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Equines  have the abilities to interbreed freely with species from the genus Equus and produce offspring even though most of them are infertile.  Mule and hinny are ideal examples of the breeding. Equines are also known for their abilities to show xenogeneic extraspecies pregnancies through ET, which means that they support pregnancy after ET and acts as recipient for exotic species of equine's embryos include Przewalski’s horse and Grant’s zebra. (Smits, et al, 2012).

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Yup, it's this small doggo looking animal.

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Yup, it's this small doggo looking animal.

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Approximately 50 million years ago, horses underwent evolution from Hyracotherium or Eohippus (the dawn horse) in the northern hemisphere. (Smits, et al, 2012).

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A warm welcome to you all for visiting the blog!!! Lets start the experience joyously with some amazing NEIGH asmr ! Because..... Why not? Do loop it during the reading process ! This blog is all about Equine and Assisted Reproductive Technique (ARTs) involved.

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The Second Generation method of ET is not used commonly since it is not able to collect more than one embryo per cycle per mare and the costs are quite high.The Third Generation are also not preferred due to the inefficiencies, high costs, and the incomplete registry recognition on breed of equine. (University of Wisconsin-Madison, n.d.).

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Scientific Classification  for Genus Equus.

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Scientific Classification  for Genus Equus.

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Kingdom: | Animalia
Phylum: | Chordata
Class: | Mammalia
Order: | Perissodactyla
Family: | Equidae
Tribe: | Equini
Genus: | Equus
Species
E. africanus—African wild 🤬
E. ferus—wild and domestic horse
E. grevyi—Grévy's zebra
E. hemionus—onager
E. kiang—kiang
E. quagga—plains zebra
E. zebra—mountain zebra

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History of Assisted Reproductive Technology(ART) in mare.

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History of Assisted Reproductive Technology(ART) in mare.

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First Generation:- Discovered six decades ago, artificial insemination with frozen fresh sperm methods is still the horse management technique in general.
Second Generation:- Comes into people sight around 4 decades ago with techniques of embryo transfer(ET) and ultrasonography. This allow the imaging for reproductive organs and sexing for fetuses.
Third Generation:- Blooming of technologies and methods where techniques and methods include oocyte recovery, zygote intrafallopian transfer (ZIFT), embryo sexing, gamete intrafallopian transfer (GIFT) and intracytoplasmic sperm injection (ICSI) were discovered not longer than 20 years ago.

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🐎Johnson Wong Xiu Yang
🐎 Lee Hui Qian
🐎 Ngu Hui En
🐎 Wiiliam Tong Meng Yang

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The cumulus cells that surrounds the egg after being removed from the ovary makes it difficult to be located under the microscope and makes it difficult to determine the maturity of the egg and as immature eggs cannot be fertilised, it will cause the failure of fertilisation.

During ICSI, the cumulus cells are stripped away, allowing the scientists to confirm the maturity of the egg. The Figure below illustrates the cells surrounding the egg.

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1. The horse embryos in a ‘holding medium’ can be transferred to the uterus of the recipient mare.

2. Transcervical transfer of equine embryos can be performed using a transfer pipette after loading the embryo into either a 0.25- or 0.50- mL straw.

3. The straw is loaded with three columns of fluid separated by air spaces.                                i. First fluid column serves to lubricate the pipette exit.                                                            ii. Second fluid column is the embryo.                                                                                          iii. Third fluid column ensure the embryo is flushed out from the pipette.

4. The pipette is enclosed with a sterile plastic sheath to prevent contamination from the recipient’s 🤬 or 🤬 when entering the uterus.

5. Upon reaching the cervix, the sheath is pulled slowly and firmly backwards while maintaining the pipette is in the correct place.

6. In addition, the pipette should manoeuvre through the constricted cervical canal without causing any excessive dilation or trauma.

7. Hence, the protruding portion of the cervix can be straightened by holding it firmly between two fingers and pulling it backwards, or using a modified rat-toothed Velsellum forceps to grasp and straighten it.

(Stout, 2010)

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What is Intracytoplasmic Sperm Injection (ICSI)?

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What is Intracytoplasmic Sperm Injection (ICSI)?

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To fertilise the egg in IVF by using ICSI, a micropipette is used to inject a single sperm into the centre of the egg (Hinrichs, 2018). The diagram below illustrates the injection of sperm into an egg.

ICSI is performed during embryo genetic testing as it can prevent sperm that failed to fertilise the egg from attaching to the exterior of the egg, causing inaccurate test results.

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Generally the Royal Family LOVES their horses (especially the queen) 

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Generally the Royal Family LOVES their horses (especially the queen) 

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Members of the Royal Family and the Monarchs have been using the Windsor Grey Horses to pull their carriages for ceremonial purposes ever since 1837.

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Fun Fact: Do you know that The Queen of England loves her horses? 

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Fun Fact: Do you know that The Queen of England loves her horses? 

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The Windsor Grey and the Cleveland Bay stallions are among the two breeds chosen to be bred by the Royal Family

The Windsor Grey has been bred since the 19th century and is preferred by the Royal Family because of its beautiful coat colour

The Windsor Grey were used in many occasions such as royal weddings, the queen's birthday, the queen's coronation and so on.

The late George Duell bred the Cleveland Bay stallion Mulgrave Supreme in North Yorkshire in 1961 and was later purchased by The Queen.

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There are more than 350 different breeds of horses !

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"It reminds you to settle down, breathe, get grounded and everything will be okay."

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Horses Help Heal Veterans' Invisible Wounds | National Geographic

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Equine therapy: How horses are helping teens' mental health | ITV News

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Equine Therapy Helps Veterans Cope With Post-traumatic Stress Disorder

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Equine Therapy Helps Veterans Cope With Post-traumatic Stress Disorder

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“Words don’t need to be said to express who you are.”

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How Equine Therapy Helps Veterans Cope With PTSD

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Increases the reproductive fitness by removing naturally occurring selective barriers (Hanevik, et al., 2016).

The use of ARTs has influenced the evolution of organisms (Hanevik, et al., 2016)

Due to the selection pressure during IVF, a study has shown that there is a significant effect of evolution as it directly influences reproductive success (Hanevik, et al., 2016)

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→ Sonogram of a dominant follicle of mare 

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→ Sonogram of a dominant follicle of mare 

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(Sertich, 2014)

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The development of IVF and ICSI has eliminated the natural breeding and natural selection during fertilisation as they are able to assist the males with dramatic sperm abnormalities to reproduce (Price, Murphy and Younglai, 2006.)

This may cause a shrinkage in the gene pool as only the few breeds that have the superior genes will be chosen to be bred.

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10% fetal bovine serum

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IVF ICSI Procedure - Important things you need to know

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DMEM/F-12 medium

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Mexico, Switzerland, Kazakhstan, Belgium, Japan, Germany, Indonesia, Poland and China are among the common horse consuming countries.

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Ensures the sustainability of the desirable phenotype to produce high quality and safe meat.

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A treatment that involves activities with horses to promote occupational, physical and emotional growth in persons.

It has been proven to be medically beneficial for conditions such as cerebral palsy, traumatic brain injury, sensory deficit in children, autism and hyperactivity (Menon, 2018).

Hippotherapy in Malaysia: Happy Farms Hippotherapy Malaysia, Green Apple Hippotherapy
Riding for Disabled Association: RDA Penang Turf Club, Selangor Turf Club and Equestrian Centre

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Equine Therapy for COVID Front Line Workers

Front line workers are getting a lot of attention, and rightly so, as they put themselves in harm's way dealing with the damage done by COVID-19. To help workers deal with the stress, a stable near Oceanside, California is offering equine therapy. IVey Ranch Park offers therapeutic riding and other equine related activities to children and adults with all sorts of special needs. And now, as they offer a bit of equine respite to those in the front lines of the pandemic.

Related Horse News

More News by Winniefield Park

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Equine Therapy for COVID Front Line Workers

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Front line workers are getting a lot of attention, and rightly so, as they put themselves in harm's way dealing with the damage done by COVID-19. To help workers deal with the stress, a stable near Oceanside, California is offering equine therapy.

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One commonality for ARTs is expensive, thus constricting the widespread commercial adoption of ARTs.

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Preservation of endangered species, such as the Przewalski's horse

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Enables breeding between animals in different geographic locations

Ovaries collected can be shipped to specialized labs in other countries to perform ICSI.

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Genetic improvement

Allow genetically valuable animals to procreate more extensively

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Embryo quality

Transfer technique

Donor-recipient synchrony

Recipient suitability (age, normal cyclicity,  no signs of reproductive tract pathology, relative size between mare and stallion, general health and body condition).

(Stout, 2010)

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04. Ramblin' In My Shoes - Hank Williams Jr. & Boxcar Willie - The Pressure Is On

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Superior sports horse mares are usually the primary choice for ET to produce offspring to ensure their competitive careers. More then one offspring can be produced in a single breeding session to ensure the economic values. Older mares that are not able to pregnant and those that have degeneration in endometrium while possessing values can look for healthier recipient mare.

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Three plastic tubes that is one inside another was guided into
the 🤬's anterior fornix. A needle in the middle tube was penetrated through vaginal wall into the lumen of uterine. Apparatus was manipulated and guided into position through rectal wall. CO2 was channeled into uterus to determine location of the tip of the needle. Rubber bulb was installed at opposite end of the hypodermic needle for transferring of eggs. Tubes with eggs were passed into uterine lumen which is the location that the eggs was deposited together with 0.2 ml of medium.

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First ET case was performed surgically between horses and donkeys, producing one mule and one hinny in 1974. For horses, 18 embryos from pony mares were collected and 15 blastocysts were transferred into uteri of 15 mares using non surgical ways. 4 off spring were successfully produced from 6 pregnancies. (Kramer, 2013).

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Then, Equine's ET techniques were improved for commercialization 10 years after the first succession.

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♩♪♫ Horses like classical and country music ♩♪♫ 

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♩♪♫ Horses like classical and country music ♩♪♫ 

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A study on the effect of different music genres on stabled horses was done by Carter and Greening (2012).

Jazz and rock are not the best genres to play for equine.

Minor key effect stimulate adverse behavioural responses.

Display alert behaviours such as stamping, snorting, head tossing and whinnying.

They prefer music with short melodies and strong rhythmic patterns such as those found in classical and country music.

Rest and relaxed behaviours.

The horses respond to the vibrations, not the words. They won’t get bored like we do. Thus, it is ideal to have music playing on repeat.

Music has also been used during post-surgery recovery of equine.
Benefits: calming effect, a sense of safety, masking jarring sounds

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Did you know that music can be a very effective tool for working with horses? Yes, horses like music. However, they aren't crazy about all kinds of music. By using music in the correct way, we can help our horses to be more docile and willing to work with us. In this article, we will discuss why you should play music for your horse, and what kind of music horses like the best.

How Horses Use Hearing

Horses need to be sensitive to their environment for survival. As prey animals, they use hearing and sight to alert them to any dangers. A warning nicker from another horse in the herd will have the whole group on high alert.

Although horses' hearing is similar to humans', they can detect sounds at higher and lower pitches than we can. A sudden grating sound can cause stress and anxious behavior. A very high-pitched sound can be painful. Take, for instance, the squealing of a microphone. If a squealing microphone hurts our ears, imagine what it is like to a horse.

The Benefits of Playing Music

Calming Effect

A calm, relaxed horse is not as likely to display dangerous behaviors, thus keeping themselves and us safer. Music can be a useful tool for keeping horses calm. If we teach our horses to associate music with being relaxed, we can use music during stressful situations to help our equine friends cope with stress.

A Sense of Safety

Taking a horse out of his home environment can be very stressful and scary for some horses. If we can keep things close to what they are at home, we can bring about a small sense of safety. For example, if you haul to a different barn for a competition, having his favorite tunes playing by his stall can help him feel more at home.

Masking Jarring Sounds

Thunderstorms or fireworks can play a number on the agitation levels of our equine friends. The loud, jarring sounds at irregular intervals keep them wondering when the next one will come and if they need to run from it. If they are in a stall, they don't have the option to run away, which can add more to their frustration. Playing music in the barn can help mask irritating sounds.

How and when to play music for your horse

You want the music to have a calming effect. Remember, they have better hearing than we do, so do not have the volume up loud. Blaring the music at top volume will have the opposite effect of calm. It is better to have the radio on a shelf that is level with the horse's head. Be sure it is out of their reach!

Play the music during certain times of the day, but not 24-7; they need a time of peace and quiet. Do you want music playing in your ear all the time? Well, some people do, but it is better to give our equine partners a break once in a while. As a rule, I turn the radio off during the night, unless a thunderstorm is forecasted.

Once your horse starts to associate music with being calm, you can use it during different situations to help them stay relaxed. Examples are farrier and veterinary visits, hauling in a trailer, etc. Music has also been used during post-surgery recovery.

The Research

Janet Marlow is a music composer who is internationally known for her role in understanding animal hearing sensitivities and how sound and music affect the behavior of dogs, cats, birds, and horses.

She has this to say about her research on Equine Wellness,

She goes on to say that horses respond best to music with short melodies and strong rhythmic patterns. She recommends classical and country music played at a low volume.
"In my clinical research over the past ten years, I've observed that horses prefer being in a barn with music as opposed to one without. Playing music helps balance equine behavior because it helps mask outside sounds and vibrations, such as tractor engines, high-pitched tools, thunder, and other intense sounds."
Clare Carter and Linda Greening at Hartpury University did a clinical trial on "Auditory stimulation of the stabled equine; the effect of different music genres on behavior."

The basics of the trial included exposing eight thoroughbreds to jazz, rock, country, and classical music. The horses were observed for behavior during the time of exposure to each genre of music. The results were that the horses were more relaxed when country and classical were played. The horses seemed to be more agitated during the jazz and rock sessions.

What Music to Play

The research tells us that jazz and rock are not the best genres to play for our equine friends. They prefer music with short melodies and strong rhythmic patterns, like those found in classical and country music.

It is ideal to have music playing on repeat. The horses are responding to the vibrations, not the words. They won't get bored like we do. If you use a radio, be sure not to tune into an all-talk radio station.

After doing the research we mentioned above, Janet Marlow started Petacoustics.com. Pet Acoustics developed a product called Pet Tunes. Pet Tunes is a speaker system with downloaded music composed specifically for horses. They also have Pet Tunes for cats, dogs, and birds. The features of this handy speaker system are as follows:

Bluetooth® Compatible

Portable

USB Charging Cable

Continuous Play

Lanyard Strap

Can order more music to download

Final thoughts

A calm, relaxed horse is safer and more enjoyable to be around. Furthermore, they are also more willing to work with us. Playing music can have a calming effect while providing a sense of safety. Also, music can mask jarring sounds that keep horses hyper-vigilant.

Although we may prefer a different genre of music, horses seem to prefer classical and country. I don't typically mention a product in my articles. However, I do think the Pet Tunes speaker is worth checking out.

Did you learn something new about what music to play for horses? Let us know in the comments below!
Next, check out our article on horse sounds and their meanings.

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Why You Should Play Music for Your Horse-And What They Like

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Did you know that music can be a very effective tool for working with horses? Yes, horses like music. However, they aren't crazy about all kinds of music. By using music in the correct way, we can help our horses to be more docile and willing to work with us.

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Quickest recorded sprinting speed: 88 km/h (55 mph)

Gallop: around 44 km/h (27mph)

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ICSI is able to assist the following infertility problems

Low sperm count

Poor sperm movement

Low quality sperm

Failure of insemination

Abnormal sperm

(Hinrichs, 2018)
The diagram below illustrates the types of abnormal sperm

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Embryo transfer is a process where one or more embryos are being removed (flushed) from the uterus located in the reproductive trait of a donor female (e.g., mare) and transferring the embryos into one or more recipient females (e.g., mare) (McCue, n.d.). However, laboratory techniques such as in vitro fertilization can also produce embryos that involve the fertilization of oocytes with sperm cells in a dish (Troxel, n.d.). Later, the embryos produced in vitro will be transferred into the uterus of a female to enable pregnancy to begin.

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1. The development of equine embryos in vitro starts with the formation of a morula (8 to 16 cell stage) which usually occurs after 3 to 4 days of fertilization and can be supported by a variety of media.

2. However, the development of equine blastocyst requires the embryos to be cultured in DMEM⁄F-12 medium with 10% fetal bovine serum, at a ratio of 1 μl medium per embryo.

3. The atmospheric air mixture should consist of 5% O₂, 5% CO₂ and 90% N₂ at 38.2℃.

4. DMEM⁄F-12 contains 17 mM glucose which acts as a predominant energy substrate in the blastocyst.

5. The embryos will start to develop to the blastocyst stage at the 7 days through 11 days of culture.

6. Hence, the embryos should be transferred to recipient mares once the blastocyst starts to develop even at an early stage.

(Hinrichs, 2010)

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Increase in production

Enables the production of foals from infertile animals

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In 1972, First report on equine and donkey offspring produced through ET was published by Allen and Rowson. 81 years after first successful mammalian case, rabbit's ET. 39 years after sheep and goat and 21 years after cattle and pigs.

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Opinion on the effects of ARTs

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Carter, C. and Greening, L., 2012, July. Auditory stimulation of the stabled equine; the effect of different music genres on behaviour. Applied Animal Behaviour Science. [online] Available at: <https://www.researchgate.net/publication/249339705_Auditory_stimulation_of_the_stabled_equine_the_effect_of_different_music_genres_on_behaviour> [Accessed 1 September 2020].

Carter, J.A., Bellow, S., Meintjes, M., Perez, O., Ferguson, E. and Godke, R.A., 2002. Transvaginal ultrasound-guided oocyte aspiration for production of embryos in vitro. Archives Animal Breeding. [online] Available at: <https://www.researchgate.net/publication/233122519_Transvaginal_ultrasound-guided_oocyte_aspiration_for_production_of_embryos_in_vitro> [Accessed 31 August 2020].

Choi, Y.-H., Ross, P., Velez, I.C., Macias-Garcia, B., Riera, F.L. and Hinrichs, K., 2015. Cell lineage allocation in equine blastocysts produced in vitro under varying glucose concentrations. Reproduction, 150(1), pp.31-41.

Hanevik, H. I., Hessen, D. O., Sunde, A., and Breivik, J., 2016. Can IVF influence human evolution? Human Reproduction, [e journal], 31(7), pp. 1397–1402. 10.1093/humrep/dew089

Hinrichs, K., 2010. In Vitro Production of Equine Embryos: State of the Art. Reproduction in Domestic Animals. [online] Available at: <https://www.researchgate.net/publication/51444033_In_Vitro_Production_of_Equine_Embryos_State_of_the_Art> [Acessed 31 August 2020].

Hinrichs, K., 2018. Assisted reproductive techniques in mares. Reproduction in Domestic Animals, [e journal], 53, pp. 4–13 10.1111/rda.13259

Kraemer, D.C., 2013. A history of equine embryo transfer and related technologies. Journal of Equine Veterinary Science, 33(5), pp.305-308.

McCue, P.M., n.d. Embryo Transfer. [online] Available at: <http://csu-cvmbs.colostate.edu/Documents/Learnmares9-ET-embtrans-apr09.pdf> [Accessed 1 September 2020].

Menon, S., 2018. Changing lives one ride at a time. The Star, [online] 5 August. Available at: <https://www.thestar.com.my/news/education/2018/08/05/changing-lives-one-ride-at-a-time> [Accessed 1 September 2020].

Price, T. M., Murphy, S. K., and Younglai, E. V., 2006. Perspectives: The Possible Influence of Assisted Reproductive Technologies on Transgenerational Reproductive Effects of Environmental Endocrine Disruptors. Toxicological Sciences, [e journal], 96(2), pp. 218–226. 10.1093/toxsci/kfl196

Sertich, P.L., 2014. Reproductive cycle in horses. [online] Available at: <https://www.msdvetmanual.com/management-and-nutrition/management-of-reproduction-horses/reproductive-cycle-in-horses> [Accessed 2 September 2020].

Smits, K., Hoogewijs, M., Woelders, H., Daels, P. and Van Soom, A., 2012. Breeding or assisted reproduction? Relevance of the horse model applied to the conservation of endangered equids. Reproduction in Domestic Animals, 47, pp.239-248.

Stout, T.A.E., 2010. Equine embryo transfer: review of developing potential. Equine Veterinary Journal, 38(5), pp.467-478.

Troxel, T.R., n.d. Embryo Transfer in Cattle. [online] Available at: <https://www.uaex.edu/publications/PDF/FSA-3119.pdf> [Accessed 1 September 2020].

University of Wisconsin-Madison, n.d. Assisted Reproductive Technology. [online] Available at:< http://www.ansci.wisc.edu/jjp1/equine/technology/intro.html> [Accessed 7 September 2020]

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Step 2: In Vitro Maturation (IVM) of oocytes

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Step 2: In Vitro Maturation (IVM) of oocytes

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1. The oocytes are cultured in a maturation medium (TCM-199) to finish the first meiotic division and stop at the metaphase II so that oocytes are ready to be fertilised.

2. The maturation medium is incubated for about 30 hours at 38.5°C in a humidified atmosphere of 5% CO₂ in air.

3. The oocytes are assessed under the microscope for the evaluation of maturity. The extrusion of the first polar body can be determined and the oocytes are ready for ICSI.

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There are 2 approaches to collect immature oocytes (cumulus-oocyte complexes) which are from live donor animals or directly from ovaries obtained from an abattoir (Carter et al., 2002).

(A) From live donor animals - The ovum pick-up technique (OPU)

1. The donor female is restrained in a suitable holding chute and sedated with detomidine. The faeces of mare are removed. The extra genital area and perineum are cleaned. The mare is then administered epidural anaesthesia.

2. The vaginal canal is inserted with the ultrasound probe for visualization of ovaries on the ultrasound. The ovary is grasped via the rectum and held against the vaginal wall.

3. Follicles are identified as black (hypo echoic) circular shapes on the monitor screen.

4. A 12 gauge double lumen needle with 60 cm long is inserted through the needle guide in the plastic handle to aspirate and inject fluid at the same time.

5. The needle is connected to a vacuum pump, passing into a 500 ml glass bottle to collect follicular fluid containing the oocytes.

6. The fluid of 1 cm or bigger follicle is aspirated and flushed repeatedly with commercial embryo flushing medium supplemented with heparin while scraping the follicular wall for the enhancement of oocyte recovery.

7. The follicular fluid is then passed through the standard bovine embryo filter.

(B) From an abattoir

1. The ovaries are removed from the mare reproductive tract after the extraction of internal organs from the carcass. The ovaries are placed into saline containers.

2. The ovaries are then transferred to the second container with saline concentration and placed in the thermos.

3. The ovaries are transported to the lab in physiological saline that consists of antibiotics to prevent bacterial contamination at 22-24°C.

4. The best results are obtained when oocytes are collected within 4-6 hrs after slaughter.

5. Oocytes can be collected by aspiration using a syringe and needle or by slashing the ovary surface with a scalpel blade and collecting it into a beaker by cell strainer method.

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Lab-based Equine Assisted Reproductive Technologies (ARTs) 

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Lab-based Equine Assisted Reproductive Technologies (ARTs) 

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In Vitro Fertilization (IVF) is the process of creating embryos from oocytes (unfertilised egg cells) by fertilising them with sperms on a petri dish (Hinrichs, 2010).

However, standard in vivo fertilization is not yet repeatable in the horse, the fertilisation process can also be performed by using Intra Cytoplasmic Sperm Injection (ICSI) which is the injection of one spermatozoa into oocyte cytoplasm.