Aggressive behaviour of male mice (Mus musculus) towards familiar and unfamiliar opponents.

Two types of colonies of laboratory mice were employed; hierarchically organized ones formed by placing five unfamiliar 8-week-old mice in a cage together and amicably organized colonies in which four male litter mates were kept together throughout the whole period of the experiment. During a 21-day pre-experimental period intra-colony aggressive behaviour was recorded. A dominant animal and ranked subordinates occurred in every hierarchical colony, whilst no aggression was recorded in any of the amicable colonies. During a 25-day period unfamiliar adult male or female mice were introduced daily into the amicable or hierarchical colonies for 10 min. In a third experiment juvenile mice 17, 24, 31 or 38 days old were introduced into hierarchically organized colonies during a 20-day period. In all hierarchical colonies the stranger was attacked irrespective of sex and age; the majority of attacks were carried out by the dominant mouse. Aggression by the dominant declined exponentially throughout the experimental period and regression analyses compared the different data. Unfamiliar adult females were the recipients of fewer attacks than unfamiliar adult males and the age of juvenile strangers was found to be an important factor. Amicably organized mice initially did not attack strangers, but over a period of 25 days the number of attacks on unfamiliar males gradually increased.     

The effects of oestrous condition and familiarity on the sexual behaviour of polecats (Mustela putorius and M. fur × M. putorius hybrids)

The effects of two factors on the sexual behaviour of polecats ( Mustela furo and M. putorius × M. furo hybrids) were investigated, in an indoor arena of 16 m², firstly the oestrous condition of the female and secondly the familiarity of the two individuals.
The initial approaches of a male polecat are similar irrespective of the oestrous condition of the female but his subsequent behaviour is dependent upon the female's response. Whereas males copulated with oestrous females they showed a wider repertoire of social acts and a greater number of changes of behaviour towards females which were not in oestrous. Unreceptive females elicited both sexual and aggressive behaviour from males.
Unfamiliar females provide a more powerful eliciting stimulus for the male's copulatory behaviour than do familiar females.     

Experimental infection of domestic ferrets (Mustela putorius furo) and Siberian polecats (Mustela eversmanni) with Yersinia pestis

Eight domestic ferrets (Mustela putorius furo) and two Siberian polecats (M. eversmanni) were inoculated subcutaneously with 12 to 1.2 x 10(7) Yersinia pestis originally isolated during an epizootic of plague in white-tailed prairie dogs (Cynomys leucurus) near Meeteetse, Park County, Wyoming (USA) in 1985. None of the ferrets or polecats developed clinical signs of disease which suggested that black-footed ferrets (M. nigripes), a congener, also would be resistant to plague. All animals receiving greater than or equal to 1.2 X 10(3) organisms produced serum antibodies detected by the passive hemagglutination test with titers peaking at 1:1,024 and remaining positive until at least 219 days postinoculation. Sera collected from 12 free-ranging black-footed ferrets near Meeteetse in 1984 and 1985 were negative for antibodies against Y. pestis. Prevalence of antibodies against Y. pestis was high in other carnivores collected from the same area in 1986.     

A panel of microsatellite markers for genetic studies of European polecats (Mustela putorius) and ferrets (Mustela furo)

We report a panel of 12 microsatellite markers, including nine novel polymorphic loci isolated in the European polecat and three loci previously developed in closely related species, for genetic studies of polecats and ferrets. We tested the panel at fine geographic scales in polecat and domestic ferret populations of Britain and Portugal and at a broad geographic scale, using 50 polecat samples from across Western Europe. At the fine geographic scales, we recorded one to five alleles per locus and observed and expected heterozygosities ranging from 0 to 0.80 and from 0 to 0.74, respectively. These values increased markedly in the European sample of polecats. No linkage disequilibrium was detected between any pair of loci and all genotypic frequencies complied with Hardy–Weinberg equilibrium expectations. The microsatellite panel allowed the molecular discrimination of polecats and domestic ferrets in Britain, where hybridisation between the two species has been documented. Thus, these loci may be useful not only in genetic studies of polecats and ferrets but also in studies to assess hybridisation between these two mustelid species.     

Comparison of black-footed ferret (Mustela nigripes) and domestic ferret (M. putorius furo) courtship activity

Because of the scarcity of the endangered black-footed ferret (Mustela nigripes) and the amount of knowledge necessary for their conservation, surrogate research can play an important role in recovery. In this paper, we investigate surrogate behavioral research potential by comparing courtship behavior of the black-footed ferret to the congeneric domestic ferret (M. putorius furo). Ten female domestic ferrets were bred to five male domestic ferrets and eight female black-footed ferrets were bred to five black-footed ferret males. Courtship activities were defined, analyzed, and quantitatively compared between both groups. Lag sequential analysis of was used to prepare the behavioral matrices, and matrix cells were compared between groups with an equality of proportions test. Courtship patterns did not differ significantly between the two closely related species, and the domestic ferret would probably be an adequate surrogate for reproductive behavior research on the black-footed ferret.     

The ecological significance of resting sites and the seasonal habitat change in polecats (Mustela putorius)

The daytime resting sites of 13 radio-tracked polecats ( Mustela putorius ) were recorded in a mountainous and a lowland study area in Switzerland. Information on breeding sites was collected from questionnaires distributed throughout the country. At all resting sites, the polecats were completely invisible from more than a metre distant. During summer, when polecats mainly live in forests, individuals used many different hiding-places (e.g. small self-dug burrows, woodpiles, heaps of branches and dry leaves, dense vegetation). Such places were used for short periods, and then abandoned. Above ground, the polecats sometimes built nests of dry grass or moss. In winter, the polecats slept mainly inside barns, stables and other buildings. These resting sites were changed less frequently. In rainy weather throughout the year, subterranean places were preferred. Breeding polecats were often found inside houses, and obviously did not avoid human presence.
The importance of different types of resting sites for polecats is discussed. In summer, the quality of a resting place is of less importance than its distance from the foraging area, but in winter, warm resting places are essential and buildings are therefore considered an important resource for polecats in Switzerland. The seasonal habitat change can more readily be explained as a consequence of thermoregulatory problems than of food availability, and the distribution of polecats in Switzerland may be affected by the availability of suitable winter resting places. The northern limit of polecat distribution and its historical changes can be explained by the changing availability of human buildings which provide winter resting sites.     

The ferret, Mustela putorius furo, as a new species in toxicology.

Comprehensive studies on the haematology, urine chemistry, serum chemistry and gross pathology have been carried out and reported. Seasonal weight changes were confirmed and were initiated, together with sexual activity, when ferrets were brought in to a constant temperature (18-19 degrees C) and 14-hour daylight cycle during the winter months. Levels of haemoglobin, erythrocytes and haematocrit were higher than in most common laboratory species. Similarly, serum electrolytes and glucose levels were higher than in Wistar rats or beagle dogs. Ovaries, uteri and testes displayed a seasonal maturation and atrophy. All other parameters were similar to common laboratory species.     

Hormonal induction of oestrus and pregnancy in anoestrous ferrets (Mustela putorius furo)

Three doses of FSH were tested for their ability to induce oestrus in ferrets. A dose of 0.25 mg, administered twice daily, induced oestrus and breeding in most females within 6-13 days. Addition of 5 i.u. hCG during the final stages of follicular development enhanced the percentage of females with implantation sites (85%), and 23% of the ferrets so treated gave birth to kits, none of which survived for more than 3 days. This may have been due to insufficient prolactin secretion and thus inadequate luteal maintenance and milk production, as these females were maintained on a short-day photoperiod.     

Stages and duration of spermatogenesis in the domestic ferret (Mustela putorius furo)

Classification of seminiferous tubules is the basis for understanding normal and abnormal spermatogenesis. The aim of the present study was to determine spermatogenic stages and the duration of the cycle in the domestic ferret using bromodeoxyuridine (BrdU) as a tracer. Eleven adult male ferrets that were maintained in a breeding condition were used. Testicular sections were stained with the periodic acid-Schiff reaction for light microscopy. To determine the cycle duration, six ferrets were injected intraperitoneally with BrdU, and testes were collected 3h later and 10 days and 3h later. BrdU was detected by immunohistochemistry. Seminiferous tubules were classified into eight stages, and frequencies of stages I-VIII were 10.6, 2.2, 7.9, 13.1, 22.3, 21.9, 14.0 and 8.0%, respectively. The most advanced BrdU-labeled cells at 3h post-injection were leptotene spermatocytes in stage VI and those at 10 days and 3h were pachytene spermatocytes in stage V. From differences in stage frequency and BrdU staining frequency between two time points, the duration of one cycle was estimated to be 13.0 days. The present observations indicate that stages and the cycle duration of the ferret spermatogenesis are similar to those reported in other carnivores.     

Courtship patterns in induced oestrous and natural oestrous domestic ferrets (Mustela putorius furo)

Ten anoestrous female domestic ferrets (Mustela putorius furo) had estrus artificially induced by injection of porcine follicle stimulating hormone. Pre-copulatory activities were defined, quantitatively analyzed, and compared to the pre-copulatory activities of 10 control females placed with the same male ferrets. Lag sequential analysis was used to prepare the behavioral matrices. Matrices were compared with an equality of proportions test. Courtship patterns between the 2 groups did not differ significantly.     

Untersuchung zum Einfluß des Erkundungs- und Spielverhaltens auf das Lernen bei Iltisfrettchen (Mustela putorius X M. furo)

This paper investigates the dependence of learning performance on previous exploration and playing with objects. The subjects are polecats (Mustela putorius X M. furo). Behaviour of social animals, who are offered changing tube systems in certain periods, is described and analysed. A maze-test shows that subjects with tube training are superior to subjects without tube training as regards acquisition and reversal learning. The learning performance in pattern recognition tests is the same in all tests with the different groups, however.     

Macroscopic and roentgenographic anatomy of the skull of the ferret (Mustela putorius furo)

Normal macroscopic and roentgenographic features of the skull of the ferret (Mustela putorius furo) were examined and described. Data were based on a sample of 100 (50 male and 50 female) adult ferrets of known body weight and age. The skull was described macroscopically according to six standard views, i.e. dorsal, lateral, ventral, caudal, cranial and midsagittal. The mandible was described separately. The roentgenographic characteristics of the ferret skull were demonstrated only in lateral and dorsoventral projections. Furthermore, the skull length and width as well as the minimum frontal width were measured, and skull indices were derived from relevant measurements. Sexual dimorphism was examined both morphologically and craniometrically. Besides the common features of a carnivore skull, the ferret skull is relatively elongated and flat with a short facial region. The skulls of adult male ferrets are about 17% longer and 22% wider than those of the females. Significant sexual dimorphism also exists regarding certain skull indices. The general features and some dimensional parameters of the adult ferret skull support the contention that the ferret would be an interesting and workable alternative animal model in craniofacial research.     

Immune Response to and Histopathology of Campylobacter jejuni Infection in Ferrets (Mustela putorius furo)

Campylobacter jejuni is 1 of the most common enteric bacterial pathogens worldwide. The mechanisms of pathogenesis remain obscure, in part because of limitations of small animal models. Young ferrets develop diarrhea when fed C. jejuni, but their pathology and the immune response after infection have not been examined in detail. In the present study, we examined the pathogenesis of C. jejuni CG8421 and associated immune responses in ferrets. After oral infection with C. jejuni CG8421, 86.7% of the animals developed diarrhea and inflammatory responses that were similar to those seen in human infection. Pronounced histopathologic changes in the colonic mucosa of infected animals were observed during the acute phase (days 1 through 3) of infection. Electron micrographs of colonic epithelium revealed disruption of the villi and internalized bacteria that were not within membrane vacuoles. During the acute phase, C. jejuni was isolated from the livers of 7 of 9 (78%) animals, and bacteria were visualized immunohistochemically in the livers from 5 of the 7 animals (71%) from which C. jejuni was isolated. A vigorous systemic and mucosal immune response to Campylobacter antigens was elicited after infection of ferrets. The data presented contribute to the current knowledge of the pathogenicity of and immunologic response to C. jejuni CG8421 in ferrets and better understanding of this model.Abbreviation: ASC, antibody-secreting cellsCampylobacter jejuni are gram-negative, spiral, microaerophilic, motile bacteria and 1 of the most common enteric bacterial pathogens globally.^15,20 The annual incidence of C. jejuni diarrheal disease varies geographically, ranging from sporadic infections in young adults in developed countries²⁸ to as frequent as 40,000 per 100,000 among children living in hyperendemic regions.¹⁵ In addition, C. jejuni is the primary cause of ‘traveler''s diarrhea’ in various regions of the world.^39,41,44 The clinical disease of C. jejuni infection has a wide spectrum of symptoms, varying from a mild, transient watery diarrhea to a bloody diarrhea with severe abdominal cramps and fever. C. jejuni infection also is associated with development of Guillain–Barré syndrome.⁵¹ Most strains of C. jejuni contain lipooligosaccharides that mimic human gangliosides structurally. Antibodies against these lipooligosaccharide structures lead to an autoimmune response, resulting in Guillain–Barré syndrome.⁵²Despite extensive study, little is understood about the mechanism by which C. jejuni causes diarrheal disease. Various Campylobacter small animal models have been reported, but these require surgical manipulation,^13,46 administration of chemicals or antibiotics (or both),^17,26,30 or inoculation by an unnatural route.⁵ None of these published models incorporate the natural route of infection. Although several immunodeficient mouse models have been reported,^35,36,49 the applicability of these models for studying the pathogenesis and immune response of natural infection is limited. An adult mouse lung model has been described that may be useful for studying pathogenesis and inflammatory response and acquired immunity, but the route of infection is not the same as that in humans.^1,5 Unlike other small animals, naïve ferrets (younger than 11 wk) intragastrically inoculated with C. jejuni developed mild to moderate diarrhea with mucus, fecal leukocytes or frank blood that lasted for as long as 3 d and remained colonized for as long as 8 d.^9,10,18 Moreover, on rechallenge with the homologous bacterial strain, ferrets asymptomatically excreted C. jejuni in their stools.¹⁰ The observation of resistance to disease, but not to infection, is similar to findings from human studies, in which protection against disease developed before colonization resistance.¹¹ Ferrets have been used to evaluate vaccines against C. jejuni¹⁴ and in molecular pathogenesis studies using strain 81-176 to confirm a role in virulence for capsular polysaccharide and flagellar glycans.^3,21 However, we evaluated the histopathology and immune response after C. jejuni infection in ferrets in more detail in the present study.C. jejuni CG8421 was isolated from a case of dysentery in Thailand. It has an lipooligosaccharide core that, as shown by chemical analyses, lacks all ganglioside mimicry.³⁸ The absence of ganglioside mimicry is predicted to reduce the risk of Guillain–Barré syndrome. We selected strain CG8421 for the current studies as part of an evaluation of a potential strain for use in future human vaccine challenge studies. Here we report the pathogenesis, histopathology, inflammatory, and immune responses after oral infection of ferrets with C. jejuni strain CG8421.     

Effects of Buprenorphine Treatment on Influenza Pathogenesis in the Ferret (Mustela putorius furo)

Ferrets are the gold-standard model for influenza A virus (IAV) research due to their natural susceptibility to human and zoonotic IAV, comparable respiratory anatomy and physiology to humans, and development of clinical signs similar to those seen in infected people. Because the presence and progression of clinical signs can be useful in infectious disease research, uncertainty in how analgesics alter research outcomes or compromise characteristics of disease progression have outweighed the concern regarding animal discomfort from these symptoms. Nonetheless, the principles of animal research require consideration of refinements for this important model for IAV research. Opioids offer a possible refinement option that would not directly affect the inflammatory cascade involved in IAV infection. Mirroring pathogenicity studies that use ferrets, 12 ferrets were inoculated intranasally with the A(H3N2) IAV A/Panama/2007/1999 and divided into 3 treatment groups (n = 4 each), of which 2 groups received buprenorphine treatments on different schedules and the third received a saline control. The duration and location of viral replication, lymphohematopoietic changes, and clinical signs were comparable across all groups at all time points. High quantities of infectious virus in nasal wash specimens were detected in ferrets from all groups through day 5 after inoculation, and peak viral titers from the upper respiratory tract did not differ between ferrets receiving buprenorphine treatments on either schedule. Compared with the saline group, ferrets receiving buprenorphine exhibited transient weight loss and pyrexia, but all groups ultimately achieved similar peaks in both of these measurements. Collectively, these findings support the continued evaluation of buprenorphine as a refinement for IAV-challenged ferrets.

Despite decades of international research and the availability of public health countermeasures, including vaccines and antivirals, influenza viruses remain a persistent threat to human and animal health.^26,35 Influenza A viruses (IAV) exhibit a diverse range of virulence, exist in several host reservoirs, and can show rapid rates of antigenic change.²⁶ As a result, IAV are associated with both seasonal epidemics and occasional pandemics in humans,³⁵ and animal infections with IAV have become key for understanding multifactorial traits that include pathogenicity, transmissibility, and vaccine efficacy. Due to their relatively small size, adaptability to the research setting, and similarities to human lung anatomy and physiology, ferrets provide an excellent model for respiratory diseases in humans and are a valuable small-animal model for such studies.^8,30 Data generated from ferrets are included in numerous risk-assessment rubrics evaluating the pandemic potential of novel and emerging influenza viruses, including those established by the Centers for Disease Control and Prevention and the World Health Organization.^14,51The study of influenza virus in ferrets dates back to the early 1930s, when this species was first found to be susceptible to influenza virus.⁴⁴ Ferrets are naturally susceptible to both human and zoonotic IAV.⁴⁷ After infection, ferrets present with clinical signs like those of humans; these signs are often not recapitulated in other species, such as mice and guinea pigs.^28,39,46 The severity and spectrum of clinical signs associated with influenza virus–inoculated ferrets can vary, depending on the virus strain, route and dose of inoculation, and various host parameters.⁵ Whereas influenza viruses with low virulence in ferrets may cause only acute pyrexia and mild to moderate weight loss, isolates with high virulence can cause severe, systemic illness with gastrointestinal and neurologic symptoms.⁴The 3Rs, replace, reduce, refine, encourage investigation of how research involving animals can be conducted in more humane ways.^2,13,37,41 Analgesia for symptoms of influenza in ferrets represents an opportunity for refinement, but this intervention could confound research assessing disease progression. NSAID and corticosteroids are often prescribed to treat the clinical signs associated with influenza in humans.⁴³ These interventions could alter the inflammatory cascade and subsequent pathophysiology of the disease, thus reducing the validity of studies designed to characterize and compare influenza viruses.^6,43 NSAID reportedly inhibit nuclear factor κB, a regulator of inflammatory processes that is involved in viral RNA synthesis.^25,27 In addition, NSAID have been found to increase survival rates in influenza virus-infected mice.⁵³ Therefore, the use of NSAID may be problematic in studies investigating the pathogenesis of influenza viruses.Buprenorphine, an opioid, is an established analgesic in ferrets that can be administered either intravascularly, intramuscularly, or subcutaneously at 0.01 to 0.05 mg/kg with an analgesic duration of 6 to 12 h.^{11,16,24,38,52} Historically buprenorphine has been described as a partial µ receptor agonist and κ and γ receptor antagonist,^22,29,40,48 but the drug recently was described to behave as a full µ agonist.³⁶ The ceiling effect of analgesia and the immunosuppressive effects reported with other opioids have not been documented to occur with buprenorphine.^15,36,42 However, the use of buprenorphine does have the possibility of adverse effects, including sedation, weight loss, constipation, and respiratory depression.^{10,15,16,22,23,34,42} Nonetheless, buprenorphine is a commonly prescribed analgesic for numerous small mammalian species used in research settings.^20,22,40Given that influenza is an ongoing threat to human and animal health and because no replacement is available for data gained with the ferret model, pain mitigation options for research conducted in this species must be addressed. To date, concerns about altering the course of the disease have precluded the evaluation of refinements options in IAV-infected ferrets. The goal of the current study was to assess the effects of buprenorphine treatments on the pathogenesis of a seasonal IAV in ferrets; this assessment was achieved by comparing virus-inoculated ferrets that were either sham-treated or that received buprenorphine according to 2 different dosing schedules. We hypothesized that buprenorphine treatments would not affect experimental readouts, including morbidity, viral shedding, lymphopenia, and seroconversion in convalescent serum; these parameters are commonly measured during IAV research. Study results indicate that buprenorphine did not uniformly or significantly modulate disease progression, peak viral titers in the upper respiratory tract, or clinical responses used to characterize viral pathogenicity in ferrets.     

Characterization of transforming growth factors beta1 and 2 in ferrets (Mustela putorius furo)

Unwanted scar tissue after surgical procedures remains a central problem in medicine. Nowhere is this problem more evident than within the pediatric airway, where excess scarring, termed subglottic stenosis, can compromise breathing. Recent advances in molecular biology have focused on ways to decrease scar formation through understanding of the wound repair process. Transforming growth factor beta (TFGbeta) plays a central role in this pathway. Ferrets serve as an ideal model for the pediatric airway, and reproduction of subglottic stenosis in ferrets is possible. However, ferret cytokine profiles have not been established. In this study, we characterized the presence and nucleotide sequence of the TGFbeta1 and 2 genes in ferrets by using total RNA isolated from airways. Amino acid sequence homology between human and ferret was determined to be 96.6% for TGFbeta1 and 99.3% for TGFbeta2. Given the nearly total homology between TGFbetas of ferret and human origin, the ferret may serve as an ideal model for future molecular studies.