Male reproductive biotechnologies in South American Camelids Part II: Semen dilution, cryopreservation and artificial insemination.

Even though South American Camelids (SAC) are a rustic species, adapted to harsh environments, their ability to reproduce is low in their natural habitat. Conception and birth rates in camelids vary from 50 % to 90 %. This depends on the mating system used, sire and dam fertility, postpartum interval, environmental conditions, and nutrition.

Male reproductive biotechnologies in South American Camelids Part I: Semen collection, evaluation and handling.

This review describes the first steps necessary to apply any reproductive biotechnology in South American camelids (SAC) semen or sperm: sample collection, evaluation and handling. In camelids, the length and position adopted for mating and the site of semen deposition have conditioned semen collection methods. The advantages and disadvantages of available collection methods are summarized.

Evaluation of DNA integrity and chromatin condensation in cat sperm collected by urethral catheterization and epididymis slicing.

The aim of this study was to evaluate the effectiveness of the original sperm chromatin dispersion (SCD) assay and the toluidine blue (TB) stain to assess DNA fragmentation and chromatin condensation, respectively, in cat sperm obtained by urethral catheterization (CT) and epididymis slicing (EP). CT and EP samples were collected from the same cat, and sperm motility, concentration, morphology, DNA integrity and chromatin condensation were evaluated. As controls, aliquots of the samples were incubated with 0.

Use of commercial extenders, with and without the addition of egg yolk, for cooling llama semen.

The objective of this study was to evaluate the effect of two commercial extenders, AndroMed® (AM) and Androstar® Plus (AS) both with and without the addition of egg-yolk (EY), for cooling llama semen. A total of sixteen ejaculates were collected from four males. Each ejaculate was divided into four aliquots and diluted with: AM, AM with 20 % EY (AM-EY), AS and AS with 20 % EY (AS-EY) and then cooled to 5 °C in an Equitainer®.

Evaluation of DNA fragmentation in dog sperm using the sperm chromatin dispersion test.

The study's objective was to adapt the Sperm Chromatin Dispersion (SCD) protocol to evaluate sperm DNA fragmentation and implement a fragmentation control in dogs. Correlation between DNA status and routine sperm parameters was also analysed. To adapt the SCD, two different mercaptoethanol (ME) concentrations were assayed (2.

Dehydration of llama sperm using different osmolarity media and temperatures for preservation.

The objective of this study was to evaluate effects of dehydration on sperm DNA with the aim of eventually using this method for preserving llama spermatozoa. Two experiments were conducted: 1) sperm preservation at 5 °C for 60 days in different hyperosmotic solutions (500, 800, 1000 and 1200 mOsmol/l) (n = 6, replications = 2) and 2) sperm preservation at 5 and -20 °C for 60 days in the same hyperosmotic solutions, with supplementary antibiotics (n = 6, replications = 2). Sperm motility, membrane functional integrity, viability and morphology were evaluated at 0 and 48 h of the preservation period (Experiment 1) and at 30 min and 24 h (Experiment 2).

Air-Drying Llama Sperm Affects DNA Integrity.

The objective of this study was to evaluate the effects of air-drying preservation on llama sperm DNA. Semen collections were carried out using electroejaculation under general anesthesia. A total of 16 ejaculates were processed from 4 males ( = 4, = 4).

Uterine and Corpus Luteum Blood Flow Evaluation Prior to Uterine Flushing in Llama Embryo Donors.

The aim of this study was to assess the uterine blood flow (UBF) and corpus luteum blood flow (CLBF) in llamas 8 days post-mating, using color-Doppler ultrasonography (CDU), to determine the possible relationship between vascularization and the presence of an embryo. Adult females ( = 25) were used to monitor ovarian dynamics by palpation and transrectal ultrasonography until detection of a ≥6 mm growing follicle. Females were randomly assigned to one of two groups: Group I ( = 19), were mated and ovulation was induced by a single dose of buserelin (GnRH analog) that same day (Day 0); and Group II ( = 6), only ovulation was induced (control).

Development of a GnRH-PGF Based Synchronization and Superstimulation Protocol for Fixed-Time Mating in Llama Embryo Donors.

The aim of the present study was to evaluate the application of a GnRH-PGF based synchronization and superstimulation protocol for fixed-time natural mating in llama embryo donors. All females ( = 8) received 8 μg IM of GnRH analog (GnRHa; buserelin) on day 0, regardless of follicular status. After eight days, another GnRHa dose was administered followed by 250 μg IM PGF (cloprostenol).

Incubation of frozen-thawed llama sperm with seminal plasma.

Seminal plasma is intimately connected to sperm physiology and particularly in South American Camelids, has demonstrated to be involved in multiple physiological reproductive events. Different percentages of seminal plasma (0%, 10% and 50%) were added to thawed llama semen samples with the objective of evaluating the interaction with cryopreserved sperm over time (0, 1.5 and 3 hr at 37°C).

Evaluation of the cryoprotective effect of seminal plasma on llama (Lama glama) spermatozoa.

In South American camelids, sperm survival is low after thawing and poor results are obtained when artificial insemination is performed with cryopreserved semen. The aim of this study was to evaluate the effect of different percentages (10% and 50%) of seminal plasma added prior to the process of cryopreservation and also to evaluate the absence of seminal plasma on llama sperm survival after freezing and thawing. A total of 15 ejaculates from five adult llama males (n = 5; r = 3) were evaluated.

Production, Preservation, and Transfer of South American Camelid Embryos.

The current review summarizes progress in the field of and production of South American Camelid embryos. Both methods require ovarian superstimulation (with FSH and eCG) to obtain multiple ovulations ( embryo production) or to induce follicle growth for oocyte collection ( embryo production). Moreover, superstimulation entails prior administration of hormones that inhibit follicular growth (progesterone, progestagens, and estrogens).

Membrane changes during different stages of a freeze-thaw protocol for equine semen cryopreservation.

Many theories have been postulated concerning the possible effects of cryopreservation on spermatozoa, including suggestions the freeze-thawing process produces membranes that have greater fluidity and are more fusogenic, thus inducing changes similar to those of capacitation. The main objectives of this study were to determine at what stage of the freeze-thaw process membrane changes occur and whether evaluation with chlortetracycline (CTC) stain could predict the freezability of stallion sperm. Sperm viability and state of capacitation were simultaneously evaluated using CTC and Hoechst 33258 (H258) techniques.