Production of Oocytes of Nile Tilapia (Oreochromis niloticus) for In Vitro Fertilization via Hormonal Treatments

Only a few studies have described hormonal treatments for induction of synchronicity and gamete collection in Nile tilapia (Oreochromis niloticus), both important for assortative matings in breeding programmes and essential for polyploidy technologies. In this study, we compared the effectiveness of carp pituitary extract (CPE), Nile tilapia pituitary extract (TPE), human chorionic gonadotropin (hCG) and gonadotropin‐releasing hormone (GnRH) protocols on the induction of spawning and egg production in Nile tilapia. Among the hormonal treatments analysed, only hCG was effective for producing viable gametes for in vitro fertilization. To verify the viability of this hormonal treatment, hCG was tested using different doses (1000, 2000, 3000, 4000 and 5000 IU/kg) in a large number of females (208 animals) from two Nile tilapia lines. The results indicated that hCG doses between 1000 and 5000 IU/kg could be used to induce final oocyte maturation in Nile tilapia with collection of stripped oocytes. This is the first study to report differential reproductive responses to hormonal treatment between tilapia lines: line 1 was more efficient at producing eggs and post‐hatching larvae after hCG induction than line 2. In conclusion, we demonstrated that the hCG protocol may be applied on a large scale to induce final oocyte maturation in Nile tilapia. The development of a protocol for in vitro fertilization in Nile tilapia may aid in breeding programmes and biotechnological assays for the development of genetically modified lines of Nile tilapia.     

Wood density,phytomass variations within and among trees,and allometric equations in a tropical rainforest of Africa

The development of tree allometric equations is crucial to accurate forest carbon assessment. However, very few allometric equations exist for sub-Saharan Africa and as a result generalized allometric equations, often established for forests in other continents, are used by default. The objectives of this study were (1) to propose a sampling methodology and calculation procedures to assess biomass for tropical tree species of contrasted tree shapes in Africa, (2) to identify factors affecting within and between trees wood density, (3) to propose an allometric model that integrates these factors and (4) to evaluate the reliability of using generalized allometric equations in this type of forests. Models were developed to predict wood density and phytomass of the trees based on the harvesting of 42 trees from 16 species, representing three guild status in the wet evergreen forest of Boi Tano in Ghana. Results indicated that the wood density was highly influenced by the tree species, guild status, size of the tree and pith to bark distance. Dry mass of a tree was influenced by diameter at breast height, crown diameter and wood density. The wood density depends on the position of the wood within the tree and the guild status considered. The use of generalized allometric models in literature is limited by the specific climate zone, the consideration of tree height and species specific wood density. In considering those factors, using generalized allometric equations could result in an error of 3%. Further research should better consider the bigger trees and the influence of the topography and ecosystem history.     

Effects of meiotic inhibitors and gonadotrophins on porcine oocytes in vitro maturation,fertilization and development

This study evaluated the effect of three reversible meiotic inhibitors (MINs) and their interaction with gonadotrophins (Gns) on the meiotic maturation and developmental competence of porcine oocytes. In experiment 1, the oocytes were matured for 22 hr in the presence or absence of dbcAMP (1 mM), cycloheximide (7 μM) or cilostamide (20 μM) with or without Gns, and for an additional 22 hr in the absence of MINs and Gns. At 22 hr of maturation, regardless of the presence of Gns, a higher proportion (p < .001) of oocytes cultured in the presence of MINs were effectively arrested at the germinal vesicle stage compared with the oocytes cultured without MINs. At 44 hr of maturation, the proportion of oocytes that reached MII was higher (p < .05) in groups with Gns compared with groups without Gns. In experiment 2, oocytes that were matured as in experiment 1 were inseminated and cultured for 7 days to evaluate fertilization parameters and blastocyst formation. Only oocytes from the dbcAMP + Gns group had higher (p < .05) efficiency of fertilization compared with the other treatment groups. The presence of dbcAMP during maturation also increased (p < .05) blastocyst formation and efficiency of blastocyst formation in both the presence and absence of Gns. These results indicate that the interaction of Gns with the tested MINs improved meiotic progression. In addition, regardless of supplementation with Gns, the presence of dbcAMP during the first maturation period increased and even doubled the capacity of oocytes to develop to the blastocyst stage.     

In Vitro assessment of the nutritive value of expanded soybean meal for dairy cattle

ABSTRACT: Little information is available about the nutritive value of expanded soybean meal, which is produced by expansion of soybeans prior to solvent extraction of the oil. During processing, expanded soybean meal is subjected to additional heat, which might increase the concentration of ruminally undegraded protein. Processing of soybeans with heat during oil extraction could affect lysine availability by increasing ruminally undegraded protein or by impairing intestinal digestion. Our objective was to compare solvent and expanded soybeans with regard to chemical composition and nutritive value for dairy cattle. Samples of expanded soybean meal (n = 14) and solvent-extracted soybean meal (n = 5) were obtained from People's Republic of China to study effects of the expansion process on nutritive value for dairy cattle. Solvent-extracted soybean meal (n = 2) and mechanically extracted (heated) soybean meal (n = 2) from the United States served as references for comparison. Samples were analyzed for crude fat, long-chain fatty acids, crude protein, amino acids, chemically available lysine, in situ ruminal protein degradation, and in vitro intestinal digestibility. No differences were found between solvent-extracted soybean meals from China and expanded soybean meals from China for crude fat, crude protein, amino acids, or chemically available lysine. In situ disappearance of nitrogen, ruminally undegraded protein content, and in vitro intestinal digestion of the ruminally undegraded protein were generally similar between solvent-extracted soybean meals made in China and expanded soybean meals made in China; variation among soybean meals was small. Results indicate that the additional heat from the expansion process was not great enough to affect the nutritive value of soybean meal protein for ruminants. Although expansion may improve the oil extraction process, the impact on the resulting soybean meal is minimal and does not require consideration when formulating ruminant diets.     

In Vitro assessment of the nutritive value of expanded soybean meal for dairy cattle

Little information is available about the nutritive value of expanded soybean meal, which is produced by expansion of soybeans prior to solvent extraction of the oil. During processing, expanded soybean meal is subjected to additional heat, which might increase the concentration of ruminally undegraded protein. Processing of soybeans with heat during oil extraction could affect lysine availability by increasing ruminally undegraded protein or by impairing intestinal digestion. Our objective was to compare solvent and expanded soybeans with regard to chemical composition and nutritive value for dairy cattle. Samples of expanded soybean meal (n = 14) and solvent- extracted soybean meal (n = 5) were obtained from People’s Republic of China to study effects of the expansion process on nutritive value for dairy cattle. Solvent-extracted soybean meal (n = 2) and mechanically extracted (heated) soybean meal (n = 2) from the United States served as references for comparison. Samples were analyzed for crude fat, long-chain fatty acids, crude protein, amino acids, chemically available lysine, in situ ruminal protein degradation, and in vitro intestinal digestibility. No differences were found between solvent-extracted soybean meals from China and expanded soybean meals from China for crude fat, crude protein, amino acids, or chemically available lysine. In situ disappearance of nitrogen, ruminally undegraded protein content, and in vitro intestinal digestion of the ruminally undegraded protein were generally similar between solvent-extracted soybean meals made in China and expanded soybean meals made in China; variation among soybean meals was small. Results indicate that the additional heat from the expansion process was not great enough to affect the nutritive value of soybean meal protein for ruminants. Although expansion may improve the oil extraction process, the impact on the resulting soybean meal is minimal and does not require consideration when formulating ruminant diets.     

Motility Characteristics and Fertilizing Capacity of Boar Spermatozoa Stained with Hoechst 33342

Flow cytometry sorting of spermatozoa using fluorescence dye Hoechst 33342 is the only effective sex selection methodology validated in numerous laboratories. This study was carried out to determine the effect of Hoechst 33342 on the motility and fertility of stained boar spermatozoa. Experiment 1 evaluated motility parameters (percentage of motile spermatozoa, velocity, angularity and oscillation) of boar spermatozoa stained with Hoechst 33342 by a computer‐aided sperm analysis (CASA) instrument. Spermatozoa (30 million/ml) were divided into five treatment groups and stained during 1 h at 35°C with 9, 18, 27, 60 and 90 μM of H33342. There were no differences in sperm motility patterns nor percentages of motile spermatozoa incubated in the presence of 9, 18 or 27 μM. Percentage of motile spermatozoa and motility parameters decreased significantly (p < 0.05) at 60 μM of Hoechst 33342. Spermatozoa were immotile at concentration of 90 μM. In experiment 2, pregnancy rates, farrowing rates and litter size from sows (n = 275) artificially inseminated (AI) with either Hoechst 33342 stained (27 μM) or unstained (control) spermatozoa were determined. Sows inseminated with stained spermatozoa had no significant lower pregnancy rate (88.33%) as compared with controls (90.32%). Staining neither affected farrowing rates (85.0 vs 87.7%) nor total number of piglets born (10.56 ± 0.32 vs 10.47 ± 0.24, stained and controls, respectively). No phenotypical abnormalities were registered among the newborn piglets. The data suggest that incubating spermatozoa with Hoechst 33342 at levels required for X‐ and Y‐bearing chromosome sperm sorting, does not impair sperm viability or their fertility after AI.     

Muscle strain injuries of the hindlimb in eight horses: diagnostic imaging,management and outcomes

Objective To describe the clinical presentation, ultrasound findings, management and outcome in horses with muscle tear injuries of the hindlimbs. Design Retrospective case series Procedure Medical records of eight horses were reviewed and information on signalment, history, presenting complaint, physical examination findings and further diagnostic tests were recorded. Diagnosis of muscle injury was determined by the presence of abnormal ultrasound findings, compared with the contralateral limb, and, when required, nuclear scintigraphy. Follow-up information was obtained via telephone interviews with owners, trainers and referring veterinarians. Results Muscle tears causing lameness were identified in the middle gluteal (3), semitendinosus (1), semimembranosus (2) and gracilis (2) muscles. Tears were classified by ultrasound imaging as partial (6) or complete (2). The degree of lameness did not appear to be indicative of the extent of injury or of completion of healing. Long-term follow-up was available for seven horses and the outcome was favourable in six cases. Conclusions and clinical relevance Ultrasonography is useful in the diagnosis and assessment of moderate to severe muscle strain injuries. The prognosis appears to be favourable in most cases, although recurrence of injury and lameness can delay the return to athletic activity and an inferior outcome with persistent gait abnormality may occur.     

The Effect of Two Antibiotic Mixtures on Haemophilus somnus, Campylobacter fetus ssp. venerealis, Mycoplasma bovis, and Ureaplasma diversum in Frozen Bovine Semen

A two-step semen-extending protocol was compared to a one-step protocol in its efficiency in inhibiting growth of Haemophilus somnus, Campylobacter fetus ssp. venerealis, Mycoplasma bovis , and Ureaplasma diversum in experimentally infected semen. In both protocols, the effect of an antibiotic mixture of 500 μg gentamycin, 100 μg tylosin, 300 μg lincomycin, and 600 μg spectinomycin (GTLS) was compared to a mixture of 500 IU penicillin, 500 IU streptomycin, 150 μg lincomycin, and 300 μg spectinomycin (PSLS). The one-step extending method was as effective as the two-step extending method. Both antibiotic mixtures were equally effective in controlling C. fetus . For H. somnus and U. diversum , the PSLS mixture was more effective than the GTLS mixture. It was striking that both antibiotic mixtures had no effect in decreasing the numbers of M. bovis .