Semen Doses Research Articles

175 Analysis of Cryopreserved Semen Quality: With the Tools Available, What Is Possible to Be Interpreted

Abstract Identifying doses of post-thawed semen with high and low fertility potential is the main objective pursued by professionals and companies involved in commercializing frozen semen. In post-thaw semen, we can evaluate the motility characteristics and the integrity of the spermatozoa membranes and their DNA so that we can identify the number of cells that have the minimum characteristics to be considered with the potential to fertilize the oocyte. However, even with these analyzes using fluorescent probes, flow cytometry, and a computerized semen analysis system, it is impossible to predict the dose’s fertilizing potential accurately. The spermatozoa and seminal plasma originate from an individual (e.g., boar, bull, stallion), and even the spermatozoa and seminal plasma from this individual are different between different ejaculates. Factors such as genetics, age, ambiance, nutrition, and semen manipulation can alter the cryotolerance capacity of a given ejaculate, thus affecting its fertility potential. However, recent studies with assessments of proteomics, lipidomics, metabolomics, and miRNAs have associated cryotolerance and semen fertility with markers that can be evaluated before the ejaculate cryopreservation process—in this way, creating the possibility of selecting high fertility doses before the freezing process. Suppose these biological markers will conclusively make it possible to inform whether post-thawed semen dose has high or low fertility potential. In that case, only future research work verifying fertility will allow us to know. In the meantime, it is highly recommended to evaluate the post-thaw semen by assessing characteristics of motility, the integrity of the plasma and acrosomal membranes, and the integrity of DNA. Thus, ensuring that inseminations are carried out with the minimum number of sperm able to provide a high potential for fertility. These minimum numbers are related to the species and type of cryopreserved semen: conventional or sexed.

PSVI-9 Effects of Mixing Prior to Automated Packaging in Extended Boar Semen

Abstract It is known that sperm cells will settle out of extender over time which could impact the concentration per dose within a batch. The objective of this study to investigate the effects of mixing prior to packaging on concentration of packaged insemination doses. At a commercial boar stud, three ejaculates were collected, extended, pooled and assigned to one of three treatment groups: pour-mix (PM) where ejaculates were immediately packaged after combining the three ejaculates, stir-mix (SM) where three ejaculates were combined and stirred thoroughly then packaged, or no mixing (NM) where pooled ejaculates were allowed to sit for 10 min then packaged. Semen was packaged with 35ml per tube creating ~350 individual doses of semen. Every fifth dose was labeled with consecutive numbers (~70/treatment) and overnighted to Purdue University. Concentration of each dose was determined using the NucleoCounter® SP-100TM. The entire study was repeated over 6 weeks. Within treatment, concentration did not differ among doses for the PM and SM treatments. However, NM doses were less concentrated (P< 0.001) in doses >300 compared to all others and doses 251–300 were less concentrated than doses 1 to 50. NM samples 1–250 averaged 36.12 million cells/ml whereas samples 251 to the end of the batch averaged 34.68 million/ml. This results in the last 20 doses having ~50.4 million less cells/dose. Across treatments, NM had lower concentrations (P=0.02) from doses >200 compared to the SM treatment. PM appeared to be an intermediate as it was numerically lower than SM, but not statistically different from either NM or SM. The difference in concentrations between the NM and SM >200 was 1.85 million/ml equaling 64.75 million fewer cells. Overall, allowing semen to settle and not mixing prior to packaging can result in 50–64 million cells fewer per dose in the last 50–100 doses packaged.

ВЛИЯНИЕ МЕСЯЦА ГОДА НА ИНТЕНСИВНОСТЬ ВЫБРАКОВКИ СПЕРМЫ БЫКОВ-ПРОИЗВОДИТЕЛЕЙ

The article analyzes the influence of the season of the year on qualitative and quantitative sperm parametres of servicing bulls, it also considers the structure of reasons for culling of ejaculates. The research was carried out on bulls of Holstein, Black Spotted, brown Swiss and Hereford breeds at the age of 2-6 years in OOO “Nizhegorodskoe” for breeding work”. Statistically significant influence of the month of the year on concentration and activity of spermium, the ejaculate volume, the proportion of rejection of ejaculates that do not meet the requirements was established. The sperm concentration was 0.592 billion / ml at the beginning of the year. In spring, there was an increase in concentration and a sharp jump in June to 0.886 billion / ml, then it gradually decreased. The average volume of ejaculate at the beginning of the year was quite high 7.69-8.45 ml, in June there is a sharp decrease in volume, in subsequent months there was a gradual increase in volume to 7.68 ml. The sperm activity parametres change slightly during the year. At the beginning of the year, sperm activity was 5.98 points, in March there is an increase to 6.33 points, and in July to 6.47 points. After July, there is a decrease in activity to 5.95 points. The month influence on sperm concentration was 28.18 %, on the average volume of ejaculate - 19.73 %, on sperm activity - 6.36 %, on the number of frozen semen doses - 10.49 %. The most intensive culling of native sperm occurs in winter-spring months - 16-18 %, in summer culling is reduced to 3-8 %. Thus, in June, the maximum number of semen doses per a servicing bull was received - 887.4. The main reason for culling throughout the year was a low concentration of sperm in the ejaculate, more than half of the culled ejaculates (54.3 %) had this cause, 26.6 % - low activity, 13.6 % - both defects at the same time, 5.5 % were culled for other reasons

High-speed centrifugation of extender of freeze-thaw boar semen.

In semen cryopreservation, egg yolk is still widely used as a non-penetrating cryoprotectant. Much has been developed in the search for alternatives for this biological product. This work aimed to evaluate the processed egg yolk through ultracentrifugation and/or sonication in the cryopreservation of swine semen. Twenty-seven semen doses were purchased from a commercial boar stud and processed for cryopreservation using egg yolk lactose 11% (control) extender, processed using two different methods: high-speed centrifugation and sonication. Then, they were submitted to freeze-thawing protocol and were assessed for kinematic and cell structural parameters. Samples in which extenders underwent centrifugation had better results in velocity parameters, meanwhile those that only sonication was performed had poorest results in this parameter. The preservation of the membrane and mitochondria structure had better results when the diluent was only centrifuged in comparison with the other treatments. Therefore, centrifugation of extender containing egg yolk is important for better cryopreservation of swine semen.

Developing a Cost Structure of Frozen Semen Production and Performance of Artificial Insemination for Sheep Breeding Program

Background: Artificial insemination is an important applicable tool in increasing productivity by genetic exploitation through the male line. But farmers or breeders adoption require to be known the detailed cost structure and the nature of the cost function of the new technology. In this paper, therefore an attempt is made to develop a cost structure for frozen semen dose and a representative artificial breeding program in ewes.Methods: Semen was frozen twice-weekly for a period of a month following traditional box-freezing. Variable and fixed costs associated with the ram management, semen collection, processing and freezing were estimated and used in frozen semen production cost calculation. Cost items estimated as per cost assumption model and used for artificial insemination cost calculation.Result: The mean production cost of a frozen semen straw was estimated as Tk. 41.66 where ram maintenance cost, semen processing and preservation cost and staff cost shared the maximum expenses were to be 53.69, 12.93 and 33.38%, respectively. The average performance cost of artificial insemination incurred in the study was approximately Tk. 217.31, there in the semen dose, AI pipette and hormone cost covered the top areas of expenses to be 38.34, 23.00 and 19.33%, respectively. The cost estimate of the current study will be of more informative for breeders, stakeholders and researchers in sheep breeding technology.

Sperm binding to hyaluronan is an excellent predictor of Nili-Ravi buffalo bull fertility.

This study reports the first evaluation of sperm hyaluronan binding assay (HBA) for predicting the fertility of Nili-Ravi buffalo bulls in relation to standard parameters of sperm quality. Cryopreserved semen doses of low (n=6), medium (n=3) and high fertility (n=8) bulls based on their respective return rates were used. Significantly, more spermatozoa bound to hyaluronan from the most fertile bulls (57.15%±1.44) compared with medium (42.46%±1.08) and low fertility bulls (29.70%±0.78). A strongly positive correlation (r=.824, p<.01) was found between HBA and fertility that predicts a 67.9% variability (r2 =.679, p<.01) in fertility. HBA was also strongly positively correlated with sperm viability (r=.679, p<.01) followed by their live/dead ratio (r=.637, p<.01), uncapacitated spermatozoa (r=.631, p<.01), normal apical ridge (r=.459, p<.01), motility (r=.434, p<.01), mature spermatozoa with low residual histones (r=.364, p<.01), high plasma membrane integrity (r=.316, p<.01) and nonfragmented DNA levels (r=.236, p<.05). It was negatively correlated with spermatozoa having reacted acrosome (r=-.654, p<.01). A fertility model built using a combination of sperm HBA and either sperm livability or viability predicts, respectively, 86.1% (r2 =.861, p<.01) and 85.9% (r2 =.859, p<.01) variability in buffalo bull fertility. In conclusion, sperm HBA may prove to be a single robust predictor of Nili-Ravi buffalo bull fertility.

Genetic improvement of cattle and buffaloes in smallholder production systems in India

Cattle and buffaloes have been supporting livelihoods of a large number of poor farmers in India. Eighty-five percent of 63 million dairy households are smallholder having less than three animals. The country has 191 million cattle and 105 million buffaloes and 37 distinct cattle and 13 buffalo breeds. During 2017–18, the country produced 115 million semen doses and carried out 100 million inseminations, but efforts towards genetic improvement were negligible prior to the launching of National Dairy Plan (NDP) by National Dairy Development Board in 2012. Under NDP, 13 progeny testing (PT) and 10 two-tier nucleus schemes (referred to as Pedigree Selection (PS)) have been initiated. This paper describes the experiences of implementing these programs. During the period between 2012 and 2018, the PT programs together have completed test mating of about 2,000 bulls and built the infrastructure to test annually more than 400 bulls and evaluate them for various traits with 70–100 daughter records per bull. Breeding values are estimated every six months. The PT and PS programs together have supplied 1,720 young bulls, replacing about 40% of the AI bulls in the country. The key learning of implementing PT and PS includes: implementing a classical PT program is not feasible under Indian conditions; producing young males using progeny tested semen and selecting them based on their breeding value is feasible; introduction of genomic selection is very relevant; unravelling environmental effects from genetics under the smallholder production system is the key challenge; developing an appropriate information system is the key to success; evolving right institutional arrangement is very important, and genetic improvement has to be treated as a public good and programs need to be publicly funded at least for the initial 15 years.

Influence of non-genetic factors on semen characteristics of Malabari bucks

A study was designed to analyse the effect of non-genetic factors on semen quality traits of Malabari bucks maintained at Frozen Semen Bank, Madras Veterinary College, Chennai, Tamil Nadu. Data on 675 ejaculates collected from four Malabari bucks spread over six years were used. The overall least-squares means for semen volume, sperm concentration, mass activity, initial sperm motility, pre-freeze motility, post-thaw motility and number of frozen semen doses per ejaculate were 0.81±0.01 ml, 3144±17.24 million per ml, 4.80±0.02, 79.60±0.00%, 76.31±0.00%, 35.16±0.00% and 27.16±0.59 doses, respectively. The bucks had significant influences (P<0.01) on all the traits. Season had highly significant influence on semen volume, sperm concentration, initial sperm motility, post-thaw motility, number of doses per ejaculate (P<0.01) and pre-freeze motility (P<0.05). Winter season was found to have the highest semen volume, post-thaw motility and number of doses per ejaculate, whereas initial sperm motility was highest in summer and sperm concentration and pre-freeze motility in south-west monsoon. The effect of ejaculate number was significant on post-thaw motility (P<0.01), sperm concentration and pre-freeze motility (P<0.05). Among ejaculates, significantly higher sperm concentration, pre-freeze and post-thaw motility were observed in second ejaculates. The semen samples obtained during winter season and in second ejaculate were superior in seminal characters and more suitable for cryopreservation.

Rumen bacterial community, indicators of reproduction and milk productivity at different levels of protein concentrate in the diets of dairy cows

The use of new sources of by-pass protein in the cow diets to optimize milk production and herd reproduction performance indicators in modern economic conditions is relevant. The article presents data on the use of a protein concentrate containing plant and animal components and its effect on the quality of the milk obtained, the state of the rumen bacterial community and reproduction indicators. The use of protein concentrate of both 1.0 kg and 1.5 kg/head/day contributed to improving the quality and increasing the yield of dry matter of cow's milk by 0.17-0.43 %. During the days in milk, with the use of protein concentrate it was observed that increase the content of Selenomonas by 1.7 % and 4.1%, which cleave the urea, indicates a more efficient use of nitrogen in the diet. The inclusion of protein concentrate in the composition of daily diets helps to improve reproduction indicators, reduce the duration of the service period and the number of semen doses spent.

INFLUENCE OF BREED ATTRIBUTE ON LENGTH OF BREEDING USE AND PRODUCTIVITY OF SIRE BOAR

Based on the results of a five-year research (2016-2020) average length of breeding use for sire boars of the planned breeds in Belarus was established: Yorkshire – 27.8, Landrace – 28, Duroc – 19, Pietrain – 45.2 months. The average yield of semen doses per 1 boar per month for the Yorkshire breed is 140.6, Landrace – 159.6, Duroc – 119.9, Pietrain – 141.1, hybrids – 166.4 units.

Effects of semen dosage, oviductal sperm storage and insemination interval on egg fertility, embryo mortality and hatchability in Nera Black breeder chickens

The biological basis of sustaining fertility in poultry is their ability to store sperm cells in the sperm storage tubules (SST) located in the uterovaginal junction. However, artificial insemination in poultry industry is haphazardly administered in Nigeria without regulation on semen dose and frequency of insemination for optimum fertility. The objective of this study was to establish a semen dosage and insemination interval for maximum fertility and embryonic survival in Nera black layer breeder chickens. A total of 80 breeder hens (52 weeks) were allotted to five (5) treatments with four (4) replicate per treatment. Semen was pooled from 10 matured breeder cocks and inseminated to four groups of hens at varied semen dose of 0.02mL (T1), 0.04mL (T2), 0.06mL (T3) and 0.08mL (T4) of undiluted semen while hens in T5 were mated naturally, both for two consecutive days. 0.02, 0.04, 0.06 and 0.08mL of pooled semen contained 20.43×10 , 40.87×106 , 61.30×106 and 81.74×106 motile spermatozoa. Eggs were collected, stored and artificially incubated weekly for 4 weeks. Fertility, embryo mortality and hatchability parameters were determined. Another 78 breeder hens were allocated into 4 treatments of 5 replicates per treatment with unequal number of hens and were inseminated with 0.02mL of raw semen containing 20.43 × 106 motile sperm cells at 3, 6, 9 and 12 days intervals. Fertility, hatchability and embryo mortality were determined. Data were analysed using descriptive statistics and ANOVA at &amp;0.05 Hatch of fertile eggs in T5 at week 2 (65.36±13.28) was significantly higher (p&lt;0.05) . than T1 (33.83±12.65), T2 (13.25±6.88), T3 (39.17±14.17) and T4 (28.21±11.37). At weeks 1 and 2, there was no significant different across the treatments. Fertility at 4 weeks in T1 (11.53±6.66) was significantly (p&lt;0.05) different from treatments T2 (0.00±0.00), T3 (0.00±0.00), T4 (1.66±1.66) and T5 (0.00±0.00). Total and early embryo mortality in week 3 was significantly higher (p&lt;0.05) in T1 (100.00±0.00, 95.00±5.00) than in treatments T2 (43.75±0.00, 43.75±25.77), T3 (66.67±23.57, 66.67±23.57), T4 (95.00±5.00, 85.00±15.00) and T5 (37.50±23.94, 22.92±15.73). Fertility was significantly (p&lt;0.05) higher in 3 days insemination interval (52.65±7.25) compared with 6 days (39.87±4.70), 9 days (22.98±5.71) and 12 days (36.14±6.89). At weeks 1 and 3, the hatch of fertile eggs across the treatments was not significantly (p&gt;0.05) different from one another. This study suggests that 6 inseminating semen dose of 0.02mL containing approximately 20.43×10 motile sperm cells in Nera black layer breeder chickens will give a maximum fertile period of 5 days, while insemination interval of 3 days using 0.02mLof semen gave highest fertility level.

Policies to Increase Calves Production in East Nusa Tenggara as a National Production Center

The determinant factors of the growth of calf cattle production in NTT Province (both technical/technological, economic, socio-cultural, investment, and related institutions) have not been specifically identified and measured. This is very important for the purpose of formulating and implementing related policies. The objectives of this study were to analyze the main factors affecting the level of calf production, and to formulate policy interventions to increase calf production. A research survey was carried out on parties related to the planning and development of cattle business. The quantitative approach is in the form of regression analysis of time series data. The economic phenomenon of calf production in Kupang Regency has been simplified into a mathematical model (response function). The result reported that the estimated value of the parameters in the total calf production equation model in NTT was: TPRAS= – 20434.1 + 0.2262 LTTSIP* – 0.0032 TLPRAns + 0.2147 TLLTP* + 0.1036 TPTRU* + 0.1877 TRKUT** + 0.2067TRDIB** + 0.1057 TRDVT** + 0.3647 LTPRAS* + 811.7645 TREND**. In conclusionl, the increased production of calves was dependent on the increase in support for the main production factors [number of productive cows, number of farmers, amount of feed, and amount of cement/vaccine] adequatly. Policy investment (through interest rates and realization farming loans), policy of productive cows control (through postponement of slaughter and the provision cash incentives), and policy of technology support (through realization AI doses of semen and livestock vaccines) significantly affect to production of calves.

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.

Rationalizing ex situ collection of reproductive materials for endangered livestock breed conservation

Improvements in ex situ storage of genetic and reproductive materials offer an alternative for endangered livestock breed conservation, but collections should be optimized cost-effectively to avoid duplication, and with reference to the sustainability of in situ breeds. We developed a multi-period chance-constrained optimization model to rationalize collections of endangered livestock breeds at risk of in situ extinction in Spain. The model configures collections by determining the least-cost optimal collection locations, timing and material quantities (semen doses). A decision variable defining an “acceptable level of risk” allows decision makers to specify tolerable levels of in situ breed endangerment when taking ex situ collection and storage decisions. Using data from 18 gene banks we demonstrate how collections can be rationalized, and derive cost curves relating marginal ex situ collection cost and accepted probability of in situ extinction covering the period 2018 to 2060. The modelling framework can be replicated in countries seeking to rationalize ex situ collections under limited conservation budgets and uncertain in situ extinction risks.

PSXI-9 CriaCerto - Breeding System Simulator App: a case of study in Brazil

Abstract CriaCerto App was launched in February 2019 by Embrapa, Brazil, as free access, available at www.criacerto.com. It simulates the costs and benefits of the main breeding systems for cattle production: Natural Service (NS), One Fixed Time Artificial Insemination plus NS (1FTAI), Two Fixed Time Artificial Insemination plus NS (2FTAI), and Three Fixed Time Inseminations (3FTAI). This work aimed to evaluate the use of the App as well as the results of the simulations carried out during the 12 months following its launch. From February 14th, 2019 to February 20th, 2020, 4,780 accesses to the platform were recorded, with an average access time of 3 min 30 s. Eighty percent of accesses were made using mobile phones, 15% from desktops, and 5% via tablets. These accesses resulted in 975 different simulations, where 51% were from NS, 24% of 1FTAI, 14% 2FTAI, and 11% 3FTAI. There was a difference between the prices the farmers were willing to pay for the bulls according to the reproduction method. Those who performed NS simulations were willing to pay less for bulls than those who simulated 1FTAI or 2FTAI. It was also observed that the price to be paid per semen dose on 1FTAI simulations was practically double of the 2FTAI. This indicates that farmers are willing to invest in genetically superior bulls, which are usually more expensive, but the decision is made based on the total amount of money spent. It also indicates that farmers take into account the trade-off between the cost of the bull, semen dose and the benefits of genetic improvement. In Brazil only 12% of the total beef cows participate in artificial insemination programs. Nonetheless, through this study, we concluded that farmers are opened to invest in FTAI, even though they do not want to pay high prices in semen doses.

Effects of the classification of boars according to progressive sperm motility and the extender type on the reproductive performance of a single fixed-time insemination

The study aimed to evaluate the fertility of boars according to the resistance of their semen to storage using dilution in either Short- or Long-term extender for single fixed-time insemination. From a total of 32 boars, twelve boars were classified during three semen collection (one collection/boar/week) as Low- (64.5%) or High-preservation (83.9%) capacity for maintaining progressive motility (PM) at 120 h of storage using Short-term extender. After the selection period, six ejaculates (weekly collected) from the Low- and High-preservation boars were diluted in Short- or Long-term extender (2 × 2 factorial design) for insemination and evaluation of fertility. A total of 519 weaned sows were submitted to induction of ovulation with triptorelin (OvuGel®) at 96 h post-weaning. Twenty-four hours later, estrus sows were single fixed-time inseminated (FTAI) with semen doses from the different groups of evaluation. The SAS® software was used for statistical analysis considering the class of boar, type of extender, and interaction as fixed effects. The GLIMMIX procedure was used, considering a binomial distribution for total motility (TM) and PM, binary distribution for pregnancy (PR), and farrowing rate (FR), and the total born (TB) was analyzed assuming a normal distribution with the comparison of means by Tukey-Kramer test. An interaction of class of boars and type of extender was observed for TM and PM at insemination (P < 0.001). Long-term extender increased TM in Low-preservation boars, with no effect in High-preservation boars. The ejaculates from High-preservation boars diluted in Short- or Long-term extender showed higher PM at insemination (86.8 and 87.8%, respectively) compared to those from Low-preservation boars in Short- or Long-term extender (73.2% and 77.9%, respectively). There was no effect of the interaction of boar preservation class and type of extender (P ≥ 0.163) on PR, FR or TB. However, Low-preservation boars presented lower TB (14.1 ± 0.2) compared to High-preservation boars (15.0 ± 0.2; P < 0.01). The PR (93.3 vs. 90.1) and FR (88.8 vs. 88.2) were not affected by class of Low- or High-preservation boars, respectively (P ≥ 0.187). The type of extender did not affect PR, FR, or TB (P ≥ 0.440). In conclusion, Low-preservation boars impaired the reproductive performance of single-FTAI sows by reducing TB with no apparent effect on PR or FR.

Purification of cryopreserved camel spermatozoa following protease-based semen liquefaction by lectin-functionalized DNA-defrag magnetic nanoparticles.

Although incorporating proteases into sperm medium is considered the most effective procedure to eliminate camel semen viscosity, it drastically affects viability, morpho-functional properties and, hence, fertilization potential of spermatozoa. The present work aimed at evaluating adequacy of employing magnetic nanoparticles-based sperm purification technique for eluting impaired and apoptotic camel spermatozoa from cryopreserved semen doses following protease-based semen liquefaction. Thirty cryopreserved semen doses (50 x 106 sperm/straw) representing the following liquefaction treatments: control (untreated), 0.1mg/ml papain or 5 U/ml bromelain were used (n=10 straws per treatment). Immediately after thawing (38°C for 40s), sperm concentration of each straw within treatment was readjusted to 15 x 106 sperm/mL by dilution in PBS (37°C). Sperm physical and cytological properties were then assessed (non-purified semen). Thereafter, each specimen was subjected to lectin-functionalized DNA-defrag magnetic nanoparticles sperm purification, and the same sperm traits were re-evaluated after undergoing purification (purified semen). Sperm DNA fragmentation level within each group, prior to and after magnetic nano-purification, was also determined by fluorescent imaging. The results showed a dramatic improvement (p<.05) in post-thaw motility (%), viability (%), normal sperm (%), intact acrosome (%) and HOST-reacted (%) spermatozoa in protease-liquefied semen following sperm magnetic nano-purification. Additionally, the highest (p<.05) DNA fragmentation level was recorded in all cryopreserved semen groups prior to purification, whereas the lowest (p<.05) was observed in the protease-treated specimens after magnetic nano-purification. These results indicate that protease-based semen liquefaction prior to cryopreservation in conjunction with magnetic nano-purification post-thawing holds potential for reducing the proportion of damaged and dead spermatozoa, hence improving camel sperm fertilization competence.

Reproductive performance in gilts following applications of different insemination doses and techniques

This study evaluated the applicability of intrauterine artificial insemination (IUAI) in gilts and the impact of age at insemination and different body characteristics of gilts on the success rate for cannula insertion. Additionally, reproductive performance was evaluated for IUAI and cervical artificial insemination (CAI), considering different semen dose sizes. A total of 636 gilts were assigned in a 2 × 2 factorial design: two artificial insemination techniques (CAI and IUAI) and two semen dose sizes (1.5 × 109 sperm cells/50 mL or 2.5 × 109 sperm cells/80 mL). In those gilts assigned to IUAI (n = 319) the success rate for intrauterine cannula insertion was evaluated according to weight at first detected estrus, body condition score (BCS), and age at insemination. Reproductive performance, occurrence of bleeding, and semen backflow during all inseminations were compared among groups (2 × 2). Two subgroups were evaluated regarding the time expended to perform insemination (n = 380), and the semen backflow collected during 1 h after insemination (n = 114). The success rate for intrauterine cannula insertion, based on a successful insertion in all inseminations performed during estrus, was 58.9%. Additionally, greater possibility (>60%; P ≤ 0.04) of cannula insertion was observed in heavier gilts (≥124 kg), as well as in older gilts (≥225 d) and those with greater BCS (>3). There were no differences among the groups in pregnancy rate (≥95.3%; P = 0.23), farrowing rate (≥93.7%; P = 0.54), total piglets born (≥14.5; P = 0.45), as well as, bleeding (P = 0.48) and backflow (P = 0.48) during insemination. However, the percentage of semen backflow volume and percentage of sperm cells in the backflow were lower in gilts inseminated by CAI with 1.5 billion sperm cells/50 mL (P < 0.01) than in the other groups. There was no expressive reduction in time expended to perform IUAI compared to CAI. However, gilts inseminated with 1.5 billion sperm cells/50 mL showed a lower total time to inseminate than all other groups (P < 0.01). In conclusion, higher weight, BCS, and age increased the success rate for cannula insertion. However, IUAI did not optimize the insemination procedure, and remains limited for gilts due to the low success rate for cannula insertion. Reproductive performance was not affected by IUAI or CAI using 1.5 or 2.5 billion sperm cells in 50 or 80 mL, respectively, suggesting the possibility of using CAI with 1.5 billion sperm cells/50 mL in gilts.

The Effects of Model IVS 2 MAP Progesterone, eCG, GnRH, and PGf2? on Pregnancy of Tail Fat Ewes Strain Sapudi Islands Subjected to A Short-term Synchronization of Estrus

The objective of this study was to verify the effects of engineering design of IVS 2 progesterone with 2cm thick sponge and 2 cm length and 5 cm silicon plate in the middle with 13 cm nylon cord ties to pullthe sponge position. Try to determine the effective dose of progesterone (MAP) with a dose of 60 mgprogesterone, eCG, GnRH and PGf2? on pregnancy of ewes subjected to a short-term synchronization ofestrus. Tail fat sheep ewes strain sapudi Islands received 60 mg MAP sponges model IVS 2 during 12 daysplus 500 IU eCG folligon and 2.5 mg d-cloprostenol capriglandin (0.5 ml) submucosa vulva 24 h prior toIVS 2. Control groups (T0) 10 Ewes were assigned to receive 0.9% NaCl solution 2 ml im only when estrussign showed waited for natural estrus. Group treatment 10 Ewes were (T1) IVS2 + 500 IU eCG folligon andgroup treatment 10 Ewes were (T2) IVS2 +500 IU eCG folligon 21.5 µg (0.5 ml) GnRH fertagyl gonadorelin,24 hours after withdrawal. Each group was assigned to intracervical insemination by Vaginoscopy (n = 30).Artificial insemination was performed with a single dose of fresh semen on egg yolk skim milk dilluters.For controlled mating, females were exposed to males 48 hours after sponges removed. Ten females pertreatment were subjected to transrectal ultrasound examination at 35 days late after AI. Estrous response(100.0%), pregnancy rates (T0 80% T1 70% and T2 70%), did not differ between T1 and T2 respectively (p&gt;0.05). Administration of GnRH 24 hours after IVS 2 removed does not improve pregnancy rate in estroussynchronization in Tail fat ewes strain sapudi Islands (p &gt;0.05) on day 35.

Effect of Sperm Concentration and Storage Temperature on Goat Spermatozoa during Liquid Storage.

Simple SummaryOne of the problems with the refrigerated storage of spermatozoa in goats is the short shelf life. The study of the effect of the different storage conditions on the different sperm parameters could help to increase sperm survival. In this work, we studied the effect of temperature and sperm concentration during refrigerated storage on sperm motility, mitochondrial membrane potential and DNA fragmentation. We observed that refrigerated storage of semen doses reduced the majority of sperm quality parameters, however, sperm DNA fragmentation was not affected. Storage at 5 °C preserved higher sperm motility than at 17 °C. Moreover, the reduction of sperm concentration below 500 × 106 sperm/mL did not seem to improve the quality of spermatozoa.The use of cooled semen is relatively common in goats. There are a number of advantages of cooled semen doses, including easier handling of artificial insemination (AI) doses, transport, more AI doses per ejaculate, and higher fertility rates in comparison with frozen AI doses. However, cooled semen has a short shelf life. The objective of this study was to examine the effect of temperature and sperm concentration on the in vitro sperm quality during liquid storage for 48 h, including sperm motility and kinetics, response to oxidation, mitochondrial membrane potential (MMP) and DNA fragmentation in goats. Three experiments were performed. In the first, the effects of liquid preservation of semen at different temperatures (5 °C or 17 °C), durations (0, 24 and 48 h) and sperm concentrations (250 × 106 sperm/mL (1:2 dilution rate), 166.7 × 106 sperm/mL (1:3 dilution rate) or 50 × 106 sperm/mL (1:10 dilution rate)) on sperm motility and kinetics were studied. In the second experiment, the effect of temperature, sperm washing and concentration on sperm motility and DNA fragmentation was studied. Finally, the effect of sperm concentration and duration of storage at 5 °C on sperm motility, response to oxidative stress and MMP was examined. We found that refrigerated liquid storage of goat sperm impaired sperm quality, such as motility, MMP and response to oxidation, as storage time increased; however, sperm DNA fragmentation index was not significantly affected. Liquid storage at 5 °C preserved higher total motility than at 17 °C. Moreover, we observed that the reduction of sperm concentration below 500 × 106 sperm/mL did not seem to improve the quality of spermatozoa conserved in milk-based extender in the conditions tested.