Use Of Ferrets Research Articles

Pharmacokinetics of hydrorphone hydrochloride after intravenous and subcutaneous administration in ferrets (Mustela putorius furo)

ObjectiveTo determine the pharmacokinetic profile of hydromorphone 0.2 mg kg–1 administered by the intravenous (IV) and subcutaneous (SC) route in ferrets. Study designRandomized, crossover study. AnimalsA group of eight adult ferrets weighting (mean ± standard deviation) 1.02 ± 0.22 kg. MethodsHydromorphone hydrochloride 0.2 mg kg–1 was administered IV or SC with a washout period of 7 days. Blood samples were collected from a jugular catheter before administration of hydromorphone and at 5, 10, 15, 20, 30, 45, 60, 90, 120, 240, 360, 480 and 720 minutes after hydromorphone administration. Plasma hydromorphone concentrations were determined by liquid chromatography/tandem mass spectrometry. Data were analyzed using a non-linear mixed effects model. ResultsThe hydromorphone effective half-life was (t1/2) 45 min–1. Systemic clearance (Cls) and the volume of distribution (Vdss) following IV administration were 84.8 mL kg–1 min–1 and 5.59 L kg–1, respectively. The maximum observed plasma concentration was 59.53 ± 14.02 ng mL–1 within 10 minutes following SC administration. The SC bioavailability was 102.0%. Conclusions and clinical relevanceAdministration of IV and SC hydromorphone (0.2 mg kg–1) was characterized by a high clearance, short terminal half-life and large volume of distribution. Hydromorphone plasma concentrations remained greater than 2 ng mL–1 for 2 hours in most ferrets, a threshold reported to provide antinociceptive effects in other species. Hydromorphone was well absorbed following SC injection, providing an alternative administration route for clinical use in ferrets.

Ferrets as a Mammalian Model to Study Influenza Virus-Bacteria Interactions.

Ferrets represent an invaluable model for the study of influenza virus pathogenicity and transmissibility. Ferrets are also employed for the study of bacterial pathogens that naturally infect humans at different anatomical sites. While viral and bacterial infection studies in isolation using animal models are important for furthering our understanding of pathogen biology and developing improved therapeutics, it is also critical to extend our knowledge to pathogen coinfections in vivo, to more closely examine interkingdom dynamics that may contribute to overall disease outcomes. We discuss how ferrets have been employed to study a diverse range of both influenza viruses and bacterial species and summarize key studies that have utilized the ferret model for primary influenza virus challenge followed by secondary bacterial infection. These copathogenesis studies have provided critical insight into the dynamic interplay between these pathogens, underscoring the utility of ferrets as a model system for investigating influenza virus-bacteria interactions.

Host Transcriptome Analysis of Ferret Tissues Following Henipavirus Infection.

Ferrets are commonly used as experimental models of infection for a variety of viruses due to their susceptibility to human respiratory viruses and the close resemblance of pathological outcomes found in human infections. Even though ferret-specific reagents are limited, the use of ferrets as a preclinical experimental model of infection has gained considerable interest since the publication of the ferret transcriptome and draft ferret genome. These advances have made it feasible to easily perform whole-genome gene expression analysis in the ferret infection model. Here, we describe methods for genome-wide gene expression analysis using RNA sequence (RNAseq) data obtained from the lung and brain tissues obtained from experimental infections of Hendra (HeV) and Nipah (NiV) viruses in ferrets. We provide detailed methods for RNAseq and representative data for host gene expression profiles of the lung tissues that show early activation of interferon pathways and later activation of inflammation-related pathways.

First serological study of Dirofilaria immitis antibodies in household domestic ferrets (Mustela putorius furo) in southern Spain - Short communication.

Dirofilaria immitis is an endemic mosquito-borne pathogen widely spread throughout Europe as well as North and South America. Infection by D. immitis has been reported in domestic ferrets, although little is known about the occurrence and the epidemiological features of this nematode in this species. Theaim of the present retrospective study was to assess the prevalence of D. immitis antibodies using an in-house enzyme-linked immunosorbent assay specifically developed for use in ferrets. One hundred and eighty-six serum samples were obtained from the Province of Valencia (Spain), an area endemic for dirofilariosis. Of the 186 serum samples included in the study, 27 (14.51%) were classified as D. immitis seropositive and 159 samples as D. immitis seronegative. The results provide valuable information on the seroprevalence of D. immitis infection in domestic ferrets in an area endemic for this vector-borne pathogen. The presence of seropositive ferrets should be taken into account and preventive measures should be implemented, including the possibility of serological screening for the early detection of Dirofilaria antibodies as a serological marker of exposure. This is the first study that demonstrates thepresence of D. immitis exposure in ferrets in Spain. Veterinarians working in endemic areas should be aware of this infection in ferrets and their susceptibility.

Cardiac Radiographic Measurements in Ferrets Using the OsiriX MD Programme.

Objectives: To adapt the vertebral heart scale (VHS) for use in ferrets and identify new scales and tools that allow to establish the normal heart size by means of radiography more quickly and effectively.Methods: Forty healthy pet ferrets (Mustela putorius furo) were used in this prospective study. The measurements were made on right lateral, left lateral, ventrodorsal, and dorsoventral projections, using OsiriX MD medical imaging software, to evaluate sex effect and variance within the different heart scales. Cardiac measurements were also correlated to VHS and the cardiac dimension in the same projection.Results: Most of the cardiac measurements were significantly different between males and females. The results for the VHS were: right lateral VHS (RL-VHS): 5.52 ± 0.28 v (vertebrae units); left lateral (LL-VHS): 5.55 ± 0.28 v; and dorsoventral VHS (DV-VHS): 6.22 ± 0.34 v for males and RL-VHS: 5.24 ± 0.2 v; LL-VHS: 5.25 ± 0.20 v; and DV-VHS: 5.97 ± 0.35 v for females. Regarding the sternebral heart scale (SHS), the values were: RL-SHS: 5.10 ± 0.20 s (sternebrae units) and LL-SHS: 5.11 ± 0.20 s for males and RL-SHS: 4.67 ± 0.24 s and LL-SHS: 4.67 ± 0.28 s for females. The new measurements based on determining the cardiac area were also marked by clear sexual dimorphism, as shown for the cardiac area-axis (AREA-AXIS): RL-AREA-AXIS: 3.82 ± 0.45 cm2; LL-AREA-AXIS: 3.87 ± 0.41 cm2; ventrodorsal (VD)-AREA-AXIS: 4.59 ± 0.64 cm2; and DV-AREA-AXIS: 4.80 ± 0.50 cm2 for males and RL-AREA-AXIS: 2.39 ± 0.23 cm2; LL-AREA-AXIS: 2.41 ± 0.26 cm2; VD-AREA-AXIS: 3.08 ± 0.45 cm2; and DV-AREA-AXIS: 3.06 ± 0.47 cm2 for females. The cardiac area open polygon (AREA-POL) values were: RL-AREA-POL: 6.78 ± 0.65 cm2; LL-AREA-POL: 6.88 ± 0.68 cm2; VD-AREA-POL: 7.20 ± 0.91 cm2; and DV-AREA-POL: 7.57 ± 0.88 cm2 for males and RL-AREA-POL: 4.28 ± 0.30 cm2; LL-AREA-POL: 4.35 ± 0.35 cm2; VD-AREA-POL: 4.72 ± 0.65 cm2; and DV-AREA-POL: 4.79 ± 0.66 cm2 for females, with similar differences noted from various radiographic projections. A good correlation was noted between VHS and SHS, and a very strongly positive correlation existed between cardiac area measurements and cardiac dimensions.Conclusion: The VHS adapted to ferrets, the SHS, as well as the cardiac area measurements presented in our study are ideal tools for the assessment of cardiac size in ferrets.

An explorative study on the effect of provision of preferred and non-preferred enrichment on behavioural and physiological parameters in laboratory ferrets (Mustela putorius furo)

Environmental enrichment is often advocated to refine animal studies. Despite the increasing use of ferrets as an animal model in biomedical research, the knowledge on effects of the provision of enrichment on these animals is limited. Additionally, it is unknown whether varying types of enrichment (i.e. preferred and non-preferred) have a different effect. Therefore, to explore the behavioural and physiological effects of providing (differently valued) enrichment to ferrets, three groups of six female ferrets were housed in standard conditions (with bedding, a flexible bucket, a food bowl and a water nipple), with additional non-preferred enrichment (with two ferret balls, a golf ball and an extra food bowl) and with additional preferred enrichment (with two hammocks, a foraging ball and a water bowl) for eight weeks. At the beginning and end of this period, behavioural (i.e. time spent on food and water intake, elimination, maintenance, inactivity, enrichment interaction, exploration, play, and agonistic behaviour) and physiological (i.e. bodyweight and Neutrophil/Lymphocyte ratio [N/L ratio]) parameters were recorded and compared. Results showed that agonistic behaviour increased in the ferrets housed in standard conditions, which was not observed in ferrets that were provided with preferred or non-preferred enrichment. In addition, the ferrets housed with preferred enrichment showed an increase in social play behaviour and a decrease in rearing behaviour (as part of the exploratory behaviours) which were not observed in the ferrets housed in standard conditions or with non-preferred enrichment. Moreover, the ferrets housed with preferred enrichment showed a clear preference for being inactive in the hammock and drinking from the water bowl. As there was only one cage per condition, the results of this explorative study should be considered preliminary and tentative. Nevertheless, the results are a first indication that providing laboratory ferrets with preferred enrichment has positive effects on their behaviour that are not observed in ferrets provided with non-preferred enrichment or housed in standard conditions. Therefore, we recommend to house laboratory ferrets with a hammock, foraging ball and water bowl as these enrichments might help to refine studies using ferrets.

Use of ferrets for electrophysiologic monitoring of ion transport.

Limited success achieved in translating basic science discoveries into clinical applications for chronic airway diseases is attributed to differences in respiratory anatomy and physiology, poor approximation of pathologic processes, and lack of correlative clinical endpoints between humans and laboratory animal models. Here, we discuss advantages of using ferrets (Mustela putorus furo) as a model for improved understanding of human airway physiology and demonstrate assays for quantifying airway epithelial ion transport in vivo and ex vivo, and establish air-liquid interface cultures of ferret airway epithelial cells as a complementary in vitro model for mechanistic studies. We present data here that establishes the feasibility of measuring these human disease endpoints in ferrets. Briefly, potential difference across the nasal and the lower airway epithelium in ferrets could be consistently assessed, were highly reproducible, and responsive to experimental interventions. Additionally, ferret airway epithelial cells were amenable to primary cell culture methods for in vitro experiments as was the use of ferret tracheal explants as an ex vivo system for assessing ion transport. The feasibility of conducting multiple assessments of disease outcomes supports the adoption of ferrets as a highly relevant model for research in obstructive airway diseases.

Parathyroid Hormone, Ionized Calcium, and 25-Hydroxyvitamin D Concentrations in the Domestic Ferret (Mustela putorius furo)

The objective of this study was to measure parathyroid hormone (PTH), ionized calcium, and 25-hydroxyvitamin D concentrations in healthy intact adult ferrets. Serum was collected from 16 clinically healthy adult ferrets (8 males and 8 females). Concentrations of PTH and 25-hydroxyvitamin D were measured via commercially available radioimmunoassays validated for humans, dogs, and cats (PTH) plus horses (25-hydroxyvitamin D). Concentrations of ionized calcium (at pH 7.4) were measured using an ion-specific electrode. Median (minimum, maximum) concentrations were as follows: PTH 8.7 (2.2, 24.4)pmol/L, ionized calcium 1.15 (1.09, 1.25)mmol/L, and 25-hydroxyvitamin D 94 (61, 138) nmol/L). Female ferrets had significantly higher concentrations of PTH than male ferrets (female median: 17.1pmol/L; male median: 6.9pmol/L). Associations among PTH, ionized calcium, 25-hydroxyvitamin D, calcium, phosphorus, magnesium, and weight were evaluated. There was a weak negative correlation between PTH concentration and total calcium concentration in female ferrets. As none of these assays have been validated for use in ferrets, results may provide a baseline for clinicians evaluating ferrets for disorders of calcium homeostasis, parathyroid glands, and paraneoplastic syndromes.

Ferret airway epithelial cell cultures support efficient replication of influenza B virus but not mumps virus.

Ferrets have become the model animal of choice for influenza pathology and transmission experiments as they are permissive and susceptible to human influenza A viruses. However, inoculation of ferrets with mumps virus (MuV) did not lead to successful infections. We evaluated the use of highly differentiated ferret tracheal epithelium cell cultures, FTE, for predicting the potential of ferrets to support respiratory viral infections. FTE cultures supported productive replication of human influenza A and B viruses but not of MuV, whereas analogous cells generated from human airways supported replication of all three viruses. We propose that in vitro strategies using these cultures might serve as a method of triaging viruses and potentially reducing the use of ferrets in viral studies.

Assessment of a blood preservation protocol for use in ferrets before transfusion

Blood transfusion has been described in ferrets as a treatment for oestrus-associated anaemia and as a life-saving therapy following trauma, iatrogenic (usually surgery-induced) anaemia, autoimmune haemolytic anaemia and pure red...

The current state of H5N1 vaccines and the use of the ferret model for influenza therapeutic and prophylactic development

Highly pathogenic avian influenza H5N1 is a threat to global public health as a natural pandemic causing agent but has recently been considered a bioterrorism concern. The evolving view of the H5N1 virus necessitates the re-evaluation of the current status of H5N1 therapeutics and prophylactics, in particular the preparation of viable H5N1 vaccination strategies as well as the use of ferrets in influenza research. Here the highly pathogenic H5N1 virus dilemma is discussed in context with the current H5N1 vaccine status and the use of the ferret model. Previously, the development of various H5N1 vaccine platforms have been attempted, many of them tested in the ferret model, including vector vaccines, adjuvant vaccines, DNA vaccines, and reverse engineered vaccines. Moreover, as ferrets are a superlative animal model for influenza investigation and vaccine testing, it is imperative that this model is recognized for its uses in prophylactic development and not only as an agent for creating transmissible influenza viruses. Elucidating the ferret immune response and creating ferret immune reagents remain important goals in conjunction with the development and manufacture of H5N1 vaccines. In summary, an efficacious H5N1 vaccine is urgently needed and the ferret model remains an appropriate model for its development.

Enteric Coronavirus in Ferrets, the Netherlands

Enteric Coronavirus in Ferrets, the Netherlands

Evaluation of a black‐footed ferret resource utilization function model

AbstractResource utilization function (RUF) models permit evaluation of potential habitat for endangered species; ideally such models should be evaluated before use in management decision‐making. We evaluated the predictive capabilities of a previously developed black‐footed ferret (Mustela nigripes) RUF. Using the population‐level RUF, generated from ferret observations at an adjacent yet distinct colony, we predicted the distribution of ferrets within a black‐tailed prairie dog (Cynomys ludovicianus) colony in the Conata Basin, South Dakota, USA. We evaluated model performance, using data collected during post‐breeding spotlight surveys (2007–2008) by assessing model agreement via weighted compositional analysis and count‐metrics. Compositional analysis of home range use and colony‐level availability, and core area use and home range availability, demonstrated ferret selection of the predicted Very high and High occurrence categories in 2007 and 2008. Simple count‐metrics corroborated these findings and suggested selection of the Very high category in 2007 and the Very high and High categories in 2008. Collectively, these results suggested that the RUF was useful in predicting occurrence and intensity of space use of ferrets at our study site, the 2 objectives of the RUF. Application of this validated RUF would increase the resolution of habitat evaluations, permitting prediction of the distribution of ferrets within distinct colonies. Additional model evaluation at other sites, on other black‐tailed prairie dog colonies of varying resource configuration and size, would increase understanding of influences upon model performance and the general utility of the RUF. © 2011 The Wildlife Society.

Cloning, expression and immunoassay detection of ferret IFN- γ

SummaryFerrets (Mustela putorius furo) develop symptoms upon influenza infection that resemble those of humans, including sneezing, body temperature variation and weight loss. Highly pathogenic strains of influenza A, such as H5N1, have the capacity to cause severe illness or death in ferrets. The use of ferrets as a model of influenza infection is currently limited by a lack of species-specific immunological reagents. Interferon gamma (IFN-γ) plays a key role in the development of innate and adaptive immunity and the regulation of Th1-type immune responses. Here we describe the cloning of the full-length cDNA for ferret IFN-γ. Multiple sequence alignment of the predicted amino acid sequence with those of other species indicates that the predicted ferret protein shares the highest identity with Eurasian badger IFN-γ. We raised two hybridoma clones expressing monoclonal antibodies against recombinant ferret IFN-γ capable of detecting IFN-γ protein derived from mitogen-stimulated ferret PBMCs by immunoblotting, ELISA and ELISPOT assay. Finally, an ELISA utilizing the ferret-specific antibodies detected elevated levels of IFN-γ in serum samples from H3N2 influenza A-infected ferrets.

Use of an adjustable restraint device for prolonged and intermittent intravenous infusion and blood sampling in ferrets

For experiments requiring repeated venipuncture or prolonged intravenous infusion or sampling techniques, the ferret can serve successfully as a surrogate nonrodent animal model. Because experimental protocols must be suitable for the test system or model used, the author describes specific restraint equipment designed to facilitate the use of ferrets in the research laboratory.

Strong local and systemic protective immunity induced in the ferret model by an intranasal virosome-formulated influenza subunit vaccine

The proliferation of influenza viruses causes costly, recurrent, annual epidemics. Current vaccines, mainly administered parenterally, have been shown to be suboptimal in terms of efficacy, particularly where local IgA responses are concerned. Recent investigations of virosomes as delivery systems for viral HA and NA antigens have demonstrated an improved immune response. This paper investigates the efficacy of a novel virosome-based intranasal influenza vaccine by its ability to reduce disease symptoms and its effect on viral shedding in nasal secretions of immunised ferrets. The use of ferrets in the study of influenza vaccines is based on the good comparability between ferret and human response to the disease. Intranasal, as opposed to parenteral, administration of a trivalent virosome-based subunit vaccine adjuvanted with HLT provides an almost total prevention of virus shedding combined with a high level of immunological protection against homologous virus challenge. The ease of application of an intranasal vaccine may have positive repercussions in the adoption of influenza vaccinations, particularly in ‘at-risk’ groups.

Assessment of development of resistance to antivirals in the ferret model of influenza virus infection.

We attempted to develop in vivo resistance of influenza virus to amantadine and to zanamivir, by use of the ferret model of influenza virus infection. Resistance of influenza virus A/LosAngeles/1/87 (H3N2) to amantadine was generated within 6 days, during a single course of treatment, and mutations in the M2 gene that are characteristic of human infections were observed. In contrast, during an identical single course of treatment with zanamivir, no evidence of reduced susceptibility was demonstrated. Pooled virus shed by zanamivir-treated ferrets was used to infect another group of ferrets. Twenty virus clones grew in plaque assays containing zanamivir, indicating possible reduced susceptibility; however, none exhibited reduced susceptibility to zanamivir in neuraminidase (NA) inhibition assays. Sequencing of the NA gene of these clones revealed only a noncoding nucleotide mutation at position 685. Sequencing of the hemagglutinin gene revealed mutations at positions 53, 106, 138, 145, 166, and 186. Similar to the situation in humans, amantadine use in ferrets rapidly produces antiviral resistance, but zanamivir use does not, although nucleotide changes were observed.

Incidence of adverse events in ferrets vaccinated with distemper or rabies vaccine: 143 cases (1995-2001).

To determine the incidence of adverse events in ferrets vaccinated with a modified-live avian cell culture canine distemper virus vaccine licensed for use in ferrets, an inactivated rabies vaccine licensed for use in ferrets, or both. Retrospective study. 143 ferrets. Medical records were reviewed to identify ferrets that had an adverse event after vaccination. Adverse events developed within 25 minutes after vaccination in 13 ferrets. One ferret developed an adverse event after receiving a distemper and a rabies vaccine simultaneously and developed a second adverse event the following year after receiving the rabies vaccine alone. Therefore, a total of 14 adverse events were identified. All adverse events were an anaphylactic reaction characterized by generalized hyperemia, hypersalivation, and vomiting. Ten of the 14 anaphylactic reactions occurred after ferrets received both vaccines, 3 occurred after ferrets received the distemper vaccine alone, and 1 occurred after a ferret received the rabies vaccine alone. Incidences of adverse events after administration of both vaccines, the distemper vaccine alone, and the rabies vaccine alone were 5.6, 5.9, and 5.6%, respectively. Ferrets that had an anaphylactic reaction were significantly older at the time of vaccination than were ferrets that did not. Results suggest that there may be a high incidence of anaphylactic reactions after vaccination of domestic ferrets. Ferrets should be observed for at least 25 minutes after vaccination, and veterinarians who vaccinate ferrets should be prepared to treat anaphylactic reactions.

Successful production of offspring after superovulation and in vitro culture of embryos from domestic ferrets (Mustela putorius furos)

In an effort to expand the use of ferrets as models for genetic disease, several experimental parameters that are required for successful genetic manipulation in this species were investigated. Optimum superovulation (19.3 +/- 0.6 oocytes and embryos per female) was achieved after injections of 100 iu equine chorionic gonadotrophin (eCG) and 150 iu human chorionic gonadotrophin (hCG). The ovulation rate achieved by the treatment was more than double that induced by mating. Mating with a male immediately after hCG treatment did not significantly alter the number of oocytes ovulated or the number of embryos present, indicating that mating is not required for superovulation in ferrets. Of embryos harvested at the one-cell stage, 64.5% and 47.1% developed into blastocysts when cultured in vitro in CZB or TCM-199 plus 10% fetal bovine serum (FBS) media, respectively. In contrast, only 17.1% of embryos cultured in vitro in NCSU-23 developed to the blastocyst stage. Both freshly retrieved and in vitro cultured embryos from cinnamon-coloured parents produced live young when transferred at the eight-cell stage into albino, pseudo-pregnant recipients. The percentage of kits delivered relative to embryos transferred was 61% for freshly retrieved embryos and 32% for embryos cultured in vitro. These results demonstrate successful embryo transfer in ferrets and provide a basis for further study of genetic modelling approaches in this species after embryo manipulation.

The Host Range of Chronic Wasting Disease Is Altered on Passage in Ferrets

Chronic wasting disease (CWD), a member of the transmissible spongiform encephalopathies (TSEs), was first identified in captive mule and black-tail deer in 1967. Due to the failure to transmit CWD to rodents, we investigated the use of ferrets ( Mustela putorius furo) as a small animal model of CWD. The inoculation of CWD into ferrets resulted in an incubation period of 17–21 months on primary passage that shortened to 5 months by the third ferret passage. The brain tissue of animals inoculated with ferret-passaged CWD exhibited spongiform degeneration and reactive astrocytosis. Western blot analysis of ferret-passaged CWD demonstrated the presence of PrP-res. Unlike mule deer CWD, ferret-passaged CWD was transmissible to Syrian golden hamsters ( Mesocricetus auratus). Increasing the passage number of CWD in ferrets increased the pathogenicity of the agent for hamsters. This increase in host range of a field isolate on interspecies transmission emphasizes the need for caution when assessing the potential risk of transmission of TSEs, such as bovine spongiform encephalopathy, to new host species.