Evaluation of sperm morphometry of rabbits (Oryctolagus cuniculus f. domesticus)

Sperm morphology and morphometry are considered parameters in fertility diagnosis. They are especially important in the case of species for which there is no standard with respect to morphometric sperm parameters. It is then crucial to apply the staining technique that has the least influence on the sperm structure and provides the most detailed image, so as to enable measurements. The aim of the research was to assess the morphometric parameters of rabbit sperm using silver nitrate staining. The staining process revealed a detailed image of the spermatozoon head and tail, thus enabling precise measurements. From these basic morphometric parameters, four additional shape indices characterizing the sperm head were calculated: ellipticity, elongation, roughness and regularity. These parameters more precisely characterize the shape of the sperm head. Silver nitrate staining can be used as an independent technique in assessment of sperm structure or to supplement routine diagnostics.     

Tolerance evaluation of overdosed dietary levels of 25‐hydroxyvitamin D3 in growing piglets

Forty‐eight, cross‐bred (GL × LW × P) piglets were used in a 42‐day tolerance trial to assess the effects of feeding diets supplemented with vitamin D or increasing levels of 25‐hydroxyvitamin D₃ (25‐OH‐D₃). Six‐week‐old piglets (24 castrate males, 24 females) were used. Two replicate groups of 6 piglets were randomized by weight and allocated to four dietary treatments. The control group (T1) was supplemented with 50 μg vitamin D₃/kg feed. The experimental groups received 25‐OH‐D₃ at the recommended dose (T2: 50 μg/kg = 1x), at 250 μg/kg (T3: 5x) or at 500 μg/kg (T4: 10x) respectively. Feed intake and daily weight gain were measured weekly, and the animals were examined by a veterinarian daily. After 42 days, body mass, blood, urine, bone and tissue samples were analysed and a pathology examination conducted. Dietary treatments had no significant effect on final body mass or daily weight gain. The 25‐OH‐D₃ plasma concentration in T1 was 17 ± 3 ng/ml (mean ± SD) while the respective values of the experimental groups were significantly increased in T2, T3 and T4. Tissue concentrations of 25‐OH‐D₃ were higher in liver and muscle for T3 and T4 and in skin for T4 than in T1. However, neither gross pathology nor histology, nor blood and urine characteristics, nor bone parameters were affected by dietary treatments. Weight of organs as well as dry matter, ash and calcium content of kidneys remained unaffected by dietary 25‐OH‐D₃ intake. Furthermore, no changes were observed for general indicators of health. The results of this study demonstrated that feeding piglets with 25‐OH‐D₃ at 5 or 10 times the recommended level had no adverse effects on any of the biological parameters measured. It was concluded that 25‐OH‐D₃ can be regarded as a supplement with a very high safety margin when used at the recommended level.     

Ultrasonographic anatomy of the atlanto‐occipital region and ultrasound‐guided cerebrospinal fluid collection in rabbits (Oryctolagus cuniculus)

Cerebrospinal fluid analyses are important for diagnosis of neurologic problems in rabbits and for translational research projects using rabbits as models. Blind puncture of the cisterna magna is the current standard technique for sampling cerebrospinal fluid in this species. However, the complexity and small size of the cisterna magna and surrounding structures are limitations of this technique. Aims of this prospective, anatomic, pilot study were to (1) describe the normal anatomy of the atlanto‐occipital region, (2) describe ultrasonographic anatomic landmarks, and (3) develop and evaluate a technique for ultrasound‐guided puncture of the cisterna magna for cerebrospinal fluid sampling in rabbits. Thirty healthy rabbits were included and the study was conducted in three stages. Three rabbit cadavers were used for the first stage of the study. Then, the second stage was completed using 13 rabbit cadavers. Finally, the third stage was completed in 14 live rabbits. The ultrasound‐guided puncture performed in 13 cadavers was successful at the first attempt in 10 cases, and at the second attempt in the remaining three cases. In the in vivo study, the ultrasound‐guided puncture was successful in all 14 cases, without signs of complications. Findings supported the use of ultrasound‐guided puncture of the cisterna magna as a safe technique that may be used routinely or when the sample of cerebrospinal fluid cannot be obtained with the blind technique in rabbits.     

Use of contrast‐enhanced ultrasonography in the diagnosis of a liver lobe torsion in a rabbit (Oryctolagus cuniculus)

A 1‐year‐old male French Lop rabbit (Oryctolagus cuniculus) was presented with a sudden onset of hyporexia. Physical examination revealed cranial abdominal discomfort. Liver enzymes were elevated on serum biochemistry profile. Abdominal radiographs showed diffusely gas‐dilated small intestinal loops. On abdominal ultrasound, a lobe in the right aspect of the liver was hypoechoic with rounded margins and was surrounded by hyperechoic fat and anechoic fluid. Contrast‐enhanced ultrasonography showed complete lack of perfusion in the abnormal right liver lobe. Exploratory laparotomy revealed torsion of the quadrate liver lobe. Diffuse coagulation necrosis was noted on histopathologic examination. The rabbit fully recovered after surgery.     

A comparison of UVb compact lamps in enabling cutaneous vitamin D synthesis in growing bearded dragons

The effect of exposure to different UVb compact lamps on the vitamin D status of growing bearded dragons (Pogona vitticeps) was studied. Forty‐two newly hatched bearded dragons (<24 h old) were allocated to six treatment groups (n = 7 per group). Five groups were exposed to different UVb compact lamps for two hours per day, with a control group not exposed to UVb radiation. At 120 days of age, blood samples were obtained and concentrations of 25(OH)D₃, Ca, P and uric acid were determined. In addition, plasma 25(OH)D₃ concentration was determined in free‐living adult bearded dragons to provide a reference level. Only one treatment resulted in elevated levels of 25(OH)D₃ compared to the control group (41.0 ± 12.85 vs. 2.0 ± 0.0 nmol/L). All UVb‐exposed groups had low 25(OH)D₃ plasma levels compared to earlier studies on captive bearded dragons as well as in comparison with the free‐living adult bearded dragons (409 ± 56 nmol/L). Spectral analysis indicated that all treatment lamps emitted UVb wavelengths effective for some cutaneous vitamin D synthesis. None of these lamps, under this regime, appeared to have provided a sufficient UVb dose to enable synthesis of plasma 25(OH)D₃ levels similar to those of free‐living bearded dragons in their native habitat.     

Effect of theatre temperature on body temperature during anaesthesia for routine neutering of domestic rabbits (Oryctolagus cuniculus)

ObjectiveTo investigate the effect of theatre temperature on body temperature in rabbits undergoing castration or ovariohysterectomy surgery during general anaesthesia.Study designProspective, clinical study.AnimalsA group of 88 rabbits presented for elective neutering.MethodsRabbits were divided into male (31/54) and female (23/54) groups and assigned to one of two theatre temperatures via coin toss. Theatre temperature was 23 °C (±2 °C) for group A (n = 37/54) and 28 °C (±2 °C) for group B (n = 17/54). During anaesthesia and recovery, theatre temperature and rectal temperature were recorded every 5 minutes. Time to resumption of feeding and passing faeces were recorded. Data are presented as median (interquartile range) or mean (± standard deviation). Statistical analyses comprised a mixed-effects model, with Sidak’s multiple comparison test for post-hoc testing and Fisher’s exact test; p < 0.05.ResultsA total of 54 rabbits completed the study, with median age 6 (4–9) months and median weight 1.53 (1.30–1.79) kg. In rabbits undergoing castration, theatre temperature did not significantly affect body temperature. Mean temperatures immediately after induction were 38.6 °C and 38.7 °C and at the end of the procedure 38.5 °C and 38.5 °C for group A and group B, respectively. In rabbits undergoing ovariohysterectomy, mean temperatures immediately after induction were 38.3 °C and 38.8 °C and at the end of the procedure 38.1 °C and 39.2 °C for group A and group B, respectively. Rabbits undergoing ovariohysterectomy at an ambient temperature of 28 °C had a significantly higher final temperature, mean ± 1.15 °C (95% confidence interval, 0.47–1.83), compared with 23 °C (p = 0.001). Theatre temperature did not affect return to feeding or defaecating.Conclusions and clinical relevanceDuring anaesthesia an ambient theatre temperature of 28 °C may reduce the risk of hypothermia in rabbits undergoing ovariohysterectomy or similarly invasive surgery.     

Spontaneous Extraskeletal Osteosarcoma in a Rabbit (Oryctolagus cuniculus): Histopathological and Immunohistochemical Findings

A spontaneously occurring subcutaneous mass in the left forelimb of a nine-year-old rabbit (Oryctolagus cuniculus) was examined histopathologically and immunohistochemically. Clinically, edema and hemorrhage were seen around the mass. No connection of the tumor mass to the appendicular skeleton was found. The tumor was arranged in a solid growth pattern and irregular bundles, and neoplastic cells were polygonal to spindle-shape. Osteoid (positive for osteocalcin) and multinucleated giant cells were diffusely or focally seen. Neoplastic cells were positive for vimentin, osterix and Ki-67, indicating the nature of osteoblasts with proliferating activity, but negative for α-smooth muscle actin, desmin or CD204. Based on these findings, a diagnosis of extraskeletal osteosarcoma was made, a very rare tumor both in laboratory and pet rabbits.     

Influence of diet on calcium metabolism, tissue calcification and urinary sludge in rabbits (Oryctolagus cuniculus)

Rabbits absorb more calcium (Ca) from their diet than they require, and excrete surplus via urine, which therefore contains a typical 'sludge'. This makes rabbits susceptible to Ca-containing uroliths. But given the Ca content of diets of free-ranging specimens, and the limited reports of urinary sludge and Ca contents in free-ranging lagomorphs, we can suspect that rabbits are naturally adapted to high urinary Ca loads. We fed four groups of New Zealand hybrid rabbits [n = 28, age at start 5-6 weeks) pelleted diets consisting of lucerne hay only (L, Ca 2.32% dry matter (DM)], lucerne:oats 1:1 (LG, Ca 1.36%), grass hay only (G, Ca 1.04%), or grass:oats 1:1 (GG, 0.83%) for 25 weeks, with water available ad libitum. Diets were not supplemented with Ca, phosphorus, or vitamin D. Rabbits on diets LG and GG had lower food and water intakes, lower faeces and urine output, grew faster and had higher body mass at slaughter (mainly attributable to adipose tissue). Apparent Ca digestibility decreased in the order L-LG-G/GG. Rabbits on L had larger and heavier kidneys, more urinary sediment at sonography, and a higher urinary Ca content than the other groups. No animal showed signs of urolithiasis/calcinosis at X-ray, sonography, or gross pathology. Kidney/aorta histology only sporadically indicated Ca deposits, with no systematic difference between groups. Under the conditions of the experiment, dietary Ca loads in legume hay do not appear problematic for rabbits, and other factors, such as water supply and level of activity may be important contributors to urolithiasis development in veterinary patients. However, due to the lower Ca content of grass hay, the significantly lower degree of urinary sludge formation, and the significantly higher water intake related with grass hay feeding, grass hay-dominated diets are to be recommended for rabbits in which urolithiasis prevention is an issue.     

Evaluation of alfaxalone total intravenous anesthesia in rabbits (Oryctolagus cuniculus) premedicated with dexmedetomidine or dexmedetomidine and buprenorphine

ObjectiveTo evaluate alfaxalone for total intravenous anesthesia (TIVA) in rabbits premedicated with dexmedetomidine or dexmedetomidine and buprenorphine.Study designCrossover study (part 1) with observational study (part 2).AnimalsA total of eight New Zealand White rabbits (Oryctolagus cuniculus), four female and four male, aged 12–16 weeks and weighing 2.8–3.5 kg in part 1. Separately, four additional rabbits in part 2.MethodsCrossover study design with eight rabbits per treatment. Rabbits were administered treatment D, dexmedetomidine (0.2 mg kg^–1), or treatment DB, dexmedetomidine (0.1 mg kg^–1) and buprenorphine (0.05 mg kg^–1) intramuscularly. Anesthesia was induced with alfaxalone intravenously until a supraglottic airway device was placed to deliver 100% oxygen. Anesthesia was maintained with alfaxalone (TIVA). Infusion rates were adjusted to achieve an absent pedal withdrawal reflex. Heart rate, respiratory rate, noninvasive blood pressure, end-tidal carbon dioxide partial pressure and peripheral hemoglobin oxygen saturation (SpO₂) were recorded every 5 minutes. Subsequently, four rabbits underwent ovariohysterectomy using treatment DB and alfaxalone TIVA.ResultsThe mean ± standard deviation alfaxalone infusion rate was 9.6 ± 2.6 and 4.5 ± 1.3 mg kg^–1 hour^–1 for treatments D and DB, respectively. In both treatments, blood pressure remained within acceptable range and SpO₂ was > 95%. Postinduction apnea and respiratory depression were observed in both treatments and managed with manual positive pressure ventilation. Four separate rabbits underwent successful ovariohysterectomy with treatment DB and alfaxalone TIVA. One rabbit required supplementation with inhalant anesthesia; three rabbits were successfully maintained using alfaxalone TIVA alone.Conclusions and clinical relevancePremedication with dexmedetomidine–buprenorphine combined with alfaxalone TIVA may be a viable alternative for performing abdominal surgery in the rabbit. The use of supplemental oxygen and ability to provide respiratory support are advised.     

Association of Vitamin D Status and Clinical Outcome in Dogs with a Chronic Enteropathy

Background

Dogs with a chronic enteropathy (CE) have a lower vitamin D status, than do healthy dogs. Vitamin D status has been associated with a negative clinical outcome in humans with inflammatory bowel disease.

Objectives

To examine the relationship between serum 25 hydroxyvitamin D (25(OH)D) concentrations at diagnosis and clinical outcome in dogs with a CE.

Animals

Forty‐one dogs diagnosed with CE admitted to the Royal Dick School of Veterinary Studies, Hospital for Small Animals between 2007 and 2013.

Methods

Retrospective review. Serum 25(OH)D concentrations were compared between dogs which were alive at follow up or had died because of non‐CE‐related reasons (survivors) and dogs which died or were euthanized due to their CE (non‐survivors). A binary logistic regression analysis was performed to determine significant predictors of death in dogs with CE.

Results

Serum concentrations of 25(OH)D at the time a CE was diagnosed were significantly lower in nonsurvivors (n = 15) (median nonsurvivors 4.36 ng/mL, interquartile range 1.6–17.0 ng/mL), median survivors (n = 26) (24.9 ng/mL interquartile range 15.63–39.45 ng/mL, P < .001). Serum 25(OH)D concentration was a significant predictor of death in dogs with CE (odds ratio 1.08 [95% CI 1.02–1.18)]).

Conclusions

Serum 25(OH)D concentrations at diagnosis are predictive of outcome in dogs with CE. The role of vitamin D in the initiation and outcome of chronic enteropathies in dogs is deserving of further study.     

Comparison of blind and endoscopic-guided orotracheal intubation on laryngeal and tracheal damage in domestic rabbits (Oryctolagus cuniculus)

ObjectiveTo compare blind and endoscopic-guided techniques for orotracheal intubation in rabbits and the number of intubation attempts with laryngeal/tracheal damage.Study designProspective, randomized experimental study.AnimalsA total of 24 healthy, intact female New Zealand White rabbits, weighing 2.2 ± 0.2 kg (mean ± standard deviation).MethodsRabbits were randomly assigned to blind (group B) or endoscopic-guided (group E) orotracheal intubation with a 2.0 mm internal diameter uncuffed tube. Intramuscular (IM) alfaxalone (7 mg kg^–1), hydromorphone (0.1 mg kg^–1) and dexmedetomidine (0.005 mg kg^–1) were administered, and additional IM alfaxalone (3–5 mg kg^–1) and dexmedetomidine (0.025 mg kg^–1) were administered to rabbits with strong jaw tone. An intubation attempt was defined as the advancement of the endotracheal tube from the incisors to the laryngeal entrance. Tracheal intubation was confirmed via capnography and anesthesia was maintained with isoflurane for 2 hours. Following euthanasia, laryngeal and tracheal tissues were submitted for histopathology. Quality of anesthesia for orotracheal intubation, intubation procedure and tissue damage were numerically scored. Data were analyzed using Poisson regression, Spearman’s correlation, t test, mixed anova, Mann–Whitney U test, Friedman and Chi square tests as appropriate.ResultsMedian (range) intubation attempts were 2 (1–8) and 1 (1–3) for groups B and E, respectively. More rabbits in group E (91.6%) required additional alfaxalone and dexmedetomidine than in group B (16.7%). Median (range) cumulative histopathology scores were 6 (3–10) and 6 (2–9) for groups B and E, respectively. Scores were highest in the cranial trachea, but there was no difference between groups and no correlation between laryngeal/tracheal damage and the number of intubation attempts.Conclusions and clinical relevanceBoth orotracheal intubation techniques were associated with laryngeal/tracheal damage. Although blind orotracheal intubation was associated with a higher number of attempts, the tissue damage was similar between groups.     

1,25(OH)2D3以双向方式影响猪前体脂肪细胞增殖分化的研究

1,25（OH）₂D₃是维生素D的主要活性形式,影响人和动物脂肪形成,为探究其在猪脂肪细胞增殖分化中的作用,试验从3~5日龄仔猪皮下脂肪组织分离培养前体脂肪细胞,并以浓度为0、0.1、1、10、100和1 000 nmol/L的1,25（OH）₂D₃分别处理,在培养的0、1、2、4、6、8和10 d采用MTT比色法检测细胞增殖活性;在诱导分化后0、1、2、4、6、8和10 d,以油红O染色提取法和实时荧光定量PCR检测细胞成脂分化及分化标志基因过氧化物酶体增殖物激活受体γ（PPARγ）和脂肪酸合成酶（FAS）表达。结果显示,0.1和1 nmol/L 1,25（OH）₂D₃显著促进猪前体脂肪细胞增殖（P<0.05）,而浓度为10~100 nmol/L时则抑制细胞增殖（P<0.05）;0.1和1 nmol/L 1,25（OH）₂D₃显著抑制猪前体脂肪细胞分化（P<0.05）,降低PPARγ和FAS mRNA表达水平（P<0.05）,但在浓度为10和100 nmol/L时,显著促进猪前体脂肪细胞分化（P<0.05）,上调PPARγ和FAS mRNA表达（P<0.05）;浓度达1 000 nmol/L时,可能对细胞有毒性作用。综合以上结果,低浓度1,25（OH）₂D₃促进猪前体脂肪细胞增殖,而通过下调PPARγ表达抑制分化;高浓度1,25（OH）₂D₃抑制猪前体脂肪细胞增殖,通过上调PPARγ表达促进分化。1,25（OH）₂D₃对猪前体脂肪细胞增殖和分化具有双向作用。     

Effects of animal type (wild vs. domestic) and diet alfalfa level on intake and digestibility of European adult rabbits (Oryctolagus cuniculus)

The aim of this study was to evaluate the effect of the level of alfalfa in the diet on feed intake and digestibility of two types of rabbits, wild (Oryctolagus cuniculus algirus) vs. domestic (O. cuniculus cuniculus). Ten wild (W; mean LW = 927 g) and 10 domestic (D; mean LW = 4,645 g) adult rabbit does were fed ad libitum two pelleted diets: a control diet (C) with 15% of dehydrated alfalfa hay (as feed basis) and a test diet (A) with 36% of dehydrated alfalfa hay (as feed basis), according to a change‐over design. Wild does dry matter (DM) intake per kg live weight (BW) was 55% higher (p < .001) than the intake of the D ones (58 g vs. 37 g DM per kg BW respectively). However, no difference (p > .05) was found when intake was expressed per kg^0.75 BW (ca. 56 g DM) and tended to be higher (p = .07) in D does when expressed per kg^0.67 BW (62 g vs. 55 g DM). Domestic does showed a higher (p < .05) DM, organic matter, crude energy and neutral detergent fibre digestibility (3; 2; 3; 3 percentage points respectively) than W does. The amount of nutrients and energy digested by D does was lower per kg BW (p < .001), similar per kg^0.75 BW (p > .05) and tended to be higher per kg^0.67 BW (p < .1) than in W does. The diet content of alfalfa did not affect (p > .05) the feed intake nor the diet digestibility. This study suggests that W rabbits exhibit a higher intake per kg BW and a lower digestibility than their D counterparts, which results in similar digestible nutrient and energy intake per kg BW powered to 0.75. The nutritive value of dehydrated alfalfa for rabbits, evaluated through intake and digestibility, seems to be equivalent to their base diets (forage plus concentrate).