Fertilizing Capacity of Frozen Epididymal Sperm Collected from Dogs

The collection of epididymal sperm may be a valuable tool for canine reproduction especially since it can enable collection of cells after death of a valuable dog. The aim of the present study was to evaluate the viability of epididymal sperm after freeze-thawing. Epididymides were obtained from four adult dogs by elective orchiectomy. The caudal portion of the epididymides and part of the deferential ducts were squeezed by means of an anatomic clamp into a Petri dish containing either 0.9% saline solution (Group 1) or Ringer solution without lactate (Group 2). Samples were centrifuged at 800 ×  g for 10 min, the supernatant was removed and the pellet was diluted in one step with a Tris/citric acid/OEP (Orvus Es Paste) extender containing 7% glycerol and subjected to semen freezing. Oocytes were obtained from canine ovaries, after ovariohysterectomy. Only oocytes that were approximately 100 μm in diameter, with a dark ooplasm surrounded by three- or four-well formed cumulus cell layers were used for sperm testing. Frozen semen samples were thawed in a water bath at 70°C for 8 s and analysed at room temperature for sperm motility and velocity. Oocytes were incubated with spermatozoa in humidified atmosphere containing 5% CO₂ at 38°C for 18 h. Morphological and functional characteristics of spermatozoa were similar in both groups. However, the percentage of sperm cells bound to oocytes was significantly higher in Group 2 than in Group 1. This result suggests that the Ringer solution without lactate was a more suitable medium for collecting epididymal canine sperm than 0.9% saline.     

Feeding Dietary Mannan Oligosaccharides to Juvenile Nile Tilapia,Oreochromis niloticus,Has No Effect on Hematological Parameters and Showed Decreased Feed Consumption

Impaired immune system by environmental stressors can lead fishes to be more susceptible to diseases that limit the economic development of aquaculture systems. This study was set out to determine the effect of six levels of mannan oligosaccharides (MOS; ActiveMOS^®; Biorigin, Lençois Paulista, São Paulo , Brazil) on the performance index and hematology of Nile tilapia, Oreochromis niloticus juveniles. Fish (13.62 g) were randomly distributed into 18 plastic aquaria (300 L; 20 fishes per aquarium) and fed during 45 d with a commercial diet supplemented with 0, 0.2, 0.4, 0.6, 0.8, and 1% dietary MOS, in a totally randomized design trial (n = 3); biometrical and hematological data were collected and analyzed. There were no significant differences in hematological parameters between fish fed control and MOS supplementation diets, and daily feed consumption (FC) decreased (P < 0.05) with increasing levels of dietary MOS. Dietary MOS did not increase leukocyte count and presented negative effects on FC of Nile tilapia. At 0.4% MOS supplementation, the individual weight gain was higher in absolute values but not different (P > 0.05) compared to control diet.     

Identification of the SP22 Sperm Protein in Santa Inês and Dorper Rams

The sperm membrane protein referred to as SP22 has been identified in different species and, at least in rats, is highly correlated with fertility. The goals of this study were to identify and to quantify the SP22 protein on spermatozoa from adult rams (Dorper and Santa Inês breeds), and to correlate its levels to morphological and kinematics parameters. SP22 on ram sperm was effectively quantified by both enzyme‐linked immunosorbent assay (ELISA) and fluorescein isothiocyanate immunostaining analysis and the two methods were significantly correlated (R² = 0.70). Clustering analysis of motility parameters obtained by computer‐assisted semen analysis system was used to establish that three distinct kinematic subpopulations with different vigour and progressiveness coexistent within ejaculate. While there were significant differences in the distribution of the three subpopulations in the rams, there was no significant correlation between the proportion of each subpopulation in the rams and the SP22 levels. Quantification of SP22 immunostaining intensity was not correlated with any of the sperm parameters. However, SP22 levels obtained by ELISA were negatively correlated with morphological abnormalities and positively correlated with membrane integrity (three variable R² = 0.47). Future breeding studies are now needed to validate that this protein is a biomarker of fertility in this species.     

In vitro Developmental Competence of Adult Sheep Oocytes Treated with Roscovitine

The efficiency of in vitro sheep embryo production is still low compared to that observed in vivo and in other species. In this context, meiotic inhibition strategies emerged as a promising alternative to improve this biotechnology. So, this study aimed to evaluate, for the first time, the effects of roscovitine on in vitro maturation of sheep oocytes and their subsequent embryo development. For this, cumulus–oocyte complexes (COCs) were cultured for 6 h in the presence (Rosco) or absence (Control) of 75 μm roscovitine and, subsequently, in vitro matured (IVM) for 18 h with gonadotropins. At 0 (Immature), 6 and 24 h of culture, the nuclear status of oocytes was evaluated by Hoechst staining. Embryo cleavage and blastocyst formation were recorded 30 h after in vitro fertilization and on day 7 of culture, respectively. Blastocyst quality was evaluated by differential staining. At 6 h, the GV rate in the Rosco treatment (93.8%) was similar to that observed in the Immature oocytes (94.9%) and significantly higher compared to Control (41.3%). After IVM for 18 h, a high and similar proportion of oocytes from Rosco (93.6%) and Control (88.4%) reached the MII stage. In both treatments, approximately 70% of oocytes cleaved and 50% of them developed up to blastocyst. The mean percentage of blastocyst cells, embryoblast, trophoblast and pyknosis did also not differ between Control and Rosco. In conclusion, roscovitine, at the studied experimental conditions, was efficient to reversibly inhibit the meiosis of adult sheep oocytes without detrimental effect on development and quality of the in vitro produced embryos.     

Protection against enzootic pneumonia of pigs: intraperitoneal inoculation with live LKR strain of Mycoplasma hyopneumoniae

Pigs obtained from a mycoplasma-free piggery were randomised into 4 groups of 9. Groups 1 and 2 were injected by the intraperitoneal route with liquid culture of the LKR strain of Mycoplasma hyopneumoniae. Group 1 was injected once and group 2 twice. Group 3 was made up of pigs inoculated by the intranasal route with the virulent Beaufort strain of M. hyopneumoniae; they served as the source of infection for the challenge. Group 4 were uninfected, uninjected controls. Six weeks after the last injection, groups from 1 to 4 were placed in contact. Seven of the pigs in the 1-dose group and 6 in the 2-dose group were free of lesions at necropsy 6 weeks after challenge. Of the two pigs with lesions in the 1-dose group one had only a small lesion but the other had extensive lesions; it had not shown an antibody response after injection of culture. The lesions in the 3 pigs in the 2-dose group were all small. All 9 control pigs had lesions which varied from medium to large in size. The difference in the incidence of pneumonia between the injected and control groups was significant (P less than 0.05) and the proportion of severely affected pigs in the vaccinated groups was significantly lower (P greater than 0.01). There was no difference between those given one dose of vaccine and those receiving 2 doses.     

Heat-treated bean (Phaseolus vulgaris) residue meal as an alternative protein source in pelleted diets for Nile tilapia fingerlings: growth,body composition,and physical characteristics of diets

Tropical Animal Health and Production - The study aimed to evaluate heat-treated bean residue meal (Phaseolus vulgaris)—BRM—as an alternative protein source in diets for Nile tilapia...     

In vitro Maturation of Oocytes from Santa Ines Ewes Subjected to Consecutive Sessions of Follicular Aspiration by Laparoscopy

The success of embryo production in vitro depends upon the use of an efficient oocyte retrieval technique, and the best results have been obtained by laparoscopic aspiration. The aim of this study was to evaluate the effect of consecutive sessions of follicular aspiration on the quantity, quality and in vitro maturation competence of oocytes obtained from ewes subjected to hormonal stimulation. Six Santa Ines ewes underwent nine sessions of follicular aspiration by laparoscopy with a 7‐day interval between sessions, totalling 56 aspirations. After 24 h of culture, oocytes were stained and classified according to the stage of nuclear and cytoplasmic maturation. Oocyte retrieval rate was 61.4 ± 2%, resulting in a total of 249 oocytes. No significant variation was observed between sessions (p > 0.05). The average number of oocytes retrieved from each ewe was 6.4 ± 2 per session and 42 ± 4 in total. No significant difference was observed between the frequencies of the different stages of nuclear maturation: 32.72% mature, 40.74% immature and 26.54% degenerated/indeterminate oocytes; however, a significant difference was observed between the frequencies of the different stages of cytoplasmic maturation: 10.7% mature, 73.25% immature and 16.05% degenerated/indeterminate oocytes. No significant difference was observed in nuclear or cytoplasmic maturation between the weeks of procedure. We conclude that after nine consecutive sessions of follicular aspiration, the quantity and quality of retrieved oocytes remained unchanged as well as the levels of nuclear and cytoplasmic maturation obtained, demonstrating the viability of this technique for repetitive follicular aspirations on the same donor.     

Double-outlet right ventricle in a 10-month-old Friesian filly

A 10-month-old Friesian filly had a presentation that was consistent with chronic left- and right-sided congestive heart failure. Clinical pathology findings included abnormal haematological and biochemical variables, abnormal blood gas values and increased serum concentration of cardiac troponin I. Echocardiography revealed cardiac chamber dilation and dextropositioning of the aorta. Radiography revealed a generally enlarged heart and pulmonary interstitial infiltration. These findings were supported at necropsy and the diagnosis of double-outlet right ventricle was confirmed. The pathological changes and physiological responses subsequent to double-outlet right ventricle have not previously been described in detail in horses. Clinical progression closely resembles that seen in humans, in whom antemortem diagnosis relies on echocardiography. In horses, complex cardiac disease presents a diagnostic challenge to the clinician. Appropriate therapy must be based on an accurate diagnosis.