Gender Identification in Great Sturgeon (Huso huso) Using Morphology,Sex Steroids,Histology and Endoscopy

Gender identification of great sturgeon during different life stages was conducted using morphology, sex steroid concentrations, histological and endoscopic techniques. One hundred and forty great sturgeons from two sturgeon farms at ages between 3‐16 years were selected for examination. Most of the morphological indices showed no significant sex‐related differences except for the ratio of fork length to distance of snout to anterior of dorsal fin (PD). The urogenital opening had no difference in shapes for both sexes. Mean sex steroid concentrations showed no significant difference in levels of testosterone, estradiol and progesterone between males and females in the stage II of maturity. 95.8% of fish were correctly sexed using endoscopy at this stage. Histological investigation indicated the ovaries of 3‐ to 9‐year‐old females were at the different stages of pre‐vitellogenic, and most males were at the early spermatogenesis for age 3. The result of this study suggests that endoscopy is a reliable tool for sex and stage determination after the age of 3 years. This method is useful in providing a quick discernment of sexes and stages compared with utilizing morphometric and blood indices. Direct observation of gonads can give us important information about the gonadal and maturity stages in sturgeon aquaculture and wild population management, with minimal damage or stress to this endangered species.     

Evaluation of an endoscopic liver biopsy technique in green iguanas

OBJECTIVE: To establish a safe and effective endoscopic technique for collection of liver biopsy specimens from lizards by use of a 2.7-mm rigid endoscope system that is commonly available in zoologic veterinary practice. DESIGN: Prospective study. ANIMALS: 11 subadult male green iguanas (Iguana iguana). PROCEDURES: Each lizard was anesthetized, and right-sided coelioscopic examination of the right liver lobe and gallbladder was performed. Three liver biopsy specimens were collected from each lizard by use of a 2.7-mm rigid endoscope and 1.7-mm (5-F) biopsy forceps. Biopsy samples were evaluated histologically for quality and crush artifact. Ten days following surgery, all iguanas were euthanatized and underwent full necropsy examination. RESULTS: For all 11 iguanas, the right liver lobe and gallbladder were successfully examined endoscopically, and 3 biopsy specimens of the liver were collected without complications. Mean +/- SD durations of anesthesia and surgery were 24 +/- 7 minutes and 6.8 +/- 1.0 minutes, respectively. At necropsy, there was no evidence of trauma or disease associated with the skin or muscle entry sites, liver, or any visceral structures in any iguana. All 33 biopsy specimens were considered acceptable for histologic interpretation; in most samples, the extent of crush artifact was considered minimal. CONCLUSIONS AND CLINICAL RELEVANCE: By use of a 2.7-mm rigid endoscope, liver biopsy procedures can be performed safely, swiftly, and easily in green iguanas. Biopsy specimens obtained by this technique are suitable for histologic examination. For evaluation of the liver and biopsy specimen collection in lizards, endoscopy is recommended.     

Paralumbar Fossa Laparoscopic Ovariectomy in Horses With Use of Endoloop Ligatures

Objective —The purposes of this study were to develop a technique of paralumbar fossa laparoscopic ovariectomy using Endoloop ligatures and to avoid a laparotomy incision for ovary removal by using a 33–mm diameter muscle spreader trocar-cannula unit.
Animals or Sample Population —Seven mares.
Methods —Bilateral laparoscopic ovariectomy was performed under general anesthesia in two horses and under neuroleptanalgesia and local anesthesia in five standing mares. Ovaries were approached from the ipsilateral paralumbar fossa through two portal sites located in the paralumbar fossa and a third between the 17th and 18th ribs. Insufflation of the abdominal cavity was achieved using an automatic carbon dioxide insufflator. Two Endoloop ligatures were placed on the mesovarium. The cannula, located in the center of the paralumbar fossa, was removed, and a 33-mm diameter trocar-cannula unit, with a cone-shaped muscle spreader extremity, was inserted through the same portal. The mesovarium was transected between the ovary and the ligatures. The ovaries were removed from the abdomen through the 33-mm diameter cannula. The abdominal wall was closed in a routine manner.
Results —No major complications occurred during or after surgery.
Conclusion —Paralumbar fossa laparoscopic ovariectomy in mares using Endoloop ligatures and the 33-mm diameter trocar-cannula unit was an effective technique for ovariectomy of normal ovaries in this study.
Clinical Relevance —Endoloop ligatures provide a viable alternative for laparoscopic ligation of ovarian pedicles in mares.     

Standing female equine urogenital endoscopic surgery.

Standing female urogenital endoscopic surgery is facilitated by the more dorsal location of the organs of the female reproduction tract. The most common reason for laparoscopic surgery on the female urogenital system is ovariectomy; however, the technique has been used to diagnose periparturient or reproductive diseases and to perform surgical embryo transfer. Standing surgical approaches avoid the risk and expense of general anesthesia, but these techniques are limited by the temperament and size of the patient and the availability of facilities for restraint. Owner acceptance of laparoscopic procedures is excellent. It is anticipated that standing endoscopic surgery may become the preferred technique for applicable procedures.     

Comparison between coelioscopy and coeliotomy for liver biopsy in channel catfish

Objective-To evaluate endoscopic liver biopsy and compare that technique with a standard coeliotomy biopsy technique in fish. Design-Randomized controlled clinical trial. Animals-30 channel catfish (Ictalurus punctatus). Procedures-10 fish were randomly assigned into control, coeliotomy, and coelioscopy groups. Anesthesia was performed with a recirculating anesthesia machine. Body weight, PCV, and total protein (TP) concentration in blood as well as plasma activities of aspartate aminotransferase, creatinine phosphokinase, lactate dehydrogenase, and sorbitol dehydrogenase were measured before and after surgery. Standard ventral coeliotomy or coelioscopy was performed, and the biopsy specimens were scored histologically. Results-Coeliotomy and coelioscopy procedures were well tolerated without acute deaths. Blood TP concentration and PCV decreased after surgery in the coelioscopy group because of intracoelomic fluid administration to aid visualization. Minor changes in activities for hepatic and muscular enzyme activities were apparent, but were not significantly different between the coelioscopy and coeliotomy groups. Coelioscopy and coeliotomy yielded biopsy specimens of similar diagnostic quality. However, coelioscopy permitted a more extensive evaluation of the viscera, and all 10 surgical wounds healed completely, compared with severe wound dehiscence in 3 of 10 fish that underwent coeliotomy. Conclusions and Clinical Relevance-Both coelioscopy and coeliotomy were capable of yielding antemortem liver biopsy specimens of diagnostic quality in catfish. Coelioscopy permitted a more detailed examination of the coelomic viscera through a smaller surgical incision, was less traumatic, and resulted in decreased wound dehiscence.     

Laparoscopic esophagopexy,fundopexy, and hiatal herniorrhaphy for refractory regurgitation in a racing Alaskan husky sled dog

A 2-year-old intact female Alaskan husky sled dog was presented with a history of chronic exercise-induced regurgitation refractory to medical management. Previous diagnostics were unremarkable except for an endoscopic examination and histopathologic evaluation of the upper gastrointestinal tract that revealed the presence of Helicobacter spp. and mild non-specific inflammation of the proximal duodenum. A laparoscopic hiatal herniorrhaphy, esophagopexy, fundopexy, and ovariectomy were performed without complications in anesthesia or surgery and clinical improvement was observed with continued follow-up for 8 months after surgery.Key clinical message:Surgical treatment for hiatal hernia may be considered in racing Alaskan sled dogs with regurgitation refractory to gastric protectant therapy.     

Laparoscopy in zoological medicine.

The technique of laparoscopy was adapted and utilized in zoological medicine for various mammals, birds, and reptiles for reproductive and diagnostic studies as well as clinically related research. It was concluded that since anesthesia was routinely required for most manipulative procedures in zoo animals, and since laparoscopy adds little additional risk, the use of this technique provides an additional diagnostic aid when indicated. Laparoscopy was found to be effective for evaluating reproductive status, particularly ovarian anatomy and function, direct visual biopsy of internal organs, sex determination in selected birds, and as a surgical means of fertility control.     

Evaluation of transcutaneous pulmonoscopy for examination and biopsy of the lungs of ball pythons and determination of preferred biopsy specimen handling and fixation procedures

OBJECTIVE: To establish a safe and effective technique for the endoscopic examination and biopsy of snake lungs by use of a 2.7-mm rigid endoscope system. DESIGN: Clinical trial. ANIMALS: 17 adult ball pythons (Python regius). PROCEDURES: The right lung of each anesthetized snake was transcutaneously penetrated at a predetermined site. Endoscopic lung examination was objectively scored, and 3 lung biopsies were performed. Tissue samples were evaluated histologically for diagnostic quality. One year later, 11 of the 17 snakes again underwent pulmonoscopy and biopsy; specimens were placed in various fixatives to compare preservation quality. All 17 snakes were euthanatized and necropsied. RESULTS: No major anesthetic, surgical, or biopsy-associated complications were detected in any snake. In 16 of 17 pythons, ease of right lung entry was satisfactory to excellent, and views of the distal portion of the trachea; primary bronchus; intrapulmonary bronchus; cranial lung lobe; and faveolar, semisaccular, and saccular lung regions were considered excellent. In 1 snake, mild hemorrhage caused minor procedural difficulties. After 1 year, pulmonoscopy revealed healing of the previous transcutaneous lung entry and biopsy sites. Important procedure-induced abnormalities were not detected at necropsy. Diagnostic quality of specimens that were shaken from biopsy forceps into physiologic saline (0.9% NaCl) solution before fixation in 2% glutaraldehyde or neutral-buffered 10% formalin was considered good to excellent. CONCLUSIONS AND CLINICAL RELEVANCE: By use of a 2.7-mm rigid endoscope, lung examination and biopsy can be performed safely, swiftly, and with ease in ball pythons. Biopsy specimens obtained during this procedure are suitable for histologic examination.     

Ovarian biopsy: a non-terminal method to determine reproductive status in giant kokopu,Galaxias argenteus (Gmelin 1789)

AIM: To establish a method of gonad biopsy for ovarian tissue collection in the declining giant kokopu Galaxias argenteus (Gmelin 1789) as an alternative to lethal sampling in order to understand the species' reproductive biology.

METHODS: Six female giant kokopu weighing between 200 and 350 g were caught from the wild in early December of 2009 and transferred to a holding facility (Department of Zoology, University of Otago, Dunedin) where they were kept under a simulated natural photo-thermal regime for 10 months. Fish were repeatedly biopsied for ovarian tissue at near-monthly intervals (mean number of days between biopsies = 33) until ovulation.

RESULTS AND CONCLUSIONS: Ovarian samples were successfully collected from giant kokopu by biopsy for use in downstream analyses. Among a total of 23 biopsy events, a single death occurred when a two-layered suturing approach was used, highlighting the value of this method for study of the reproductive biology of valuable fish.

CLINICAL RELEVANCE: This biopsy method may have implications for veterinary research on fish physiology, pathology, conservation and development, when repeated tissue samples need to be collected over a prolonged period of time or for general surgical manipulations on fish when accessing the coelom. Furthermore, this approach allows the implementation of a more powerful experimental design, as repeated measures reduces the variability of estimates due to the removal of inherent stage differences among individuals.     

Use of ketamine, xylazine, and diazepam anesthesia with retrobulbar block for phacoemulsification in dogs

Objective  The study was undertaken to evaluate the use of ketamine, xylazine, and diazepam along with a local retrobulbar nerve block for routine phacoemulsification in the dog.
Animals  Ten clinically healthy mixed-breed dogs of either sex, weighing between 10 and 15 kg.
Procedures  Ten mixed-breed dogs were selected for unilateral cataract removal by phacoemulsification. Standard preoperative preparations for cataract surgery were followed. Pre-anesthetic medication consisted of atropine sulfate (0.02 mg/kg, SC). Anesthesia was induced by xylazine HCl (1.0 mg/kg, IM) followed by ketamine (5.0 mg/kg, IM). Anesthesia was maintained subsequently with IV ketamine and diazepam to effect and depth of anesthesia was assessed clinically by pedal reflex and jaw reflex. After induction of anesthesia, a retrobulbar nerve block was performed using 2 mL of 2% lignocaine. Eye position was graded after retrobulbar block and IOP was examined preoperative, post-anesthetic, 6 h postoperative and 24 h after surgery. Phacoemulsification was performed using the phaco-chop technique and an intraocular lens was placed. Anesthetic recovery and postoperative recovery following surgery was recorded.
Result  The exposure of the globe in all the dogs was adequate; the desired central fixation of the eye was obtained and surgery could be performed uneventfully. The mean IOP recorded after induction of anesthesia was 15.75 ± 0.82, which was not significantly ( P  > 0.01) different from pre-anesthetic values (14.85 ± 0.85).
Conclusion  Phacoemulsification was successfully performed with this anesthetic regimen without encountering major intraoperative or anesthetic complications.     

Standing abdominal surgery.

Safe, effective surgery can be performed in the standing, conscious horse using a combination of mechanical and chemical restraint. Clear indications for performing standing abdominal surgery exist, but many procedures are best performed under general anesthesia. The preferred approach involves a modified grid incision of the left paralumbar fossa. Flank celiotomy allows the surgeon to thoroughly explore the abdominal cavity, but few structures can be exteriorized, and visibility of abdominal contents is poor. Indications for standing abdominal surgery include diagnosis of abdominal masses, drainage and biopsy techniques, correction of left dorsal displacement of the large colon, and evaluation of rectal injuries, and performing loop colostomy techniques, laparoscopy, removal of retained testicles, correction of uterine torsions, surgical embryo transfer, ovariectomy in normal mares, and some experimental procedures. Standing surgical techniques are most useful and appropriate for surgical exploration, to correct uterine torsions, and to perform loop colostomy and surgical embryo transfer techniques. Perioperative antimicrobial and antiinflammatory therapy is recommended. Mild discomfort and ventral incisional swelling after surgery are expected.     

Standing Laparoscopic Ovariectomy Technique with Intraabdominal Dissection for Removal of Large Pathologic Ovaries in Mares

Objective: To describe a standing laparoscopic ovariectomy technique with intraabdominal ovarian dissection inside a specimen retrieval bag for removal of large pathologic ovaries through small incisions. Study Design: Case series. Animals: Mares (n=43) aged 2–21 years and weighing 380–680 kg. Methods: Unilateral laparoscopic ovariectomy was performed on 43 standing sedated mares. Ovaries were approached via 3 portal sites, 2 in the paralumbar fossa and a 3rd between the 17th and 18th ribs on the ipsilateral side. Ovaries were dissected free using either a LiNA Tripol‐bipolar laparoscopic forceps or a Ligasure™ Vessel Sealing Device and removed by enlarging the proximal portal site ventrally using a grid technique. Use of a plastic specimen retrieval bag and cannula suction device facilitated intraabdominal dissection of very large ovaries into 2 or more pieces, before removal through small incisions. Results: Regardless of size, all ovaries were removed successfully through small incisions (range, 5–10 cm), with no major complications and an excellent cosmetic result. All sport horses returned to previous levels of work or higher, with 93% of breeding mares successfully bred in the 1st season after surgery. Conclusions: Large pathologic ovaries can be easily and safely removed by standing laparoscopic ovariectomy on the mare. Clinical Relevance: Standing laparoscopic ovariectomy combined with a small flank incision is a safe and highly effective technique for removal of large pathologic ovaries in the mare, negating the requirement for general anesthesia or large incisions.     

Evaluation of rhinoscopy and rhinoscopy-assisted mucosal biopsy in diagnosis of nasal disease in dogs: 119 cases (1985-1989).

The case records of 149 dogs examined from 1985 to 1989 with clinical signs of nasal disease were reviewed. Gross rhinoscopy was performed in 119 dogs, and rhinoscopy-assisted pinch biopsy was performed in 109. Rhinoscopy was performed by use of a 2.7-mm rigid fiberoptic endoscope. Mucosal biopsy specimens were obtained with rhinoscopic guidance by use of a 2 x 3-mm biopsy forcep. Gross, cytologic, and histologic findings are summarized. Ninety-four of 119 cases could be evaluated on the basis of diagnostic and follow-up criteria established by the authors. The diagnostic success rate of gross rhinoscopy with rhinoscopy-assisted biopsy was 83% (78 of 94 evaluated cases). Protracted hemorrhage was a complication in 2 of 109 cases in which rhinoscopy-assisted biopsy was performed. It was concluded that rhinoscopy with rhinoscopy-assisted biopsy contributes important diagnostic information in dogs with nasal disease without the relative invasiveness, expense, and risk of surgery.     

DETERMINATION OF FISH GENDER USING FRACTAL ANALYSIS OF ULTRASOUND IMAGES

The gender of cod Gadus morhua can be determined by considering the complexity in their gonadal ultrasonographic appearance. The fractal dimension ( D _B) can be used to describe this feature in images. B-mode gonadal ultrasound images in 32 cod, where gender was known, were collected. Fractal analysis was performed on these images and D _B was determined using the box counting method. A receiver–operating curve (ROC) was drawn for D _B as a test for male fish. Using a range of D _B values, the maximum accuracy for this test was calculated and compared with the accuracy for identifying male fish by subjective analysis alone. The mean (and standard deviation) of the fractal dimension D _B for male fish was 1.554 (0.073) while for female fish it was 1.468 (0.061); the difference was statistically significant ( P =0.001). The area under the ROC curve was 0.84 indicating the value of fractal analysis in gender determination in cod. Maximum accuracy (0.84) for D _B as a test for male fish was obtained using the threshold value D _B=1.5058 compared with an accuracy of 0.78 for subjective image evaluation. The use of two thresholds, D _B<1.4475 (females) and D _B>1.5054 (males) gives an 80% certainty in the classification result. Fractal analysis is useful for gender determination in cod. This or a similar form of analysis may have wide application in veterinary imaging as a tool for quantification of complexity in images.     

Ventral Abdominal Approach for Laparoscopic Ovariectomy in Llamas

Objective —To evaluate a technique for laparoscopic ovariectomy by a ventral abdominal approach in the llama.
Study Design —Prospective evaluation.
Animal Population —Six healthy adult female llamas, aged 2 to 6 years.
Methods —Laparoscopic ovariectomy was performed with the llamas in a 30-degree Trendelenburg position. The mesovarium and proper ligament of the ovary were isolated, and two Hulka clips were placed in opposite directions across each mesovarium and proper ligament of the ovary. Ovaries were removed through either the instrument cannula or instrument portal.
Results —The ovaries, mesovarium, and proper ligaments of the ovaries were easily identified using laparoscopy by a ventral abdominal approach. The Hulka Clip Applicator was easy to use, and the Hulka Clips provided secure hemostasis of the vessels within the mesovarium. Mean duration of surgery was 35 minutes (range, 17 to 85 minutes). Two llamas were depressed after surgery but improved within 24 hours. No other complications occurred during an 18-month follow-up period.
Conclusions —Laparoscopic ovariectomy in the llama was facilitated by the use of Hulka Clips placed across the mesovarium and proper ligament of the ovary.
Clinical Relevance —Laparoscopic ovariectomy in the llama can be accomplished easily with few complications.     

Laparoscopic adrenalectomy for treatment of unilateral adrenocortical carcinomas: technique, complications, and results in seven dogs

Objective— To investigate the feasibility of, and outcome after, laparoscopic adrenalectomy in dogs with unilateral adrenocortical carcinoma.
Study Design— Case series.
Animals— Dogs (n=7) with Cushing's syndrome caused by unilateral adrenocortical carcinoma.
Methods— Laparoscopic adrenalectomy with the dog in lateral recumbency on the unaffected side. Three 5-mm portals (1 laparoscopic portal, 2 instrument portals) were placed in the paralumbar fossa. A fourth instrumental portal (5–12 mm) was placed above the kidney. After dissection and hemostatic control of the phrenicoabdominal vein, the adrenal gland was carefully dissected or when there was capsule fragility, necrotic content was partially aspirated. The remaining glandular tissue was removed through the 12-mm trocar site.
Results— Dogs with unilateral adrenocortical carcinoma (3 right-sided, 4 left-sided) without invasion of the caudal vena cava were successfully operated by laparoscopic approach. There were no significant intraoperative complications; 2 dogs died within 48 hours of surgery because of respiratory complications. Five dogs were discharged 72 hours after surgery, and signs of hyperadrenocorticism disappeared thereafter (survival time ranged from 7 to 25 months).
Conclusions— Laparoscopic adrenalectomy is feasible in dogs with either right- or left-sided adrenocortical carcinoma not involving the caudal vena cava.
Clinical Relevance— When performed by experienced surgeons, laparoscopic adrenalectomy offers a minimally invasive alternative to open laparotomy or retroperitoneal surgery for the treatment of unilateral adrenocortical carcinoma in dogs.     

Evaluation of the harmonic scalpel for laparoscopic bilateral ovariectomy in standing horses

OBJECTIVE: To evaluate use of the Harmonic Scalpel (Ethicon Endo-Surgery Inc., Cincinnati, OH) for performing laparoscopic bilateral ovariectomy in standing horses. STUDY DESIGN: Experimental study. ANIMALS: Eight mares aged 2-20 years and weighing 410-540 kg. METHODS: Standing laparoscopic bilateral ovariectomy was performed in 8 mares with normal reproductive tract anatomy. The Harmonic Scalpel (an ultrasonically activated instrument) was used to simultaneously transect and obtain hemostasis of the ovarian pedicle. Necropsy was performed on 4 mares 3 days after surgery and 4 mares 30 days after surgery. Gross and histopathologic evaluation of the ovarian pedicles was performed to characterize tissue reaction. RESULTS: Complete hemostasis of the ovarian pedicles was obtained in all mares. Median transection time for the ovarian pedicle was 28 minutes. Postoperative complications included transient fever, moderate subcutaneous emphysema, and incisional seroma formation. On necropsy examination, there were no signs of generalized peritonitis, postoperative hemorrhage, or adhesion formation. Mild to moderate acute inflammation and scar formation with moderate chronic inflammation at the ovarian pedicle was found at 3 and 30 days. Median depth of coagulation necrosis at 3 days was 2.87 mm. CONCLUSIONS: The Harmonic Scalpel appears to provide reliable hemostasis of the ovarian pedicle during elective laparoscopic ovariectomy in horses. Clinical Relevance-The Harmonic Scalpel represents a safe alternative to other methods of hemostasis during elective laparoscopic ovariectomy in horses.     

Endoscopic removal of esophageal and ruminal foreign bodies in 5 Holstein calves

Endoscopic removal of esophageal and ruminal foreign bodies was successfully performed in 5 Holstein-Friesian calves under sedation or general anesthesia by using an electrocautery snare or a wire-guided Dormi basket. This report describes the endoscopic manipulations, treatment, and outcomes of esophageal foreign body removal in these calves.     

Microscopic Evidence of the Opportunistic Reproductive Strategy and Early Sexual Maturation of the Small‐Sized Characin Knodus moenkhausii (Characidae,Pisces)

We investigated the reproductive biology of Knodus moenkhausii, an abundant small‐sized characin fish with broad occurrence in the Paraná River basin, Brazil. Specimens were collected monthly to determine fecundity, length at first maturity, reproductive period and spawning type. Gonads were macroscopically classified according to their form, size and texture in three different stages (immature, maturing or mature). Histological procedures were conducted to confirm gonadal developmental stages, and it was possible to notice that maturing females actually presented atretic oocytes, and all males that were macroscopically classified as immature, maturing and mature actually presented abundant spermatozoa in their gonads. Because of these discrepancies, a reclassification of gonadal maturations stages was needed after histological analysis, reinforcing its importance to studies on the reproduction of small characins. Reproduction occurred throughout the year though with two peaks. The length of the smallest mature individuals was 13 mm SL for males and 24 mm SL for females. Despite presenting relatively small batch fecundity, some life history traits such as early reproduction, multiple spawning throughout the year, in association with known opportunistic feeding habits, explain the high abundance of this species in locations where it occurs.     

Ovariectomy in a brook trout (Salvelinus fontinalis).

A 1.7-kg mature, female brook trout (Salvelinus fontinalis) was diagnosed with egg retention. Eggs could not be manually stripped and so were surgically removed. To prevent future complications, bilateral ovariectomy was performed. Complications were minimal, and the fish resumed eating within 2 days. Sutures were removed 21 days after surgery. Ovariectomy in display aquarium fish is a viable option for the resolution of ovarian disease.