Insemination Process Research Articles

Gene Sequence of Rubella Virus Infection in Serum of Wives That Correlate to Intrauterine Insemination Success Rate in Baghdad's Population Couples

The current study focused on the prevalence of Rubella virus disease among wives who underwent the process of artificial insemination (IUI) and its relationship to the elevated proportion of free radicals in the blood plasma and its correlation with water pollution in the spreading of the disease for the period from 15/2/2022 to 1/2/2023 in private laboratory for IUI procedure (AL- Amal). The study included 60 women who underwent (IUI) procedure and have positive results for Rubella virus (IgG and IgM) compared to wives who tested negative for Rubella virus for ages groups from 20 to 35 years.

Effect of L-carnitine and Alpha lipoic acid on some quality parameters of buck semen preserved by cooling

Fertility is the key of reproduction and it is affected by many factors especially artificial insemination process (AI), the quality of semen used is an important key for the success of AI process. In this study, we aim to evaluate the effect of adding different concentrations of different antioxidants such as L-carnitine and Alpha lipoic acid to the diluted buck semen during cooling in summer season. Four mature healthy Zaraibi bucks were used in the study. Semen was collected twice weekly during summer .Semen samples were collected by an electro- ejaculator device. Samples of good quality semen were pooled and diluted by extender and divided according to experimental groups. Group 1: L-carnitine : 1 ml mol , 5 ml mol , 10 ml mol. Group 2: Alpha lipoic acid : 0.025 ml mol, 0.05 ml mol. Group 3: is the control group .After cooling by 1 hr, 2hr,12 hr , 24 hr and 48 hr. Semen parameters were examined as motility, livability, acrosome integrity and cell membrane integrity .The results indicated that L-carnitine 5 ml mol has a favorable effect on the quality of cooled diluted buck semen more than other groups of L-carnitine and 0.025 ml mol of Alpha lipoic acid is better than other concentration. In conclusion, L-Carnitine and alpha lipoic acid as antioxidants improve buck semen quality cooled under low temperature especially 5 ml mol and 0.025 ml mol concentration respectively.

Study on the Effect of Bee Strain and Insemination Method on the Hygienic Behavior of Bee Colonies Tested Using the Pricking Method

The process of artificial and natural insemination was carried out on the queen bees of the local and Carniolan strains in order to study their hygienic behavior and the Pricking method for the strains, as well as the cross-insemination results between them. The study results showed that the local honey bee strain T1 outperformed the other studied strains in its hygienic behavior, as the detection rate of the dead brood by Pricking was (44.16, 93.16, and 100) % respectively, after (1, 6, and 24) hours of conducting the study. The same strain also outperformed all other strains in removing the dead brood with a percentage ranging from (0.16, 22.66, and 100) % respectively, after (1, 6, and 24) hours of conducting the study. Moreover, the artificial insemination method generally outperformed the natural insemination method in the hygienic behavior of the bee colonies, as the detection rate of the brood in the artificially inseminated bee strains A2 was (40.91, 91.00, 97.33, and 100) % respectively, after (1, 6, 24, and 36) hours of conducting the study. A2 also outperformed the A1 insemination method in terms of the percentage of removing the dead brood by Pricking, with a percentage ranging from (0.00, 17.08, 96.91, and 100) % respectively, after (1, 6, 24, and 36) hours of conducting the study.

ERGONOMIC STUDIES OF THE PROCESS OF ARTIFICIAL INSEMINATION OF PIGS

Goal. To research the comparison of artificial insemination processes in a number of pig farms based on the developed theoretical principles of ergonomic assessment. Methods. Experimental research was carried out at “Agrofirma “Khlibne ” LLC, FG “Shubske ”, SE EE “Hontarivka” of Institute of Animal Science NAAS, and FOP “Syvoplias” of Kharkiv region based on traditional approaches. As a basis for dividing the process of artificial insemination of pigs into technological operations, the farm of FG "Shubske" was taken, as such, where the process of artificial insemination of pigs is the most saturated with technological operations. According to the determined technological operations of the process of artificial insemination of sows, more generalized characteristics of the labor process, the number, sequence and duration of individual component movements were determined, with the classification of labor movements by determination, severity, naturalness, purpose, direction and reach in accordance with the methods, based on the basic systems of trace elements time standards, normative documents and recommendations. The indexes obtained experimentally were worked out by the method of variational statistics, using MS Excel software. The results. The process of artificial insemination in different farms has certain differences. The process of artificial insemination from a number of evaluated farms is the most saturated with technological operations in the breeding farm of FG "Shubske". Regarding different farms, there are differences in the use of stimulating devices, the load on the operator and the way of housing sows during insemination. According to the evaluated farms, the test boar is used twice a day in FG “Shubske ”, once in the morning in the SE EE “Hontarivka” of Institute of Animal Science NAAS, in FOP “Syvoplias V.V.” the detection of sows in estrus occurs without the involvement of a test boar. Also, the anthropometric, physiological and individual characteristics of the artificial insemination operator were evaluated for two operators for each of the evaluated farms. Weight indexes between operators in different farms differed more significantly (р<0.05). At the same time, no discrepancies were found in terms of arm length. Regarding disagreements between operators within individual farms, the greatest consolidation in terms of height and live weight was found in FOP "Syvoplias V. V.". The largest discrepancies in the length of the hands of the operators were found in SE “Hontarivka” of Institute of Animal Science NAAS. In the vast majority of cases, unconsolidated values were obtained for pulse and blood pressure indexes. Conclusions. On the basis of ergonomic studies of the process of artificial insemination of pigs in the conditions of three different farms, the differences have been determined, which indicate the need to adjust individual technological approaches and carry out the conversion of premises for artificial insemination of pigs in order to further increase the efficiency of the work of technicians and the procedure of artificial insemination as a whole in the SE EE "Hontarivka" » of Institute of Animal Science NAAS and in FOP "Syvoplias V. V.". Key words: sow, artificial insemination, ergonomics, machine, reproduction, sperm production.

A novel method to collect sperm from brood parasitic cuckoos: Urodeum stimulation (UroS) method

Sperm competition has been studied in numerous species as a representative example of postcopulatory sexual selection, where sampling sperm from male is the most basic and important step. Sperm collection can be tricky in birds, however, because unlike mammals, the genitals of birds are generally latent in the cloacal region and their characteristics vary among species. Various methods to collect sperm from different birds have been tested, such as cloacal massage, feces collection, and electro-stimulation, but their applicability varies depending on species. In this study, we introduced the urodeum stimulation method (UroS method) to collect sperm from Cuculus cuckoos, such as the Common Cuckoo (C. canorus). These species are expected to have interesting patterns of pair bonding and sperm competition because of their unique breeding strategy called brood parasitism; however, it remains unexplored. In this study, we described the application of our new method to expel semen from male common cuckoos, measured the volume of semen collected, checked the presence of sperm in the semen sample, and finally estimated its success rate among 82 males. Samples were successfully collected from 76 cuckoos (approximately 93%) and the colors and volumes of the samples were very diverse. Sperm was present in 43 of these samples (57%), showing a sperm observation rate approximately twice as high as that of the conventional cloacal massage method. We believe that this novel method will contribute to a better understanding of postcopulatory sexual selection in avian brood parasites and facilitate the process of sperm collection and artificial insemination in other medium-sized birds.

Morphology of goat oviducts

The oviduct is a vital organ for the process of capacitation, sperm survival and insemination. Gametes are transported, and the embryo is formed in it. The article has described the morphological features and has studied the morphometric parameters of goats oviduct during the newborn period, in 5-, 6-, 7-months-old and in adult animals. Since the newborn period to the age of 5 months, the oviducts of female goats grow at a relative rate of 109.6%. From this age to adulthood, the relative growth rate averages 20.9%. The high epithelium in newborn goats compared to other individuals may be related with unadjusted folds of the mucous membrane, cell layering on top of each other due to lack of area, or it may be associated with hormonal stimulation.

Religious and cultural interpretations of artificial insemination in South-West Nigeria

In a constantly changing and increasingly globalized world, religion and cultural practices still play a significant role in influencing people's behaviour towards assisted reproductive technologies. This study examined the religious, cultural and personal interpretations of artificial insemination. The research methodology was triangulated using qualitative and quantitative methods of data collection. A stratified and simple random sampling technique was used to select the study participants for quantitative data, while, Key Informant Interviews (KIIs) respondents were purposively sampled. The findings showed that the majority of the respondents were aware of artificial insemination, 75.3% of the respondents had adequate knowledge of artificial insemination. About 54.2% of the respondents agreed that their religious denominations supported any form of artificial insemination, For instance, in contrast to the 61.1% of Roman Catholic respondents' who acknowledged that their religious denomination did not support artificial insemination, 75% of Shia Muslims, and 65.0% of Pentecostal respondents' acknowledged religious support for artificial insemination. For 32.8% of the respondents, the process of artificial insemination by a donor negates the will of God, while 32.8% of the respondents, posited that artificial insemination is a deviation from cultural norms. The study also revealed that 57.0% of respondents personally perceived artificial insemination to be a very good medical breakthrough. Regardless of the technological or educational prowess leading to artificial insemination, certain religious denominations and cultural practices do not support artificial insemination. Medical ethics should take cognizance of the religious and cultural ideas and principles of the society in which they are practiced.

Application of Nanoparticles and Melatonin for Cryopreservation of Gametes and Embryos.

Cryopreservation of gametes and embryos, a technique widely applied in human infertility clinics and to preserve desirable genetic traits of livestock, has been developed over 30 years as a component of the artificial insemination process. A number of researchers have conducted studies to reduce cell toxicity during cryopreservation using adjuvants leading to higher gamete and embryo survival rates. Melatonin and Nanoparticles are novel cryoprotectants and recent studies have investigated their properties such as regulating oxidative stresses, lipid peroxidation, and DNA fragmentation in order to protect gametes and embryos during vitrification. This review presented the current status of cryoprotectants and highlights the novel biomaterials such as melatonin and nanoparticles that may improve the survivability of gametes and embryos during this process.

PERANCANGAN ELEKTRO EJAKULATOR SEBAGAI ALAT BANTU INSEMINASI BUATAN PADA KAMBING TERNAK DI KOTA BENGKULU

Semen collection is one of the activities contained in the artificial insemination program in animals. Semen collection can be done in several ways, namely using an artificial vagina, massage and using an electro ejaculator module. The use of the electro ejaculator module is a safe and appropriate method for male livestock. Electro ejaculator module sold in the market has a relatively expensive price and is still difficult to find, especially in Indonesia. Therefore, an electro ejaculator module or device for goats is needed that has an affordable price so that it can be used by small to medium-sized farms. This electro ejaculator device consists of 3 main parts, namely Switch Mode Power Supply, function generator and an electro ejaculatory probe. At the testing stage of the switch mode power supply (SMPS) section, the output voltage variation is obtained from 0 - 15 volts. While in the function generator section, the output signal is in the form of a sinusoidal which can be adjusted for the amplitude and the output frequency from 0 - 1 Mhz. Based on these results, the designed Electro Ejaculator module can be applied to assist the process of artificial insemination in animals.

P-768 Families' experience of insemination fraud : a qualitative study

Abstract Study question What is the experience of parents and their offspring when they discover they were victims of insemination fraud, i.e. a voluntary substitution of sperm samples? Summary answer Participants reported experiencing shock, sideration and disbelief, feelings of being betrayed by the medical institution, and concerns about the repercussions on family bonds. What is known already Despite growing concerns about third-party assisted human reproduction, several loopholes remain in the legislation governing sperm donation. For example, the lack of consistency across clinics in donation tracking and record-keeping procedures has led to cases of accidental or voluntary substitution of sperm donations. Such substitutions imply that the sperm of the donor selected by the parents or that of the father of the child is not the sperm used for conception. With the advent of DNA kits, more and more cases of substitution are being uncovered. However, this phenomenon has not received the scientific attention it deserves. Study design, size, duration This is an exploratory study using a qualitative methodology. Data collection was conducted in Canada in the winter of 2019 and spring of 2021 using a purposive sampling method. Participants/materials, setting, methods In total, 13 people made up our sample: seven parents (two fathers and five mothers) and six donor-conceived people conceived (four teenagers, aged between 13 and 16 years and two adults aged 38 and 39 years). Donor conception was used in the context of male infertility (N = 1), vasectomy (N = 3), sperm freezing after cancer (N = 1), and celibacy (N = 1). The data was collected using semi-structured individual interviews. Main results and the role of chance A thematic analysis shows that the experience varies according to one’s position in the insemination process (whether one uses it or is the product of it), to the nature of the substitution and to the suspicions one has (or not) prior to the discovery. However, all the participants described the announcement of the substitution as a shock that strongly undermines trust in the assisted reproduction industry. In cases where the father’s sperm was substituted, this leads to having to renounce biological paternity and the ideal of a child born to both parents. In cases where the chosen donor was substituted, this forces the parents and their offspring to give up (temporarily or permanently) the possibility of tracing the man who conceived the child and/or the donor siblings. The substitution generates then for the parents a feeling of worry about the medical and psychological repercussions for their child and, for the children, a feeling of incomplete identity. On the other hand, in cases where the doctor’s sperm was substituted for that of an unknown donor, the persons conceived by donation seemed to have a better reaction because it provided them with answers as to their origins. Limitations, reasons for caution Despite the innovative nature of the study, the size of the sample and the variability of the experiences do not allow for definitive conclusions to be drawn about this phenomenon and limit the understanding that can be gained from it to this particular fraud, emanating from the same doctor. Wider implications of the findings With the democratization of DNA testing, this type of phenomenon could become more widespread. It is therefore important to document it to better understand its consequences on the disintegration of the filial or family bond, the possibility of redefining the biological family, and the redefinition of the original parental project. Trial registration number N/A

Reproductive Indices of Naturally Mated and Artificially Inseminated Quail Hens (Coturnix japonica): Is Artificial Insemination of Japanese Quail Hens Feasible in a Local Setting?

Naturally mated and artificially inseminated Japanese quail (Coturnix japonica) hens’ eggs reproductive indices were investigated to evaluate natural mating or artificial insemination techniques for breeding quails in local farms. A total of 141 (108 hens and 33 cocks) matured quails of about 11 weeks of age were used for the experiment, of which 54 hens were inseminated with Citric-quail egg albumen extended semen collected from 15 fertile cocks. Another 54 hens were naturally mated by another 18 fertile cocks in a ratio of one cock to three hens (1:3). Eggs collected from both naturally mated and artificially inseminated birds were incubated for 18 days. The percentage of incubated eggs that were fertile when candled on the 7th and 12th day and hatch-out analysis after the 18th day of incubation tested fertility. Hatchability was tested by the percentage number of fertile eggs that hatched. Naturally mated and artificially inseminated hens’ eggs had a fertility of 84.0 % and 73.3 %, and a hatchability of 82.5 % and 85.5 %, respectively. The study revealed no significant difference in the frequency of fertile or hatched Japanese quail eggs (χ2 cal < χ critical at 5% or 1% level of probability). Naturally mated and artificially inseminated birds had relative fertile egg rates of 53.4 % and 46.6 %, respectively, and relative egg hatching rates of 52.5% and 47.5 %, respectively. Artificial insemination had the advantage of introducing the same volume of semen to more quail hens than a single cock in a natural mating process. However, natural mating was recommended over artificial insemination in the study area because of the technical constraints that required more workforce and amenities in the insemination process. The authors recommended research on developing restraint instruments and non-electrical semen storage methods for quail birds. The authors also proposed a restraint model for quail birds

Characteristics of Bali Bull (Bos Sondaicus) Sexed Sperms with Freeze Dry Egg White at Different Incubation Time

The development of reproductive technology for the separation of spermatozoa with X and Y chromosomes, which is better known as sexing, can be carried out in vitro using various methods, one of which is using freeze dry egg whites. This study aims to determine the quality of semen based on incubation time. This study used four types of treatment, namely P0 = No Incubation, P1 = Incubation 30 minutes, P2 = Incubation 45 minutes, P3 = Incubation 60 minutes with six replications. Parameters observed were the quality of spermatozoa macroscopically (volume, odor, color and consistency) and microscopically (motility, abnormality, concentration, intact plasma membrane, intact acrosome cap). Y using descriptive statistical test. The results of analysis of variance that sexing spermatozoa using freeze dry egg whites at different incubation times showed a significant effect (P<0.05) on intact plasma membrane (IPM), intact acrosomal membrane (IAM), but had no significant effect P>0.05) on concentration, motility, and abnormalities. Evaluation of fresh Bali cattle spermatozoa showed results that were in accordance with the standards for further examination of semen sexing. Incubation time of 30 minutes in the sexing process of spermatozoa caused a significant decrease in motility and concentration, meanwhile, IPM and IAM of sexed spermatozoa experienced a significant decrease in incubation time of 60 minutes, but the decrease was still within the normal range for spermatozoa. to be used in the artificial insemination process. Keywords: Sexing, incubation, concentration, motility, abnormality, intact plasma membrane, intact acrosome caps.

Molecular sexing in Bos taurus using quantitative polymerase chain reaction (qPCR) method

Cattle are animals that are widely found all over the world which is the domestication of bulls. In Indonesia, domesticated cattle are used as livestock, could be meat cattle or dairy cattle. The sex of the cattle is very influential in terms of selling, especially the type of bull. Calf bull is a product of the process of artificial insemination with spermatozoa resulting in sexing. Mammals have X and Y chromosomes to distinguish their sex. Each chromosome has a special gene such as the SRY gene on the Y chromosome and the PLP gene on the X chromosome. Molecular methods are effective methods of determining the chromosomes carried by spermatozoa. Both special genes are found in the home box (GADPH) of mammals. Primer is made from the mRNA SRY (sex-determining region Y) sequence from Bos taurus with a size above 200bp (SRY B) and below 200bp (SRY A) so that optimal results are obtained between the two. The type of sample that has been used in this research was sexing semen from limousin bulls that are treated from 4 pairs of primers each. The results of the qPCR from both primers are viewed from the melt-peak graph and comparison analysis. The differences between SRY A and SRY-B primer indicates that SRY-A produces a more stable melt-peak graph than SRY B. This indicates that the SRY A primer is more stable when used for molecular verification processes using qPCR.

Detection of Bovine Herpesvirus 1 from Semen by Real-time PCR to Prevent the Spread of Infectious Bovine Rhinotracheitis Infection

Infectious Bovine Rhinotracheitis (IBR) can be transmitted by livestock seeds and semen, through the process of artificial insemination. Therefore, it is necessary to detect the presence of the Bovine Herpesvirus 1 (BHV1) in semen through artificial insemination and breeding centers of Indonesia. The current study aimed to detect the presence of the virus in semen as a source of IBR disease transmission in Indonesia. A total of 27 semen samples from artificial insemination and breeding centers (Sembawa, Lembang, Ungaran, and Sleman) in Indonesia have been examined and identified using the real-time PCR (qPCR) technique. The result showed that all samples were negative to BHV1. This indicated that semen from Ungaran, Sembawa, Sleman, and Lembang was safe to be used as a semen source for artificial insemination.

Precopulatory oral sex contact plays an important role in copulatory success in a cryptic desert beetle.

Precopulatory courtship plays an essential role in the insemination process and influences postcopulatory behavior between males and females. Male precopulatory oral stimulation of female genitals is rare for invertebrates. Here, we describe an intriguing oral sexual courtship in a cryptic desert beetle Platyope mongolica Faldermann. The males repeatedly contact the female's genitals using their mouths to gain consent to mate. Furthermore, the rate at which males contact the female's genitals relates to the copulation success in a series of observations. However, interference in oral sexual contacts decreased the proportion of successful copulation. Further no‐choice tests found homosexual behavior between males with antenna removed. We report the precopulatory oral sexual behavior and its important role for copulation success in P. mongolica for the first time. These findings highlight the significance of oral sexual courtship in sexual selection.

Systemic changes caused by artificial insemination in beef cows (Bos indicus) and their impact on animal welfare

This study aimed to verify if the process of artificial insemination (AI) characterized here as animal immobilization, the passage of the semen applicator through the cervix, and deposition of the semen in the uterus, affected cows’ welfare. For this, 18 beef calved cows were selected and divided into two groups: inseminated cows (AIG, n = 9), and not inseminated cows, the control group (CG, n = 9). Body condition score, uterus, and ovary evaluation were performed. Later, both groups were submitted into an estrus synchronization protocol and only the AIG group was inseminated. Blood components of urea, creatinine, AST, GGT, CK, glucose, triglycerides, cholesterol, HDL, LDL, VLDL, NEFA, BHB, cortisol, estradiol, progesterone, albumin, and total protein were measured 30 h before AI, and 4, 24, 48 and 168 h after AI. Statistical differences were considered when P <0.05. No differences between AIG and CG were observed. On the other hand, when the moment of insemination was evaluated, differences were observed for urea, creatinine, AST, GGT, CK, glucose, triglycerides, NEFA, BHB, albumin, and total protein. There was an oscillation of metabolic profiles depending on the time and procedures to which animals were exposed, even though it could be inferred that the AI process was incapable of altering those metabolic components on animals that were inseminated. Still, we can affirm that artificial insemination cannot be categorized as a negative reproduction tool on animal welfare. However, the containment and management procedures for AI may alter the metabolic profile of cows, especially the increase of CK.

Review of domestic and international legislative trends related to surrogate mother contracts, and differences in perspective between medicine and law

The medical development of Assisted Reproductive Technology (ART) significantly assists infertile couples who are unable to conceive or carry a child as a result of congenital or acquired diseases. However, the process of artificial insemination by surrogacy arrangements or surrogate pregnancy has caused continued controversy surrounding legal and ethical issues. While surrogacy arrangements/surrogate pregnancy are considered invalid in the court, such measures are in practice performed with the assistance of doctors. From the legal perspective, although the measures are not recognized as the form of pregnancy and birth, the practice of surrogacy has already existed from the medical & practical viewpoints, indicating a significant gap between two sides. There are growing concerns about problems with surrogate pregnancy. The absence of stated regulations on such practice could lead to the failure in fulfilling duty by mutual parties, therefore having difficulty in securing the legal status of a baby. The health of a surrogate mother can be possibly damaged. Additionally, the intervention of illegal brokers could lead to the dangers of excessive commercialization. In fact, there are few statistics on how many children are formally born through such practice, therefore causing social problems for a long time, such as the related fraud, refusal to surrender a child, etc. In spite of such problems, it is hard to come up with active countermeasures or prepare statistics. Accordingly, this paper explores the differing views between the court and the medical community on surrogate pregnancy. By conducting a comparative study on the latest social status and the systems at home and abroad, this paper is aimed to arrive at a conclusion on desirable improvements in the system based on in-depth interviews with surrogacy-related experts in Korea. To accomplish this, the paper provides a change in the current system and a review of intervention. In addition, to secure safety of surrogate mothers and contracting parties and prevent any harm, the paper presents a viewpoint on whether a social consensus can be reached as to what issues should be considered top priority regarding the progress of discussions in the future.

Effect of the Refrigeration System on In Vitro Quality and In Vivo Fertility of Goat Buck Sperm.

Simple SummaryArtificial insemination of goats is mostly performed with sperm that has been chilled to 4 °C and stored for up to 12 h. Because the chilling of samples to 4 °C must be done slowly, artificial inseminations have to be performed late in the day, especially in herds located far from artificial insemination centers, which impedes the extensive use of this reproductive technology in goats. In this study, we explored the possibility of chilling sperm doses at a controlled rate (close to −0.18 °C/min) during transportation and compared the quality and fertility of these samples with those chilled in the lab. We observed that the in vitro quality of doses chilled by the prototype procedure developed for transportation was higher than that of samples chilled in the lab, and the fertility of the sperm was similar. Therefore, it is possible to chill doses during transportation without affecting their quality or fertility.Cooling goat sperm insemination doses to 4 °C causes a delay in their delivery. However, chilling these doses during the transportation period could expedite their delivery and the insemination process. In this study, an economical and simple apparatus for chilling goat semen doses in itinere was developed, and the in vitro quality and in vivo fertility of these doses were compared with those chilled by means of a programmable water bath in the laboratory at a rate of −0.18 °C/min. Of the tested prototypes, the one that provided an optimal combination of the chilling rate (average of −0.09 °C/min) and time required to reach 4 °C (3 h 45 min) was selected for further testing. Immediately after chilling and 24 h later, the doses chilled in the prototype were determined to be of higher quality than the samples chilled in the programmable water bath. Finally, the kidding rate was similar between the doses chilled in the programmable water bath (61.7% ± 7.1%) and in the prototype (56.1% ± 5.9%). In conclusion, successful chilling of goat sperm doses during transport is possible, thereby accelerating the delivery of insemination doses.

71 Imitating the dynamic bovine cervix with 3D printing technology to teach artificial insemination in cattle

Abstract The process of artificial insemination in cattle is commonly taught using excised reproductive tracts. Excised reproductive tracts can be difficult to collect, and often require freezing and thawing prior to use; however, they are an effective tool to teach artificial insemination (AI) techniques. As an alternative tool, 3D printed cervixes were created using NinjaFlex filament in a Flashforge Creator Pro Dual Extrusion Printer. Designs of cervixes were created with a 3D CAD software while MATLAB was used to generate the random placement of the cervical ring openings to model biological diversity. The objective was to determine the effectiveness of using a 3D printed cervix compared with an excised reproductive tract to teach AI in cattle. Data were collected via surveys for 120 students from 2016 through 2019. Students were divided in two groups prior to entering live animals: 1) Excised reproductive tract and 2) 3D Cervix. The excised reproductive tracts were previously collected, preserved, and confirmed passable by instructors. The 3D cervix group included 3D cervixes mounted in a bovine pelvis inside a box. Overall, 88% of students had success passing the cervix in the cow by the third day. Eighty-seven percent of students thought the excised tracts felt somewhat similar to a live cow, compared with 54% who thought the 3D cervixes alone felt similar to a live cow. When the 3D cervixes were mounted in the box, 17/20 students thought they felt similar to a live cow. For learning AI techniques, 12/21 students thought the excised tracts were more helpful than the 3D cervixes alone (5/21); however, when the 3D cervixes were mounted in the pelvic boxes, 11/21 students preferred this setup compared with the excised tracts (6/21 students). Student feedback suggests the 3D cervixes mounted in pelvic boxes are a useful tool for teaching artificial insemination techniques.

Spotted wolffish (Anarhichas minor) sperm cryopreservation in 5-mL cryovials

In spotted wolffish Anarhichas minor aquaculture, cryopreservation is used to secure sperm availability throughout the entire spawning season. Under current protocols, sperm is cryopreserved in 0.5-mL straws. This implies thawing a considerable number of straws for insemination with cryopreserved sperm. In this work, we scale up the spotted wolffish sperm cryopreservation procedure through the development of a protocol for sperm cryopreservation in 5-mL cryovials. Different freezing (distances from the liquid nitrogen surface) and thawing rates were tested. The best results were obtained with cryovials frozen at a distance of 1.5 cm from the liquid nitrogen surface and thawed either at 15 or 10 °C for 4 and 6 min, respectively. Under these conditions, similar percentage of motile cells, sperm velocity and percentage of viable cells were obtained in comparison with the sperm cryopreserved in the traditional 0.5-mL straws. This protocol will facilitate the process of insemination with cryopreserved sperm in the spotted wolffish hatcheries.