Nicola Di Girolamo

Evaluation of intraocular pressure in conscious Hermann's tortoises (Testudo hermanni) by means of rebound tonometry

OBJECTIVE To determine intraocular pressure (IOP) in healthy Hermanns tortoises (Testudo hermanni). ANIMALS 26 outdoor-housed Hermanns tortoises (13 males and 13 females); body weight ranged from 255 to 2,310 g, and age ranged from 4 to > 50 years. PROCEDURES After a preliminary ophthalmic evaluation was performed, IOP was measured by means of a rebound tonometer in both eyes of each tortoise. Three measurements were obtained for each eye; successive measurements were obtained from alternate eyes. Each measurement was based on the mean of 6 values automatically provided by the rebound tonometer. Statistical analysis was used to evaluate correlations between variables and to identify sex- or size-related IOP variations, and changes in IOP over multiple measurements. RESULTS Mean ± SEM IOP of the 52 eyes was 15.74 ± 0.20 mm Hg (range, 9 to 22 mm Hg). Results for t tests did not reveal significant differences in IOP between the right and left eyes or between males and females. A significant moderate negative correlation (r = -0.41; r(2) = 0.169) between IOP and body weight was detected. Results of repeated-measures ANOVA revealed a significant increase in IOP over multiple measurements. CONCLUSIONS AND CLINICAL RELEVANCE Rebound tonometry was a practical and rapid means of determining IOP in small- to medium-sized tortoises that required minimal manual restraint of the animals. Establishing IOP values in healthy Hermanns tortoises will provide a reference frame for use during complete ophthalmic examinations, thus allowing clinicians to diagnose a broader spectrum of ocular pathological conditions in tortoises.

Plasma 25-hydroxyvitamin D(3) concentrations in Hermann's tortoises (Testudo hermanni) exposed to natural sunlight and two artificial ultraviolet radiation sources.

Objective-To determine the effect of various UVB radiation sources on plasma 25-hydroxyvitamin D(3) concentrations in Hermanns tortoises (Testudo hermanni). Animals-18 healthy Hermanns tortoises. Procedures-Tortoises were exposed to sunlight in an outdoor enclosure located in the natural geographic range of Hermanns tortoises (n = 6 tortoises) or a self-ballasted mercury-vapor lamp (6) or fluorescent UVB-emitting lamp (6) in an indoor enclosure for 35 days. Plasma samples were obtained from each tortoise on the first (day 0) and last (day 35) days of the study, and concentrations of 25-hydroxyvitamin D(3) were determined. Amount of UVB radiation in enclosures was measured. Results-Mean ± SD plasma 25-hydroxyvitamin D(3) concentrations for tortoises exposed to the mercury-vapor and fluorescent lamps were significantly lower on day 35 (155.69 ± 80.71 nmol/L and 134.42 ± 51.42 nmol/L, respectively) than they were on day 0 (368.02 ± 119.34 nmol/L and 313.69 ± 109.54 nmol/L, respectively). Mean ± SD plasma 25-hydroxyvitamin D(3) concentration for tortoises exposed to sunlight did not differ significantly between days 0 (387.74 ± 114.56 nmol/L) and 35 (411.51 ± 189.75 nmol/L). Mean day 35 plasma 25-hydroxyvitamin D(3) concentration was significantly higher for tortoises exposed to sunlight versus those exposed to mercury-vapor or fluorescent lamps. Sunlight provided significantly more UVB radiation than did the mercury-vapor or fluorescent lamps. Conclusions and Clinical Relevance-Plasma 25-hydroxyvitamin D(3) concentrations differed between tortoises exposed to sunlight and those exposed to artificial UVB sources. Exposure to sunlight at a latitude similar to that of the natural geographic range is recommended for healthy and calcium-deficient tortoises.

Reference values for hematology and plasma biochemistry variables, and protein electrophoresis of healthy Hermann's tortoises (Testudo hermanni ssp.)

BACKGROUND Hermanns tortoise, Testudo hermanni, is currently on the International Union for Conservation of Nature (IUCN) red list of endangered species. Reptile medicine relies also on laboratory analyses to evaluate health status, but reference ranges for hematology and biochemistry variables and protein electrophoresis in plasma of healthy tortoises are not available. OBJECTIVE The purposes of this study were to establish reference ranges for select hematologic and biochemical variables in clinically healthy Hermanns tortoises, and evaluate the impact of sex and season. METHODS Blood samples were collected from 34 healthy tortoises at the end of September and beginning of July. Blood smears, HCT, concentrations of HGB and select plasma biochemical analytes, select enzyme activities, and plasma protein fractions were evaluated. Reference ranges were determined and checked for influence of sex and sampling time point. RESULTS Typical reptilian RBC and WBC were observed in blood smears. HCT and concentrations of HGB, uric acid and urea, and ALT and AST activities were significantly higher in males than in females. Concentrations of glucose, uric acid, and phosphate, and AST activity were significantly higher at the beginning of July, whereas concentrations of urea and Cl were higher at the end of September prior to hibernation. The electrophoretic protein fractions included albumin, and α, β, and γ globulins. CONCLUSIONS The reference ranges defined in the present study are useful for clinical tortoise medicine and conservation. Sex and seasonal sampling were identified as factors significantly affecting hematology and blood chemistry analytes; they should be taken into consideration when assessing tortoise health status.

Cystoscopic sex identification of posthatchling chelonians

OBJECTIVE To describe a noninvasive technique for sex identification of posthatchling chelonians and to assess its safety in Hermanns tortoises (Testudo hermanni). DESIGN Validation study and clinical trial. ANIMALS 15 recently dead posthachling chelonians and 25 healthy posthatchling Hermanns tortoises. PROCEDURES Cystoscopy was performed on both dead and anesthetized live chelonians. Dead chelonians ranged in body weight from 32.4 to 75.1 g (0.07 to 0.17 lb; median, 45.7 g [0.10 lb]). Dead chelonians were dissected immediately after cystoscopy, and gonads were collected for histologic examination. Urinary bladder was macroscopically evaluated in situ to assess its integrity after retrograde injection of saline (0.9% NaCl) solution. Hermanns tortoises ranged in body weight from 27.3 to 57.8 g (0.06 to 0.13 lb; median, 37.0 g [0.08 lb]). Cystoscopic examination of live tortoises was performed following induction of general anesthesia with a mixture of morphine, dexmedetomidine, and ketamine administered IM. The Pearson coefficient was used to assess the consistency between procedure time and body weight; κ statistic was used to evaluate agreement between sex identified by cystoscopy and histologic examination beyond that expected by mere chance. RESULTS Visualization of gonads was feasible through the thin, transparent urinary bladder wall in all the animals evaluated in this study. Blinded histologic examination confirmed the results of cystoscopic gonad identification in all dead chelonians (κ = 1.0). The urinary bladder did not have evidence of macroscopic leakage or microscopic alterations of normal tissue architecture within the representative sections chosen for histologic examination. In live tortoises, median procedure time (range) was 90 (39 to 345) seconds. No significant correlations were found between procedure time and body weight. CONCLUSIONS AND CLINICAL RELEVANCE Cystoscopy performed by means of rigid endoscopy with fluid instillation was found to be an effective method for sex identification of immature chelonians. Furthermore, no complications were observed when this procedure was performed in vivo.

Deficiencies of effectiveness of intervention studies in veterinary medicine: a cross-sectional survey of ten leading veterinary and medical journals

The validity of studies that assess the effectiveness of an intervention (EoI) depends on variables such as the type of study design, the quality of their methodology, and the participants enrolled. Five leading veterinary journals and 5 leading human medical journals were hand-searched for EoI studies for the year 2013. We assessed (1) the prevalence of randomized controlled trials (RCTs) among EoI studies, (2) the type of participants enrolled, and (3) the methodological quality of the selected studies. Of 1707 eligible articles, 590 were EoI articles and 435 RCTs. Random allocation to the intervention was performed in 52% (114/219; 95%CI:45.2–58.8%) of veterinary EoI articles, against 87% (321/371; 82.5–89.7%) of human EoI articles (adjusted OR:9.2; 3.4–24.8). Veterinary RCTs were smaller (median: 26 animals versus 465 humans) and less likely to enroll real patients, compared with human RCTs (OR:331; 45–2441). Only 2% of the veterinary RCTs, versus 77% of the human RCTs, reported power calculations, primary outcomes, random sequence generation, allocation concealment and estimation methods. Currently, internal and external validity of veterinary EoI studies is limited compared to human medical ones. To address these issues, veterinary interventional research needs to improve its methodology, increase the number of published RCTs and enroll real clinical patients.

Osseointegration is improved by coating titanium implants with a nanostructured thin film with titanium carbide and titanium oxides clustered around graphitic carbon

Titanium implants coated with a 500nm nanostructured layer, deposited by the Ion Plating Plasma Assisted (IPPA) technology, composed of 60% graphitic carbon, 25% titanium oxides and 15% titanium carbide were implanted into rabbit femurs whilst into the controlateral femurs uncoated titanium implants were inserted as control. At four time points the animals were injected with calcein green, xylenol orange, oxytetracycline and alizarin. After 2, 4 and 8weeks femurs were removed and processed for histology and static and dynamic histomorphometry for undecalcified bone processing into methylmethacrylate, sectioned, thinned, polished and stained with Toluidine blue and Fast green. The overall bone-implant contacts rate (percentage of bone-implant contacts/weeks) of the TiC coated implant was 1.6 fold than that of the uncoated titanium implant. The histomorphometric analyses confirmed the histological evaluations. More precisely, higher Mineral Apposition Rate (MAR, μm/day) (p<0.005) and Bone Formation Rate (BFR, μm2/μm/day) (p<0.0005) as well as Bone Implant Contact (Bic) and Bone Ingrowth values (p<0.0005) were observed for the TiC coated implants compared to uncoated implants. In conclusion the hard nanostructured TiC layer protects the bulk titanium implant against the harsh conditions of biological tissues and in the same time, stimulating adhesion, proliferation and activity of osteoblasts, induces a better bone-implant contacts of the implant compared to the uncoated titanium implant.

Performance of two portable meters and a benchtop analyzer for blood glucose concentration measurement in rabbits

OBJECTIVE To evaluate performance of a human portable blood glucose meter (PBGM), a veterinary PBGM, and a veterinary benchtop analyzer for measuring blood glucose concentration in rabbits and to evaluate the effect of sample characteristics on their performance. DESIGN Observational prospective cross-sectional study. SAMPLE Blood samples from 89 pet rabbits. PROCEDURES Blood glucose concentration was measured with a human PBGM (n = 89 rabbits), a veterinary PBGM (89), and a benchtop analyzer (32) and compared with results obtained with plasma in a laboratory analyzer (hexokinase method). RESULTS The human PBGM underestimated blood glucose concentration, had decreased accuracy at high Hcts, and had the lowest total error observed (11.4%). The veterinary PBGM overestimated blood glucose concentration, had decreased accuracy at low Hcts and at high blood glucose concentrations, and had the highest total error (15.5% and 29.8% for canine and feline settings, respectively). The benchtop analyzer had good accuracy and was not influenced by Hct or glucose concentrations. Clinical errors would have occurred in 0% of cases with the human PBGM and with the benchtop analyzer and in 9% (canine setting) to 6.7% (feline setting) of cases with the veterinary PBGM. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that use of the human PBGM evaluated in this study would be acceptable for point-of-care testing of blood glucose concentration in rabbits when benchtop analyzers are not available. The use of the veterinary PBGM evaluated in this study may alter both treatment and diagnostic decisions because of the overestimation of glucose concentrations in some rabbits.

Pharmacokinetics of cefovecin sodium after subcutaneous administration to Hermann's tortoises (Testudo hermanni)

OBJECTIVE To determine the pharmacokinetics of cefovecin sodium after SC administration to Hermanns tortoises (Testudo hermanni). ANIMALS 23 healthy adult Hermanns tortoises (15 males and 8 females). PROCEDURES Cefovecin (8.0 mg/kg) was injected once in the subcutis of the neck region of Hermanns tortoises, and blood samples were obtained at predetermined time points. Plasma cefovecin concentrations were measured via ultraperformance liquid chromatography coupled to tandem mass spectrometry, and pharmacokinetic parameters were calculated with a noncompartmental model. Plasma protein concentration was quantified, and the percentage of cefovecin bound to protein was estimated with a centrifugation technique. RESULTS Cefovecin was absorbed rapidly, reaching maximum plasma concentrations between 35 minutes and 2 hours after administration, with the exception of 1 group, in which it was reached after 4 hours. The mean ± SD time to maximum concentration was 1.22 ± 1.14 hours; area under the concentration-time curve was 220.35 ± 36.18 h•μg/mL The mean protein-bound fraction of cefovecin ranged from 41.3% to 47.5%. No adverse effects were observed. CONCLUSIONS AND CLINICAL RELEVANCE Administration of a single dose of cefovecin SC appeared to be well-tolerated in this population of tortoises. Results of pharmacokinetic analysis indicated that the 2-week dosing interval suggested for dogs and cats cannot be considered effective in tortoises; however, further research is needed to determine therapeutic concentrations of the drug and appropriate dose ranges.

Insertion torque recordings for the diagnosis of contact between orthodontic mini-implants and dental roots: protocol for a systematic review

BackgroundHitting a dental root during the insertion of orthodontic mini-implants (OMIs) is a common adverse effect of this intervention. This condition can permanently damage these structures and can cause implant instability. Increased torque levels (index test) recorded during the insertion of OMIs may provide a more accurate and immediate diagnosis of implant-root contact (target condition) than radiographic imaging (reference standard). An accurate index test could reduce or eliminate X-ray exposure. These issues, the common use of OMIs, the high prevalence of the target condition, and because most OMIs are placed between roots warrant a systematic review. We will assess 1) the diagnostic accuracy and the adverse effects of the index test, 2) whether OMIs with root contact have higher insertion torque values than those without, and 3) whether intermediate torque values have clinical diagnostic utility.MethodsThe Preferred Reporting Items for Systematic review and Meta-Analysis Protocols (PRISMA-P) 2015 statement was used as a the guideline for reporting this protocol. Inserting implants deliberately into dental roots of human participants would not be approved by ethical review boards and adverse effects of interventions are generally underreported. We will therefore apply broad spectrum eligibility criteria, which will include clinical, animal and cadaver models. Not including these models could slow down knowledge translation.Both randomized and non-randomized research studies will be included. Comparisons of interest and subgroups are pre-specified. We will conduct searches in MEDLINE and more than 40 other electronic databases. We will search the grey literature and reference lists and hand-search ten journals. All methodological procedures will be conducted by three reviewers. Study selection, data extraction and analyses, and protocols for contacting authors and resolving conflicts between reviewers are described. Designed specific risk of bias tools will be tailored to the research question. Different research models will be analysed separately. Parameters for exploring statistical heterogeneity and conducting meta-analyses are pre-specified. The quality of evidence for outcomes will be assessed through the Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach.DiscussionThe findings of this systematic review will be useful for patients, clinicians, researchers, guideline developers, policymakers, and surgical companies.

Reptile Soft Tissue Surgery

The surgical approach to reptiles can be challenging. Reptiles have unique physiologic, anatomic, and pathologic differences. This may result in frustrating surgical experiences. However, recent investigations provided novel, less invasive, surgical techniques. The purpose of this review was to describe the technical aspects behind soft tissue surgical techniques that have been used in reptiles, so as to provide a general guideline for veterinarians working with reptiles.