Assessment of Suitable Porcine Semen for Freezing,According to the Ejaculate Characteristics in the Iberico × Landrace Breed

A total of 35 ejaculates were studied in order to assess the suitability of porcine semen for freezing according to the ejaculate characteristics. The effects of the freezing procedure were identified; a decrease in motility and acrosome quality was found after thawing. The best results on motility were linked to the ejaculates with a volume of less than 100 ml of the sperm‐rich fraction, a concentration lower than 450 × 10⁶ spermatozoa/ml and an agglutination score below 2. However, the best normal apical ridge (NAR) was found when the volume of the sperm‐rich fraction was greater than 150 ml. For this reason, an intermediate volume of the sperm‐rich fraction of the ejaculate for the best motility and the best NAR, a concentration lower than 450 × 10⁶ spermatozoa/ml and a rate of agglutination below 2 should provide the best quality after freezing. This study also attempted to determine whether a positive effect of ejaculate selection on the overall freezing performance might be expected.     

Laparoscopic-assisted cystotomy for urolith removal in geldings

OBJECTIVE: To describe a technique for laparoscopic-assisted removal of cystic calculi in geldings and report outcome. STUDY DESIGN: Clinical report. ANIMALS: Four geldings with cystic calculi. METHODS: Laparoscopic-assisted cystotomy and urolith retrieval was performed in 4 anesthetized geldings positioned in dorsal recumbency. With a laparoscope portal located at the umbilicus, the abdomen was insufflated and then the surgical table was tilted (30 degrees head-down position) before an instrumental portal was created parallel and 2-3 cm medial to the left external inguinal ring. Laparoscopic grasping forceps were inserted to grasp the cranial aspect of the bladder and elevate it to the ventral abdominal wall. With the instrumental portal as mid-point, the parainguinal skin incision was longitudinally extended cranial and caudal (approximately 8-10 cm) to accommodate the size of the urolith. The apex of the bladder was exteriorized and sharply incised, the urolith extracted, and after cystotomy closure, the bladder was repositioned. The mini-laparotomy and trocar incisions were closed in layers. RESULTS: There were no intra- or post-operative complications. All horses had minor incisional swelling for 3-4 days. No signs of abdominal or incisional pain were observed. Hematuria and slight stranguria occurred until the 3rd or 4th day. Surgical time (skin incision to skin closure) was 35-40 minutes. On long-term follow-up (up to 12 months) no recurrence of clinical signs associated with cystic calculi occurred. CONCLUSION: Uroliths (6-8 cm diameter) can be removed by laparoscopic-assisted cystotomy in geldings. CLINICAL RELEVANCE: Laparoscopic-assisted cystotomy combines the advantages of the parainguinal laparocystotomy with laparoscopic technique for removal of cystic calculi while avoiding their disadvantages.     

Implementation of two high through-put techniques in a novel application: detecting point mutations in large EMS mutated plant populations

Background  

The establishment of mutant populations together with the strategies for targeted mutation detection has been applied successfully to a large number of organisms including many species in the plant kingdom. Considerable efforts have been invested into research on tomato as a model for berry-fruit plants. With the progress of the tomato sequencing project, reverse genetics becomes an obvious and achievable goal.     

Interactions between different media and follicle‐stimulating hormone supplementation on in vitro culture of preantral follicles enclosed in ovarian tissue derived from collared peccaries (Pecari tajacu Linneaus, 1758)

The aim was to verify the effect of follicle‐stimulating hormone (FSH) supplementation to α‐MEM+ or TCM199+ media on the in vitro development of ovarian preantral follicles (PFs) derived from collared peccaries. Ovaries (n = 5 pairs) were collected and divided into fragments destined to control group (non‐cultured) or treatments that were cultured for 7 days. The PFs morphology, growth and activation were evaluated by classical histology. The immunohistochemistry markers Ag‐NOR and PCNA were used for nuclear proliferation analysis, and the picrosirius red labelling was used for ovarian extracellular matrix (ECM) evaluation. After 7‐day culture, only the TCM199+ treatment maintained the proportion of intact PFs similar to day 1 (63.2%), but no differences were found among treatments (p > .05). In addition, a significant increase in the growing follicles proportion was verified for all the treatments, indicating follicular activation (p > .05). By the Ag‐NOR analysis, only the TCM199+/FSH maintained the nuclear proliferation similar to the first day (p > .05). The picrosirius red staining revealed that the ECM remained intact in all the treatments (p > .05). We suggest the use of TCM199+ medium supplemented of FSH for the in vitro development of peccaries PFs under 7‐day culturing conditions.