Sperm Doses Research Articles

Infl uence of the exercise regime on the quality indicators of sperm production in breeding boars

In the pig breeding industry, the quality of sperm obtained from breeding boars plays an important role in the issue of reproduction. The purpose of the research was to investigate the eff ect of exercise regime on the quality indicators of sperm production in breeding boars. The research was carried out at the Tyumensky Pig Complex LLC in the Tyumen Region. For the research 3 groups of boars were formed with 10 heads in each. The control group did not receive exercise, the 1st experimental group received passive exercise, the 2nd experimental group received active exercise. It was found that the rectilinear motility of sperm, which aff ects the fertilizing ability of semen was in the 1st experimental group of boars 87.0 %, which was by 2.8 abs.% higher than in other groups. Also, more sperm doses were obtained from semen with high rectilinear motility with the same volume and concentration indicators In the groups of boars without exercise and with active exercise, 5.7 % of dead sperm were found, which was by 0.8 abs.% more than in the analogs that received passive exercise (p < 0,05). In the 1st experimental group of boars with passive exercise, the lowest percentage of sperm with defects was found, which aff ects the efficiency of insemination and the quality of the off spring. It was found that passive exercise had a more favorable eff ect on the qualitative indicators of sperm of the breeding boars of the 1st experimental group compared to active exercise (2nd experimental group) and its absence (control group). The use of passive exercise regime for breeding boars had a positive eff ect not only on the quantitative indicators of semen, but also on the quality of sperm and can be recommended at the enterprise when housing breeding boars.

ИЗУЧЕНИЕ БИОЛОГИЧЕСКОЙ ПОЛНОЦЕННОСТИ СПЕРМАТОЗОИДОВ В СПЕРМЕ С БАКТЕРИАЛЬНОЙ ОБСЕМЕНЕННОСТЬЮ

The aim of the study is to investigate the relationship between bacterial contamination of bull semen and the biological value of spermatozoa, as well as reproductive performance. The object of the study was Holstein bulls (n = 17). The material for the study was frozen-thawed semen of bulls in the amount of 61 samples. Microbiological studies of sperm doses were carried out using standard nutrient media in accordance with GOST 32222-2013. The total number of microorganisms was calculated by the number of grown colony-forming units (CFU/ml) in 1 cm3, with subsequent determination of their morphological pro-perties. Sperm motility and morphology were determined using the Argus CASA program. The state of DNA in spermatozoa was studied by the acridine orange test (AO test) using a fluorescence microscope. The study revealed a high positive correlation (r = +0.940**) between the number of microorganisms in sperm and pathology in individual sperm segments. Analysis of variance confirmed a statistically significant effect of the number of microorganisms on sperm morphology F = 123.2 (p=0.000). The correlation relationship between the fragmentation index and the content of microorganisms has a high value (r = +0.965**). Sperm motility negatively correlates (r = –0.768**) with the number of microorganisms (CFU/ml). In the presence of Mycoplasma spp in the samples, the number of cows with fruitful insemination was 35.5 % due to reduced motility, morphology and high fragmentation of sperm DNA (58 %). Sperm motility in samples containing Mycoplasma spp averages 24.5 %, while in samples without these microorganisms it is 48.9 %. Thus, it can be assumed that the presence of Mycoplasma spp in samples and high bacterial contamination rates lead to to lower reproductive performance in bulls.

Sow reproductive performance following artificial insemination with semen doses processed using Single Layer Centrifugation without antibiotics in the tropics

Single Layer Centrifugation (SLC) through a low density colloid offers an alternative solution to antibiotic use in boar semen extenders, with lower costs compared to high density colloids. The aim of this study was to explore the reproductive performance of sows when using SLC-prepared semen doses without antibiotics, employing low density Porcicoll to prepare semen doses for artificial insemination in a commercial swine herd in Thailand. Ejaculates were divided into two equal parts to create insemination doses, with each dose containing 3000 × 106 sperm/80 ml for intra-uterine insemination in individual sows. The sows were inseminated twice, with the interval between the two inseminations ranging from 8 to 16 h. The CONTROL group consisted of 206 semen doses treated with antibiotics, prepared for insemination in 103 sows, while the SLC group comprised 194 SLC-prepared semen doses without antibiotics for inseminating 97 sows. Fertility and fecundity traits, including non-return rate, conception rate, farrowing rate, and litter traits (i.e., the total number of piglets born per litter, number of piglets born alive per litter, number of stillborn piglets, and number of mummified fetuses), were compared between groups. Furthermore, data on piglet characteristics, including live-born and stillborn piglets (i.e., the prevalence of stillbirth (yes, no), birth weight, crown-rump length, body mass index (BMI), and ponderal index (PI)), were determined. No significant differences in non-return rate (75.7 % vs. 77.3 %), conception rate (73.8 % vs. 73.2 %), and farrowing rate (71.8 % vs. 73.2 %) were observed between the CONTROL and SLC groups, respectively (P > 0.05). Nevertheless, the total number of piglets born per litter in the SLC group was higher than in the CONTROL group (14.6 ± 0.9 vs. 12.3 ± 0.6, respectively, P = 0.049). Interestingly, the prevalence of stillbirth in the SLC group was lower than in the CONTROL group (6.2 % vs. 11.6 %, respectively, P < 0.001). Moreover, the newborn piglets in the SLC group exhibited higher birth weight and BMI compared to those in the CONTROL group (1.36 ± 0.03 vs. 1.26 ± 0.02 kg, P = 0.005, and 18.3 ± 0.3 vs. 17.3 ± 0.2 kg/m2, P = 0.003). In conclusion, employing sperm doses after SLC through a low density colloid in artificial insemination within a commercial breeding operation did not have a detrimental impact on either fertility or fecundity traits but showed potential benefits in increasing the total number of piglets born per litter. Moreover, improvements were observed in the birth weight and body indexes of piglets, and the percentage of stillbirths was reduced. Our findings introduce new possibilities for antibiotic alternatives in semen extenders to reduce the risk of antimicrobial resistance in the swine industry. Additionally, they provide compelling reproductive outcomes supporting the integration of SLC-prepared semen doses into artificial insemination practices.

Comparative effectiveness of different methods of using the analogue of gonadotropin-releasing hormone for ovulation stimulation in rabbit does

Modern reproductive technologies require artificial insemination (AI) protocols that are safe for the health of animals and do not violate their welfare. Stress factors arising in the reproduction process of industrial rabbit breeding negatively affect the sexual function and reproductive performance of rabbits and their offspring. In recent years, researchers have focused on alternative methods of ovulation stimulation, including intravaginal introduction of hormonal means of its induction. Therefore, the aim of our study was a comparative evaluation of the effectiveness of the intramuscular method of administration of the gonadotropin-releasing control (GnRH) analogue and its addition to the sperm dose for AI on Hyplus rabbit (n = 30), which were randomly divided into three groups. For this, buserelin acetate was administered intramuscularly at a dose of 1 μg per rabbit immediately after artificial insemination (control group). Experimental animals were inseminated with sperm doses with the addition of a GnRH analogue in the amount of 25 μg (experimental group 1) and 15 μg (experimental group 2). The influence of the method of administration of GnRH analogue on reproductive function was evaluated by indicators of the effectiveness of AI, fertility, and the dynamics of the progesterone level during pregnancy. A decrease in fecundity was observed in females; for example, in experimental group 1, the effectiveness of AI in terms of the number of rabbits was 64.3 %; in animals of experimental group 2 – 41.2 %, and the number of inseminations in this group of rabbits was 41.7% higher compared to control data. The fertility of rabbits of all groups was high, reaching the maximum value in the control group (10.42 ± 0.52 heads/female). Fertility according to live newborns in experimental group 1 was the highest among all groups of rabbits and was 9.67 ± 0.49 heads/female, which showed an increasing trend compared to the control. Also, in the animals of experimental group 1, an increase in the level of progesterone in blood serum was observed (on the 7th day of pregnancy by 13.2 %, on the 14th day by 14.3 %, on the 21st day by 16.4 %). In contrast, in experimental group 2, a significant decrease in the progesterone level was noted. Thus, 25 μg of GnRH analogue added to the extender is optimal. It is worth noting that there were no advantages of intravaginal administration over intramuscular administration in terms of reproductive performance. However, the number of live births and progesterone levels were improved.

Application of Flow Cytometry Using Advanced Chromatin Analyses for Assessing Changes in Sperm Structure and DNA Integrity in a Porcine Model.

Chromatin status is critical for sperm fertility and reflects spermatogenic success. We tested a multivariate approach for studying pig sperm chromatin structure to capture its complexity with a set of quick and simple techniques, going beyond the usual assessment of DNA damage. Sperm doses from 36 boars (3 ejaculates/boar) were stored at 17 °C and analyzed on days 0 and 11. Analyses were: CASA (motility) and flow cytometry to assess sperm functionality and chromatin structure by SCSA (%DFI, DNA fragmentation; %HDS, chromatin maturity), monobromobimane (mBBr, tiol status/disulfide bridges between protamines), chromomycin A3 (CMA3, protamination), and 8-hydroxy-2'-deoxyguanosine (8-oxo-dG, DNA oxidative damage). Data were analyzed using linear models for the effects of boar and storage, correlations, and multivariate analysis as hierarchical clustering and principal component analysis (PCA). Storage reduced sperm quality parameters, mainly motility, with no critical oxidative stress increases, while chromatin status worsened slightly (%DFI and 8-oxo-dG increased while mBBr MFI-median fluorescence intensity-and disulfide bridge levels decreased). Boar significantly affected most chromatin variables except CMA3; storage also affected most variables except %HDS. At day 0, sperm chromatin variables clustered closely, except for CMA3, and %HDS and 8-oxo-dG correlated with many variables (notably, mBBr). After storage, the relation between %HDS and 8-oxo-dG remained, but correlations among other variables disappeared, and mBBr variables clustered separately. The PCA suggested a considerable influence of mBBr on sample variance, especially regarding storage, with SCSA and 8-oxo-dG affecting between-sample variability. Overall, CMA3 was the least informative, in contrast with results in other species. The combination of DNA fragmentation, DNA oxidation, chromatin compaction, and tiol status seems a good candidate for obtaining a complete picture of pig sperm nucleus status. It raises many questions for future molecular studies and deserves further research to establish its usefulness as a fertility predictor in multivariate models. The usefulness of CMA3 should be clarified.

ЗООРОБОТ ДЛЯ ПЛЕМЕННОГО СКОТОВОДСТВА

A study of the daily time budget on the country's agricultural enterprises showed that the longest production processes are: animal’s rest is 12 hours or 50% of the daily budget; bulls’ sperm collecting - 7 hours or 29,2%; feeding - 3 hours or 12,5%; manure removal - 2 hours or 8,3%. Most of agroindustry processes on enterprises are carried out manually. By means of simulation modeling, this study was conducted on the possibilities of zoorobot using to increase efficiency and reduce the cost of the daily time budget for sire seed’s collecting material. Currently, a nonmechanical or mechanical mannequin with an adjustable tilt angle or a dummy animal is used to sperm collecting. The vagina’s preparation also manually is carried out, but at the several preassembled vaginas use it can be done mechanically. These studies have shown that zoorobots with an automatically adjustable tilt angle as mannequins for sperm collection can be used. Automation of these processes reduces the seed collection process from a bull in 3,8 times. The international practice of cattle breeding enterprises for seed collection material from breeding bulls is such that the main 40 thousand sperm doses from the manufacturer must be obtained in the first 2-2,5 years of its operation. At the same time, it should be borne in mind that the equipment preparation for seed’s harvesting of one bull it takes at least for 15-20 minutes of working time, i. e. during 8 hours of working time, 57 bulls can be used, at a doublet cage - 29 bulls, and with a triplet cage - 19 bulls, if there are 140 main bulls at the breeding plant. At a zoo robot using, the queue for bull's sperm collecting will decrease: in one cage - once every 2 days, in a doublet cage - every 3 days, in a triplet cage - every 5 days, or m/2n times, where m is the number of bulls, n is the number of minutes spent on the seed collecting from one bull.

Association of coenzyme Q9 (COQ9) gene polymorphism with traits of reproductive qualities of holstein cows

The paper presents the results in 2019-2022 of studying the associations of polymorphism of the coenzyme Q9 (COQ9) gene, which a ects the energy and metabolic processes in the body, with traits of the reproductive qualities of Holstein heifers. The experimental population was selected from animal analogues (251 heads).DNA testing of allelic polymorphism G → A (guanine → adenine) of the COQ9 gene, which results in changes in oxidative phosphorylation, that a ects the respiratory function of mitochondria, was carried out by PCR-RFLP. Two alleles (A - 0.604; G - 0.396) and three genotypes (AA - 36.6 %; AG - 48.2 %; GG - 15.5 %) have been identi ed, in the COQ9 - BstMB I gene locus, that indicates genetic biodiversity populations. The chi-square takes on a value well below the critical value (0.05). The population is in genetic equilibrium according to the Hardy- Weinberg law. The traits of the reproductive ability of cows (251 animals) of di erent genetic groups of the COQ9 gene have been studied. An analysis of the relationship between fertility rates and genotypes of the COQ9 gene showed that individuals with the COQ9AA genotype were characterized by increased live weight during insemination and at calving. Animals with the COQ9AG genotype were characterized by a later age of the rst fruitful insemination, while the number of sperm doses spent on them was minimal. The same group of heifers had the shortest calving interval, open day and service period and, as a result, a greater yield of live calves, a high fertility index of Dohi and a reproductive ability coe cient. The data presented in our research prove the in uence of genetic factors on the reproductive qualities of cattle, which makes it possible to develop systems for managing the productivity of farm animals based on the use of molecular genetics methods

Optimization of the Equine-Sperm Freeze Test in Purebred Spanish Horses by Incorporating Colloidal Centrifugation.

The Purebred Spanish Horse, according to our clinical experience, is characterized by having a high number of stallions that do not meet the international commercial recommendations for equine-sperm cryopreservation. This means that artificial insemination with frozen semen from these stallions is less widespread than in other breeds. In this study, we investigated if the incorporation of single-layer colloidal centrifugation prior to cryopreservation in clinical conditions could increase the number of ejaculates of Purebred Spanish stallions suitable for this processing, observing the influence of centrifugation and freezing extender protocol on post-thawed sperm motility. Using colloidal centrifugation, the percentage of ejaculates available to be frozen was increased from 35% (6/17) to 71% (12/17), doubling the number of samples that could have been subjected to cryopreservation. We only found significant differences in linearity (LIN) and lateral head displacement (ALH) after 5 min of incubation at 37 °C between colloidal and simple centrifugation processing techniques. No significant differences were found between the two different colloidal protocols in any of the variables considered. Colloidal centrifugation allowed us to obtain, from worse fresh-quality ejaculates, thawed sperm doses with similar quality to that of good-quality ejaculates. BotuCrio® produced, in general, higher motility parameters and its characteristics than the other extenders analyzed, with significant differences found in comparison to Inra-Freeze® and Lac-Edta in both total (MOT) and progressive motility (PMOT) when using colloidal centrifugation and only in PMOT when applying simple centrifugation. Colloidal centrifugation optimized the efficiency of cryopreservation, as it allowed us to increase the number of ejaculates of Purebred Spanish Horses suitable to be frozen. Including these semen processing techniques in the freeze test could help to optimize equine-sperm cryopreservation protocols, especially when dealing with individuals or breeds for which initially low sperm quality prevents or limits their inclusion in sperm cryopreservation programs.

Analysis of artificial insemination center management factors that contribute to sperm parameters and boar longevity with a major focus on PGF2α treatment.

Artificial insemination (AI) centers' economic profitability is directly impacted by the amount of high-quality sperm doses. Many internal and external factors contribute to the quality of ejaculates. To enhance the libido and to reduce the time that is required to train boars for semen collection, prostaglandin products are used routinely at AI centers. The objectives of this study were to analyze sperm parameters of 40,765 ejaculates and the survival of 406 PGF2α-treated and 417 untreated AI boars of five breeds (Duroc [DU], Large White [LW], Landrace [LR], Pietrain [PI], and Large White Sire Line [LS]). Data were assessed in one AI center in Switzerland from 2018 to 2022 and sperm parameters were determined by computer-assisted sperm analysis immediately after semen collection. For further calculations, the ejaculates were divided into four groups depending on the boar's age at semen collection day: 7 to 11, 12 to 18, 19 to 24, and 25 to 87 mo. Along with the boar age, breed, season, and semen collection frequency even the treatment with PGF2α had significant influence on sperm parameters. Overall, 19.5% ejaculates were collected after PGF2α treatment. For every age and breed group, higher ejaculate volume was found for untreated boars than for PGF2α-treated boars (P < 0.001). Higher values for total and progressive sperm motility, total sperm number, and the proportion of morphologically normal sperm were observed in untreated boars across several age and breed groups. Only for sperm concentration, higher values were found for treated boars older than 11 mo (P < 0.001) and the breeds DU, LW, and LR (P < 0.001). The chance of ejaculates being rejected for AI purposes because of low sperm quality was 21% greater for treated boars. There was no difference in survival between treated and untreated boars. The hazard for boar removal increased with decreasing age at first semen collection and decreasing semen collection frequency. The results reveal better sperm quality for untreated boars than for PGF2α-treated boars, which might indicate that the underlying fertility of boars necessitating treatment is inferior compared to boars that can be collected without further treatment. Therefore, AI boar stations that are aiming to reduce PGF2α treatment should pay special attention to high libido and fertility of young prospective AI boars.

ECONOMIC EFFICIENCY OF THE INCLUSION OF VARIOUS DOSES OF THE FEED ADDITIVE NANOPLANT CHROME (K) IN THE DIET OF SIRE BULLS

As a result of the studies, it was established that the use of the feed additive Nanoplant Chrome (K) (0.2 mg per 1 kg of dry matter) in feeding sire bulls contributes to an increase in the economic efficiency of obtaining sperm production by 11.0% due to an increase in the volume of ejaculate by 4.6%, sperm activity – by 2.5%, sperm concentration – by 7.9%, the number of frozen sperm doses – by 10.1%, sperm fertilization capacity – by 4.1 pp, and a decrease in the number of rejected frozen sperm doses – by 0.6-0.8 pp.

ФОРМИРОВАНИЕ ГЕНЕАЛОГИЧЕСКОЙ СТРУКТУРЫ ПОГОЛОВЬЯ БЫКОПРОИЗВОДЯЩИХ КОРОВ И ПЛЕМЕННЫХ БЫКОВ ГОЛШТИНСКОЙ ПОРОДЫ МОЛОЧНОГО СКОТА ОТЕЧЕСТВЕННОЙ СЕЛЕКЦИИ В ПЛЕМЕННЫХ СЕЛЬСКОХОЗЯЙСТВЕННЫХ ПРЕДПРИЯТИЯХ МИНСКОЙ ОБЛАСТИ С УЧЕТОМ ГЕНОТИПОВ ПО β -КАЗЕИНУ

For the first time in the Republic of Belarus, following the example of agricultural enterprises of the Minsk region, there has been formed the genealogical structure in herds of bull-producing cows and breeding bulls of the Holstein dairy cattle breed of domestic selection, considering the β -casein genotypes. It has been established that animals with the A2A2 β -casein genotype belong to all six planned genealogical complexes that make up the structure of the Holstein breed. The RUAE Minsk Pedigree Breeding Enterprise possesses a sufficient stock of sperm doses taken from breeding bulls with the A2A2 β -casein genotype to obtain replacement bulls of all complexes and develop selection herds with the desired genotype, producing A2 milk.

Influence of the exercise regime on the quantitative indicators of the semen of breeding boars

Currently, domestic pig breeding faces the problem of maintaining high productivity of animals due to the increase in the number of breeding farms. The most important problem in pig breeding is the problem of the correct selection of breeding boars, the providing of better feeding and housing conditions for them and obtaining more high-quality sperm. The purpose of the research was to study the e˜ect of the exercise regime on the quantitative indicators of the semen of breeding boars. The research was carried out under the conditions of OOO “Pig complex “Tyumensky” in the Tyumen region. Three groups of breeding boars with 10 heads in each were selected for research. The control group did not receive exercise, the 1st experimental group received passive exercise, and the 2nd experimental group received active exercise. The analysis of quantitative indicators of semen showed that boars of the 1st experimental group had the volume of ejaculate higher by 7,64 ml compared with similar indicators characterizing the reproductive traits of boars of the control and 2nd experimental groups. Due to the larger volume and concentration of sperm in the ejaculate by 3–6 more doses of sperm were obtained from one boar of the 1st experimental group than from the boar of the control and 2nd experimental groups. Thus, the use of passive exercise for breeding boars contributes to a larger volume of ejaculate and a larger number of sperm doses received from the boar, which undoubtedly improves herd reproduction and increases the e›ciency of the pig industry.

Sperm Quality in Young Bull Semen Can Be Improved by Single Layer Centrifugation.

Simple SummaryGenomic selection enables bulls with desirable genes to be identified early in life. Livestock producers need to use the semen from young bulls as early as possible for efficient milk and meat production with fewer greenhouse gas emissions. However, semen from young bulls is often of lower quality than needed for freezing for commercial artificial insemination. Colloid centrifugation selects spermatozoa with the desirable characteristics needed for fertilization from the rest of the ejaculate. In this study, split ejaculates from young bulls were prepared with or without colloid centrifugation. Using this technique, sperm doses of acceptable quality for artificial insemination could be produced from ejaculates that would otherwise be discarded. Thus, the semen from young bulls would be usable for artificial insemination sooner than is currently the case.Interest in using semen from young bulls is increasing due to identifying promising animals by genomic selection. However, sperm quality in these ejaculates may not reach currently accepted standards for the cattle breeding industry. The purpose of this study was to determine if centrifugation of semen from young bulls through the Bovicoll colloid could improve sperm quality sufficiently for the frozen semen to be acceptable for artificial insemination. Ejaculates from 19 young bulls were split and either processed by Single-Layer Centrifugation (SLC) or not (CON) before freezing. After thawing, sperm quality was evaluated by determination of membrane integrity, mitochondrial membrane potential, DNA integrity, production of reactive oxygen species, sperm morphology and motility. Approximately half of the CON samples reached acceptable post-thaw quality (membrane integrity ≥ 40%) despite being below the breeding company´s desired sperm concentration threshold pre-freezing. In the remaining samples, sperm quality was improved by SLC such that 45% of them reached acceptable quality post-thaw. Almost 75% of the young bull sperm samples could have produced usable frozen semen doses by adjusting the breeding company´s current processing protocols. Since lowering the generation interval has a direct effect on the genetic gain per year, SLC could aid genetic progress in cattle breeding.

Influence of feed sorbents on the productive and reproductive traits of cows of Black-and-White breed

In the composition of feed for cattle, the presence of toxins that are formed as a result of violations of the technology of harvesting and storage of feed is recorded. Aflatoxins, ochratoxins, zearalenone, T-2 toxin, DON (deoxynivalenol), fumonisin are isolated. Mycotoxins entering the blood have a rather negative effect on the body of cows, reducing their productivity and fertilizing ability. Under conditions of intensive milk production technology, the determination of toxins in feed when feeding highly productive cows is of particular importance. The purpose of the research was to establish the degree of influence of different dosages of feed sorbents on the productive and reproductive traits of cows. The research was carried out under the conditions of the breeding farm of Black-and-White breed in OOO “Barmino” in the Nizhny Novgorod region. The objects of research were cows of the first lactation of Black-and-White breed, and the subject of research was the dairy productivity of cows and their reproductive traits. The objectives of the research included a comparative study of the influence of feed additives “Provitol” and “Vetosporin-active” on the productivity and reproductive traits of cows. In the concentrate part of the feed, the excess of the permissible norm of the mycotoxins zearalenone by 2,5 times and ochratoxin by 4,2 times was revealed. It was found that the dairy productivity of cows and higher indicators of reproductive abilities were animals in the 4th experimental group, who have been received the largest amount of the feed sorbent “Vetosporin-active” 2 kg per ton of concentrated feed. In comparison with the analogues of other experimental groups, they have the best ease of calving, the absence of problems with the separation of the afterbirth and endometriosis, the optimal duration of the open days period and the minimum expenditures of sperm doses for effective insemination. It was recommended to use the feed sorbent “Vetosporin-active” at the dose of 2,0 kg per ton of feed as a feed additive in the main ration of highly productive cows to improve their reproductive and productive traits.

Minimum sperm dose for optimal fertility after artificial insemination in ostriches

Artificial insemination (AI) in ostriches may present potential solutions to high proportions of infertile eggs commonly recorded on commercial farms and assist in reducing the number of males for breeding purposes thereby leading to a more economical and efficient farming system. Although non-invasive methods to collect semen and to artificially inseminate female ostriches have been developed, the insemination dose for maximum fertility of eggs remains unknown. This study was thus conducted to determine the minimum sperm dose that would promote fertility of eggs following AI in female ostriches. A total of 22 South African black ostriches (7 males and 15 females) aged between 2 and 9 years old were used. Semen samples were collected using the dummy female method and diluted 1:4 (semen: diluent) with an ostrich specific semen diluent. Females were inseminated with various sperm doses of diluted semen from the same male three times a week, every second day resulting in a total sperm dose of A: <2.5 × 109, B: 2.5–5 × 109, C: 5–7.5 × 109 and D: 7.5–9.6 × 109 sperm/week. Eggs produced after insemination were opened to determine the fertilization status of the germinal disc (GD) with an unaided eye. The fertile period was then calculated as the number of days fertilized eggs were laid after the last AI. Furthermore, the number of sperm trapped in the outer perivitelline membrane (spermOPVL) above the GD region was counted under fluorescent light, following staining with 4′,6-diamidino-2-phenylindole to determine the rate of sperm loss and the number of days up to when the last egg containing sperm was laid. On average, a mean (±sd) of 35.34 ± 25.72% eggs produced after AI were fertilized. Fertility was lower (mean ± se) when sperm dose A was used (6.71 ± 9.40%), as compared to sperm dose B (46.01 ± 6.71%), C (37.34 ± 6.60%) and D (37.75 ± 8.36%) (P < 0.05). No significant difference was recorded in the latter three doses (P > 0.05). Furthermore, the length of the fertile period and the rate of sperm loss did not differ significantly between sperm doses (P > 0.05). Hence, a sperm dose of between 2.5 and 5 × 109 sperm/week is recommended to optimize fertility after AI in ostriches, as increasing the sperm dose would not benefit fertility. Further studies are, however, needed to determine the frequency of insemination that would maintain fertility throughout the breeding season as well as hatchability of eggs laid after AI.

Fertility prediction in dairy goats from Murciano-Granadina breed: The role of sperm evaluation and female traits

Fertility is one of the most economically important traits in farm animals, due to the direct and indirect costs associated to low pregnancy rates. Thus, one of the priority goals in animal reproduction is to predict the performance that the semen doses will have in vivo based on the quality values obtained in laboratory assays. Attempts have been made for getting a predictive model of fertility of frozen-thawed sperm in dairy goats, but similar studies have not been conducted for chilled goat buck sperm doses that are mostly used for artificial insemination in many countries including Spain. We study how parameters of in vitro sperm quality and characteristics of Murciano-Granadina dairy goats may affect the in vivo fertility obtained after artificial insemination with semen doses chilled at 4 °C. Moreover, this information was used for obtaining predictive models of the fertility. Sixty-three ejaculates from 13 males were used to prepare chilled doses for the insemination of 495 goats over 13 sessions. Fresh and chilled sperm were evaluated for motility and plasma membrane integrity with a computer-assisted sperm analysis system and flow cytometry, respectively. Fertility was determined at parturition, according to the kidding goats. Overall fertility was 59.6%. Pearson’s correlation coefficients between in vivo fertility and quality variables of fresh sperm were not significant and were low (below 0.34 in absolute value) for chilled sperm. Females’ characteristics had a low negative impact on fertility (correlation coefficients of −0.19 with age, −0.20 with parturitions and −0.11 with total milk yield obtained in the best lactation). Fixed and mixed logistic regression procedures were used trying to explain the fertility results. None of the models accurately predicted fertility, but the best models included the percentage of total motile sperm or average path velocity from fresh semen, age of the females and the session effect (uncontrolled environmental effects). These analyses showed that primiparous goats were 2.42 times more likely to get pregnant than goats that had kidded four or more times. Our field assay data on fertility in Murciano-Granadina dairy goats highlighted the importance of making quality controls of sperm, of choosing the doses presenting high percentages of motile sperm exhibiting regular trajectories and of selecting the youngest goats for AI, after their first kidding. Efforts should continue to obtain better predictive models for improving fertility in goat dairy herds.

20 Trials and Tribulations of Countering Insufficient Sperm Quantity with Sperm Quality, Fertility, Synchrony and Deeper Site of Insemination

Abstract Superior tools and techniques for boar semen evaluation, semen preservation and artificial insemination (AI) have helped build industry confidence and competence to reduce sperm per female inseminated without compromising reproductive results. Over the last 30 years, the average number of sperm per AI dose has decreased from about 5 to 2 billion in the US and doses containing 1.5 billion sperm have become common in some other countries. While even more genetic lag can be eliminated by further reduction of sperm dose, the level at which fecundity and fertility loss occurs is not a constant and likely varies between different farms and production systems. In general, uterine delivered sperm doses near or lower than 1 billion tend to reduce litter size and further sperm dose reduction eventually reduces pregnancy rate. Deeper catheter deposition of semen, beyond the uterine body, does not seem to effectively compensate for a limiting quantity of sperm. The exception is that surgical injection of sperm into the oviducts requires much less sperm than intrauterine insemination. Reduction of the AI-to-ovulation interval, from 24 to 8 h for example, does not seem to ameliorate the negative effects of a limiting number of sperm when sperm quality is high. The exception may be that a shorter AI-to-ovulation interval can increase reproductive results when both sperm quantity and quality are limiting. Examples include low boar fertility, advanced semen age, or sperm quality compromised by cryopreservation or sex-sorting. A better understating of boar fertility, the interaction of sub-populations of sperm with the oviductal reservoir and their participation in fertilization is likely requisite to continued reduction of the number of sperm required per female. Technologies that predict or regulate sperm quality, fertility and ovulation could prove useful in the quest to use sperm more efficiently and add value to swine AI.

The Quality of Buffalo Sperm Following Preservation Using Different Diluents and Sperm Concentrations

Artificial Insemination (AI) success depends on the quality of the frozen semen. The quality of the frozen semen of swamp buffalo in Indonesia is still low. The study was conducted to determine the quality of buffalo sperm following freezing using three different diluents and three different doses. The study used buffalo semen from the Tuah Sakato Artificial Insemination Center, Payakumbuh (n = 3). The semen collecting was carried out once a week for 10 weeks (replication). The research method used was 3x3 factorial randomized block design. The first factor was diluent (Triladyl®, Andromed® and Tris egg- yolk) and the second factor was the dose of spermatozoa (10 and 15 and 20 x 106 sperm/ml). Data were analyzed using variant analysis, while the differences between treatments were tested by Duncan Multiple Range Test. The results showed that the plasma membrane integrity of buffalo sperm was found in Andromed® diluent, while tris egg-yolk diluent gave better motility, viability, plasma membrane integrity and recovery rate at a sperm concentration of 20 x106 sperm/mL compared to triladyl® diluent and a sperm concentration of 10 and 15 x106 sperm/mL. It was concluded that andromed® diluent and tris egg-yolk gave better motility, viability, plasma membrane integrity and recovery rate at a sperm concentration of 20 x106 sperm/mL compared to triladyl® diluent and a sperm concentration of 10 and 15 x106 sperm/mL. of Buffalo of sperm abnormalities not sicnificantly by the type of diluent but are influenced by sperm concentration.

Is Mitochondrial Uncoupler Dinitrophenol Effective in Counterbalancing the Dilution Effect in Buffalo Semen?

Semen ejaculates were collected from Murrah buffalo (Bubalus bubalis) bulls and were diluted with low density lipoprotein (LDL) based extender into 20 million sperm/0.25 mL and 2 million sperm/0.25 mL. Dinitrophenol (DNP) was added (@ 0, 1, 10 and 50 µM) to 20 million and 2 million sperm concentration cryopreserved. After thawing, the parameters studied were plasma membrane integrity by HOST, sperm motility and kinetics by CASA, the thermal resistance of sperm by incubation test, mitochondrial superoxide status by MitoSOX through flow cytometry, mitochondrial membrane potential (MMP) evaluation by JC-1 through flow cytometry. There was no significant (P&gt;0.05) change in plasma membrane integrity, sperm motility and kinematics; thermal resistance of sperm, mitochondrial superoxide status and hMMP in comparison to control within the 2 and 20 million sperm doses. Two million sperm doses resulted in low plasma membrane integrity, low thermal resistance of sperm, high mitochondrial superoxide status and decreased high MeMP and no significant (P&gt;0.05) change in sperm motility and kinematics, but reduced total motility, beat cross frequency and no change in progressive motility, straight linear velocity, average path velocity, curvilinear velocity, the amplitude of lateral head displacement, straightness, linearity, and wobble in comparison to 20 million sperm doses. Supplementation of DNP (0, 1, 10 and 50 μM) in extender failed to improve semen quality in both 2 and 20 million sperm doses in buffalo.

КРИОРЕЗИСТЕНТНОСТЬ СПЕРМЫ ЖЕРЕБЦОВ В РАЗНЫЕ МЕСЯЦЫ ГОДА

It is known that the seasonality of reproductive function is much more pronounced in mares than in stallions. The mare goes through an anestrus period in autumn and winter, while spermatogenesis in stallions continues throughout the year. Therefore, studies of cryoresistance of stallion sperm in different months of the year are relevant. The aim of the study was to study the cryoresistance of stallion sperm in different months of the year when creating cryopreservation of genetic material with intensive breeding use of sires. The cryoresistance of the sperm of stallions with their intensive breeding use was the best from September to December during sexual rest. The highest incidence of sperm was in December, which is 0.44% more than in October, 5.12% more than in November, 7.62% more (P&lt;0.05) than in September, 20.53% more (P&lt;0.01) than in January, 27.3% more (P&lt;0.01) compared to February, 52.7% more (P&lt;0.001) than in March, 137.9% more (P&lt;0.05) than in April, 34.12% more (P&lt;0.001) than in May, 23.5% more (P&lt;0.001) than in June and 59.7% more (P&lt;0.001) than in July. The survivability of sperms after thawing in a thermostat at 37°C for less than 3 hours was observed in March, April and July. The lowest sperm motility after defrosting was observed in April, which is 36.4% less (P&lt;0.001) than in July, 38.6% less (P&lt;0.001) than in May, 45.12% less (P&lt;0.001) than in June, 50.1% less (P&lt;0.001) than in in January, 52.6% less (P&lt;0.001) than in February, 43.75% less (P&lt;0.001) than in March, 53.8% less (P&lt;0.001) than in September, 55% less (P&lt;0.001) than in October, 54.4% less (P&lt;0.001) than in November and 54.1% less (P&lt;0.001) than in December. Based on the data obtained on the mobility of deconserved sperm doses, we mat expect the highest fertility from the genetic material that is harvested from September to December.