Alternatives to improve a prostaglandin-based protocol for timed artificial insemination in sheep

The objective was to improve the reproductive performance of a prostaglandin (PG) F_2α-based protocol for timed artificial insemination (TAI) in sheep (Synchrovine®: two doses of 160 μg of delprostenate 7 d apart, with TAI 42 h after second dose). Three experiments were performed: Experiment 1) two doses of a PGF_2α analogue (delprostenate 80 or 160 μg) given 7 d apart; Experiment 2) two PGF_2α treatment intervals (7 or 8 d apart) and two times of TAI (42 or 48 h); and Experiment 3) insemination 12 h after estrus detection or TAI with concurrent GnRH. Experiments involved 1131 ewes that received cervical insemination with fresh semen during the breeding season (32/34 °S–58 °W). Estrous behaviour, conception rate, prolificacy, and fecundity (ultrasonography 30–40 d), were assessed. In Experiment 1, ewes showing estrus between 25 and 48 h or at 72 h after the second PGF_2α did not differ between 80 and 160 μg of delprostenate (73 vs 86%, P = 0.07; and 92 vs 95%, P = NS, respectively). Conception rate and fecundity were lower (P < 0.05) using 80 vs 160 μg (0.24 vs 0.42, and 0.27 vs 0.47, respectively). In Experiment 2, giving PGF_2α 7 d apart resulted in higher (P < 0.05) rates of conception (0.45 and 0.51) and fecundity (0.49 and 0.53) than treatments 8 d apart (conception: 0.33 and 0.29; fecundity: 0.33 and 0.34) for TAI at 42 and 48 h, respectively. In Experiment 3, rates of conception, prolificacy and fecundity were similar (NS) between Synchrovine® with TAI at 42 h (0.50, 1.13, and 0.56) and AI 12 h after estrus detection (0.47, 1.18, and 0.55), and Synchrovine® plus GnRH at TAI (0.38, 1.28, and 0.49). However, all TAI treatments had lower (P < 0.05) prolificacy and fecundity compared to AI following detection of spontaneous estrus (1.39 and 0.83, respectively). In conclusion, the Synchrovine® protocol was: a) more successful using 160 vs 80 μg delprostenate; b) more successful with a 7 d than 8 d PGF_2α interval; c) similarly effective for TAI versus AI 12 h after estrus detection; and d) not improved by giving GnRH at TAI.     

Diseases in swine transmitted by artificial insemination: an overview

Artificial insemination (AI) of swine is widely practiced in countries with an intensive pig production. It is a very useful tool to introduce superior genes into sow herds, with minimal risk for disease transmission. However, the impact of semen that is contaminated with pathogens can be enormous. Most of the micro-organisms that have been detected in boar semen are considered non-pathogenic, but some are known pathogens (e.g. porcine reproductive and respiratory syndrome virus) that can cause major economic losses. Microbial contamination of semen can be due to systemic and/or urogenital tract infections of the boar, or can occur during collection, processing and storage. It can result in reduced semen quality, embryonic or fetal death, endometritis and systemic infection and/or disease in the recipient female. Conventional techniques for isolation of bacteria and viruses from the semen do not always provide optimal results for various reasons, including lack of sensitivity and speed of testing, and difficult interpretation of the outcome. More recently, PCR tests are commonly used; they have a high sensitivity, the outcome is quickly obtained, and they are suitable for monitoring a large number of samples. The best strategy to prevent AI-transmitted diseases is to use boars that are free of specific pathogens, to monitor the animals and semen regularly, and to maintain very high biosecurity. Additional measures should be directed at treating semen with appropriate antimicrobials, and at reducing contamination during semen collection, processing, and storage.     

Ram-induced ovulation to improve artificial insemination efficiency with frozen semen in sheep.

Ram effect, defined as shortening of seasonal anestrus in ewes by exposure to the ram, is now well recognized but the underlying mechanisms are still unclear. Little information also exists whether the ram is able to influence the estrus cycle and ovulation. Three experiments were conducted to investigate endocrine response, time of ovulation and pregnancy rate of ewes in proestrus, exposed to the ram (treated) or an adult ewe (control). In the first experiment, ewes (n = 20) were treated with fluorgestone acetate pessaries for 12 days and were given eCG and cloprostenol one day before withdrawal of pessaries. On the day after removal of the pessaries ewes in the treated group (n = 10) were exposed to the ram and those in the control group (n = 10) were exposed to an adult ewe. Blood samples were taken for LH assay every 20 min from 2 h before to 24 h after ram exposure. In the second experiment, ewes (n = 120) were induced into proestrus and on the day after removal of the pessaries were exposed to either a ram (n = 60) or a ewe (n = 60) as described above and were laparoscoped 50, 60 or 70 h after pessary withdrawal (n = 20 at each time interval). In the third experiment ewes (n = 90) were induced and exposed to the ram (n = 45) or an adult ewe (n = 45) and inseminated via a laparoscope whit frozen-thawed semen at 50 or 60 h after pessary removal, respectively. Exposure to the ram was followed in 2 h by a marked rise in LH, equivalent to a preovulatory surge in duration and amplitude. It was also followed by concentrated ovulation within 25 to 30 h and by an increased pregnancy rate in exposed ewes (73.3 vs. 53.3%).     

Reproductive performance of early-weaned female swine according to their estrus profile and frequency of artificial insemination

We determined the estrus profile (weaning-to-estrus interval (WEI), estrus duration (ED), and frequency of estrus per detection period) in 184 female swine and estimated the effect of the WEI, ED and frequency of artificial insemination (AI) on farrowing rate (FR) and litter size. Estrus detection was done at 8:30 a.m. and 5:00 p.m. The WEI was categorized as short (<100 h), medium (100-120 h) and long (>120 h). The ED was categorized as short (<60 h), medium (60-72 h) and long (>72 h). Mean lactation length was 14.6 days, mean WEI was 124.5 h and mean ED was 69 h. In each weaning group, females received either one or two AI, following a breeding schedule based on the estrus profile. In single-mated females, Al was performed 36 h after the beginning of estrus. In double-mated females, the first AI was done 24 h after the beginning of estrus and the second AI occurred 12 h later. The period of estrus detection had no effect (P > 0.05) on WEI, ED, FR, total born (TB) and live born litter size (LB). Mean FR was 82.6%, mean TB was 10.0% and mean LB was 9.2%. Mean ED was shorter (P < 0.03) for females having medium and long WEI (67.0 and 65.4 h, respectively) than for those having short WEI (72.2 h). A linear regression analysis identified a weak (R2 = 0.02) but significant negative association between ED and WEI (P = 0.05). The WEI did not influence FR (P > 0.05). Total litter size for females having short WEI (9.4) was lower (P < 0.03) than for those having long WEI (10.4). Also, LB for females having medium and long WEI (9.7-9.8) was higher (P < 0.05) than for those having short WEI (8.7). AI frequency had no effect on FR (P > 0.05). TB and LB litter size were lower (P < 0.05) for single-mated females (9.6 and 9.0, respectively) than for double-mated females (10.7 and 9.6, respectively). Double Al was associated with higher subsequent litter size. However, breeding schedules based only on estrus profile may not be precise in determining ideal breeding time, since females having short WEI had the longest ED and produced the lowest litter size.     

Caffeine and artificial insemination

We studied the reactivation of cells and the repair of photomutagenic damage induced by xanthotoxin and visnagin plus NUV in arg-1 cells of Chlamydomonas reinhardii. Maintenance of liquid cultures in the dark resulted only in a slight reactivation of cells, even after 24 h. Repair of photomutagenic damage was more efficient: within 24 h the number of Arg⁺ revertants was reduced by 50% in cells cultured in the dark at 20°C. The repair was more efficient at 30°C. At the beginning of dark cultivation an after-effect could be observed. Cultivation in standard white light instead of dark after treatment resulted in a very strong after-effect. Therefore it was not possible to detect any photoreactivation.After treatment with xanthotoxin plus standard white light (24 h) neither reactivation of cells nor repair of photomutagenic damage was found. The after-effect was higher than after xanthotoxin plus NUV. It is possible that a small amount of repair could be masked by the after-effect.Treatment with visnagin yielded similar results. The photomutagenic effect of visnagin is described for the first time in this paper. The drug is a much less effective photomutagen than xanthotoxin. The photomutagenesis of visnagin may be attributable to photoproducts similar to those formed after treatment with furocoumarins.No definite conclusion can be drawn from the present results regarding the basis for the observed lack of repair (or reduced repair) after standard white light treatment; a possible cause might be a preferential formation of bi-adducts under these conditions.     

Strategies to improve fertility in Bos indicus postpubertal heifers and nonlactating cows submitted to fixed-time artificial insemination

Two experiments were designed to evaluate strategies to increase fertility of Bos indicus postpubertal heifers and nonlactating cows submitted to a fixed-time artificial insemination (TAI) protocol consisting of an intravaginal device containing 1.9 g of progesterone (CIDR) insertion + estradiol benzoate on Day 0, CIDR withdrawal + estradiol cypionate on Day 9, and TAI on Day 11. In Experiment 1, heifers (n = 1153) received a new or an 18-d previously used CIDR and, on Day 9, prostaglandin F_2α (PGF_2α) + 0, 200, or 300 IU equine chorionic gonadotropin (eCG). Heifers treated with a new CIDR had greater (least squares means ± SEM) serum concentration of progesterone on Day 9 (3.06 ± 0.09 ng/mL vs. 2.53 ± 0.09 ng/mL; P < 0.05) and a smaller follicle at TAI (11.61 ± 0.11 mm vs. 12.05 ± 0.12 mm; P < 0.05). Heifers with smaller follicles at TAI had lesser serum progesterone concentrations on Day 18 and reduced rates of ovulation, conception, and pregnancy (P < 0.05). Treatment with eCG improved (P < 0.05) follicle diameter at TAI (11.50 ± 0.10 mm, 11.90 ± 0.11 mm, and 12.00 ± 0.10 mm for 0, 100, and 200 IU, respectively), serum progesterone concentration on Day 18 (2.77 ± 0.11 ng/mL, 3.81 ± 0.11 ng/mL, and 4.87 ± 0.11 ng/mL), and rates of ovulation (83.8%, 88.5%, and 94.3%) and pregnancy (41.3%, 47.0%, and 46.7%). In Experiment 2, nonlactating Nelore cows (n = 702) received PGF_2α treatment on Days 7 or 9 and, on Day 9, 0 or 300 IU eCG. Cows receiving PGF_2α on Day 7 had lesser serum progesterone concentrations on Day 9 (3.05 ± 0.21 ng/mL vs. 4.58 ± 0.21 ng/mL; P < 0.05), a larger follicle at TAI (11.54 ± 0.21 mm vs. 10.84 ± 0.21 mm; P < 0.05), and improved (P < 0.05) rates of ovulation (85.4% vs. 77.0%), conception (60.9% vs. 47.2%), and pregnancy (52.0% vs. 36.4%). Treatment with eCG improved (P < 0.05) serum progesterone concentration on Day 18 (3.24 ± 0.14 ng/mL vs. 4.55 ± 0.14 ng/mL) and the rates of ovulation (72.4% vs. 90.0%) and pregnancy (37.5% vs. 50.8%). In conclusion, giving PGF_2α earlier in the protocol in nonlactating cows and eCG treatment in postpubertal heifers and nonlactating cows improved fertility in response to a TAI (progesterone + estradiol) protocol.     

Training large macaws for artificial insemination procedures

For some endangered parrot species, captive breeding may be the only insurance for their survival. However, many individuals in captivity do not reproduce. Artificial insemination (AI) may help overcome reproductive failures or geographic distance. For semen collection in birds, massage is the most commonly used method. However, this process, which usually requires capture and restraint, involves risk of stress and injuries. The aim of this experiment was to train large macaws to accept the physical manipulations of their body parts needed for the artificial insemination process. Within 15 weeks, a male and a female Buffon's macaw (Ara ambiguus) learned to accept handling without apparent stress. A pair of green-winged macaws (Ara chloropterus) progressed more slowly and displayed some signs of stress. This stress highlights the need to monitor the birds' possible signs of discomfort during the training in order to adapt the working protocol. These results demonstrate that it is possible to train the AI behaviors, thus avoiding the capture, restraint, and anesthesia. An added benefit to this is the potential for the training to provide a form of behavioral enrichment.     

Production and storage of goat semen for artificial insemination

Environmental influences on reproduction and semen production in the buck, the problem of interaction between seminal plasma and egg yolk or milk constituents in diluent, liquid storage and processing of semen for freezing are discussed. A review is given on the use of frozen-thawed semen for artificial insemination (AI) in spontaneous and induced oestrus and factors influencing the fertility.     

A model for economic comparison of swine insemination programs

Optimal artificial insemination schedules are those that result in a high farrowing rate and litter size, while minimizing costs of semen and labor by avoiding unnecessary inseminations. A simulation model programmed in a commercial spreadsheet was developed to permit comparison of alternative schedules. Farrowing rate and litter size for a particular schedule were dependent on the timing of insemination relative to the time of ovulation. Economic return was calculated by multiplying the number of pigs born per bred sow by $33.00 and subtracting the cost of producing a litter of pigs and raising them to weaning ($222.88 per sow plus $2.44 per pig born) and the cost of detection of estrus and breeding. Seven insemination schedules combined with once versus twice per day detection of estrus were simulated in 500 herds of 100 sows each. Inseminations were simulated to occur on schedules of: 1) 0, 12, 24 and 36 h; 2) 12, 24 and 36 h; 3) 0 and 24 h; 4) 12 and 36 h; 5) 12 h; 6) 24 h; and 7) 36 h after first detection of estrus. Schedule 1 was predicted to yield the highest farrowing rate and litter size. Economic return was highest for Schedule 2 with twice per day detection of estrus followed closely by Schedule 1 with once per day detection of estrus at $14.90 and $13.75 per bred sow, respectively. High performance was dependent on insuring that inseminations occurred at an optimum time in as great a proportion of sows as possible.     

Perspectives of genomics for genetic conservation of livestock

Genomics provides new opportunities for conservation genetics. Conservation genetics in livestock is based on estimating diversity by pedigree relatedness and managing diversity by choosing those animals that maximize genetic diversity. Animals can be chosen as parents for the next generation, as donors of material to a gene bank, or as breeds for targeting conservation efforts. Genomics provides opportunities to estimate diversity for specific parts of the genome, such as neutral and adaptive diversity and genetic diversity underlying specific traits. This enables us to choose candidates for conservation based on specific genetic diversity (e.g. diversity of traits or adaptive diversity) or to monitor the loss of diversity without conservation. In wild animals direct genetic management, by choosing candidates for conservation as in livestock, is generally not practiced. With dense marker maps opportunities exist for monitoring relatedness and genetic diversity in wild populations, thus enabling a more active management of diversity.     

Harvesting sperm and artificial insemination of mice

Rodents of the genus Peromyscus (deer mice) are the most prevalent native North American mammals. Peromyscus species are used in a wide range of research including toxicology, epidemiology, ecology, behavioral, and genetic studies. Here they provide a useful model for demonstrations of artificial insemination. Methods similar to those displayed here have previously been used in several deer mouse studies, yet no detailed protocol has been published. Here we demonstrate the basic method of artificial insemination. This method entails extracting the testes from the rodent, then isolating the sperm from the epididymis and vas deferens. The mature sperm, now in a milk mixture, are placed in the female's reproductive tract at the time of ovulation. Fertilization is counted as day 0 for timing of embryo development. Embryos can then be retrieved at the desired time-point and manipulated.Artificial insemination can be used in a variety of rodent species where exact embryo timing is crucial or hard to obtain. This technique is vital for species or strains (including most Peromyscus) which may not mate immediately and/or where mating is hard to assess. In addition, artificial insemination provides exact timing for embryo development either in mapping developmental progress and/or transgenic work. Reduced numbers of animals can be used since fertilization is guaranteed. This method has been vital to furthering the Peromyscus system, and will hopefully benefit others as well.     

Frpm artificial individuals to global patterns

Artificial Life is a model of biological systems that describes lives archived by computer simulation, chemical substrates or any other non-biological substrates. Artificial Life simulation adopts a bottom-up approach in which behavior of lower-level entities (e.g. molecules, cells and individuals) is all that is programed; global patterns (e.g. evolutionary patterns observed at the level of the population and the community) can emerge as a result of interaction among lower-level entities. Artificial Life simulations will be used not only to test ecological and evolutionary hypotheses explaining real organisms but also to show the validity of general theories, processes and concepts such as natural selection, theories of complexity, hierarchical relations and self-organization.     

Improved method for artificial insemination in the great apes

Artificial insemination in the great apes has not achieved its potential as a tool in maintenance of the endangered captive population. Three factors can influence the success rate of artificial insemination: sperm preparation, site of insemination, and timing of insemination. We have tried to optimize methods regarding these three steps. A modified method for insemination is described which has resulted in a 21% success rate (six term pregnancies from 29 inseminations) in the chimpanzee and which has successfully initiated a pregnancy in a gorilla.     

Genetics and genomics to improve fertility in high producing dairy cows

Improving dairy cow fertility by means of genetic selection is likely to become increasingly important, since it is now well established that declining fertility cannot only be arrested by improved management. Profit margins per kg milk produced are decreasing, therefore farmers need to reduce cost and increase herd size. This restricts the labor input per cow and the disposable cost of getting a cow pregnant, whilst at the same time hormone treatments have become less acceptable. This makes it unlikely that additional management interventions will maintain fertility at acceptable levels in the near future. Genetic improvement seems the obvious solution. Effective selection tools are available in most Western countries using traditional breeding value estimation procedures. Also, in addition to gene assisted selection using individual genes or QTL, high throughput Single Nucleotide Polymorphism (SNP) technology allows genetic improvement of fertility based on information from the whole genome (tens of thousands SNP per animal), i.e. genomic selection. Simulation studies have shown that genomic selection improves the accuracy of selecting juvenile animals compared with traditional breeding methods and compared with selection using information from a few genes or QTL only. Research in the areas genomics and proteomics promise to make genetic selection even more effective. The genomic and proteomics technologies combined with the bioinformatics tools that support the interpretation of gene functioning and protein expression facilitate an exciting starting point for the development of new management strategies and tools for the improvement of reproductive performance.