A case of a dog with mandibular extraskeletal osteosarcoma after long-term puncture extirpation of the salivary gland cyst in the mandible.

Background: Extraskeletal osteosarcoma, unlike skeletal osteosarcoma, is a rare malignant mesenchymal tumor with a soft tissue primary that has been reported to occur in a variety of soft tissues. Case Description: The case is a 14-year-old, unneutered male Miniature Pinscher, weighing 6.7 kg, who had been treated medically for more than 5 years with a management strategy of puncture extirpation of a salivary gland cyst in the mandible; 1 month earlier, the fluid retention could not be removed, and after a computerized tomography scan showed no lesion in the mandible adjacent to the mass lesion, surgical resection was performed. Conclusion: Previous reports of extraskeletal osteosarcoma from the salivary glands in dogs have been rare. However, treatment of a salivary gland cyst in the mandible by long-term puncture extirpation may be a potential predisposing factor for the development of extraskeletal osteosarcoma around the mandible.

Feasibility and safety of three-dimensional electroanatomical cardiac mapping, mapping-guided biopsy and transseptal puncture in dogs.

Three-dimensional electroanatomical mapping (3D EAM) has expanded radiofrequency catheter ablation applications in humans to almost all complex arrhythmias and has drastically reduced fluoroscopy use, yet its potential in dogs is poorly investigated. The objectives of the current study were to assess the feasibility and safety of 3D EAM of all four heart chambers, 3D EAM-guided biopsies and transseptal puncture in dogs. Eight healthy purpose-bred Beagle dogs. Electroanatomical mapping was performed under general anaesthesia during sinus rhythm using a 22-electrode mapping catheter. Left heart catheterisation was achieved by either retrograde transaortic access (n = 4) or transseptal puncture (n = 4). Successful 3D EAM of the right atrium and ventricle was achieved in all dogs at a median time of 33 (13-40) min and 17 (3-52) min, respectively. Left atrial and ventricular 3D EAM was successful in six and seven dogs, at a median time of 17 (4-27) min and 8 min (4-19 min), respectively. Complications requiring intervention occurred in one dog only and were a transient third degree atrioventricular block and pericardial effusion following transseptal puncture, which was treated by pericardiocentesis. All dogs recovered uneventfully. Fluoroscopy time was limited to a median of 7 min (0-45 min) and almost exclusively associated with transseptal puncture. Three-dimensional EAM of all cardiac chambers, including mapping-guided biopsy and transseptal puncture is feasible in small dogs. Complications are similar to those reported in human patients. This suggests a potential added value of 3D EAM to conventional electrophysiology in dogs with arrhythmias.

Incidence of dural sac puncture during neuraxial anesthesia in cats: an observational, retrospective study.

OBJECTIVES: The aim of this study was to determine the occurrence of dural puncture, indicated by cerebrospinal fluid (CSF) outflow, in cats receiving neuraxial anesthesia through a lumbosacral injection guided by a pop sensation method. METHODS: This was an observational, retrospective study. Cats that were scheduled for lumbosacral neuraxial anesthesia were included. Medical records were analyzed to investigate: (1) demographic data; (2) neuraxial anesthesia performed (epidural/spinal); (3) type of needle used, including gauge and length; (4) presence of CSF (yes/no) and/or blood (yes/no) in the hub of the needle; and (5) flicking of the tail during needle advancement (yes/no). RESULTS: A total of 94 medical records were analyzed. A 22 G 50 mm Tuohy needle was used in all cats scheduled for an epidural injection (n = 60), whereas a 22 G 40 mm Quincke needle was used in all cats scheduled for an intrathecal injection (n = 34). CSF outflow was detected in 55/60 (91.7%) cats in which a Tuohy needle was used, and 34/34 (100%) of the cats in which a Quincke needle was used (P = 0.15). Flicking of the tail was detected in 41/60 (68.3%) and in 24/34 (70.6%) injections with Tuohy and Quincke needles, respectively (P >0.99). Traces of blood, but not active blood outflow, were detected via staining of the first drops of CSF in 2/34 cats in which Quincke needles were used and in none of the cats in which Tuohy needles were used (P = 0.12). CONCLUSIONS AND RELEVANCE: This study shows that the lumbosacral approach for neuraxial anesthesia in cats may result in a dural sac puncture when 22 G Quincke or Tuohy needles are used. The pop sensation method should be deemed effective in predicting intrathecal but not epidural needle placement.

Puncture of the Equine Embryonic Capsule and Its Repair In Vivo and In Vitro.

Vitrification of embryos >300 µm in diameter requires puncture of the glycoprotein capsule, although the size of the hole compatible with embryo survival is unknown. Forty-five day-7 or -8 embryos were punctured using a 30-µm glass biopsy pipette mounted on a micromanipulator (n = 20) or manually with either an acupuncture needle (∼100-µm diameter -hole; n = 10) or a microneedle with a <1 µm tip to produce a ∼30-µm diameter hole (n = 15) before transferring to recipient mares; further 12 embryos were punctured with either the acupuncture needle or microneedle before being cultured in vitro for 48 hrs (n = 3 per puncture group) or transferred to recipient mares and recovered 48 hrs later (n = 3 per puncture group). No pregnancies resulted from the 10 embryos punctured with the acupuncture needle, whereas 15 of 20 (75%) and 10 of 15 (67%) punctured on the micromanipulator or manually with the microneedle resulted pregnancies. Neither acupunctured nor microneedle-punctured embryos repaired their capsules in vitro. The acupunctured embryos also failed to repair their capsule after 48 hrs in vivo and subsequent uterine flushing yielded numerous capsular vesicles. The microneedle-punctured embryos did repair their capsule in vivo. Puncture with the microneedle opens the way for development of a manual method to vitrify equine embryos.

Percutaneous posterolateral approach for the simulation of a far-lateral disc herniation in an ovine model.

PURPOSE: This work describes a minimally invasive damage model for ovine lumbar discs via partial nucleotomy using a posterolateral approach. METHODS: Two cadavers were dissected to analyze the percutaneous corridor. Subsequently, 28 ovine had their annulus fibrosus punctured via awl penetration under fluoroscopic control and nucleus pulposus tissue removed via rongeur. Efficacy was assessed by animal morbidity, ease of access to T12-S1 disc spaces, and production of a mechanical injury as verified by discography, radiography, and histology. RESULTS: T12-S1 were accessible with minimal nerve damage morbidity. Scar tissue sealed the disc puncture site in all animals within 6 weeks, withstanding 1 MP of intradiscal pressure. Partial nucleotomy led to a significant reduction in intervertebral disk height and an increased histological degeneration score. CONCLUSION: Inducing a reproducible injury pattern of disc degeneration required minimal time, effort, and equipment. The posterolateral approach allows operation on several discs within a single surgery and multiple animal surgeries within a single day.

Clinical Practice of Epidural Puncture in Dogs and Cats Assisted by a Commercial Acoustic Puncture Assist Device-Epidural Locator: Preliminary Results.

The objective of this study was to compare an Acoustic Puncture Assist Device-Epidural Locator (APAD-EL) with the "pop sensation" (POP) and "lack of resistance" (LOR) commonly used to confirm penetration of the ligamentum flavum and to ensure correct epidural placement in dogs and cats. We recruited 38 dogs and cats undergoing surgery and receiving epidural analgesia. Two anesthetists performed epidural puncture using the POP and LOR signs. Simultaneously, APAD-EL was used to collect visual and acoustic confirmation during advancement and placement of the needle tip for post hoc evaluation. A positive APAD-EL sign consists of a sudden pressure drop at the needle tip visible on a display and a concomitant pitch change of an acoustic signal. Failure to record a sudden pressure drop is considered a negative APAD sign. Descriptive statistics were used. In 32 patients with positive POP and LOR, the APAD was also positive. In one patient, POP was positive with a negative LOR and APAD result. Five patients had negative POP but positive LOR. Four patients had APAD positive and one (a dog) APAD negative. The study results showed that the APAD-EL information supports the subjective signs of correct needle placement suggested by positive POP and LOR experienced by trained anesthetists. The technique can be useful to assist difficult epidural puncture and as a training and teaching tool.

Impact of fine needle aspiration (FNA) and of the number of punctures on the feline testis: clinical, gross anatomy and histological assessment.

The safety of testicular fine needle aspiration (FNA) has been proven in dogs but has not been fully established in men, while studies in rats have given contradictory results. Furthermore, the extent of damage inflicted by multiple punctures is unknown. The aim of this study was to determine the impact of FNA and of the number of punctures on the feline testis with clinical, gross anatomy and histological examinations. Twenty-seven sexually mature healthy laboratory Domestic Shorthair cats were randomly assigned to two groups: 5 cats in which no FNA was performed (control group), and 22 cats which had their left and right testis punctured with a 26 ga needle towards 3 and 8 directions, respectively (experimental group). Two cats at a time were orchiectomized 5 or 30 min, 1, 2, 4, 7 or 14 days or 1, 2, 3 or 4 mo post-aspiration. The cats of the control group were also orchiectomized. During the first week post-aspiration clinical examination revealed vaginal cavity hematoma (8/44 testes), while the histological findings were focal hemorrhagic areas (20/24 testes), erythrocytes inside the seminiferous tubules' lumen (9/24 testes), and germinal cell degeneration in <1.94% of the seminiferous tubules (15/24 testes). After the first week the histological findings were germinal cell degeneration in <2.14% of the seminiferous tubules (19/20 testes) and enlargement of the lumen of <5.16% of the seminiferous tubules (7/20 testes). The germinal epithelium and interstitium had an overall normal appearance. No significant differences were observed between the left and right testis. The results of the study indicate that testicular FNA should be considered a safe procedure in the cat when up to 8 punctures are performed.

Effect of needle size and type, reuse of needles, insertion speed, and removal of hair on contamination of joints with tissue debris and hair after arthrocentesis.

OBJECTIVE: To assess joint contamination with tissue and hair after arthrocentesis of equine fetlock joints. STUDY DESIGN: Experimental. ANIMALS: Limb specimens from 8 equine cadavers. PROCEDURES: Soft tissues including the joint capsule were harvested from the dorsal aspect of the fetlock joints and mounted on a wooden frame. Needles inserted through the joint tissue preparation were flushed into tissue culture plates that were examined for tissue and hair debris. Variables evaluated were gauge and type of needle (16, 18, 20, and 22 G sharp disposable needles and 20 G disposable spinal needles with stylet), number of times each needle was used (1, 2, 3, 4), length of hair (unclipped, clipped, shaved with razor), and needle insertion speed (fast, slow). Descriptive and statistical evaluations were performed. RESULTS: Tissue contamination was identified in 1145 of 1260 wells and hair contamination was identified in 384 of 1260 wells. Twenty gauge needles inserted through unclipped hair resulted in the least amount of hair contamination. Compared with 20 G needles with fast insertion 1 time through unclipped hair the odds ratios for contamination with hair were significantly greater for 16 G sharp disposable needles, 20 G spinal needles, clipped hair, shaved hair, and reuse of the needles. Spinal needles inserted through unclipped hair transferred many long hairs into the joint space. CONCLUSION: Reuse of needles for arthrocentesis should be avoided. Removal of hair is not indicated for arthrocentesis with sharp injection needles but is recommended when using spinal needles with stylets. CLINICAL RELEVANCE: Joint contamination with hair and tissue debris will be decreased by specific needle insertion techniques. Decreased contamination of joints may reduce the frequency of joint infections after arthrocentesis.

Effects of twice-weekly follicular punctures of ovaries with or without the corpus luteum on follicular and luteal dynamics.

We investigated the effects of twice-weekly follicular punctures of ovaries with or without corpus luteum (CL) on follicular and luteal dynamics. A cross-over design was used, with each cow (seven Japanese Black beef cows) being assigned to one of the three groups at 2-month intervals. Follicular punctures were performed twice weekly for three consecutive weeks until day 20 (oestrus = day 0). All visible follicles (diameter >3 mm) in the ovaries bearing CL (ipsilateral group) or those in the contralateral ovaries (contralateral group) were aspirated. As a control, all visible follicles in both ovaries were aspirated (bilateral group). Follicular development, CL formation and progesterone concentrations in each cow were monitored from days 0 to 30. Follicular growth profiles in the punctured ovaries during/after puncture treatment were similar, irrespective of the presence of follicles in the unpunctured ovary and the CL in the punctured or unpunctured ovaries. After puncture, two cows (28.6%) each in the ipsilateral and bilateral groups did not exhibit behavioural oestrus until day 30, whereas all cows in the contralateral group exhibited oestrus. CL growth and increase in progesterone concentrations after the last follicular puncture in the bilateral group were delayed when compared with those in the ipsilateral group. Our results indicate that the presence of follicles in the unpunctured ovary and the CL in the punctured or unpunctured ovaries does not significantly influence follicular growth in punctured ovaries during/after puncture treatment. However, follicular puncture in ovaries bearing CL may disturb or delay oestrus occurrence after treatment.

Percutaneous external jugular vein catheterization in piglets using a triangulation technique.

Chronic jugular vein or central venous cannulation is routinely performed in human and animal patients for access to blood circulation. In mature swine, chronic catheter placement techniques have typically involved venous isolation via extensive cut-down, blunt dissection and manipulation of ventral neck tissues prior to catheter placement. More recently, guide-wire-assisted percutaneous techniques have become standard practice in human and veterinary medicine due to the minimization of soft tissue and vessel damages. Laboratory animal piglets are becoming more popular research models because of their immature immunological system, ease of handling and costs. However, external jugular veins are very difficult to catheterize in paediatric animals including freshly weaned piglets. The objective of this study was to develop a simple, safe and efficient method for external jugular vein cannulation in young piglets. In total, 20 piglets were anaesthetized and percutaneously catheterized with a guide-wire technique using palpable anatomical landmarks and triangulation. With this minimally invasive catheterization, it has allowed our veterinarians and veterinary technicians to quickly and easily obtain central venous access in piglets undergoing operative procedures.

[Subperitoneal cyst in a Friesian mare]. / Subperitoneale cyste bij een Friese merrie.

The case of a 5-year-old Friesian mare with a mass in her abdomen and a gait irregularity of the left hind limb is discussed. At rectal ultrasonogrphic examination the mass appeared to be a thin-walled, anechogenic cavity in the left abdomen. By means of laparoscopy the mass could be visualized as a smooth structure, covered by peritoneum and fat. Under laparoscopic guidance the mass was punctured and fluid was aspirated. Cytological and bacteriological findings of the fluid were indicative of a cyst. After drainage of the cyst the locomotion disorder of the left hind limb disappeared. It is concluded that this horse suffered from a subperitoneal cyst, a rare disorder in humans and never previously described in horses. The case emphasizes the value of laparoscopy in the diagnosis and treatment of (intra)abdominal masses and further clearly shows that in the differential diagnosis of locomotion disorders not only structures related to the locomotion system should be taken into account.

Ovarian function and morphology in the mare after multiple follicular punctures.

REASONS FOR PERFORMING STUDY: In the mare, ultrasound-guided transvaginal oocyte recovery and transfer might offer a way to circumvent the demanding procedures of in vitro embryo production. Before clinical application, the possible consequences for subsequent fertility have to be considered. OBJECTIVES: To examine ovarian function and morphology in mares after repeated follicular punctures. METHODS: A total of 14-26 follicular puncture sessions were conducted on each of 4 Norwegian pony mares over a period of 8 years. The ovaries of these mares were recovered by bilateral ovariectomy or at post mortem and subjected to macroscopic inspection and histology. For comparison, ovaries were collected from 7 nonaspirated control mares and processed for histology. RESULTS: In all experimental mares, ovarian function, defined as the ability regularly to ovulate preovulatory follicles and develop corpora lutea, remained normal during their last breeding season. Gross examination and histology showed that normal follicular and corpus luteum development was accompanied by the formation of condensed reparative fibrosis and normal local haemosiderosis of the ovarian stroma in all experimental mares. In one mare, an ovary contained several foci of chronic apostematous oophoritis, while a cystic structure lined with a single layer of epithelial-like cells and surrounded by a cartilaginous capsule was present in the other ovary. CONCLUSIONS AND POTENTIAL RELEVANCE: Repeated follicular aspirations do not hamper future folliculogenesis, ovulation and corpus luteum formation. However, ovarian puncture induces reparative fibrosis in the ovarian stroma and involves a risk of inducing abscess formation within the ovarian tissue which may impair fertility.

Evaluation of dorsal aorta cannulation for immunological studies of grouper (Epinephelus malabaricus).

Blood is often withdrawn to study the immune responses of fish. However, netting, handling and anaesthetising the experimental fish, and drawing blood samples cause severe stress that may alter the effects of immune study protocols and treatments. We evaluated the effect of aorta cannulation, for use in immune studies, on grouper (Epinephelus malabaricus) plasma cortisol, total red and white blood cell counts and phagocytosis. Plasma cortisol increased from 30 to 88 ng/ml 1 day after insertion of the cannula, to a maximum of 951 ng/ml 3 to 5 days after surgery, indicating the groupers were stressed by cannulation and post-cannulation inflammation. Total RBC count decreased, and total WBC count increased after surgery. Following cannulation, the phagocytic index of peripheral blood leukocytes decreased from 100% to 46%. The adverse effects of cannulation were mitigated by continuously immersing groupers in oxytetracycline (OTC), which decreased the recovery period for treated fish. In contrast, OTC-treatment did not markedly improve the recovery of groupers subjected to caudal vessel puncture. Cortisol levels in OTC-treated grouper with caudal vessel puncture were significantly higher than in OTC-treated, cannulated grouper, and remained at a high level until day 13 of the experiment. From day 7 to 13, total RBC and WBC counts in OTC-treated, cannulated groupers were significantly different from those in OTC-treated groupers with caudal vessel puncture. OTC treatment improved the phagocytic index of groupers subjected to caudal vessel puncture, but the phagocytic index was lower than that of groupers subjected to cannulation. Cannulation minimises visual and handling disturbances, and facilitates standardisation of experimental conditions and quick and easy sampling via the dorsal aorta cannula. Therefore, dorsal aorta cannulation minimises the stress of blood sampling and should prove useful for immune studies in fish.

Effects of repeated follicular punctures on ovarian morphology and endocrine parameters in dairy heifers.

Three experimental and three control heifers were included in the study. After a control period (CP), the experimental heifers were subjected to 4 and 5 weeks of twice-weekly follicular punctures (FPP1 and FPP2) with two oestrous cycles in between. The follicular punctures were performed using the ovum pick-up (OPU) technique. During the CP and FPPs, ovarian activity was monitored by transrectal ultrasonography and analyses of plasma progesterone, prostaglandin F2alpha (PGF2alpha) metabolite, luteinizing hormone (LH), oestradiol and follicle stimulating hormone (FSH) levels. The animals were slaughtered 7 days after cessation of FPP2 and their ovaries examined. The control animals were monitored for their oestrous cyclicity and were then ovariectomized and the ovaries examined. Despite the absence of ovulation, the formation of corpus luteum (CL)-like structures could be ultrasonographically observed during FPPs. These structures either had characteristics comparable to those of the CLs during the CP (group 1) or were smaller and had a shorter life span and (or) luteal active phase with a lower progesterone production (group 2). In group 1, CL-like structures emanated from punctures of large-sized follicles on or close to the days on which the animals showed oestrus and had a rise in oestradiol and LH. In group 2, the structures developed from punctures of smaller-sized follicles and no oestrus nor a detectable 1.14 rise was revealed around punctures. During most FPPs, the basal level of LH was at zero. During all FPPs, a significant rise in FSH level occurred on the day following the day of puncture. To conclude, repeated follicular puncture appeared to alter slightly endocrine profiles and cause minor morphological changes in the ovaries. Both the macro- and microscopic examinations revealed the presence of luteal structures and follicles of various sizes in the ovaries of all heifers studied. The connective tissue in the ovarian tunica albuginea of the experimental heifers was significantly thicker than that in controls (P < 0.001).

The effect of repeated follicular puncture on ovarian function in dairy heifers.

Three dairy heifers were examined during three consecutive oestrous cycles (control period, CP). Subsequently, the animals were subjected to 4 and then 5 weeks of twice-weekly ovum pick-up (OPU) (FPP1 and FPP2, respectively) with a recovery period (RP) of two consecutive oestrous cycles between FPP1 and FPP2. After FPP2, the animals were slaughtered and the ovaries were macroscopically examined. Throughout, ovarian activity was monitored by transrectal ultrasonography and concentrations of plasma progesterone. During CP, all the heifers showed normal cyclicity. During FPPs, the heifers occasionally presented oestrous activity. Corpus luteum (CL)-like structures developed from punctured follicles with diameters and life-spans varying from smaller and shorter than those in the CP (P > 0.05) to equal to those in the CP. There was a tendency for a lower number of emerging and punctured follicles in the presence of a CL-like structure. Subsequently to FPP1, all heifers regained normal cyclicity. A thickening of the ovarian tunica albuginea and a slight hardening of the ovaries were found postmortem. In conclusion, dairy heifers can occasionally show cyclic activity and form CL-like structures during twice-weekly OPU. Further, OPU did not seem to cause any major negative effects on ovarian structure and subsequent ovarian function.

[Diagnosis, therapy and endocrinologic parameters of persistent follicles in mares in comparison with preovulatory follicles]. / Diagnose, Therapie und endokrinologische Parameter persistierender Follikel bei der Stute im Vergleich zu präovulatorischen Follikeln.

During the 1997 breeding season persistent follicles were diagnosed in 17 mares. In 16 of these mares a total of 17 follicles were transabdominally punctured and the steroids oestradiol, progesterone and testosterone were measured in the follicular fluid and in blood serum. In ten mares serving as a control group preovulatory follicles were punctured. The follicular fluid of the persistent follicles revealed a very high variability of the steroid concentrations. Depending on the steroid ratio within the follicles, eight follicles were rated as being intact, three follicles were undergoing atresia and five follicles were luteinized. Because of the high oestradiol levels of the follicular fluid within the control group, all of these follicles were considered to be intact. In both groups, no correlation of the steroid concentration between serum and follicular fluid was detectable. This fact argues against a passive diffusion of the steroids through the follicular wall. By puncturing the persistent follicles it was possible to bring the affected mares back into a physiological oestrus cycle within a normal dioestrus period.

Incomplete oblique sagittal fractures of the dorsal cortex of the third metacarpal bone in six horses.

OBJECTIVE: To describe incomplete oblique sagittal dorsal cortical fractures of the equine third metacarpal bone, their surgical repair, and subsequent performance of the horses. STUDY DESIGN: Retrospective examination of medical records and racing performance. ANIMAL POPULATION: Six Thoroughbred race horses, 2 to 4 years of age. METHODS: Radiographic confirmation of all fractures preceded general anesthesia and surgical correction. Three fractures were treated by intracortical compression using screws placed in lag fashion, and five fractures were treated by osteostixis. Race records were reviewed for each horse to determine performance after surgery. RESULTS: Fractures were best observed on palmarodorsal radiographic projections. Three horses treated by intracortical compression returned to racing, but fracture recurred in one horse and was treated by osteostixis. This horse and the other three horses treated by osteostixis raced after surgery. CONCLUSIONS: Horses with incomplete oblique sagittal fractures of the dorsal cortex of the third metacarpal bone can race after surgical management of the fracture by screws placed in lag fashion or osteostixis. The authors' preferred surgical procedure for managing this fracture is osteostixis. CLINICAL RELEVANCE: Palmarodorsal radiographic projections of the third metacarpal bone are recommended in young Thoroughbred race horses suspected of having dorsal metacarpal stress fractures.

Alteration of follicular dynamics and superovulatory responses by gonadotropin releasing hormone and follicular puncture in cattle: a field trial.

A field experiment was conducted to determine the influence of follicular alteration on superovulatory responses. Ultrasonography was performed once daily over 4 d prior to gonadotropin treatment (Day 0), on the day of estrus during superstimulation, and on the day of embryo collection to monitor follicular development. Animals were superstimulated between Days 8 and 12 of the estrous cycle. Follicular status was altered 2 d prior to initiation of superstimulation (Day 0) with GnRH (Cystorelin, 200 micrograms i.m.) administered with (GnRH-puncture group, n = 31) or without (GnRH-no puncture group, n = 52) concomitant removal of the largest follicle by follicular aspiration. Responses were compared with those of an untreated control group superovulated 8 to 12 d after estrus (n = 102). The proportion of animals with a high number (> or = 2) of large follicles (> = 7 mm) on Day 0 was lower (P < 0.001) in the 2 GnRH-treated groups than in the control group, while the increase in the number of medium size follicles (4 to 6 mm) on Day 0 was greater (P < 0.02) in the GnRH-puncture group. During superstimulation, the proportion of superovulatory cycles with a high follicular (> or = 10 follicles) response was similar in the control and GnRH-no puncture groups. Within the GnRH-treated animals, follicular and ovulatory responses were greater in the GnRH-puncture than in the GnRH-no puncture group (P < 0.001 to P < 0.02). Despite these changes in follicular and ovulatory responses, however, the mean number of embryos produced did not differ (P < 0.1) among treatments (4.3 +/- 0.4, 3.7 +/- 0.7, and 5.4 +/- 0.8 in control, GnRH-no puncture, and GnRH-puncture groups, respectively). This was due primarily to an increase in the mean numbers of unfertilized ova (P < 0.005) and in degenerated embryos (P < 0.06) in the GnRH-puncture group. Results indicate that the beneficial effects of treatment with GnRH and follicular puncture 2 d prior to superstimulation on follicular and ovulatory responses were limited by an increase in the number of unfertilized ova and degenerated embryos.