Thesis (Ph.D. (Animal Production)) --University of Limpopo, 2018 / Pigs are of high economic importance, especially among the smallholder pig farmers as they contribute to human nutrition, food security, poverty alleviation, enhanced livelihood and creation of employment for the rural community. However, reproductive inefficiency is the main limiting factor due to inaccessibility to superior germplasm. Therefore, advances in reproductive technologies such as oestrus synchronization and artificial insemination (AI) offers unprecedented opportunities for livestock improvement for smallholder pig farmers. The first objective determined the status of pig productivity in smallholder farms of Gauteng Province prior to the introduction of advanced reproductive technologies (ARTs). The population was divided into four strata, namely West Rand, Ekurhuleni, Tshwane and Sedibeng district municipalities. A proportional stratified random sampling procedure was used to select 71 smallholder pig farmers with the assistance of extension officers from Gauteng Department of Agriculture and Rural Development (GDARD). The majority of the respondents were males (67%) and were above 50 years of age (67%), whilst 56% of the respondents had high school education. Majority of the respondents privately owned the farms (62%) and the farm infrastructure had facilities with low cost housing and modern facilities. Additionally, 47% of the respondents fed their pigs with feed swill. A large proportion of the respondents did not vaccinate their pigs (81%). Majority of respondents did not identify their pig herds (63%). Interestingly, majority of the respondents did not have breeding boars (73%) and sold their pigs at auctions (70%). The second objective determined semen characteristics evaluated by a Computer Aided Sperm Analyser® (CASA®) as a measure of boar fertility to be used for artificial insemination (AI). Sixteen ejaculates were collected from three Large White boars that are routinely used for semen collection purposes using the gloved-hand technique. The semen was extended with a commercial semen extender; Beltsville Thawing Solution, and the AI dose used consisted of 80mL semen sample (3×109 spermatozoa/mL). Aliquots of diluted semen were evaluated for spermatozoa motility using CASA®. Spermatozoa viability was evaluated using Synthetic Binding CD-14 (SYBR+)/Propidium Iodide (PI-), whereas spermatozoa morphology was evaluated using Eosin Nigrosin stain. The average semen volume, concentration and pH were 210 mL, 264.8 x 106 spermatozoa/mL and 7.1, respectively. The average values for total spermatozoa motility was 95.1%, ranging from 82.7 and 98.5%. However, XIX there were lower values found for progressive spermatozoa motility, ranging from 13.6 to 39.0%. The mean values for morphologically normal spermatozoa ranged from 47.8-60.9% and live spermatozoa ranged from 71.8-77.7%. The third objective determined sow fertility following AI at smallholder farms A total of 73 multiparous sows were artificially inseminated. Conception rates, farrowing rates, litter size and number born alive were recorded. The average conception and farrowing rates were 78.1 and 57.5%, respectively. Furthermore, AI resulted in acceptable fecundity (i.e., 11.8 litter size and 10.0 number of piglets born alive). The fourth objective determined the relationship between spermatozoa quality characteristics and sow fertility at smallholder farms in Gauteng Province. Of all fertility characteristics studied, conception rate was significantly related to total spermatozoa motility rate (r= 0.37, P<0.01), progressive motility (r= 0.31, P<0.01) and rapid motility (r= 0.40, P<0.01), although relatively low. There was a low negative relationship between spermatozoa morphological characteristics and fertility (P>0.05). The fifth objective determined the pre-weaning growth performance of piglets born following AI at smallholder farms of Gauteng province. Individual piglets were weighed using an electronic weighing scale. Litter size, number of piglets born alive, number of piglets weaned, the average piglet birth weight and average piglet weaned weight were recorded. The average litter size was 11.8 ± 0.2. The average birth weight and weaning weights were 1.9 and 6.2 kg, respectively. No significant differences were found between male and female piglets for all the growth performance characteristics. Piglets born during winter had a significantly higher (P<0.05) birth and weaning weight as compared to autumn and summer months. Season had a significant effect on birth and weaning weight (P<0.01). However, sex of piglets had no effect on all the characteristics recorded (P>0.05). The interaction between sex and season was only observed on the total number of weaned piglets (P<0.01). A highly significant positive correlation was found between litter size and number of piglets born alive (r= 0.86) and total number of piglets weaned (r= 0.50). A highly significant correlation was found between total number of piglets born alive and total number of piglets weaned (r= 0.55). In conclusion, the study demonstrated the potential benefit of adopting AI technology under smallholder production systems to disseminate superior genetic material to smallholder pig farmers in Gauteng Province. The total spermatozoa motility, progressive and rapid spermatozoa motility were the only spermatozoa motility characteristics significantly correlated XX with conception rate. Conversely, litter size and number born alive were not correlated with CASA® spermatozoa motility attributes. No relationships existed between spermatozoa morphological characteristics and fertility. The sex ratio percentage of piglets born following AI was 52:48% (females: males). The number of piglets born alive was 10.2 and 9.5 for number piglets weaned. Season influenced birth to weaning weight. However, sex had no significant influence at birth and weaning weight. Litter size affects the number of piglets born alive and weaned. The study showed that the introduction of advanced reproductive technologies improved productivity of pigs at smallholder pig farms in Gauteng Province. / Gauteng Department of Agriculture and Rural Development (GDARD) and Southern African Science Service Centre for Climate Change and Adaptive Land Management (SASSCAL

La transferencia nuclear de células somáticas (SCNT) en la especie porcina se ha convertido en una herramienta muy útil para para la elaboración de modelos genéticos de enfermedades humanas y para el uso en xenotransplantes. Aunque el número de cerdos clonados aumenta cada año, la eficiencia total de esta tecnología es todavía muy baja. Uno de los pasos más difíciles de la SCNT en porcino es la enucleación del ovocito, principalmente debido a que su citoplasma contiene numerosas gotas lipídicas. El principal objetivo de la tesis fue evaluar el efecto de diversas técnicas para visualizar la placa metafásica y el corpúsculo polar sobre la capacidad de desarrollo de ovocitos porcinos madurados in vitro. / Somatic cell nuclear transfer (SCNT) technology in porcine has become a very useful tool for the elaboration of genetic models for human diseases and the use in xenotransplantation. The efficiency of SCNT is still very low, although the number of cloned pigs increases each year. One of the hardest steps of porcine SCNT is the enucleation of the oocyte because its cytoplasm contains many lipid droplets. The main objective of this thesis was to assess the effect of several approaches to visualize the metaphase II plate and the first polar body on the developmental ability of in vitro mature porcine oocytes.

Artificial insemination using cooled, transported semen has become a popular practice in the equine industry. However, equine sperm are assumed to show a decline in their fertilizing ability after 24 to 48 hours of cooled storage. Two measures that are commonly used to estimate the fertility of an ejaculate are sperm motility and sperm membrane integrity (SMI). Recently, it has been suggested that SMI may have a better correlation with fertility of an inseminate than sperm motility. The effect of cooled-storage on sperm quality over an extended time period was evaluated to illustrate changes in sperm characteristics that might be related to an ejaculate’s fertility. Semen was stored at 4°C in INRA 96 extender containing 10% seminal plasma for a period of 10 days. Data were collected daily on sperm motion characteristics, SMI, mitochondrial membrane potential, and DNA quality. To measure daily changes in SMI in stallion sperm, two fluorescent vital-staining protocols used in flow cytometric analysis were compared – a combination of SNARF-1, Yo-Pro-1, and Ethidium Homodimer 1 (SYE) and a combination of lectin from Pisum sativum and propidium iodide (PSA/PI). We hypothesized that the SYE protocol adapted for use with stallion sperm could detect more subtle, and perhaps earlier, damage to the sperm plasma membrane than the PSA/PI protocol. A combination of SYBR 14, propidium iodide, and JC-1 (SYPIJC) was used to measure mitochondrial membrane potential, as well as SMI. A computer-assisted sperm motion analysis (CASA) instrument was used to evaluate sperm motion characteristics; the sperm chromatin structure assay (SCSA) was used to measure the degree of DNA fragmentation. In this study, with the exception of sperm motility, the measures of sperm quality retained values consistent with “viability” after 10 days of cooled-storage. This suggests that the fertility of some stallions may last considerably longer than previously assumed, which could ultimately alter the time-table used for artificial insemination using cooled, transported semen.