Microsatellite markers for the endangered European mink (Mustela lutreola) and closely related mustelids

The European mink (Mustela lutreola L., 1761) is an endangered carnivore species whose populations suffered a severe decline during the last century. The genotyping of eight polymorphic microsatellite loci revealed a relatively low number of alleles per locus (two to eight), as well as low levels of polymorphism (observed and expected heterozygosity values per locus were 0.49 and 0.54, respectively). Cross‐specific polymerase chain reaction amplifications were successful in seven closely related mustelid species suggesting that these loci may be useful not only for assessing genetic variability in European mink populations but also for determining potential hybridization events between M. lutreola and other mustelid species.     

Conservation genetics and population history of the threatened European mink Mustela lutreola, with an emphasis on the west European population

In species of great conservation concern, special attention must be paid to their phylogeography, in particular the origin of animals for captive breeding and reintroduction. The endangered European mink lives now in at least three well-separated populations in northeast, southeast and west Europe. Our aim is to assess the genetic structure of these populations to identify 'distinct population segments' (DPS) and advise captive breeding programmes. First, the mtDNA control region was completely sequenced in 176 minks and 10 polecats. The analysis revealed that the western population is characterized by a single mtDNA haplotype that is closely related to those in eastern regions but nevertheless, not found there to date. The northeast European animals are much more variable (pi = 0.012, h = 0.939), with the southeast samples intermediate (pi = 0.0012, h = 0.469). Second, 155 European mink were genotyped using six microsatellites. The latter display the same trends of genetic diversity among regions as mtDNA [gene diversity and allelic richness highest in northeast Europe (H(E) = 0.539, R(S) = 3.76), lowest in west Europe (H(E) = 0.379, R(S) = 2.12)], and provide evidences that the southeast and possibly the west populations have undergone a recent bottleneck. Our results indicate that the western population derives from a few animals which recently colonized this region, possibly after a human introduction. Microsatellite data also reveal that isolation by distance occurs in the western population, causing some inbreeding because related individuals mate. As genetic data indicate that the three populations have not undergone independent evolutionary histories for long (no phylogeographical structure), they should not be considered as distinct DPS. In conclusion, the captive breeding programme should use animals from different parts of the species' present distribution area.     

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.     

The first linkage map of the American mink (Mustela vison)

Described herein, the first microsatellite linkage map for the American mink consists of 85 microsatellite markers resolved into 17 linkage groups. The map was constructed using 92 F(1) progeny from five sire families created by crossing mink with different colour types. The linkage groups ranged from 0 to 137 cM. These linkage groups were assigned to 12 of the 14 mink autosomes using a somatic cell hybrid panel. The total map covered 690 sex-averaged Kosambi units with an average marker spacing of 8 cM. This map will facilitate further genetic mapping of monogenic characters and QTL.     

Genetic neighbourhood and effective population size in the endangered European mink <Emphasis Type="Italic">Mustela lutreola</Emphasis>

Genetic neighbourhood and effective population size (N _e) are critical factors when determining the potential survival of threatened species. Carnivores have intrinsically small effective numbers, because, as top predators, they show low densities. The European mink, Mustela lutreola, is one of the most endangered carnivores in the world and has suffered continual decline and local extinctions. The genetic neighbourhood, area within which adults could randomly mate, averaged N _a = 31.7 km diameter, allowing that population size within the neighbourhood area only ranged from N _b = 6.1 to 22.8 animals. Although the population size was assessed in one of the main mink populations in the world, this neighbourhood size is far below the values regarded as critical in literature. However, in contrast with recent propositions, the ratio N _e /N only ranged between 0.09 and 0.19, estimates close to the average recognised by Frankham [(1995) Genetic Research 66: 95–107] for wildlife populations. In the context of the challenge to conserve this endangered carnivore, the studied neighbourhood provided crucial information suggesting both a low neighbourhood size and severe disturbance of breeding exchanges, emphasising the dramatically threatened status of the European mink.     

Immunocytological analysis of meiotic recombination in the American mink (Mustela vison)

Using immunolocalization of MLH1, a mismatch repair protein that marks crossover sites along synaptonemal complexes, we estimated the total length of the genetic map, the recombination rate and crossover distribution in the American mink ( Mustela vison ). We prepared spreads from 130 spermatocytes of five male minks and mapped 3320 MLH1 foci along 1820 bivalents. The total recombination length of the male mink genome, based on the mean number of MLH1 foci for all chromosomes, was 1327 cM. The overall recombination rate was estimated to be 0.48 cM/Mb. In all bivalents, we observed prominent peaks of MLH1 foci near the distal ends and a paucity of them near the centromeres. This indicates that genes located at proximal regions of the chromosomes should display much tighter genetic linkage than physically equidistant markers located near the telomeres.     

Adaptations to fasting in the American mink (<Emphasis Type="Italic">Mustela vison</Emphasis>): nitrogen metabolism

The aim of this study was to investigate the adaptations of protein metabolism to seasonal fasting in an actively wintering boreal carnivore. Fifty farm-bred male American minks Mustela vison were divided into a fed control group and four experimental groups fasted for 2, 3, 5 or 7 days. The responses of nitrogen metabolism to wintertime food deprivation were determined by measuring the rate of weight loss, the tissue total protein concentrations and the plasma amino acid, urea, ammonia, uric acid and total protein levels. The mink has relatively poor adaptations to food deprivation, as it is not able to prolong phase II of fasting with fat as the major metabolic fuel. Instead, the species has to derive a part of its energy requirements from the breakdown of body proteins. The end product of protein catabolism—urea— accumulates in its circulation, and the mink may not be able to recycle urea-N. Although the mink can still have a high body fat percent at the end of the 7-day fast, it appears to enter phase III of fasting with stimulated proteolysis during this period.     

Gonadotropin-releasing hormone neurons and pathways in the brain of the female mink (Mustela vison)

Summary The distribution of gonadotropin-releasing hormone-immunoreactive neurons and processes was mapped in the female mink brain using coronal, horizontal and sagittal sections. Perikarya were found along a ventral continuum including the olfactory tubercle, the diagonal band of Broca, the lateral septum, the preoptic and anterior hypothalamic area and the mediobasal hypothalamus; 80% of the perikarya were counted in the mediobasal hypothalamus. Fibres were mainly observed in the organum vasculosum of the lamina terminalis and the median eminence. A few processes terminated in the ependymal cells lining the third and lateral ventricles. The total number of immunoreactive perikarya was the highest in the brains of females sacrificed in July; it then significantly decreased until December. This variation is discussed in relation to the annual breeding cycle.     

基于高通量测序技术研究水貂远端肠道细菌群落组成

【目的】研究水貂远端肠道微生物群落组成及其多样性。【方法】通过高通量测序技术研究水貂远端肠内容物细菌的组成和多样性。【结果】从10只健康水貂远端肠道内容物样品中得到146 287高质量序列代表17个菌门、167个细菌属。水貂肠道细菌以厚壁菌门(59.99%)、拟杆菌门(16.2%)、梭杆菌门(11.5%)、放线菌门(5.9%)和变形菌门(5.3%)为主,其中厚壁菌门最为丰富。厚壁菌门中的梭菌目是最丰富的目,而梭菌目中的链球菌占有50%以上的OTUs,是最大的细菌属。【结论】水貂肠道内存在复杂的微生物区系,这对进一步研究水貂对营养物质吸收利用提供了理论基础。     

Space use and habitat preferences of the invasive American mink (<Emphasis Type="Italic">Mustela vison</Emphasis>) in a Mediterranean area

Space use, intra-territorial habitat preferences, and factors affecting both were studied in an invading population of American mink, Mustela vison, in two rivers of a Mediterranean region of Spain. Average linear home range was 1.19 ± 0.73 km (±SD) and core area was 0.21 ± 0.08 km for resident males (n = 10); while for females (n = 5) they were 0.54 ± 0.14 and 0.19 ± 0.11 km, respectively. Overlapping between the home ranges of residents was low. In no case their core areas overlapped. Home ranges were small in comparison to other study areas and in general the resident minks were territorial. Linear home range length was related to individual weight and to the river. Weight had a positive effect indicating a potential body condition effect, while river may be showing a habitat quality effect. Habitat preferences were positively affected by the abundance of helophytic vegetation and negatively by the presence of human activity. Helophytic vegetation offers both food and refuges, while human activity may represent a potential danger. Percentage of captures was higher inside the core areas and was slightly influenced positively by abundance of helophytic vegetation. All this information should be considered when designing and implementing measures to control the expansion of American minks. We recommend keeping going with the trapping sessions but, given the results obtained, reducing the distance between traps down to 200 m to maximize capturability (i.e., about doubling the trapping effort), and, when available, placing them near helophytic vegetation. In the absence of helophytic vegetation, traps should be located near any kind of vegetation providing coverage for mink and far from human activity.     

Experimental evaluation of the reliability of placental scar counts in American mink (Mustela vison)

Counting of placental scars in the uterus is a valuable method to estimate female fecundity in wild-ranging mammalian populations, but the reliability of placental scar counts (PSC) has only been thoroughly assessed for a few species. Here we evaluate the reliability of PSC in American mink (Mustela vison) through examination of farm animals with known reproductive history. PSC estimated pregnancy rate and litter size accurately in individuals up to 3 months postpartum. Placental scars were detectable in most female American mink sampled 7–8 months postpartum, but the proportion of minks misclassified as barren was overestimated, and mean litter size was underestimated. We found a large variation in the intensity of pigmentation in placental scars with similar age. Differentiation of scars according to shading as indicating either successful full-term pregnancies or post-implantation loss of embryos was not possible. We propose that these results are applicable for fecundity analysis in American mink and closely related mustelids.