The Importance of Having Zinc During In Vitro Maturation of Cattle Cumulus–Oocyte Complex: Role of Cumulus Cells

The aim of this study was to investigate the influence of zinc (Zn) on the health of cumulus–oocyte complex (COC) during in vitro maturation (IVM). Experiments were designed to evaluate the effect of Zn added to IVM medium on: DNA integrity, apoptosis, cumulus expansion and superoxide dismutase (SOD) activity of cumulus cells (CC). Also, role of CC on Zn transport during IVM was evaluated on oocyte developmental capacity. DNA damage and early apoptosis were higher in CC matured with 0 μg/ml Zn compared with 0.7, 1.1 and 1.5 μg/ml Zn (p < 0.05). Cumulus expansion did not show differences in COC matured with or without Zn supplementation (p > 0.05). Superoxide dismutase activity was higher in COC matured with 1.5 μg/ml Zn than with 0 μg/ml Zn (p < 0.05). Cleavage and blastocyst rates were recorded after IVM in three maturation systems: intact COCs, denuded oocytes with cumulus cells monolayer (DO + CC) and denuded oocytes (DO). Cleavage rates were similar when COC, DO + CC or DO were matured with 1.5 μg/ml Zn compared with control group (p > 0.05). Blastocyst rates were significantly higher in COC than in DO + CC and DO with the addition of 1.5 μg/ml Zn during IVM (p < 0.01). Blastocyst quality was enhanced in COC and DO + CC compared with DO when Zn was added to IVM medium (p < 0.001). The results of this study indicate that Zn supplementation to IVM medium (i) decreased DNA damage and apoptosis in CC; (ii) increased SOD activity in CC; (iii) did not modify cumulus expansion and cleavage rates after in vitro fertilization; (iv) improved subsequent embryo development up to blastocyst stage; and (v) enhanced blastocyst quality when CC were present either in intact COC or in coculture during IVM.     

Effects of supplemental dietary vitamin C on quality of semen from Nile tilapia (Oreochromis niloticus) breeders

The objective of this study was to evaluate the effects of vitamin C on growth and quality of semen from Oreochromis niloticus breeders. One hundred and sixty males were fed with different levels of vitamin C (0, 261, 599 and 942 mg/kg diet). The higher weight values were recorded for 599 (166 g) and 942 (175 g) mg of vitamin C/kg diet. Sperm motility, vigour and concentration were higher with 599 and 942 mg of vitamin C/kg diet. The semen volume, gonadosomatic index and plasma protein data from the last week showed a direct relationship with increasing levels of vitamin C. No changes were observed in the hepatosomatic index and blood glucose. The haematocrit and erythrocyte showed higher values estimated by equations derived at 850 and 638 mg vitamin C/kg diet, respectively. The leucocytes were inversely proportional to the increasing levels of vitamin C. After 100 days of feeding, animals fed the diet containing 942 mg vitamin C/kg diet had higher sperm motility, linearity, curvilinear velocity, straight line velocity and average path velocity (p < .05). Higher values of beat cross‐frequency were observed in broodfish fed diets containing 942 and 599 mg vitamin C/kg. The different vitamin C levels did not cause differences in straightness, lateral head displacement and sperm morphology. For Nile tilapia males on intensive rearing and handling conditions, vitamin C levels between 599 and 942 mg/kg may be used for a better performance and quality of semen.     

Social and Breed Effects on the Expression of a PGF2α Induced Oestrus in Beef Cows

Social organization and breed effects following PGF2αwere studied in mature Angus, Brahman and Senepol cows allocated into two groups (each A = 5, B = 5 and S = 5). Variables including interval to oestrus onset (IEO), oestrous duration (DE), total mounts received (TMR), and oestrous intensity (IE) were derived via HeatWatch®. Breed‐type influenced IEO (B = 42.6 ± 6.7 h; S = 54.6 ± 6.0 h; and A = 27.8 ± 5.8 h; p < 0.003). Within breeds, dominant B (69.4 ± 13.3 h) and S (65.5 ± 7.4 h) cows were slower (p < 0.05) to be detected in oestrus than subordinate (38.1 ± 4.4 h) and intermediate (40.6 ± 6.0 h). However, within A, dominant cows (16.4 ± 12.5 h) were detected in oestrus earlier (p < 0.05) than intermediate (44.3 ± 9.2 h) and subordinates (32.7 ± 5.1 h). Angus (21.5 ± 2.4 h) and B (22.1 ± 3.0 h) cows had longer (p < 0.01) DE than S (9.1 ± 2.8 h). Dominants (20.4 ± 3.0) and intermediates (20.2 ± 2.3 h) cows had longer DE (p < 0.04) than subordinates (12.1 ± 2.1 h) although the interaction breed × social order showed that dominant S had shorter DE than dominant A and B (10.1 ± 3.3; 34.8 ± 6.0 h; and 20.0 ± 6.4 h, respectively; p < 0.001). Angus cows had less TMR than B (p < 0.02) and tended to be less than S cows (p < 0.06). Overall, greatest (p < 0.008) IE occurred in the first 9 h after onset of oestrus with no breed effect (p > 0.05). Dominant cows tended (p < 0.10) to have less TMR (3.2 ± 0.7 mounts) than subordinate (4.1 ± 0.4 mounts) and intermediate (4.7 ± 0.6 mounts) throughout, especially 3–6 h after oestrus onset (p < 0.07). Breed and social order both influence PGF2α‐induced oestrus behaviour.     

Chemical Activation with a Combination of Ionomycin and Dehydroleucodine for Production of Parthenogenetic,ICSI and Cloned Bovine Embryos

The aim of this study was to evaluate the potential of dehydroleucodine (DhL), a new drug isolated from a medicinal herb used in Argentina, for activation of bovine oocyte. Several DhL concentrations and exposure times after ionomycin (Io) treatment were tested. The optimal DhL treatment, found for parthenogenetic development, was employed to produce bovine embryos by intracytoplasmic sperm injection (ICSI) and somatic cell nuclear transfer (SCNT). The best parthenogenic embryo developments were observed with 5 μm Io for 4 min followed by 5 μm DhL concentration and after 3‐h exposure time (52.3% cleavage; 17.4% morulae; 7.3% blastocyst; n = 109). This treatment generated no significant differences with standard Io plus 6‐dimethylaminopurine (DMAP) treatment in preimplantation embryo development. In our conditions, the embryo development reached after ICSI and SCNT assisted by the DhL treatment did not differ in terms of cleavage and blastocyst development from activation with standard Io plus DMAP treatment (p > 0.05). In conclusion, DhL utilization to activate oocytes and induce development of parthenogenotes, ICSI‐embryos or SCNT‐embryos is reported here for first time.     

Comparison of cassava,yam and potato dextrose agars as fungal culture media

In experiments in vitro at 20 and 30°C the fungiPhytophthora palmivora, Aspergillus melleus, Thielaviopsis paradoxa, Pestalotiopsis versicolor andCurvularia pallescens showed a better mycelial growth on cassava dextrose agar than on potato dextrose agar and yam dextrose agar. This also applied to sporulation, except for the last two fungi at 20°C, which sporulated best on potato dextrose agar.Samenvatting Bij verggeljking van de invloed van aardappelglucose-agar (PDA), cassaveglucoseagar (CaDA) en yamglucose-agar (YDA) op de mycelium groei en sporulatie vanCurvularia pallescens, Phythophthora palmivora, Aspergillus melleus, Pestalotiopsis versicolor enThielaviopsis paradoxa, bleek CaDa in het algemeen de beste groei (Tabel 1) en sporulatie (Tabel 2) te geven. Cassaveglucose-agar kan beschouwd worden als een goede algemene voedingsboden voor het kweken van schimmels. Vooral in de tropische landen kan men hiervan een nuttig gebruik maken. De geschiktste cassavevariëteit moet langs empirische weg geselekteerd worden.     

Fibrin–alginate hydrogel supports steroidogenesis,in vitro maturation of oocytes and parthenotes production from caprine preantral follicles cultured in group

This study aimed to establish a culture system that improves the in vitro development of caprine preantral follicles. In a first experiment, follicles were encapsulated as a single unit per bead and cultured singly or in groups or with five follicles in the same alginate (ALG) bead for 18 days. In a subsequent experiment, the “five follicles per bead” design was chosen to culture in ALG, fibrin–alginate (FA) or hyaluronate (HA) for 18 days. In a third experiment, we chose the five follicles per bead in FA to culture for 30 days. The culture set‐up of five follicles per ALG bead increased antrum formation and follicle diameter compared to the other culture designs (p < .05). Moreover, under this condition, 44.44% of the oocytes from in vitro cultured preantral follicles reached meiotic resumption. A significant increase of follicle diameter occurred in attachment system and FA (p < .05), but the ALG condition reached the highest among all groups on day 18 (p < .05). Follicles encapsulated in matrix produced more estradiol and progesterone than attachment system (p < .05). The expression of MMP‐9 mRNA was higher in FA than in other groups (p < .05) and similar to antral follicles from in vivo control (p > .05). Only FA group resulted in oocytes matured. After 30 days, oocytes from preantral follicles in vitro grown in FA developed to eight‐cell parthenotes. In conclusion, a culture system using FA supported the development of caprine preantral follicles cultured in group and included in the same bead of hydrogel, improving the oocyte maturation and producing parthenotes.     

Viability of OPS Vitrified Sheep Embryos After Direct Transfer

The aim of this study was to evaluate the viability in the effect of open pulled straw (OPS) vitrification procedure of sheep embryos after direct transference. Embryos were produced in vivo and cryopreserved in slow freezing or OPS vitrification. The survival rates of cryopreserved embryos were compared to non-frozen standard pattern. In a first set of experiments, embryos at morula and blastocyst stages were dived in ethylene glycol (1.5 M) and frozen in an automatic freezer. After being thawed, they were directly or indirectly transferred to ewes recipient. A second group of embryos were drawn into OPS and plunged into liquid nitrogen after being exposed at room temperature for 1 min and 45 s in 10% EG plus 10% dimethyl sulphoxide (DMSO), then again for 30 s in 20% EG + 20% DMSO + 0.5 M sucrose. After being warmed, embryos were also directly transferred using a French mini straw as the catheter for the transplantation process or after in vitro dilution of cryoprotectants (two-step-process). No significant difference was observed among fresh, frozen or vitrified embryos on pregnancy rate (50.0%, 38.6% and 55.8%). However, when we evaluated only the direct transference, the pregnancy rate of OPS vitrified embryos was higher than that of frozen embryos (57.1% vs 34.8%) (p = 0.07). In addition, vitrified morulae had a higher pregnancy rate than the one with frozen embryos (64.0% vs 38.9%) (p = 0.07). Finally, our results indicate that OPS vitrification technique in association with direct transference improves the viability of sheep embryos with potential applications to field conditions.     

Toxicity of four commonly used chemotherapeutic compounds to fry of the African catfish, Clarias gariepinus (Burchell)

Acute toxicity tests were carried out for four compounds commonly used against various fish pathogens to determine their 24-h LC₅₀ values to fry of the African catfish, Clarias gariepinus (Burchell). Compounds tested and their calculated 24-h LC₅₀ values (mg l^?1) were as follows: praziquantel (13.4); malachite green oxalate (0.14); acriflavin neutral (10.0); and mebendazole (315). The results show that dosage levels of malachite green oxalate, praziquantel and acriflavin neutral recommended in the literature for the control of most fish ectoparasitoses caused substantial mortalities and cannot be applied to Clarias gariepinus fry.     

Effect of Post‐Thaw Addition of Seminal Plasma on Motility,Viability and Chromatin Integrity of Cryopreserved Donkey Jack (Equus asinus) Spermatozoa

Pregnancy rates in donkeys after artificial insemination with cryopreserved semen are still low, compared to the horse species. Addition of autologous seminal plasma to frozen‐thawed semen appeared to improve pregnancy rates. The aims of this study were to evaluate (1) sperm motility and plasma membrane integrity after thawing (T0) and after one and 2 h (T1 and T2) of post‐thaw incubation in either 0% (SP0) or 70% (SP70) autologous seminal plasma and (2) sperm motility, plasma membrane integrity and DNA quality (%COMP‐αt) after thawing (T0) and after 2 and 4 h (T2 and T4) of post‐thaw incubation in either 0% (SP0), 5% (SP5) or 20% (SP20) homologous seminal plasma. In experiment 1, seminal plasma decreased total and progressive sperm motility and plasma membrane intact spermatozoa immediately after dilution and at all following time points (p < 0.05). In experiment 2, total and progressive motility did not differ between treatments immediately after dilution and between SP0 and SP5 at T2, while they were lower in both SP5 and SP20 than in SP0 at T4. Plasma membrane intact sperm cells did not differ between SP0 and SP5 and were lower in SP20 at all time points. DNA quality was not affected by treatment immediately after dilution and was significantly worse for SP20 after 4 h of incubation (p < 0.05). The post‐thaw addition of seminal plasma at the tested concentrations did not improve donkey frozen semen characteristics in vitro over time.     

Farmers’ Fruit Tree-growing Strategies in the Humid Forest Zone of Cameroon and Nigeria

Many studies have stressed the importance of trees to rural households. Few, however, have focused on actual numbers and densities of trees in different land-use systems. Based on community-level participatory research in six communities, semi-structured household interviews and full-farm fruit tree inventories, this study aims to understand farmers’ tree-planting strategies. Relationships between the diversity, number and density of fruit trees and farm size, land-use system, land tenure, distance from the homestead, proximity to the forest, market access and household characteristics are investigated. The key factors determining the differences in tree-growing strategies between communities appear to be market access, land use and access to forest resources. Within communities, differences between individual households were less easy to explain but tenure was important as was farm size. Smaller farms had higher fruit tree densities, a relationship that was particularly strong in communities with good market access. Overall there was a great deal of variability both within and between communities and many of the factors affecting tree-planting decisions were found to be highly inter-related. Despite this complexity, trees on farm play an important role in rural household's livelihoods. Therefore, expansion of tree cultivation should be recognized as a promising pathway to achieve increased income and food production by policy makers and extensionists alike. In addition to improved tree propagation and management techniques, farmers should be strengthened in the processing and marketing of agroforestry tree products and more emphasis should be placed on the development of tree enterprises. By doing so, farmers will be able to earn a more important and consistent income from fruit trees, contributing to the Millennium Development Goals.