating to create offspring, and induce the embryonic stem cells into primordial germ cell-like cells (PGCLCs). Male PGCLCs are further induced into spermatogonial stem cell-like cells, and then spermatid-like cells. Female PGCLCs are induced into oocytes, which are then matured. The resulting gametes can then be mixed with each other or with opposite sex gametes from animals with desirable genetics to create the next generation of embryos, which can then be run through the process again. This method of in vitro breeding can be used to increase the speed of genetic progress in livestock.
A method and related apparatus for confirming whether a kill laser successfully destroys an undesired population of cells includes introducing fluorescent dye into cells, exciting the cells with a detection laser or a light emitting diode to cause the cell to fluoresce for a first time, measuring the amount of fluorescence in the cells with a detector capable of emitting a detection pulse, classifying the cells via embedded processing as undesired or desired cells based on the amount of fluorescence, firing a kill beam with a kill laser at any undesired cells, measuring the amount of fluorescence in the cells a second time to determine whether a fluorescent event was generated from the kill beam striking the cells, and providing feedback to an operator of the kill laser as to whether any fluorescent events were generated from the kill beam striking the cells.
Disclosed is an approach to differentiating between different particle types in samples flowing through microfluidics chips. A sample may have an initial proportion of a first cell type to a second cell type. An illuminating light source may emit a coherent light at the sample, and light leaving the chip in a first direction may be detected using a first light detector, and light leaving the chip in a second direction (e.g., orthogonal to the first direction) may be detected using a second light detector. The detected light may be fluorescence. An orientational feature of a plurality of cells in the sample may be determined based on the light detected by the detectors. Based on the orientational features and the detected light, a biasing operation may be performed for each cell in the sample to obtain a modified proportion of cell types in the sample.
Disclosed is an approach to differentiating between different particle types in samples flowing through microfluidics chips. A sample may have an initial proportion of a first cell type to a second cell type. An illuminating light source may emit a coherent light at the sample, and light leaving the chip in a first direction may be detected using a first light detector, and light leaving the chip in a second direction (e.g., orthogonal to the first direction) may be detected using a second light detector. The detected light may be fluorescence. An orientational feature of a plurality of cells in the sample may be determined based on the light detected by the detectors. Based on the orientational features and the detected light, a biasing operation may be performed for each cell in the sample to obtain a modified proportion of cell types in the sample.
A cytometer includes an avalanche photodiode, a switching power supply, a filter, and voltage adjustment circuitry. The switching power supply includes a feedback loop. The filter is electrically connected between the switching power supply and the avalanche photodiode. The voltage adjustment circuitry adjusts a voltage on the feedback loop based at least in part on a voltage measured between the filter and the avalanche photodiode.
H01L 31/107 - Devices sensitive to infrared, visible or ultraviolet radiation characterised by only one potential barrier or surface barrier the potential barrier working in avalanche mode, e.g. avalanche photodiode
H01L 31/024 - Arrangements for cooling, heating, ventilating or temperature compensation
G01N 15/14 - Optical investigation techniques, e.g. flow cytometry
Provided are genetically edited animals, particularly dairy animals such as cows or heifers, that lactate without having first been pregnant. Also provided are methods and reagents for creating genetically modified animals. Specifically provided are genetically edited dairy animals that have a histidine to arginine substitution in the prolactin receptor gene.
A microfluidic system configured to focus particles suspended in a fluid. One general aspect includes a microfluidic system comprising one or more substrates and a focusing channel formed in the one or more substrates and spanning a length from an inlet to an outlet for receiving a flow of particles suspended in fluid, wherein the particles have a diameter (a) and the focusing channel has a hydraulic diameter (dh).
Modular flow cytometry systems and methods for processing samples are described herein. The systems include automated or semi-automated modules that are replaceable and removable. A sample pathway module may be removed and placed in a microfluidic device cleaning module for cleaning, and then reinstalled or stored for later use. The systems further include optical modules, electronics modules, and mixing and collection modules. The optical module includes a photo-damaging assembly and detection laser assembly that may be on the same side relative to a plane or surface of a flow cytometry device and opposite of a detection assembly. The laser beam may have a beam waist that is wider in a direction perpendicular to a flow direction than in the flow direction. The mixing and collection module can automatically mix a sample being collected in a sample tube and switch to another sample tube when the other tube is full.
Modular flow cytometry systems and methods for processing samples are described herein. The systems include automated or semi-automated modules that are replaceable and removable. A sample pathway module may be removed and placed in a microfluidic device cleaning module for cleaning, and then reinstalled or stored for later use. The systems further include optical modules, electronics modules, and mixing and collection modules. The optical module includes a photo-damaging assembly and detection laser assembly that may be on the same side relative to a plane or surface of a flow cytometry device and opposite of a detection assembly. The laser beam may have a beam waist that is wider in a direction perpendicular to a flow direction than in the flow direction. The mixing and collection module can automatically mix a sample being collected in a sample tube and switch to another sample tube when the other tube is full.
44 - Medical, veterinary, hygienic and cosmetic services; agriculture, horticulture and forestry services
Goods & Services
Animal breeding, artificial insemination of animals, animal
semen extraction, animal sperm bank services; animal stud
services; veterinary services; genetic testing of animals;
advisory, information and consultancy services related to
all the aforesaid services.
44 - Medical, veterinary, hygienic and cosmetic services; agriculture, horticulture and forestry services
Goods & Services
Animal breeding, artificial insemination of animals, animal
semen extraction, animal sperm bank services; animal stud
services; veterinary services; genetic testing of animals;
advisory, information and consultancy services related to
all the aforesaid services.
52.
METHODS AND SYSTEMS FOR PROCESSING GENETIC SAMPLES TO DETERMINE IDENTITY OR DETECT CONTAMINATION
Methods and systems for processing semen samples from straws for determining genetic identity, testing the purity of the sample, detecting errors or contamination, calculating an amount of contamination, and determining the identity of the contaminant. The methods herein can detect low levels of contamination, such as contamination of about 1%, 2%, etc.
Methods and systems for processing semen samples from straws for determining genetic identity, testing the purity of the sample, detecting errors or contamination, calculating an amount of contamination, and determining the identity of the contaminant. The methods herein can detect low levels of contamination, such as contamination of about 0.5%, 1%, 2%, etc.
The present disclosure relates generally to methods for using porcine sex-sorted sperm cells for the efficient dissemination of desirable traits in multi-level swine production systems. The methods include using sex-sorted sperm cells for skewing offspring gender at the commercial farm level, producing porcine herds having improved growth performance traits, producing pathogen-resistant porcine herds, and disseminating desirable traits from a genetic nucleus to commercial farms using low dose artificial insemination techniques. The methods also provide a means for reducing costs at the production level by increasing the ratio of female offspring and improving animal welfare at all levels of production by reducing or eliminating male castration. In addition, the methods of the present technology may be employed to develop production flows for specialized pork products.
The present disclosure relates generally to methods for using porcine sex-sorted sperm cells for the efficient dissemination of desirable traits in multi-level swine production systems. The methods include using sex-sorted sperm cells for skewing offspring gender at the commercial farm level, producing porcine herds having improved growth performance traits, producing pathogen-resistant porcine herds, and disseminating desirable traits from a genetic nucleus to commercial farms using low dose artificial insemination techniques. The methods also provide a means for reducing costs at the production level by increasing the ratio of female offspring and improving animal welfare at all levels of production by reducing or eliminating male castration. In addition, the methods of the present technology may be employed to develop production flows for specialized pork products.
44 - Medical, veterinary, hygienic and cosmetic services; agriculture, horticulture and forestry services
Goods & Services
Animal breeding, artificial insemination of animals, animal semen extraction for animal breeding purposes, animal sperm bank services; animal stud services; veterinary services; genetic testing of animals for diagnostic or breeding planning purposes; advisory, information and consultancy services related to all the aforesaid services
44 - Medical, veterinary, hygienic and cosmetic services; agriculture, horticulture and forestry services
Goods & Services
(1) Agricultural services, namely animal breeding, artificial insemination of animals, animal semen extraction, animal sperm bank services; stud services for cattle; veterinary services; genetic testing of animals; advisory, information and consultancy services related to the breeding of cattle.
44 - Medical, veterinary, hygienic and cosmetic services; agriculture, horticulture and forestry services
Goods & Services
(1) Agricultural services, namely animal breeding, artificial insemination of animals, animal semen extraction, animal sperm bank services; stud services for cattle; veterinary services; genetic testing of animals; advisory, information and consultancy services related to the breeding of cattle.
44 - Medical, veterinary, hygienic and cosmetic services; agriculture, horticulture and forestry services
Goods & Services
Animal breeding, artificial insemination of animals, animal semen extraction for animal breeding purposes, animal sperm bank services; animal stud services; veterinary services; genetic testing of animals for diagnostic or breeding planning purposes; advisory, information and consultancy services related to all the aforesaid services
60.
SYSTEM AND METHODS FOR SUB-POPULATION IDENTIFICATION
Systems and methods are provided for identification and discrimination of subpopulations within a mixture of particles. The systems and methods implement continuous calibration of the classification of particles within the mixture to provide consistency in operation and to reduce inter- and intra-batch processing variation. The systems and methods produce advantageously sorted particle products.
A method of inseminating an animal including flowing a stream of a population of sperm cells through a channel, differentiating the sperm cells into two subpopulations of X-chromosome containing sperm cells and Y-chromosome containing sperm cells, selecting a desired subpopulation, ablating an undesired subpopulation, and collecting both the subpopulations of sperm cells including the desired subpopulation and the ablated undesired subpopulation together, wherein the collected population of sperm cells is used to fertilize an egg.
A microfluidic chip orients and isolates components in a sample fluid mixture by two step focusing, where sheath fluids compress the sample fluid mixture in a sample input channel in one direction, such that the sample fluid mixture becomes a narrower stream bounded by the sheath fluids, and by having the sheath fluids compress the sample fluid mixture in a second direction further downstream, such that the components are compressed and oriented in a selected direction to pass through an interrogation chamber in single file formation for identification and separation by various methods. The isolation mechanism utilizes external, stacked piezoelectric actuator assemblies disposed on a microfluidic chip holder, or piezoelectric actuator assemblies on-chip, so that the actuator assemblies are triggered by an electronic signal to actuate jet chambers on either side of the sample input channel, to jet selected components in the sample input channel into one of the output channels.
Microfluidic devices and methods for focusing components in a fluid sample are described herein. The microfluidic devices feature a microfluidic chip having a micro-channel having a constricting portion that narrows in width, and a flow focusing region downstream of the micro-channel. The flow focusing region includes a positively sloping bottom surface that reduces a height of the flow focusing region and sidewalls that taper to reduce a width of the flow focusing region, thereby geometrically constricting the flow focusing region. The devices and methods can be utilized in sex-sorting of sperm cells to improve performance and increase eligibility.
Microfluidic devices and methods for focusing components in a fluid sample are described herein. The microfluidic devices feature a microfluidic chip having a micro-channel having a constricting portion that narrows in width, and a flow focusing region downstream of the micro-channel. The flow focusing region includes a positively sloping bottom surface that reduces a height of the flow focusing region and sidewalls that taper to reduce a width of the flow focusing region, thereby geometrically constricting the flow focusing region. The devices and methods can be utilized in sex-sorting of sperm cells to improve performance and increase eligibility.
Technologies for assessing, quantifying and isolating sperm cell populations and/or subpopulations having specific genetic signatures are provided, as well as methods and systems to assess the efficacy of chromosomal differentiation processes. Compositions for identification and differentiation of X-chromosomes and Y-chromosomes in DNA are also provided.
The present teachings provide for and include methods of preparing semen for in vitro fertilization (IVF). Provided are methods of removing dead cells from semen preparations by applying the semen to a colloidal silica gradient and centrifuging the gradient. In some configurations, the gradient is a 45-90% colloidal silica gradient and the centrifugation is at 419 x g. Further provided are use of the method of the present teachings for processing sex selected semen. Also provided are ova and zygotes matured therefrom that have been fertilized by semen that has been processed by a method of the present teachings.
A cytometer includes an avalanche photodiode, a switching power supply, a filter, and voltage adjustment circuitry. The switching power supply includes a feedback loop. The filter is electrically connected between the switching power supply and the avalanche photodiode. The voltage adjustment circuitry adjusts a voltage on the feedback loop based at least in part on a voltage measured between the filter and the avalanche photodiode.
H01L 31/107 - Devices sensitive to infrared, visible or ultraviolet radiation characterised by only one potential barrier or surface barrier the potential barrier working in avalanche mode, e.g. avalanche photodiode
H01L 31/024 - Arrangements for cooling, heating, ventilating or temperature compensation
G01N 15/14 - Optical investigation techniques, e.g. flow cytometry
Bos taurus variety JE840003146074527. Included in the present disclosure are cells comprising the Bovine variety JE840003146074527. Also provided by the present disclosure are tissue cultures of cells, animals obtained from said cells, and parts thereof, including F1 spermatozoa. The disclosure further provides for methods of breeding, selecting, and using the germplasm to improve existing commercial cattle herds generated from in vitro fertilization methods and progeny cattle obtained from in vitro fertilization and implantation and artificial insemination methods.
A method of choosing which undesired cell to destroy in a multi-cell fluorescent event includes detecting fluorescence of cells, converting photons detected in the fluorescence into an analog voltage output signal, and identifying at least two discernable peaks associated with the cells. By looking solely at properties measured within the multi-cell fluorescent event, a decision of which cell to target for elimination can be made. Using this method with large population sizes can result in an effective sex skewed product. The sex skewed product can, for example, be formed from bull semen which is then later used to inseminate cows which results in an increased likelihood of giving birth to female cattle.
An integrated system and method for preparing sperm cells to improve their survivability during cryopreservation are described herein. The system features a vessel, a controlled dispenser, and a dispense tube. The dispense tube has a first end fluidly connected to the dispenser and a second end disposed inside the vessel. The second end can be submerged in the sperm cell fluid. The controlled dispenser may be a syringe pump that includes a syringe for containing a cryoprotectant and a pushing mechanism for displacing the syringe. The syringe pump is configured to discharge the cryoprotectant through the dispense tube and into the vessel, thereby dispensing the cryoprotectant into the sperm cell fluid. Mixing of the fluids is achieved using a shaker table agitation.
A61B 18/02 - Surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body by cooling, e.g. cryogenic techniques
C12M 1/00 - Apparatus for enzymology or microbiology
C12N 5/00 - Undifferentiated human, animal or plant cells, e.g. cell linesTissuesCultivation or maintenance thereofCulture media therefor
75.
System and process for continuous addition of cryoprotectant
An integrated system and method for preparing sperm cells to improve their survivability during cryopreservation are described herein. The system features a vessel, a controlled dispenser, and a dispense tube. The dispense tube has a first end fluidly connected to the dispenser and a second end disposed inside the vessel. The second end can be submerged in the sperm cell fluid. The controlled dispenser may be a syringe pump that includes a syringe for containing a cryoprotectant and a pushing mechanism for displacing the syringe. The syringe pump is configured to discharge the cryoprotectant through the dispense tube and into the vessel, thereby dispensing the cryoprotectant into the sperm cell fluid. Mixing of the fluids is achieved using a shaker table agitation.
The disclosure relates to Bovine germplasm of Bos taurus variety HO840M003150607238. Included in the present disclosure are cells comprising the genome of Bovine variety HO840M003150607238 characterized by the presence of homozygous loci and spermatozoa obtained from said cells. Also provided by the present disclosure are tissue cultures of cells, animals obtained from said cells, and parts thereof, including F1 spermatozoa. The disclosure further provides for methods of breeding, selecting, and using the germplasm to improve existing commercial cattle herds generated from in vitro fertilization methods and progeny cattle obtained from in vitro fertilization and implantation and artificial insemination methods.
A cytometer includes an avalanche photodiode, a switching power supply, a filter, and voltage adjustment circuitry. The switching power supply includes a feedback loop. The filter is electrically connected between the switching power supply and the avalanche photodiode. The voltage adjustment circuitry adjusts a voltage on the feedback loop based at least in part on a voltage measured between the filter and the avalanche photodiode.
G01N 21/00 - Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
H01L 31/024 - Arrangements for cooling, heating, ventilating or temperature compensation
H01L 31/107 - Devices sensitive to infrared, visible or ultraviolet radiation characterised by only one potential barrier or surface barrier the potential barrier working in avalanche mode, e.g. avalanche photodiode
A microfluidic chip orients and isolates components in a sample fluid mixture by two-step focusing, where sheath fluids compress the sample fluid mixture in a sample input channel in one direction, such that the sample fluid mixture becomes a narrower stream bounded by the sheath fluids, and by having the sheath fluids compress the sample fluid mixture in a second direction further downstream, such that the components are compressed and oriented in a selected direction to pass through an interrogation chamber in single file formation for identification and separation by various methods. The isolation mechanism utilizes external, stacked piezoelectric actuator assemblies disposed on a microfluidic chip holder, or piezoelectric actuator assemblies on-chip, so that the actuator assemblies are triggered by an electronic signal to actuate jet chambers on either side of the sample input channel, to jet selected components in the sample input channel into one of the output channels.
Microfluidic devices and methods for focusing components in a fluid sample are described herein. The microfluidic device has at least one flow focusing channel where the components are focused or re-oriented by the geometry of the channel. From an upstream end of the flow focusing channel to a downstream end of the flow focusing channel, at least a portion of the flow focusing channel has a reduction in height and at least a portion of the flow focusing channel narrows in width, thereby geometrically constricting the flow focusing channel. The devices and methods can be utilized in sex-sorting of sperm cells to improve performance and increase eligibility.
B01L 3/00 - Containers or dishes for laboratory use, e.g. laboratory glasswareDroppers
G01N 35/00 - Automatic analysis not limited to methods or materials provided for in any single one of groups Handling materials therefor
G01N 33/48 - Biological material, e.g. blood, urineHaemocytometers
G01N 15/06 - Investigating concentration of particle suspensions
G01N 15/14 - Optical investigation techniques, e.g. flow cytometry
B65G 51/00 - Conveying articles through pipes or tubes by fluid flow or pressureConveying articles over a flat surface, e.g. the base of a trough, by jets located in the surface
A method and related apparatus for confirming whether a kill laser successfully destroys an undesired population of cells includes introducing fluorescent dye into cells, exciting the cells with a detection laser or a light emitting diode to cause the cell to fluoresce for a first time, measuring the amount of fluorescence in the cells with a detector capable of emitting a detection pulse, classifying the cells via embedded processing as undesired or desired cells based on the amount of fluorescence, firing a kill beam with a kill laser at any undesired cells, measuring the amount of fluorescence in the cells a second time to determine whether a fluorescent event was generated from the kill beam striking the cells, and providing feedback to an operator of the kill laser as to whether any fluorescent events were generated from the kill beam striking the cells.
A microfluidic system configured to focus particles suspended in a fluid. One general aspect includes a microfluidic system comprising one or more substrates and a focusing channel formed in the one or more substrates and spanning a length from an inlet to an outlet for receiving a flow of particles suspended in fluid, wherein the particles have a diameter (a) and the focusing channel has a hydraulic diameter (dh).
A microfluidic system configured to focus particles suspended in a fluid. One general aspect includes a microfluidic system comprising one or more substrates and a focusing channel formed in the one or more substrates and spanning a length from an inlet to an outlet for receiving a flow of particles suspended in fluid, wherein the particles have a diameter (a) and the focusing channel has a hydraulic diameter (dh).
Applicants have identified that three critical phenotypic/genetic measures are highly correlated with transition period health and may be used in selection and breeding protocols and/or in combination with traditional breeding and marker assisted selection methods to improve predictability of transition period health. According to the invention genetic evaluations for mastitis, ketosis, and metritis have been found to be highly predictive of overall transition health. The genetic evaluations are produced by directly measuring thousands of clinical cases of mastitis, ketosis, and metritis in ancestors of a particular animal and using this data in selection. Applicant's selection criteria and quickly impact a breeders population by reducing transition cow disease incidence in the initial population and in progeny.
G16B 20/40 - Population geneticsLinkage disequilibrium
G16B 20/20 - Allele or variant detection, e.g. single nucleotide polymorphism [SNP] detection
G16H 50/30 - ICT specially adapted for medical diagnosis, medical simulation or medical data miningICT specially adapted for detecting, monitoring or modelling epidemics or pandemics for calculating health indicesICT specially adapted for medical diagnosis, medical simulation or medical data miningICT specially adapted for detecting, monitoring or modelling epidemics or pandemics for individual health risk assessment
G16B 20/00 - ICT specially adapted for functional genomics or proteomics, e.g. genotype-phenotype associations
A61D 19/04 - Instruments or methods for reproduction or fertilisation for embryo transplantation
84.
Avalanche photodiode and cytometer sperm sex sensing apparatus
A cytometer includes an avalanche photodiode, a switching power supply, a filter, and voltage adjustment circuitry. The switching power supply includes a feedback loop. The filter is electrically connected between the switching power supply and the avalanche photodiode. The voltage adjustment circuitry adjusts a voltage on the feedback loop based at least in part on a voltage measured between the filter and the avalanche photodiode.
H01L 31/107 - Devices sensitive to infrared, visible or ultraviolet radiation characterised by only one potential barrier or surface barrier the potential barrier working in avalanche mode, e.g. avalanche photodiode
G01N 15/14 - Optical investigation techniques, e.g. flow cytometry
A method of inseminating an animal including flowing a stream of a population of sperm cells through a channel, differentiating the sperm cells into two subpopulations of X-chromosome containing sperm cells and Y-chromosome containing sperm cells, selecting a desired subpopulation, ablating an undesired subpopulation, and collecting both the subpopulations of sperm cells including the desired subpopulation and the ablated undesired subpopulation together, wherein the collected population of sperm cells is used to fertilize an egg.
A cytometer includes an avalanche photodiode, a switching power supply, a filter, and voltage adjustment circuitry. The switching power supply includes a feedback loop. The filter is electrically connected between the switching power supply and the avalanche photodiode. The voltage adjustment circuitry adjusts a voltage on the feedback loop based at least in part on a voltage measured between the filter and the avalanche photodiode.
H01L 31/107 - Devices sensitive to infrared, visible or ultraviolet radiation characterised by only one potential barrier or surface barrier the potential barrier working in avalanche mode, e.g. avalanche photodiode
G01N 15/14 - Optical investigation techniques, e.g. flow cytometry
H01L 31/024 - Arrangements for cooling, heating, ventilating or temperature compensation
Technologies for assessing, quantifying and isolating sperm cell populations and/or subpopulations having specific genetic signatures are provided, as well as methods and systems to assess the efficacy of chromosomal differentiation processes. Compositions for identification and differentiation of X-chromosomes and Y-chromosomes in DNA are also provided.
A cytometer includes an avalanche photodiode, a switching power supply, a filter, and voltage adjustment circuitry. The switching power supply includes a feedback loop. The filter is electrically connected between the switching power supply and the avalanche photodiode. The voltage adjustment circuitry adjusts a voltage on the feedback loop based at least in part on a voltage measured between the filter and the avalanche photodiode.
H01L 31/107 - Devices sensitive to infrared, visible or ultraviolet radiation characterised by only one potential barrier or surface barrier the potential barrier working in avalanche mode, e.g. avalanche photodiode
This disclosure concerns a method of providing a hydrophobic coating on a microfluidic chip that promotes the discrete flow of at least one liquid. It includes applying the hydrophobic coating onto an area of the microfluidic chip. The disclosure further includes a microfluidic chip that provides discrete flow of at least one liquid.
This disclosure concerns a method of providing a hydrophobic coating on a microfluidic chip that promotes the discrete flow of at least one liquid. It includes applying the hydrophobic coating onto an area of the microfluidic chip. The disclosure further includes a microfluidic chip that provides discrete flow of at least one liquid.
Applicants have identified that three critical phenotypic/genetic measures are highly correlated with transition period health and may be used in selection and breeding protocols and/or in combination with traditional breeding and marker assisted selection methods to improve predictability of transition period health. According to the invention genetic evaluations for mastitis, ketosis, and metritis have been found to be highly predictive of overall transition health. The genetic evaluations are produced by directly measuring thousands of clinical cases of mastitis, ketosis, and metritis in ancestors of a particular animal and using this data in selection. Applicant's selection criteria can quickly impact a breeders population by reducing transition cow disease incidence in the initial population and in progeny.
