How was genetic diversity transferred with translocations from ex situ to in situ? A case study of the European mink translocation to Hiiumaa Island in Estonia.

The European mink (Mustela lutreola) is one of the most threatened small carnivores, listed as Critically Endangered on the IUCN Red List. Tallinn Zoo started a conservation breeding operation in 1980, which in 1992 was shaped into the European mink EEP Program to maintain a demographically and genetically healthy population in captivity. Since 2000, mink have been translocated on a yearly basis from the breeding facility in the zoo to Hiiumaa Island (Estonia) until the formation of the wild island population in 2016. Maintaining a healthy genetic structure in a captive population was a priority, so genetically the least valuable animals, according to calculations made by a population management program, were used for translocation. This study aims to assess the amount of genetic diversity passed from an ex situ population on to the island population. Comparisons of the genetic diversity were made by mitochondrial, microsatellite and nuclear markers. In addition, our results were combined with the pedigree data from the European mink EEP Studbook to further evaluate the flow of genetic diversity from the founder population to the established wild population. According to the findings, the island population's allelic richness was comparable to that of the founder population, and no evidence was found that its genetic structure had diverged from that of the founder population. It seems that the formation of the island population has been a gradual process of no more than the last seven yearly translocations.

Reproductive parameters of critically endangered European mink (Mustela lutreola) in captivity.

Founding captive populations is often the last chance for saving endangered species from extinction. Ensuring successful reproduction is typically most critical for the maintenance of captive populations, with purposeful selection of individuals for breeding being one of the crucial aspects. Comparable cross-species data on the determinants of reproduction success are most useful for solving problems in captive species programs. In the present study, we provide an overview of a 20-year captive breeding program of the critically endangered European mink. The mating season starts in March, reaching its peak in the middle of April. The average gestation length was 43.8days (mode 43), the mean litter size being 4.4 (mode 4). Litter size and cub survival were negatively correlated with maternal age but this effect was entirely due to the lower performance of the females over 4 years of age. Female body weight also showed a positive correlation with litter size, with the weight itself having increased by 10% during the 20- year period. We did not find any signs of a cost of reproduction: the number of litters the female had delivered earlier in her life did not have an effect on her litter size in the focal year. Beyond the effect of age and size, individual females did not differ in litter sizes. Consistently, we found the heritability of litter size to be low. We conclude that, when selecting females for breeding, there is little need to consider aspects other than genetic relatedness crucial for avoiding progressive inbreeding.

The influence of spatiotemporal conditions and personality on survival in reintroductions-evolutionary implications.

Personality exists in non-human animals and can impact fitness. There is, however, a shortage of empirical studies in certain areas within the field, and fundamental evolutionary theory on personality remains largely untested. For example, little is known on how variation in personality is maintained over evolutionary time. Theory suggests that fluctuating selection pressures due to spatiotemporal variation in conditions, e.g. food availability, is a possible mechanism and a few studies have shown that the success of different personality types varies with spatiotemporal conditions. However, it remains unknown whether different mechanisms can maintain personality within a species. Here we use a reintroduction programme for the critically endangered European mink (Mustela lutreola) to test whether multiple personality trait domains (boldness, exploration and sociability) affected survival in two different years and islands. This was done through pre-release personality tests and post-release radio-tracking monitoring. Survival was positively correlated with boldness, whereas the relationship with exploration was either negative or positive depending on year/island. The results show a complex relationship between personality and survival and suggest that exploration can be maintained over evolutionary time via spatiotemporal variation in conditions. However, in contrast to exploration, boldness did not vary spatiotemporally and sociability had no impact on survival. This indicates that different personality trait domains might be maintained by different mechanisms. To date, personality has been studied primarily within behavioural sciences, but through empirical findings we highlight the importance of personality also in ecology and conservation biology.

Noninvasive monitoring of female reproductive hormone metabolites in the endangered European mink (Mustela lutreola).

This study examined the reproductive physiology of female European mink (Mustela lutreola) to augment the available information on estrus, ovulation, and pregnancy with the long-term goal of supporting ex situ breeding management of this highly endangered species. Fecal reproductive hormone metabolites were measured using EIAs for estrogen and 20-oxo-pregnane metabolites. Seasonal hormone profiles were established. A comparison of hormone fluctuations in pregnant and nonpregnant females showed that both estrogen and 20-oxo-pregnane metabolites were significantly elevated during gestation, which is 42 days in length. Delayed implantation or embryonic diapause does not occur in this species. Litter size was correlated with 20-oxo-pregnane levels but not with estrogen concentrations. During lactation, 20-oxo-pregnane metabolite levels remained higher than in nonpregnant females. The breeding season was characterized by peaks in vaginal cornified cells and fecal estrogen metabolite levels. Up to four peaks in estrogen levels were identified and confirmed that European mink are seasonally polyestrous. The results of 20-oxo-pregnane measurements indicated that hCG can be applied to induce ovulation. With the establishment of this noninvasive method, we present a new tool to support population management of this species.

Molecular epidemiology of Aleutian mink disease virus (AMDV) in Estonia, and a global phylogeny of AMDV.

Aleutian mink disease virus (AMDV) causes a severe disease called Aleutian disease (AD). AMDV infects primarily mustelids, but also other mammal species. Recent evidence suggests that AMDV may also affect humans. To examine AMDV in different wild animals and in farmed mink in Estonia, we collected 203 blood samples from eight mammal species in 2007-2010, of which 152 were from species living in the wild (American mink, European mink, pine marten, polecat, raccoon dog, badger, otter, and stone marten) and 51 were from farmed mink. AMDV was tested by PCR amplification of NS1 and VP2 gene fragments, and was only detected in 4 free-ranging (14.8%) and 11 farmed (21.6%) American mink. No other species was positive for AMDV. In addition, the VP2 gene fragment was sequenced for 14 farmed mink isolates from Finland for which NS1 sequences were already publicly available. None of the four Estonian AMDV isolates found in free-ranging mink had identical sequences with farmed mink. In fact, isolates from free-ranging and farmed mink belonged to different clades, suggesting that the analyzed virus isolates circulating in nature are not from escapees of current farms. Two global phylogenies were built: one based on NS1 (336 bp, 151 taxa from nine countries); the other based on a combined NS1-VP2 dataset (871 bp, 40 taxa from six countries). AMDV genotypes did not cluster according to their geographic origin, suggesting that transport of farm mink from multiple source farms has been intense. Nevertheless, one subclade in both phylogenies was comprised solely of isolates from farmed mink, while several subclades comprised isolates only from free-ranging mink, indicating that some isolates may circulate more in the wild and others among farm animals.

Situation and context impacts the expression of personality: the influence of breeding season and test context.

Non-human animal personality is defined as consistent behavioural differences across time and situations/contexts. Behaviours are, however, often plastic and to explain how plasticity and personality may coexist an adaptive framework has been developed. Still, there is little information on how personality is impacted by situations and contexts on an individual level. We investigated this in the European mink (Mustela lutreola) by performing a set of five experiments in two situations consisting of non-breeding and breeding season, and by using different test contexts. Three personality trait domains were identified; boldness, exploration and sociability. The levels of boldness and exploration changed between seasons but remained repeatable, which implies behavioural reaction norms and supports that the concept of personality remained applicable despite plasticity. Whilst males became bolder and more explorative in the breeding season females became shyer, which reflects European mink breeding behaviour. Furthermore, behaviours performed in mirror stimulus tests fell into different domains depending on whether, the test was conducted in the own territory or not, suggesting plasticity in the response towards conspecifics. To conclude, our results highlight the importance of situation and context for the expression of personality, and the significance of measuring multiple personality trait domains with several methods.

The causes of the low breeding success of European mink (Mustela lutreola) in captivity.

High among-individual variation in mating success often causes problems in conservation breeding programs. This is also the case for critically endangered European mink and may jeopardize the long-term maintenance of the species' genetic diversity under the European mink EEP Program. In this study, breeding success of wild and captive born European minks at Tallinn Zoological Garden are compared, and the mating behavior of the males is analyzed. Results show that wild born males successfully mate significantly more often than captive born males (89% and 35%, respectively). On the basis of an extensive record of mating attempts, both male aggressiveness and passivity are identified as primary causes of the observed mating failures. All other potential determinants have only a minor role. Mating success as well as a male's aggressiveness and passivity are shown to depend more strongly on the male than the female partner. We did not find any evidence that the behavior of an individual is dependent on the identity of its partner. We suggest that aggressiveness and passivity are two expressions of abnormal behavior brought about by growing up in captivity: the same individuals are likely to display both aggressive and passive behavior. The results point to the need to study and modify maintenance conditions and management procedures of mink to reduce the negative impact of the captive environment on the long-term goals of the program.