Evaluation of sources cited by an exotic animal formulary for supporting drug dosages and reference intervals in mammals.

OBJECTIVES: Formularies are intended to simplify clinical decision-making by collecting evidence-based information on drugs and their dosages. This study assessed the characteristics of sources used to support drug dosages and reference intervals for mammals in a specific exotic animal formulary, and how the sources had changed over five editions. METHODS: Each reference supporting drug dosages and reference intervals in the sections for ferrets, rabbits, rodents, hedgehogs and miniature pigs in all five editions of the formulary was evaluated and classified by two independent investigators in terms of the type of source cited. Univariable and multi-variable logistic regression models were built to evaluate changes between editions and sections. RESULTS: In total, 1338 references supporting drug dosages and 180 references supporting reference intervals were included from all editions of the formulary. Primary sources were cited by 525 (39.2%) and 39 (21.7%) of the drug and reference interval references, respectively. For drug dosages, the current edition of the formulary (2018) cited a higher proportion of primary rather than secondary sources compared with the first edition (odds ratios 3.4, 95% confidence interval 2.1 to 5.6), while for reference intervals there were no significant changes between editions. In the current edition of the formulary, the 168 secondary sources cited for drug dosages included 78 (46.4%) textbooks, 63 (37.5%) reviews, 14 (8.3%) personal communications and 7 (4.2%) other formularies. CLINICAL SIGNIFICANCE: A large proportion of references supporting drug dosages and reference intervals in the evaluated sections cited secondary sources. Although modest improvements have been observed over time, practitioners should be aware that the evidence supporting several drugs and dosages was limited, and assess the information within the formulary critically.

Rectal temperature is a prognostic indicator in client-owned guinea pigs.

OBJECTIVES: To determine whether rectal temperature was associated with patient mortality in client-owned guinea pigs upon presentation to a veterinary hospital. MATERIALS AND METHODS: The medical record database at a veterinary teaching hospital was searched for records of guinea pigs from January 2016 through June 2019. Guinea pigs were included in the study if a rectal temperature was measured at presentation and there was data on survival status 7 days post-presentation. If survivor status was not documented in the medical record, follow-up information was obtained from the client via telephone or email. The data was ultimately collected from 201 client-owned guinea pigs who presented for 388 independent examinations. Univariable, multivariable and sensitivity analyses were performed. RESULTS: Guinea pigs with hypothermia (<37.9°C) at presentation had a relative risk of mortality within 7 days of presentation almost 3 times greater than guinea pigs without hypothermia (relative risk: 2.88; 95% confidence interval: 1.86 to 4.48). For each 0.55°C decrease in rectal temperature, the odds of death increased 1.6 times (odds ratio: 1.64; 95% confidence interval: 1.42 to 2.89). Sensitivity analyses confirmed the robustness of the finding. CLINICAL SIGNIFICANCE: Rectal temperature was a predictor of death for guinea pigs presenting for care at a veterinary hospital. Obtaining a rectal temperature recording should be considered for patient guinea pigs.

Surgical resolution of uterine prolapse in three pet rabbits.

This report describes a surgical technique for resolution of uterine prolapse in rabbits. Three pet rabbits presented within 24 hours of parturition with a red mass protruding from the vagina, which was diagnosed as uterine prolapse. In the first case, an attempt to reduce the prolapse by manual compression was ineffective. A laparotomy was used to apply internal uterine traction while simultaneously using gentle external pressure with cotton-tip applicators and resulted in successful resolution. After repositioning, an ovariohysterovaginectomy was performed in all three rabbits. All rabbits recovered uneventfully. Laparotomic repositioning of the uterus and ovariohysterovaginectomy, not previously described in rabbits, was easy to perform and permitted resolution of uterine prolapse.

Evaluation of point-of-care analysers for blood gas and clinical chemistry in Hermann's tortoises (Testudo hermanni).

OBJECTIVE: To assess the agreement between point-of-care and laboratory analysers in measuring biochemical and blood gas analytes in venous samples from tortoises and to define preliminary reference intervals for venous blood gas analysis in Hermann's tortoises (Testudo hermanni). MATERIALS AND METHODS: Jugular venous blood samples from 47 Hermann's tortoises underwent paired analysis with a portable gas analyser (i-STAT 1, Abaxis), a portable chemical analyser (VetScan VS2, Abaxis), and with the respective reference analysers. Agarose gel electrophoresis was used to determine albumin concentrations on 12 specimens. Agreement was evaluated with Bland-Altman plots and regression analysis using the Passing-Bablok method. RESULTS: Point-of-care analysers had variable agreement with the reference analysers, presenting constant or proportional bias depending on the analyte. Relevant analytes in reptiles, such as ionised and total calcium, had acceptable agreement. The method for determining albumin concentration currently available in both point-of-care and laboratory analysers significantly overestimated albumin concentrations as compared to protein electrophoresis. CLINICAL SIGNIFICANCE: While the use of POC analysers is extremely advantageous in small animal primary care facilities, agreement between point-of-care and laboratory analysers varies depending on the analyte. For certain analytes, interchangeability of results is limited and specific reference intervals for point-of-care analysers are required. Veterinarians should be aware of the size and the direction of the bias of each analyte.

Vertebral heart size in chinchillas (Chinchilla lanigera) usingradiography and CT.

OBJECTIVE: To determine the vertebral heart size in chinchillas using right and left lateral radiographic views and CT images. To evaluate the agreement between radiographic and CT modalities. METHODS: Twenty-one clinically healthy chinchillas and seven chinchillas with cardiovascular abnormalities underwent cardiovascular examination before thoracic radiographs and thoracic CT obtained under dexmedetomidine-ketamine anaesthesia. Two observers calculated vertebral heart size for radiographic and CT studies. Reference intervals were calculated with the robust method. Agreement between radiographic and CT-derived vertebral heart size was evaluated with Bland-Altman plots and Deming regression. RESULTS: Mean ±sd vertebral heart size for lateral radiographs was 8·9 ±0·62 (reference interval: 7·5 to 10·2) and for CT-derived vertebral heart size was 8·2 ±0·55 (reference interval: 7·1 to 9·4). CT significantly underestimated the radiographic vertebral heart size by 0·66 vertebrae. There was no significant difference between vertebral heart size for right and left lateral radiographic views, or between female and male chinchillas. CLINICAL SIGNIFICANCE: Radiographic vertebral heart size for chinchillas is larger than that reported for similar rodents. Vertebral heart size can be calculated using radiography or CT in chinchillas, but these techniques are not interchangeable.

In veterinary trials reporting and communication regarding randomisation procedures is suboptimal.

To evaluate randomisation mechanisms in the veterinary literature, all trials defined as 'randomised' were extracted from five leading veterinary journals for the year 2013. Three blinded investigators evaluated (1) if the random sequence generation was actually non-random, and (2) whether method (CONSORT item 8A) and (3) type of randomisation (CONSORT item 8B) were reported. Trialists were contacted via email to establish (1) willingness to respond to questions on randomisation procedures, (2) whether reporting of randomisation improved following a suggestion to use the CONSORT 2010 guideline. Seven per cent ((95 per cent CI 2 to 12 per cent); 8/114) of the trials defined as 'randomised' explicitly used methods that are considered non-random. Almost half of the trials (49 per cent (40 to 59 per cent); 52/106) did not report any mechanism of randomisation. Only 13 trials (12.3 per cent (6 to 19 per cent); 13/106) reported both items. 39 of 114 (34.2 per cent) trialists contacted were willing to respond to further questions on randomisation mechanisms; 4 (3.5 per cent) trialists were unwilling and 71 (62.3 per cent) trialists did not respond. Email correspondence resulted in a mean clarification of 0.7 items (95 per cent CI 0.4 to 1.0) for the 15 trials for trialists that replied. Improved adherence to CONSORT guidelines and trialists communication is imperative to increase the quality of published evidence in veterinary medicine and to reduce research waste.

Epidemiology of bacterial conjunctivitis in chinchillas (Chinchilla lanigera): 49 cases (2005 to 2015).

OBJECTIVES: To evaluate the anamnesis, clinical signs, diagnostic test results, treatment and outcome of chinchillas diagnosed with bacterial conjunctivitis. METHODS: Medical records of 49 chinchillas diagnosed with bacterial conjunctivitis were retrospectively reviewed. Association between clinical signs and type of bacteria involved was determined by means of univariate logistic regression. RESULTS: 61·5% of the isolated bacteria were Gram-negative, and the most common bacterial species was Pseudomonas aeruginosa (50%), followed by Staphylococcus species (26·9%). Chinchillas with acute conjunctivitis (1 to 3 days) were much more commonly affected by Gram-negative organisms. The majority of chinchillas that presented with concurrent respiratory signs were diagnosed with P. aeruginosa. Clinical resolution of conjunctivitis was reported in 87·8% chinchillas with a median time to clinical resolution of 17·5 days. Susceptibility of P. aeruginosa isolates to potentiated sulphonamides, enrofloxacin, ciprofloxacin, gentamicin, amikacin and polymyxin B was 8·3, 36, 62·5, 88·5, 100 and 100%, respectively. CLINICAL SIGNIFICANCE: P. aeruginosa is the predominant bacterial species associated with bacterial conjunctivitis in chinchillas. With the exception of duration of clinical signs, information on the anamnesis or physical examination findings cannot aid in distinguishing conjunctivitis caused by P. aeruginosa or other Gram-negative bacteria from the ones caused by Gram-positive bacteria. Gentamicin- or polymyxin B-containing antibiotic formulations are recommended for empirical topical therapy.

Polyostotic Lymphoma in a Ferret (Mustela putorius furo).

An 8.5-year-old, neutered female ferret (Mustela putorius furo) was presented with tachypnoea, polyuria, polydipsia, anorexia and depression. Radiographs revealed multiple osteolytic lesions of the bone, characterized cytologically by the infiltration of medium- to large-sized lymphocytes. The animal was humanely destroyed and post-mortem examination revealed multifocal masses obliterating the bone marrow of the mandible, right and left humeri and femur, and consisting of an infiltrative population of neoplastic lymphocytes. Immunohistochemical labelling for CD3 and CD79a revealed a CD3-positive neoplastic population. A diagnosis of polyostotic T-cell lymphoma was made, which is the first report of this condition in a ferret.

Porous CS based membranes with improved antimicrobial properties for the treatment of infected wound in veterinary applications.

Recently, much attention has been given to the use of innovative solution for the treatment of infected wounds in animals. Current applied treatments are often un-effective leading to infection propagation and animal death. Novel engineered membranes based on chitosan (CS) can be prepared to combine local antimicrobial effect, high flexibility and easy manipulation. In this work, CS crosslinked porous membranes with improved antimicrobial properties were prepared via freeze-drying technique to promote wound healing and to reduce the bacterial proliferation in infected injuries. Silver nanoparticles (AgNPs) and gentamicin sulfate (GS) were incorporated into the CS matrices to impart antibacterial properties on a wild range of strains. CS based porous membranes were tested for their physicochemical, thermal, mechanical as well as swelling and degradation behavior at physiological condition. Additionally, GS release profile was investigated, showing a moderate burst effect in the first days followed by a decreasing release rate which it was maintained for at least 56 days. Moreover, porous membranes loaded with GS or AgNPs showed good bactericidal activity against both of Gram-positive and Gram-negative bacteria. The bacterial strains used in this work were collected in chelonians after carapace injuries to better mimic the environment after trauma.

Tracing the fate of limbal epithelial progenitor cells in the murine cornea.

Stem cell (SC) division, deployment, and differentiation are processes that contribute to corneal epithelial renewal. Until now studying the destiny of these cells in a living mammal has not been possible. However, the advent of inducible multicolor genetic tagging and powerful imaging technologies has rendered this achievable in the translucent and readily accessible murine cornea. K14CreER(T2)-Confetti mice that harbor two copies of the Brainbow 2.1 cassette, yielding up to 10 colors from the stochastic recombination of fluorescent proteins, were used to monitor K-14(+) progenitor cell dynamics within the corneal epithelium in live animals. Multicolored columns of cells emerged from the basal limbal epithelium as they expanded and migrated linearly at a rate of 10.8 µm/day toward the central cornea. Moreover, the permanent expression of fluorophores, passed on from progenitor to progeny, assisted in discriminating individual clones as spectrally distinct streaks containing more than 1,000 cells within the illuminated area. The centripetal clonal expansion is suggestive that a single progenitor cell is responsible for maintaining a narrow corridor of corneal epithelial cells. Our data are in agreement with the limbus as the repository for SC as opposed to SC being distributed throughout the central cornea. This is the first report describing stem/progenitor cell fate determination in the murine cornea using multicolor genetic tracing. This model represents a powerful new resource to monitor SC kinetics and fate choice under homeostatic conditions, and may assist in assessing clonal evolution during corneal development, aging, wound-healing, disease, and following transplantation.

Human limbal epithelial progenitor cells express αvß5-integrin and the interferon-inducible chemokine CXCL10/IP-10.

Stem cell (SC) therapy is the main treatment modality for patients with limbal stem cell deficiency. If limbal epithelial stem cells (LESC) can be more readily identified, isolated and maintained ex vivo, patients could be treated with better quality grafts. With prior knowledge that vitronectin (VN) is present within the LESC niche and that it supports LESC in vitro, we postulated that VN receptors (integrins αvß3/5) are expressed by, and can be used to identify and isolate LESC. Immunolocalization studies were conducted on human corneas. Corneas were also used to expand limbal epithelial cells from either biopsies or enzyme-dissociated tissue and αvß3/5 expression determined by flow cytometry. Integrin expressing cells were isolated by magnetic activated cell sorting then assessed by immunocytology, colony forming efficiency, RT-PCR and microarray analysis. Integrin αvß5(+) cells co-localized to N-cadherin(+)/CK-15(+) putative LESC. αvß5 was restricted to less than 4% of the total limbal epithelial cells, which expressed higher levels of CK-15 and formed more colonies compared to αvß5(-) cells. Transcriptional profiling of αvß5(+/-) cells by microarray identified several highly expressed interferon-inducible genes, which localize to putative LESC. Integrin αvß5 is a candidate LESC marker since its expression is restricted to the limbus and αvß5(+) limbal epithelial cells have phenotypic and functional properties of LESC. Knowledge of the niche's molecular composition and the genes expressed by its SC will facilitate isolation and maintenance of these cells for therapeutic purposes.

Evaluation of intraocular pressure in conscious ferrets (Mustela putorius furo) by means of rebound tonometry and comparison with applanation tonometry.

To determine normal intraocular pressure (IOP) values by means of rebound tonometry in unanaesthetised ferrets, and to compare rebound and applanation tonometry, 55 clinically healthy ferrets were included in an observational, prospective, blinded study. On 52 ferrets, IOP was measured by means of rebound and applanation tonometry. On 3 ferrets, rebound tonometry was performed every two hours over a 24-hour period. Mean IOPs of the 104 eyes obtained with the rebound tonometer were 14.07±0.35 (95% CI 13.37 to 14.77) tonometer-units (TU). The IOP was significantly higher in males than in females, controlling for age and weight. A study-ordered decrease in coefficient of variation (CV) was observed (measurement 1-52=21.2±1.4 per cent v measurement 53-104=14.4±1.1 per cent) and high CVs (>30 per cent) were significantly less frequent in measurement 53-104. A significant difference in IOP during the 24- hour measurements was found, with the lowest IOP recorded at 22:00. The tonometers presented poor agreement, and IOP values were not correlated. The difference in IOP estimation increased with the magnitude of the measurements. Applanation tonometry presented a significant higher frequency of per-eye IOP values exceeding 25 and 30 TU, and a significant lower repeatability (CV=37.1±2.6 per cent v 17.8±1.2 per cent) compared with rebound tonometry. In conclusion, several factors need to be considered when measuring IOP in ferrets.

Multiple congenital malformations in a dicephalic spur-thighed tortoise (Testudo graeca ibera).

A 22-day-old dicephalic spur-thighed tortoise (Testudo graeca ibera) died following a history of lethargy, anorexia and absence of defecation. The two heads were anatomically similar with independent reaction to external stimuli. The carapace showed doubled first and extra second vertebral scutes. Radiography and transplastronal ultrasonography, performed when the animal was alive, revealed two symmetrical stomachs and two asynchronous hearts. These findings were confirmed by necropsy examination. Oesophagus, liver, gallbladder and trachea were also duplicated. Other malformations included pyloric valve atresia of the left stomach, focal stenosis of the transverse colon and liver hypoplasia. Dicephalism rarely occurs in Testudinidae and its pathogenesis, still unclear, is discussed.

Ventricular septal defect in a ferret (Mustela putorius furo).

A four-year-old, castrated male ferret (Mustela putorius furo) was evaluated because of a one-year history of sporadic cough. On physical examination a grade 5 of 6 holosystolic murmur was audible over the right apex of the heart. Radiographic findings included the presence of air bronchograms in apical lobes accompanied by pulmonary venous congestion. Colour Doppler echocardiography revealed a left-to-right shunting compatible with a ventricular septal defect. Medical therapy was initiated at the time of the diagnosis. The ferret was presented again 2 months after the initial examination for coughing and respiratory distress. Echocardiographic findings included tricuspid regurgitation, relative enlargement of left-atrial diameter and decreased systolic function, with presence of pleural effusion. Thoracocentesis was performed and the therapeutic plan was revised. In the following months the symptoms did not recur. In the authors' opinion this is the first report to describe the clinical findings of isolated ventricular septal defect in the ferret. Congenital heart defects are rare in this species, the present ferret being only the second case described.

Association of human papilloma virus with pterygia and ocular-surface squamous neoplasia.

There are more microorganisms that colonize the human body than resident cells; some are commensal whereas others are pathogenic. Pathogenic microorganisms are sensed by the innate or adaptive immune system, an immune response is initiated, and the infection is often cleared. Some microorganisms have developed strategies to evade immune defenses, ensuring their long-term survival with potentially devastating consequences for the host. Approximately 18% of all cancers can be attributed to infective agents; the most common being Helicobacter pylori, Human papilloma virus (HPV) and Hepatitis B and C virus in causing stomach, cervical and liver carcinoma, respectively. This review focuses on whether HPV infection is necessary for initiating pterygia, a common benign condition and ocular-surface squamous neoplasia (OSSN), a rare disease with metastatic potential. The search engine PubMed was used to identify articles from the literature related to HPV and pterygium or conjunctival neoplasia. From 34 investigations that studied HPV in pterygia and OSSN, a prevalence rate of 18.6% (136/731) and 33.8% (144/426), respectively, was recorded. The variation in HPV prevalence (0-100%) for both disease groups may have arisen from study-design faults and the techniques used to identify the virus. Overall, the data suggest that HPV is not necessary for initiating either condition but may be a co-factor in susceptible hosts. Currently, over 60 million people worldwide have been immunized with HPV vaccines, but any effect on pterygium and OSSN development may not be known for some time as these lesions can evolve over decades or occur in older individuals.

Ultraviolet radiation-induced cytokines promote mast cell accumulation and matrix metalloproteinase production: potential role in cutaneous lupus erythematosus.

OBJECTIVE: To examine the role of mast cells (MCs), cytokines, and matrix metalloproteinases (MMPs) following ultraviolet B (UVB) radiation in cutaneous lupus erythematosus (CLE). METHODS: Immunohistochemistry was used to determine the presence of MCs and the expression of MMP-1, MMP-9, interleukin (IL)-15, and CCL5/RANTES in skin from patients with CLE. Human keratinocytes were exposed to varying doses of UVB and supernatants were collected and assessed for IL-15, CCL5, MMP-1, and MMP-9 by protein assays. MC migration was determined against supernatants from UVB-treated keratinocytes. RESULTS: MCs in the skin of patients with CLE were significantly increased. MMP-1 and MMP-9 expression was abundant in these lesions. Intense reactivity for IL-15 and CCL5 was found in skin, particularly in epidermal keratinocytes, from patients with CLE. UVB irradiation induced IL-15, CCL5, MMP-1, and MMP-9 production from keratinocytes in a dose- and time-dependent manner. Supernatants obtained from UVB-treated keratinocytes induced MC migration, which was attenuated by anti-IL-15 and anti-CCL5 neutralizing antibodies. IL-15 induced MC-derived MMP production. CONCLUSIONS: Our results indicate that MCs and MMPs may play a role in the skin inflammation in CLE. MC recruitment as well as MMP production may be perpetuated by UV irradiation through locally released mediators.