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Concept: Artificial insemination


Unlike current chemical trace detection technology, dogs actively sniff to acquire an odor sample. Flow visualization experiments with an anatomically-similar 3D printed dog’s nose revealed the external aerodynamics during canine sniffing, where ventral-laterally expired air jets entrain odorant-laden air toward the nose, thereby extending the “aerodynamic reach” for inspiration of otherwise inaccessible odors. Chemical sampling and detection experiments quantified two modes of operation with the artificial nose-active sniffing and continuous inspiration-and demonstrated an increase in odorant detection by a factor of up to 18 for active sniffing. A 16-fold improvement in detection was demonstrated with a commercially-available explosives detector by applying this bio-inspired design principle and making the device “sniff” like a dog. These lessons learned from the dog may benefit the next-generation of vapor samplers for explosives, narcotics, pathogens, or even cancer, and could inform future bio-inspired designs for optimized sampling of odor plumes.

Concepts: Fluid dynamics, Aerodynamics, Olfaction, Odor, Artificial insemination, Dog, Aroma compound, Body odor


Cryopreservation has been widely utilized in livestock and human embryos, which allows for storage of worthy embryos for a long period of time, although it is still uncertain as how long embryos can be cryopreserved in liquid nitrogen. The aims of this study were to evaluate the effects of long-term cryopreservation on birth rate of transferred sheep embryos at morula or blastocyst stage, and to investigate growth performance and viability of their offspring. A total of 373 sheep embryos from the same batch, which had been cryopreserved by conventional procedure for 0.5yr (n=259) or 7.5yr (n=114), respectively, were transferred to 373 recipient ewes. In parallel, artificial inseminations, acting as controls, were conducted in the same month in both years (n=81 and n=110) that embryo transfers were performed. Results showed that there were no significant differences in birth rate between short-term cryopreservation group (cryopreserved for 0.5yr in 2003) and long-term cryopreservation group (cryopreserved for 7.5yr in 2010) either at the morula or blastocyst stage (p>0.05). No specific differences in birth weight were observed among short-term cryopreservation, artificial insemination 1 (performed in 2003), long-term cryopreservation and artificial insemination 2 (performed in 2010) group (p>0.05). The weaning weights were similar between the short-term cryopreservation and long-term cryopreservation group (p>0.05). The mortality rates of the offspring were similar in both groups as well (p>0.05). We concluded that the long-term cryopreservation did not appear to adversely affect birth rate of the embryos, growth performance and viability of their offspring. Our results indicated that the cryopreserved sheep embryos should be stable in liquid nitrogen for at least 7.5 years.

Concepts: Time, Embryo, Artificial insemination, Livestock, Liquid nitrogen, Cryobiology, Cryopreservation, Ex-situ conservation


The aim of this study was to evaluate the efficiency of presynchronization with GnRH and PGF(2α) or with progesterone on overall Ovsynch (OVS) outcomes in noncyclic dairy cows. Cows were scanned 7d apart with ultrasonography to determine cyclicity. Noncyclic cows (n=281; no corpus luteum on ovaries at both examinations) were randomly divided into 3 groups. In the GP group (n=108), the cows received GnRH and PGF(2α) (PGF) administrations 7d apart, and OVS was started 11d after PGF (GnRH-7 d-PGF-11 d-OVS). In the P4 group (n=90), the cows were treated for 7d with an intravaginal progesterone (P4) implant (PRID), and then OVS was started 11d after removal of the implant (7d PRID-11 d-OVS). The control group (CON, n=83) did not receive any presynchronization, and OVS was started at the same time as in the other groups (18 d-OVS). The percentage of cows that became cyclic at the beginning of OVS was lower in the CON group (38.6%; 32/83) than in the presynchronization groups (66.7%, 72/108 in GP; 71.1%, 64/90 in P4). The response to the first GnRH of OVS did not differ among groups (63.9%, 53/83 in CON; 67.6%, 73/108 in GP; 63.3%; 57/90 in P4), and synchronization rates were similar among the groups (74-82%). The cows that responded to presynchronization treatments (GP or P4) had higher pregnancy per artificial insemination (P/AI) than did nonresponding cows. Pregnancy per AI at 31d did not differ between groups (30.1%, 25/83 in CON; 43.5%, 47/108 in GP; and 35.6%, 32/90 in P4). However, CON cows (24.1%, 20/83) had lower P/AI at 62d than GP cows (41.7%, 45/108). Embryonic loss was higher in CON (20%, 5/25) compared with the P4 group (3%, 1/32). The administration of GnRH followed by PGF or exogenous progesterone (PRID) similarly increased the percentage of cows that became cyclic before Ovsynch in noncyclic cows, but fertility did not improve. However, the cows that responded to presynchronization had higher fertility rates than the nonresponding cows.

Concepts: Pregnancy, Milk, Fertility, Menstrual cycle, Endometrium, Corpus luteum, Ovulation, Artificial insemination


Ineffective estrus detection is the foremost limiting factor in the fertility of farmed cattle worldwide. Failure to detect estrus or erroneous diagnosis of estrus results in great economic losses in Korea each year. This study was carried out in order to comprehensively describe the estrus behaviors and conception rates of different estrus synchronization protocols applied to 40 cycling native Korean cattle (Hanwoo). The cows were grouped into four (n = 10) and treated with the following protocols: (1) Day -15: controlled intravaginal drug-releasing device (CIDR) for 12 days; Day -5: prostaglandin F2α (PGF2α), (2) ovulation synchronization (OVS): Day -15: GnRH; Day -6: PGF2α; Day -4: GnRH, (3) Day -15: progesterone-releasing intravaginal device for 12 days; Day -5: PGF2α; and (4) Day -15: PGF2α; Day -4: PGF2α. Artificial insemination was performed 12 hours after the detection of estrus using frozen-thawed semen. Estrus signs were compared using a charge-coupled device camera (CCDC) and a control method (direct visual observation). The pregnancy of the cows was determined by transrectal ultrasonography at Days 25 to 30 postinsemination. The results indicated that the day of estrus return was significantly earlier using the CCDC method compared with direct visualization (P < 0.05). Mounting of other cows was the most predominant sign of estrus among the flock (P < 0.05), as analyzed using the CCDC. In the OVS group, a lower rate of mounting was observed than in the other three groups. Moreover, significantly fewer estrus behaviors were noticed in the OVS protocol group (P < 0.05). Both first service conception and overall conception rates were significantly higher (P < 0.05) in the CIDR and OVS treatment groups. In conclusion, the CIDR and OVS protocols appear to be the best practice for the synchronization of estrus for reproductive competence through the CCDC in Hanwoo cows. However, CIDR has a practical advantage over OVS with respect to estrus detection.

Concepts: Observation, Spermatozoon, Cattle, Dairy cattle, Transrectal ultrasonography, Ovulation, Artificial insemination, Amphetamine Reptile Records


Objectives were to determine associations between percentage pregnancy loss (PPL) in dairy cattle and: (i) pregnancy diagnosis by ultrasonography; (ii) pregnancy diagnosis by serum pregnancy-specific protein B (PSPB) concentrations, with or without serum progesterone concentrations; and (iii) production and environmental factors. This study included 149 822 pregnancy diagnoses conducted over 13 years in Holstein-Friesian cows in Hungarian dairy herds. The following were determined: PPL in cows diagnosed pregnant by transrectal ultrasonography 29-42 days after artificial insemination (AI; n = 11 457); PPL in cows diagnosed pregnant by serum PSPB 29-35 days after AI (n = 138 365); and PPL and its association with serum progesterone concentrations, PSPB and production/environmental variables. The definition of PPL was percentage of cows initially diagnosed pregnant based on ultrasonography or PSPB, but not pregnant when examined by transrectal palpation 60 -70 days after AI. The PPL was lower (p < 0.001) in cows following ultrasonographic vs PSPB diagnosis of pregnancy at 29-35 days (8.1 vs 19.3%, respectively), but was higher in cows following ultrasonographic pregnancy diagnosis on 29-35 vs 36-42 days (8.1 vs 7.1%, respectively, P < 0.05). Furthermore, 72.9% of pregnancies with ultrasound-detected morphological abnormalities resulted in pregnancy loss. As a subset of PSPB data, a fully quantitative PSPB assay was used for 20 430 samples; PPL in cows with a high PSPB concentration (>1.1 ng/ml) was lowest (15.0%), whereas cows with low concentrations of both PSPB and progesterone (0.6-1.1 and <2 ng/ml, respectively) had the highest PPL (76.3%; p < 0.0001). Furthermore, PPL was higher in cows with advanced parity and with high milk production, when ambient temperatures were high, although body condition score (BCS) had no effect on PPL. Finally, there were no significant associations between serum PSPB and environmental temperatures or number of post-partum uterine treatments.

Concepts: Pregnancy, Milk, Cattle, Dairy farming, Dairy cattle, Medical ultrasonography, Human chorionic gonadotropin, Artificial insemination


Despite excellent published results, the lack of well-designed, multicentre, randomized clinical trials results in an absence of general consensus on the efficacy of autologous endometrial cells coculture (AECC) in Assisted Reproductive Technology (ART). An open, multicentre, prospective, randomized controlled trial was designed to compare the pregnancy rate (PR) after the transfer of one blastocyst on day 5 after AECC to the transfer of one embryo on day 3 (control group). Patients were women aged 18 to 36, undergoing an ART cycle with no more than 1 embryo transfer failure. Sample size was calculated at 720 for a superiority trial involving an intermediate analysis at 300 patients. We present the results of the intermediate analysis that resulted in the study ending considering the observed difference. Three hundred thirty nine patients were randomized: 170 in the AECC group and 169 in the control group. The clinical PR per transfer was 53.4% with AECC and 37.3% in the control group (p=0.025). The quality of embryos was improved with AECC. These results suggest that implementation of the AECC technique to a large number of In-Vitro Fertilization (IVF) centres could lead to a substantial improvement in the proportion of successful assisted reproduction. The study was supported by the Laboratoires Genévrier, France.

Concepts: Pregnancy, Reproduction, Embryo, Randomized controlled trial, In vitro fertilisation, Preimplantation genetic diagnosis, Artificial insemination, Embryo transfer


Artificial insemination (AI) with cryopreserved semen has a pivotal role in wider dissemination of germplasm of elite livestock and also for conservation of various endangered species. Cryopreservation allows storage of semen for a prolonged period of time and facilitates greater exchange of genetic material among distant populations. Cryopreservation, however, leads to certain deleterious effects on sperm including premature induction of the acrosome reaction, reduced sperm motility and viability, and impaired sperm DNA integrity and fertility. During cooling procedures, membrane phase transitions take place, which result in micro-domain formation from aggregation of lipids, leading to impaired functions of the sperm membrane, and gap formation between gel and fluid domains. Cyclodextrins are produced by enzymatic degradation of starch and possess a unique feature, that when added alone to sperm cyclodextrins facilitate the removal of cholesterol from the membrane. When preloaded with cholesterol, however, cyclodextrins stimulate the insertion of cholesterol into the sperm membrane due to presence of a hydrophobic core in addition to an outer hydrophilic face. Treating sperm of various species with cholesterol-loaded cyclodextrin improves the quality of sperm during cryopreservation. It is still not clearly known how cholesterol-loaded cyclodextrin functions at sperm cells to enhance the survival during cryopreservation. The present review, therefore, highlights possible mechanisms of cholesterol-loaded cyclodextrin action on sperm during cryopreservation.

Concepts: DNA, Sperm, Fertility, Spermatozoon, Semen, Lipid bilayer, Artificial insemination, Ex-situ conservation


To study the effectiveness of four cycles of intrauterine insemination (IUI) with ovarian stimulation (OS) by follicle-stimulating hormone (FSH) or by clomiphene citrate (CC), and adherence to strict cancellation criteria.

Concepts: Reproduction, Randomized controlled trial, Menopause, Assisted reproductive technology, Follicle-stimulating hormone, Artificial insemination, Citrates, Clomifene


In dairy cattle, the widespread use of artificial insemination has resulted in increased selection intensity, which has led to spectacular increase in productivity. However, cow fertility has concomitantly severely declined. It is generally assumed that this reduction is primarily due to the negative energy balance of high-producing cows at the peak of lactation. We herein describe the fine-mapping of a major fertility QTL in Nordic Red cattle, and identify a 660-kb deletion encompassing four genes as the causative variant. We show that the deletion is a recessive embryonically lethal mutation. This probably results from the loss of RNASEH2B, which is known to cause embryonic death in mice. Despite its dramatic effect on fertility, 13%, 23% and 32% of the animals carry the deletion in Danish, Swedish and Finnish Red Cattle, respectively. To explain this, we searched for favorable effects on other traits and found that the deletion has strong positive effects on milk yield. This study demonstrates that embryonic lethal mutations account for a non-negligible fraction of the decline in fertility of domestic cattle, and that associated positive effects on milk yield may account for part of the negative genetic correlation. Our study adds to the evidence that structural variants contribute to animal phenotypic variation, and that balancing selection might be more common in livestock species than previously appreciated.

Concepts: Genetics, Milk, Cattle, Dairy farming, Dairy cattle, Meat, Artificial insemination, Livestock


Genetic variants underlying reduced male reproductive performance have been identified in humans and model organisms, most of them compromising semen quality. Occasionally, male fertility is severely compromised although semen analysis remains without any apparent pathological findings (i.e., idiopathic subfertility). Artificial insemination (AI) in most cattle populations requires close examination of all ejaculates before insemination. Although anomalous ejaculates are rejected, insemination success varies considerably among AI bulls. In an attempt to identify genetic causes of such variation, we undertook a genome-wide association study (GWAS). Imputed genotypes of 652,856 SNPs were available for 7962 AI bulls of the Fleckvieh (FV) population. Male reproductive ability (MRA) was assessed based on 15.3 million artificial inseminations. The GWAS uncovered a strong association signal on bovine chromosome 19 (P = 4.08×10(-59)). Subsequent autozygosity mapping revealed a common 1386 kb segment of extended homozygosity in 40 bulls with exceptionally poor reproductive performance. Only 1.7% of 35,671 inseminations with semen samples of those bulls were successful. None of the bulls with normal reproductive performance was homozygous, indicating recessive inheritance. Exploiting whole-genome re-sequencing data of 43 animals revealed a candidate causal nonsense mutation (rs378652941, c.483C>A, p.Cys161X) in the transmembrane protein 95 encoding gene TMEM95 which was subsequently validated in 1990 AI bulls. Immunohistochemical investigations evidenced that TMEM95 is located at the surface of spermatozoa of fertile animals whereas it is absent in spermatozoa of subfertile animals. These findings imply that integrity of TMEM95 is required for an undisturbed fertilisation. Our results demonstrate that deficiency of TMEM95 severely compromises male reproductive performance in cattle and reveal for the first time a phenotypic effect associated with genomic variation in TMEM95.

Concepts: DNA, Gene, Genetics, Reproduction, Genome-wide association study, Spermatozoon, Semen, Artificial insemination