Transverse and vertical incisions affect the viability of in vitro-produced embryos submitted to a simplified microsurgery approach.

Integrating in vitro embryo production with embryonic microsurgery facilitates the generation of monozygotic twins. However, despite their potential benefits, these methods have not been widely adopted in commercial settings because of their substantial costs. Hence, there is a need to streamline the bisection procedure while ensuring efficient production of viable demi-embryos. In this study, we investigated the impact of different orientations of microsurgical incisions in relation to inner cell mass on embryonic development, morphology, viability, and expression of cell fate protein markers using a simplified microsurgery approach. Ovaries were transported from the slaughterhouse to the laboratory and aspirated to obtain oocytes that were selected and subjected to in vitro embryo production. The selected expanded blastocysts (n = 204) underwent microsurgery. The blastocysts were immobilized to facilitate incision using an adapted microblade, yielding demi-embryos (vertical incision) and viable embryonic fragments (transverse incision). The structures were then re-cultured for 12 h. Viability was assessed by measuring the re-expansion rate after re-culture, followed by immunofluorescence analysis of proteins (CDX2 and NANOG) and apoptosis analysis using terminal deoxynucleotyl transferase dUTP nick end-labeling (TUNEL). Microsurgically derived embryos exhibited remarkable plasticity, as evidenced by a slight reduction (P < 0.05) in the re-expansion rate (transverse 64.2 % and vertical 57.2 %) compared to that of the control group (blastocysts without microsurgery) (86.7 %). They also demonstrated the ability of morphological reconstitution after culturing. Despite the anticipated decrease (P < 0.05) in the total number of cells and embryo volume, microsurgery did not result in a significant increase (P > 0.05) in the number of apoptotic cells. Furthermore, microsurgery led to higher (P < 0.05) expression of markers associated with pluripotency, indicating its efficiency in preserving regenerative capacity. Moreover, microsurgery, whether followed by immunosurgery or not, made the isolation of embryonic cells easier. In conclusion, both transverse and vertical microsurgery incisions enabled the production of identical demi-embryos and served as tools for isolating embryonic cells without compromising the resumption of development and the apoptotic index.

Comparison of surgical invasiveness between micro-endoscopic discectomy/microscopic discectomy and conventional hemilaminectomy in dogs.

Micro-endoscopic discectomy (MED) or microscopic discectomy (MD) have been performed for disc herniation in humans. The purpose of this study was to compare the invasiveness of the hemilaminectomy in dogs between the approach using a cylindrical retractor for the MED/MD and a conventional open surgical approaches in dogs. First, as preliminary studies, we analyzed the suitability of the cylindrical retractor for the vertebral body of small to medium-sized dogs on the X-ray computed tomographic images using the three-dimensional analysis software, and confirmed that it was possible to open a bone window of an approximate length of 1.72 clto the spinal canal with the cylindrical retractor with a diameter 17 mm using two medium-sized canine cadavers. Next, to determine difference in the invasiveness of hemilaminectomy, the magnitude of tissue damage, surgical stress and postoperative pain were compared between the conventional open approach (hemilaminectomy group: HL group, n=6) and the surgical approach using the cylindrical retractor (MD group, n=6) in 12 beagle dogs. The plasma creatine phosphokinase, C-reactive protein and cortisol concentrations, incision length and University of Melbourne Pain Scale scores after the hemilaminectomy were significantly lower in the MD group than in the HL group. There were no significant differences between the durations of surgery and the other evaluated indices. The approach using the MD can provide a less invasive hemilaminectomy than the conventional approach in dogs.

Microsurgical arterial anastomosis in young and adult rats: an evolutive and comparative study

Purpose: To evaluate the caliber of an arterial micro-anastomosis in the young growing animal using a continuous suture technique. Additionally, late morphological changes and blood flows distal to the anastomosis were evaluated. Methods: Seventy-four Wistar rats were submitted to laparotomy to access the aorta for blood flow measurement. The aorta was sectioned using microsurgery technique and an end-to-end anastomosis with continuous suture. After a period of six months to one year, the anastomosis was checked. Results: Regarding the size of the aortas, comparing the pre- and postoperative values, there was an increase of 13.33% in adult animals and 25% in young animals, without any difference in the blood flows. Conclusions: The arteries of young rats show signs of growth at the site of the anastomosis performed with continuous suture.

Anatomical description of the extratemporal facial nerve under high-definition system: a microsurgical study in rats

Purpose: To describe the microsurgical anatomical aspects of the extratemporal facial nerve of Wistar rats under a high-definition video system. Methods: Ten male Wistar rats (12­15 weeks old), without veterinary diseases, weighing 220­280 g, were used in this study. All animals in this study were submitted to the same protocol and by the same surgeon. A 10-mm incision was made below the bony prominence of the right or left ear, and extended towards the angle of the mandible. The dissection was performed and the main branches of the facial nerve were dissected. Results: The main trunk of the facial nerve has a length of 0.88 ± 0.10 mm and a length of 3.81 ± 1.03 mm, measured from its emergence from the stylomastoid foramen to its bifurcation. Seven branches originating from the facial nerve were identified: posterior auricular, posterior cervical, cervical, mandibular, buccal, temporal, and zygomatic. Conclusions: The anatomy of the facial nerve is comparable to that of humans, with some variations. The most observed anatomical division was the distribution in posterior auricular, posterior cervical, cervical, mandibular, buccal, temporal, and zygomatic branches. There is no statistical difference between the thickness and distance of the structures compared to the contralateral side.

Minimally invasive microsurgical decompression of an intervertebral disc protrusion in a dog.

OBJECTIVE: To report the successful treatment of intervertebral disc protrusion with minimally invasive microsurgery in a large breed dog. STUDY DESIGN: Retrospective case report. ANIMAL: A 4-year-old, neutered male, German shepherd dog presented with chronic progressive ambulatory paraparesis and thoracolumbar pain; lumbar intervertebral disc protrusion and severe spinal cord compression at L2-L3 were diagnosed. METHODS: A minimally invasive approach was used to access the target surgical area by using a muscle splitting technique and retractors. Intraoperative fluoroscopy confirmed correct placement. Magnification and illumination through a surgical microscope were used (microsurgery) to perform the spinal cord decompression by means of a foraminotomy and lateral corpectomy. RESULTS: Technically, the combination of fluoroscopy and muscle splitting approach offered adequate minimally invasive access. Microsurgery allowed for precise and efficient spinal cord decompression. Clinically, no immediate postoperative neurological deterioration was observed. Opioid usage was limited to 24 hours postoperatively. Focal muscle swelling was observed postoperatively for 2 days, and hospital stay was 3 days. At 7 weeks postoperatively, neurological examination results were normal, and postoperative MRI confirmed spinal cord decompression. No complications were reported. CONCLUSION: The procedure was associated with a small incision, limited muscle trauma, early improvement of the neurological status, a short hospital stay, and limited postoperative pain. CLINICAL SIGNIFICANCE: This case provides evidence that minimally invasive microsurgery is an effective treatment for intervertebral disc protrusion in large breed dogs and may offer benefits regarding postoperative pain and functional recovery.

Mast cell stabilization with ketotifen decreases il-13 and cgrp in the ascitic fluid of rats with microsurgical cholestasis

Ascites is one of the most severe complications of cirrhosis with portal hypertension. Since mast cells by type 2 immunity could be involved in the production of portal hypertensive ascites, we are studying the effectiveness of Ketotifen administration, a mast cell stabilizer, to modulate the production of interleukin-13, a type-2 associated cytokine, as well as calcitonin gene-related peptide (CGRP), one important regulator of its production, in the ascitic fluid of microsurgical extrahepatic cholestatic rats. The increased IL-13 and CGRP release in the ascitic fluid of the rats with obstructive cholestasis and its significant reduction after both, prophylactic plus therapeutic and delayed therapeutic oral administration of Ketotifen, allows for proposing that mast cells could play an important role in the etiopathogeny of portal hypertensive ascites

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Urethral prolapse corrected by microsurgery in a cat: a case report.

Urethral prolapse is a rare condition in dogs and cats. A 7-month-old male cat presented with intermittent penile bleeding and a red mass protruding from the tip of the penis. Urethral prolapse was diagnosed by using a surgical microcamera to examine the genital area. Urethroplasty and preputial advancement surgery were performed using a surgical microscope, to successfully correct the urethral prolapse.

Avian Soft Tissue Surgery.

Basic surgical instrumentation for avian soft tissue surgery includes soft tissue retractors, microsurgical instrumentation, surgical loupes, and head-mounted lights. Hemostasis is fundamental during the surgical procedures. The indications, approach, and complications associated with soft tissue surgeries of the integumentary (digit constriction repair, feather cyst excision, cranial wound repair, sternal wound repair, uropygial gland excision), gastrointestinal (ingluviotomy, crop biopsy, crop burn repair, celiotomy, coelomic hernia and pseudohernia repair, proventriculotomy, ventriculotomy, enterotomy, intestinal resection and anastomosis, cloacoplasty, cloacopexy), respiratory (rhinolith removal, sinusotomy, tracheotomy, tracheal resection and anastomosis, tracheostomy, pneumonectomy) and reproductive (ovocentesis, ovariectomy, salpingohysterectomy, cesarean section, orchidectomy, vasectomy, phallectomy) systems are reviewed.

Video Telescope Operating Microscopy.

Exotic pet veterinarians frequently have to operate on small animals, and magnification is commonly used. Existing endoscopy equipment can be used with a mechanical arm and telescope to enable video telescope operating microscopy. The additional equipment items and their specifics are described, and several case examples are provided.

Reducing the number of animals used for microsurgery training programs by using a task-trainer simulator.

To master the skills needed for microsurgery techniques, residents must enrol in a long and complex training program that includes manipulations on simulators, on ex vivo tissues and finally in vivo training. This final step consists of performing vascular anastomoses on murine models. We propose here a simulation program designed to decrease the number of rats used during the final in vivo training. Our study presents the materials used, the various exercises proposed and their evaluations. Two identical student groups were compared in the framework of the University Diploma of Microsurgery. Group A (seven students) followed a classic training program, all of whom achieved permeable vascular anastomoses. A total of 149 rats were needed for this group. Group B (seven students) first validated their manipulations on the task-trainer simulation program. A mean of 6 h was necessary to obtain this validation. All these students achieved the required permeable vascular anastomoses but only 77 rats were used for this group. This simulation program spared 72 rats, abiding by the Russell and Burch concept of a humane experimental technique, namely the 3R principles. This home-made, cost-efficient and easy-to-use task trainer included various exercises with increasing difficulty levels and a progressive scoring system. We believe that microsurgery training needs to include both simple and sophisticated tools in order to reduce the number of animals used to master these surgical skills.

A jugular bleeding technique in rabbits.

When studying pharmacokinetics in rabbits, researchers must often take multiple blood samples from conscious rabbits. Researchers usually collect these samples via the auricular vein, typically through a port or an indwelling catheter. The authors have developed an easy and efficient alternative method for obtaining multiple blood samples from conscious rabbits via the external jugular vein. This jugular bleeding technique serves as a refinement to blood sampling methods that require rabbits to undergo surgery (e.g., to insert a port) because it requires no alleviation of pain. During a 2-year period, the authors have taken multiple blood samples from more than 400 rabbits and have seen no adverse events attributed to this procedure.

Characterization of a rodent model for the study of arterial microanastomoses with size discrepancy (small-to-large).

Microsurgical autotransplantation of tissues is employed clinically to reconstruct defects following burns, trauma and surgical cancer ablation, and to correct congenital abnormalities. Transplant vessels of <3 mm are anastomosed by hand under the microscope. Experimentally, anastomotic patency rates decrease with increasing vessel diameter mismatch, and clinically, ratios of 3:1 or greater lead to unacceptably low arterial patency rates. A number of surgical techniques for dealing with size mismatch are described, but no one method has found favour, and few controlled studies of technique are reported. In this report, a rodent superficial caudal epigastric artery (SCEA)/femoral artery (FA) model for the study of these techniques is described in detail. The diameter ratio between these vessels lies in the clinically relevant range of 1:1.5-1:2.5. In the male Wistar rat, external vessel diameters were not found to increase markedly in size between animal weights of 300 and 500 g. The length of FA distal to the origin of the SCEA, which is important in allowing undisturbed distal run-off, was found to be negatively associated with animal body weight, implying that a smaller animal would be better for this model. Mean femoral arterial flow rate, measured by transit-time ultrasound, was noted to be statistically and physiologically significantly higher in the right artery when compared with the left. This model has advantages over interposition vein graft models in that it minimizes vessel compliance mismatch, and avoids the need for a second anastomosis.

Surgical treatment of an elbow hygroma utilizing microvascular free muscle transfer in a newfoundland.

Microvascular free muscle transfer utilizing the rectus abdominis muscle, in combination with a mesh skin graft and transarticular external fixator, were used following the surgical excision of an elbow hygroma in a Newfoundland dog. Postoperatively, the dog did well with minimal complications and has yet to have a recurrence 10 months following surgery. This report describes the utility and successful use of the rectus abdominis free tissue transfer in the surgical correction of a difficult elbow hygroma in the dog.

Bandaging, endoscopy, and surgery in the emergency avian patient.

This article is divided into three parts and describes procedures used in avian medicine after initial stabilization. The first part includes the application of bandages and splints for fractures. The second part describes the use of endoscopy to examine the choana, oral cavity, trachea, and, to a lesser extent, internal organs. The last part discusses equipment required and techniques used for esophagostomy tube placement, air sac cannulation, and surgery, with approaches to the coelomic cavity and a select number of surgical procedures. This article is intended to provide the emergency animal clinician with knowledge necessary to provide basic stabilization for avian patients and knowledge of procedures that can be performed by the experienced clinician.

Training protocol for feline renal transplantation, using rat vascular anastomosis.

Renal transplantation (RTx) has been a potential treatment for renal failure in pet cats. We developed a training protocol for the microsurgical skills required for feline RTx using rat vessels, which are the same size as the feline renal artery and vein. Using interrupted sutures, the transected abdominal aorta was reestablished in an end-to-end fashion. Venous anastomosis was performed with a continuous running suture in an end-to-side fashion between the portal vein and inferior vena cava. In the arterial anastomotic model, technical errors were checked by postoperative hemorrhaging. Those failures in the venous anastomotic model, technical errors were confirmed by the rat's death. Histological examinations of the epithelialization at the anastomotic site were evaluated in both groups. After training, nine cases of feline RTx were performed safely, using an adequate microsurgical technique.

Preliminary tests of a new reversible male contraceptive in bush dog, Speothos venaticus: open-ended vasectomy and microscopic reversal.

Open-ended vasectomies were performed on four male bush dogs (Speothos venaticus), with three having microscopic reversal surgery (vasovasostomy) between 10 and 20 mo post-vasectomy. The key to ease of reversal is leaving the distal (testicular) end open to allow leakage, resulting in a pressure-relieving granuloma. The proximal (abdominal) end is cauterized, providing an effective seal. This technique prevents the buildup of pressure in the epididymis, therefore limiting damage to the male's reproductive capacity. Described here are detailed procedures for both surgeries. One of the three males that underwent vasovasostomy has successfully impregnated his female partner. This study demonstrates that these techniques can be successfully applied to animals. With the two remaining pairs, none of the four individuals were proven breeders prior to the study, so it is not possible to eliminate the possibility of previously existing infertility. This technique may have limited application for carnivores, because vasectomy does not prevent potential adverse effects to females from prolonged, cyclic exposure to endogenous progesterone. In other taxonomic groups (e.g., primates, ungulates, marsupials, and rodents) in which multimale groupings are common, this reversible male sterilization technique could provide managers with the ability to control which males reproduce without eliminating their future reproductive capacity or social interaction.

Chronic implantation of transit-time flow probes on the ascending aorta of rodents.

This study describes the implantation of transit-time flow probes on the ascending aorta of rats while minimizing the risk of postoperative complications. Special emphasis is placed on our new method of rat intubation as well as the production of materials necessary for the implantation procedure such as endotracheal tubes and heparin bonded vessel catheters. The effects of these devices on the response to acute hypoxia were studied in rats following a 5-7 day recovery from the implantation procedure. Systemic and microvascular measurements were made on instrumented rats (n = 5) and non-instrumented controls (n = 3) that were ventilated with 21%, 15%, 10%, 8% and 5% oxygen. Arterial pressure, PO(2), lactate, and base deficit were not different between the implanted and control animals at any inspired oxygen concentration. Microvascular flow in the primary arterioles of the spinotrapezius muscle was also similar between the two groups at all inspired oxygen concentrations. We conclude that this novel methodology facilitates the measurement of whole body oxygen delivery in resting and haemodynamically-stressed rats.

Applying the "3 Rs": training course in surgical techniques.

As the use of surgical procedures in rodents becomes increasingly common in biomedical research, institutions face the challenge of ensuring that personnel are properly trained to perform these procedures. The author describes a microsurgery training course in use at Columbia University.

Microvascular free tissue transfer of the rectus abdominis muscle in dogs.

OBJECTIVE: To assess donor-site morbidity and survival of the rectus abdominis muscle with an overlying skin graft after free tissue transfer to a medial femorotibial defect in dogs. STUDY DESIGN: Experimental study. SAMPLE POPULATION: Phase 1: 6 canine cadavers; phase 2: 7 adult mixed-breed dogs. METHODS: Phase 1: The rectus abdominis muscle was removed from canine cadavers, muscular and vascular dimensions were recorded, and angiography was performed. Phase 2: Muscular transfer was performed through anastomosis of the caudal epigastric artery and vein to the saphenous artery and medial saphenous vein. Transferred tissues were evaluated on postoperative days 3, 6, 10, and 13. Animals were examined daily until euthanasia between postoperative days 31 and 42. Postmortem angiograms were performed and tissues collected for histopathologic evaluation. RESULTS: Phase 1: Appropriate vascular dimensions for microvascular anastomosis were confirmed and surgical technique perfected. Phase 2: Muscular excision produced minimal donor-site morbidity. All muscles survived after microvascular transfer and angiography confirmed vascular patency. All of the skin grafts survived, with one graft undergoing partial necrosis. CONCLUSIONS: The rectus abdominis muscle can be successfully transferred to a medial femorotibial defect and can serve as a bed for acute skin grafting. No significant donor-site morbidity is associated with its removal. CLINICAL RELEVANCE: Microvascular free tissue transfer of the canine rectus abdominis muscle has not been previously described. This technique provides a new alternative for repair of appropriate wounds. Additional studies are needed to define its utility in clinical patients.