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Future of reproductive biotechnologies in water buffalo in Southeast Asian countries

Hufana-Duran D, Chaikhun-Marcou T, Duran PG, Atabay EP, Nguyen HT, Atabay EC, et al.

Theriogenology, 2025 Feb;233:123-130.
PMID: 39613496 DOI: 10.1016/j.theriogenology.2024.11.016

The future of reproductive biotechnologies in water buffalo in Southeast Asian countries holds significant promise for enhancing genetic quality and productivity. Fixed-time artificial insemination remains the commonly used technology, with advances in assisted reproductive technologies (ART) such as in vitro embryo production (IVEP), embryo transfer (ET), and the use of sex-sorted sperm increasingly adopted to improve breeding efficiency. These technologies overcome traditional breeding limitations, such as low reproductive rates, genetic diversity constraints, and the production of sex-predetermined offspring. The application of multiple ovulation and embryo transfer (MOET) is constrained by poor embryo recovery in this livestock species. Somatic cell nuclear transfer (SCNT) offers great potential for producing sex-predetermined and genetically superior buffalo but requires further research to increase efficiency. Cryopreservation of buffalo genetics is bolstered by the establishment of Gene Banks. Challenges such as high costs, the need for skilled personnel, and infrastructure development remain constraints. Integration of genomic selection, automation, and expansion of ET programs are clear directions. Strengthening research and collaboration among Southeast Asian countries is essential to fully realize the benefits of these biotechnologies and ensure sustainable and profitable buffalo farming.

Matched MeSH terms: Insemination, Artificial/veterinary
Fulltext Associated factors affecting the successful pregnancy rate of intrauterine insemination at International Islamic University Malaysia (IIUM) Fertility Centre

Azantee YW, Murad ZA, Roszaman R, Hayati MY, Norsina MA

Med J Malaysia, 2011 Aug;66(3):195-8.
PMID: 22111439 MyJurnal

The aim was to determine pregnancy rate and its associated factors in Intrauterine Insemination (IUI) at IIUM Fertility Centre.

Matched MeSH terms: Insemination, Artificial*
Unexplained subfertility: active or conservative management?

Ahmad SM, Mat Jin N, Ahmad MF, Abdul Karim AK, Abu MA

Horm Mol Biol Clin Investig, 2023 Dec 01;44(4):379-384.
PMID: 38124670 DOI: 10.1515/hmbci-2022-0087

OBJECTIVES: Unexplained subfertility (UEI) describes a couple whose standard subfertility workout consider acceptable but unable to conceived.
METHODS: This retrospective study was conducted in the Advanced Reproductive Centre, UKM Hospital, Kuala Lumpur, from January 2016 to December 2019. The data of 268 UEI couples were obtained from the clinical database. Women aged 21-45 years old was included and further divided into four groups according to the female partner's age and subfertility duration: group A (age <35 years and subfertility <2 years), group B (age <35 years and subfertility >2 years), group C (age >35 years and subfertility <2 years), and group D (age >35 years and subfertility <2 years). All statistical analyses were performed using SPSS 22.0 for Windows.
RESULTS: A total of 255 cases were included in this study. The mean age of the women was 32.9 ± 4.04 years, and the mean subfertility duration was 5.04 ± 2.9 years. A total of 51 (20 %) cases underwent timed sexual intercourse, 147 (57.6 %) cases had intrauterine insemination (IUI), whereas 57 (22.4 %) cases opted for in vitro fertilization (IVF). A total of 204 cases underwent active management (IUI/IVF), which showed a significant difference (p<0.05). Out of eight clinical pregnancies, half of them were from group B.
CONCLUSIONS: Active management in younger women with a shorter subfertility duration revealed a better pregnancy outcome. Otherwise, individualized treatment should be considered in selecting a suitable treatment plan.

Matched MeSH terms: Insemination, Artificial*
Fulltext Predictors of intrauterine insemination success

Zainul MR, Ong FB, Omar MH, Ng SP, Nurshaireen A, Rahimah MD, et al.

Med J Malaysia, 2006 Dec;61(5):599-607.
PMID: 17623962 MyJurnal

Intrauterine insemination (IUI) remains a therapeutic option within means of the majority of infertile couples in Malaysia. Therefore additional information on predictors of IUI success in the local context would provide a more concrete basis for counseling patients on expectations and treatment options. A retrospective analysis of 297 couples who underwent 445 IUI cycles from Jan 2005-Mar 2006 was undertaken. Four fifths were Malay with a mean paternal and maternal age of 35.53 +/- 5.82 (range 24-59) and 33.02 +/- 4.69 (range 21-46) years respectively. Causes of infertility were idiopathic (50%), endometriosis (17%) and anovulation/polycystic ovarian syndrome (15%). Almost 10% were oligoastenoteratozoospermic with another 23% oligozoospermic or astenozoospermic. Combined male and female factors occurred in 26%. A pregnancy rate (PR) of 9.4% per cycle; 14.1% per couple with a cumulative PR of 36.7% per 4 cycles was achieved. Those who became pregnant were significantly younger (31.29 +/- 4.43 vs. 33.21 +/- 4.68 years, p = 0.011) and had more follicles (13.95 +/- 9.72 vs. 11.43 +/- 6.67, p = 0.029) at the time of insemination. PR depreciated with maternal age and semen quality. Maternal and paternal age was inversely correlated to the number of follicles recruited (r = -0.30, p < 0.0005) and progressive sperm motility (r = -0.125, p = 0.013) respectively.

Matched MeSH terms: Insemination, Artificial/methods*; Insemination, Artificial/standards
Fulltext Prognostic factors influencing pregnancy rate after stimulated intrauterine insemination

Tay PY, Raj VR, Kulenthran A, Sitizawiah O

Med J Malaysia, 2007 Oct;62(4):286-9.
PMID: 18551930 MyJurnal

To determine the prognostic factors such as age, diagnosis, number of cycle attempts and semen parameters on the pregnancy rate of controlled ovarian hyperstimulation (COH) /intrauterine insemination (IUI). Three hundred and seventeen women who underwent 507 consecutive COH/IUI cycles were recruited from 1st January 2002 to 31st December 2005 inclusively. This retrospective study was done in University Malaya Medical Centre, a tertiary care academic centre. The main outcome measure was pregnancy rate according to age, infertility diagnosis, duration of infertility, semen parameters, and the number of treatment cycles. The overall pregnancy rates were 16.9% per cycle and 25.9% per couple. Pregnancy rates decreased with advancing maternal age. Pregnancy rate was also significantly lower in patient with postwash total motile sperm count (TMSC) < or = 20 million/ml compared to those with TMSC >20 million/ml. The cumulative pregnancy rates varied greatly by diagnosis from 16% for patients with male factor infertility to 60% for patients with ovulatory disorder. Pregnancies among patients with male infertility, tubal factors infertility and endometriosis were achieved during the first three cycles. There is a clear age-related decline in fecundity associated with COH/IUI treatment. Women of > 40 years old, couple with postwash TMSC < or = 20 million/ml, severe endometriosis and tubal factors have a particularly poor prognosis.

Matched MeSH terms: Insemination, Artificial, Homologous/methods*
Fulltext A rare case of heterotopic quintuplets pregnancy

Wu D, Hii LY, Shaw SWS

Taiwan J Obstet Gynecol, 2019 Sep;58(5):684-687.
PMID: 31542093 DOI: 10.1016/j.tjog.2019.07.018

OBJECTIVE: We report a rare case of heterotopic pregnancy and high-order pregnancy occurring simultaneously following the use of the assisted reproductive technique (ART).
CASE REPORT: A 29-year-old woman, Gravida 2 Para 1, became pregnant after receiving intrauterine insemination (IUI). She came to our emergency room due to diffuse low abdominal pain at seven weeks of gestational age. Transabdominal sonography (TAS) revealed a quadruplet intrauterine pregnancy with an enlarged left adnexa and intrapelvic fluid accumulation. Simultaneous occurrence of high-order pregnancy and left tubal pregnancy with internal hemorrhage was suspected. The patient received an emergent laparoscopic resection of the affected Fallopian tube and recovered well for the remaining hospitalization course. Afterwards, she received fetal reduction procedure and eventually gave birth to twin babies.
CONCLUSION: Gynecologist should increase the awareness of heterotopic pregnancy in patients receiving ART. On the other hand, reproductive endocrinologist should reduce the risk of high-order pregnancy without compromising pregnancy rate.

Matched MeSH terms: Insemination, Artificial/adverse effects*
Fulltext Effects of timed artificial insemination following estrus synchronization in postpartum beef cattle

Malik A, Wahid H, Rosnina Y, Kasim A, Sabri M

Open Vet J, 2012;2(1):1-5.
PMID: 26623282

The objectives of this study were to evaluate estrus response and pregnancy rates resulting from timed artificial insemination (AI) following estrus synchronization using CIDR in postpartum beef cattle. A total of 100 cows were randomly divided into three groups. Groups 1, 2 and 3 were artificially inseminated at 48-50 h (n=30), 53-55 h (n=30) and 58-60 h (n=40) after CIDR removal, respectively. Estrus synchronization was carried out using a CIDR containing 1.38 mg progesterone. All cows were given 2 mg estradiol benzoate, intramuscularly on the day of CIDR insertion (D 0). The CIDR was removed after 8 days and 125 μg of prostaglandin F2α (PGF2α) was injected intramuscularly. One day after CIDR removal all cows were given 1 mg of estradiol benzoate intramuscularly (D 9). Cows were observed visually for estrus after removal of CIDR. Between 30 and 32 days after timed AI, pregnancy was determined using transrectal ultrasonography. The first estrus observation which is approximately 32 h after CIDR removal showed no significant difference (P>0.05) among the three groups. The onset response of estrus after 32 h removal of CIDR was less than 10% in all three groups 6.6% (G1), 6.8% (G2) and 7.3% (G3). Furthermore, percentages of estrus response (D 10) following CIDR removal were 76.6%, 75.0% and 77.5%. The difference between on D 9 and D 10 estrus response were statistically significant (P<0.05). The pregnancy rates were 23.3% (G1), 26.6% (G2) and 37.5% (G3), which were not significant (P>0.05).

Matched MeSH terms: Insemination, Artificial
Fulltext Semen characteristics, extension, and cryopreservation of Rusa deer (Rusa timorensis)

Fitri WN, Wahid H, Rosnina Y, Jesse FFA, Aimi-Sarah ZA, Mohd-Azmi ML, et al.

Vet World, 2017 Jul;10(7):779-785.
PMID: 28831222 DOI: 10.14202/vetworld.2017.779-785

AIM: The objective of this research is to report parameters for breeding soundness evaluation, semen extension, and cryopreservation in Rusa timorensis.
MATERIALS AND METHODS: Seven healthy stags were chosen for semen collection using an electroejaculator. The collections were performed twice in a breeding season between February and June 2016. Samples were collected between 2 and 3 weeks interval, collected twice for each animal. Semen was evaluated, extended, and cryopreserved using four different extenders; Andromed®, BioXcell®, Triladyl®, and a modified Tris-egg yolk combined with Eurycoma longifolia Jack.
RESULTS: R. timorensis semen characteristics according to volume (ml), color, sperm concentration (106/ml), general motility (%), progressive motility (%), and % morphology of normal spermatozoa are 0.86±0.18 ml, thin milky to milky, 1194.2±346.1 106/ml, 82.9±2.8%, 76.1±4.8%, and 83.9±4.8%, respectively.
CONCLUSION: Semen characteristics of R. timorensis collected by electroejaculation is good allowing for cryopreservation and future artificial insemination work. The most suitable extender for Rusa deer semen is Andromed®.

Matched MeSH terms: Insemination, Artificial
Fulltext Assessing the status of intrauterine insemination before forming a medically assisted conception unit

Omar MH, Ong FB, Adeeb NN, Sharif JM, Nasri N, Yassin MJ

Med J Malaysia, 1999 Mar;54(1):65-71.
PMID: 10972007

A survey in 1996 of our female patients suggest that the three commonest causes of infertility were endometriosis, anovulation and idiopathic which comprises of about 70% of all the patients. In the male patients, sperm morphology evaluation by critical criteria showed that abnormal morphology was present in 71% while 87% of all the semen analysis were abnormal. The objective of this study was to assess the status of IUI before proceeding to formulate patient protocols for IVF in a tertiary infertility referral unit. A retrospective study of patients data was done from Jan 1995 to Dec 1996. Ovarian stimulation by clomiphene for anovulatory and idiopathic patients was performed on couples with at least one patent fallopian tube. Ovulation induction was by an intramuscular injection of 5000 i.u of HCG after follicular maturation. IUI was performed approximately 36-40 hours later. A total of 74 couples received 114 treatment cycles producing a total of 9 conceptions. The conception rate of IUI was therefore 7.89% per cycle and 12.16% per couple with the cumulative pregnancy rate of 28.21%. Associated success features of IUI found in this study were the age of the woman and the semen parameters of the husband.

Matched MeSH terms: Insemination, Artificial, Homologous*
Fulltext No evidence for successful interspecific cross-mating of transgenic Aedes aegypti (L.) and wild type Aedes albopictus Skuse

Lee HL, Aramu M, Nazni WA, Selvi S, Vasan S

Trop Biomed, 2009 Dec;26(3):312-9.
PMID: 20237445

The natural and artificial mating of laboratory bred Aedes albopictus and transgenic Aedes aegypti RIDL-513A-Malaysian strain was conducted. The experiment consisted of crossmating of homologous Ae. aegypti RIDL female symbol X Ae. aegypti RIDL male symbol and reciprocal Ae. aegypti RIDL female symbol X Ae. albopictus WT male symbol. The other set comprised homologous Ae. albopictus WT female symbol X Ae. albopictus WT male symbol and reciprocal Ae. albopictus WT female symbol X Ae. aegypti RIDL male symbol. This study demonstrated that reproductive barriers exist between these two species. Cross insemination occurred between A. albopictus male and Ae. aegypti female and their reciprocals. There was 26.67% and 33.33% insemination rate in Ae. aegypti RIDL female cross-mating with A. albopictus WT male and Ae. albopictus female cross-mating with Ae. aegypti RIDL male, respectively. There was 0% hatchability in both directions of the reciprocals. There was also no embryonation of these eggs which were bleached. Although none of the female Ae. albopictus WT was inseminated in the cross-mating with Ae. albopictus WT female symbol X Ae. aegypti RIDL male symbol, a total of 573 eggs were obtained. The homologous mating was very productive resulting in both high insemination rate and hatchability rates. Generally there was a significantly higher insemination rate with artificial mating insemination of homologous than with artificial mating of reciprocal crosses. Interspecific mating between Ae. aegypti RIDL and Ae. albopictus wild type was not productive and no hybrid was obtained, indicating absence of horizontal transfer of introduced RIDL gene in Ae. aegypti to Ae. albopictus.

Matched MeSH terms: Insemination, Artificial
Effect of antioxidants on post thaw microscopic, oxidative stress parameter and fertility of Boer goat spermatozoa in Tris egg yolk glycerol extender

Memon AA, Wahid H, Rosnina Y, Goh YM, Ebrahimi M, Nadia FM

Anim. Reprod. Sci., 2012 Dec;136(1-2):55-60.
PMID: 23182473 DOI: 10.1016/j.anireprosci.2012.10.020

This study was conducted to determine the effect of antioxidants on standard semen parameters, lipid peroxidation and fertility of Boer goat semen after cryopreservation. Ejaculates from four bucks were collected, evaluated and pooled at 37°C. The pooled semen was diluted with Tris citric acid fructose for washing. Semen samples, which were diluted with a Tris-based extender containing the antioxidant ascorbic acid (8.5mg/ml), butylated hydroxytoluene (2mM), cysteine (5mM) and hypotaurine (10mM) and an extender without antioxidant supplementation were cooled to 4°C and frozen in 0.25 straws with programmable freezer and finally stored in liquid nitrogen. Data (10 replicates) were analyzed by one-way analysis of variance. Mean (±SEM) progressive motility was significantly higher in ascorbic acid than other supplement groups and control samples (P>0.05). Best values were observed in ascorbic acid followed by BHT, cysteine, and hypotaurine. Antioxidant supplementation in extender showed significant (P<0.05) better values than the control group for sperm membrane integrity, acrosome integrity and viability. The ability of antioxidants to reduce the lipid peroxidation (LPO) after freeze thawing was measured by the formation of malondialdehyde (MDA) using the thiobarbituric acid method. Results showed that addition of antioxidants significantly reduced the rate of LPO in comparison to control (P<0.05). Ascorbic acid exhibited better values (1.27±0.28), than butylated hydroxytoluene, cysteine and hypotaurine 1.32±0.42, 2.27±0.16 and 2.38±0.17 respectively, which are significantly better than control (3.52±0.54). Higher pregnancy rate was observed with ascorbic acid followed by butylated hydroxtolune, hypotaurine and cysteine. However, differences in the fertility rate were non-significant with hypotaurine, cysteine and control groups.

Matched MeSH terms: Insemination, Artificial
Successful pregnancy outcome following gamete intra-Fallopian transfer in a patient with Müllerian dysgenesis

Lee CS, Lie AT

Reprod Biomed Online, 2012 May;24(5):547-9.
PMID: 22410277 DOI: 10.1016/j.rbmo.2012.01.021

A 29-year-old lady with Müllerian dysgenesis was keen to have a baby. Clinically, she was medium built with well-developed secondary female sexual characteristics. There was a short and blind vagina. She had undergone surgery for an imperforated hymen. Her FSH and LH concentrations were normal. Laparoscopy revealed a patent right Fallopian tube, a rudimentary right uterus and extensive pelvic endometriosis. She subsequently underwent gamete intra-Fallopian transfer (GIFT). Oocyte retrieval was carried out laparoscopically and a total of nine oocytes were retrieved. Four of the oocytes were transferred together with motile spermatozoa into the right Fallopian tube and the remaining five oocytes were inseminated with spermatozoa for IVF. Three embryos resulted and were frozen. She subsequently developed moderate ovarian hyperstimulation syndrome. Serum β-human chorionic gonadotrophin concentration 14 days after GIFT was 1612 IU/l. Her antenatal care was relatively uneventful until 31 weeks of gestation when she was diagnosed to have intrauterine growth retardation and oligohydramnios. She then underwent an emergency Caesarean section at 32 weeks of pregnancy delivering a normal baby. This case study describes a successful pregnancy outcome following gamete intra-Fallopian transfer (GIFT) in a woman with malformation of the vagina (Müllerian dysgenesis). A 29-year-old lady with Müllerian dysgenesis diagnosed at 16 years of age was keen to become pregnant. Upon examination, a decision was made for a William's vulvovaginoplasty but as the patient was indecisive the surgery was deferred. Clinically, she is a medium-built lady with well-developed secondary female sexual characteristics. There was a short and blind vagina. Her serum FSH and LH concentrations were normal. Laparoscopy revealed a patent right Fallopian tube, a rudimentary right uterus and extensive pelvic endometriosis. She subsequently underwent GIFT. Nine oocytes were retrieved through laparoscopy. Four of the oocytes were transferred together with motile sperm into the right Fallopian tube and the remaining five oocytes were inseminated with sperm for IVF. Three embryos resulted and were frozen. Serum β human chorionic gonadotrophin concentration measured 14 days after GIFT was 1612 IU/l. An abdominal ultrasonography performed at 5 weeks showed one intrauterine gestational sac. Her antenatal care was uneventful until 31 weeks of gestation when she developed a deficiency of amniotic fluid in the amniotic sac. She then underwent an emergency Caesarean section at 32 weeks of pregnancy. She delivered a healthy, normal 1.24 kg baby boy. Her post-natal care was uneventful.

Matched MeSH terms: Insemination, Artificial
A pilot study to determine whether progestogen supplementation using dydrogesterone during the first trimester will reduce the incidence of gestational hypertension in primigravidae

Zainul Rashid MR, Lim JF, Nawawi NH, Luqman M, Zolkeplai MF, Rangkuty HS, et al.

Gynecol Endocrinol, 2014 Mar;30(3):217-20.
PMID: 24552449 DOI: 10.3109/09513590.2013.860960

Gestational hypertension (GH) remains one of the main causes of high maternal and perinatal morbidity and mortality worldwide with the highest incidence among primigravidae of about 10%-15%. However, it was noted that the incidence of GH in primigravidae who conceived following assisted reproductive technique (ART) or intrauterine insemination (IUI) supplemented with dydrogesterone during the first trimester was low.

Study site: Obstetrics and Gynecology
Department, Pusat Perubatan Universiti Kebangsaan Malaysia PPUKM

Matched MeSH terms: Insemination, Artificial

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