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Context: New animal welfare legislation and ethical guidelines encourage alternative approaches for canine contraception, instead of surgical gonadectomy which is considered invasive and unjustified in healthy dogs. Aims: Reversible contraception might be achieved by inhibition of aromatase (CYP19), an enzyme catalysing the conversion of androgens to oestrogens. This study provides insights into the spatio-temporal expression and distribution of aromatase in canine ovarian tissue. Methods: Ovarian tissue was collected from 39 healthy and sexually mature bitches during different stages of the oestrous cycle: pro-oestrus (n = 8), oestrus (n = 12), dioestrus (n = 9) (luteal phase) and anoestrus (n = 10). Localisation of cytochrome P450 aromatase was determined by immunohistochemistry. Key results: Aromatase activity in the dog is high during pro-oestrus, ovulation and early dioestrus. Comparing types of follicles and corpora lutea, the highest aromatase abundance was found in antral follicles and luteinising follicles, whereas corpora lutea and early antral follicles showed an intermediate presence of the enzyme. Interesting was the high abundance of aromatase in luteinising theca interna cells, prevailing over granulosa cells. Conclusions and implications: Understanding of cells involved in oestradiol production is important for targeted inhibition of oestradiol synthesis, possibly offering an approach for contraception and suppression of oestrus. In clinical practice, contraception in female dogs is traditionally achieved by surgical gonadectomy. As this method is discouraged by new animal welfare legislation in some parts of the world and is also related to several side effects, alternative possibilities for contraception are needed. Here, we report the distribution of aromatase in the canine ovary during different stages of the oestrous cycle. In-depth knowledge of oestrogen synthesis in the female dog is mandatory for targeted inhibition of oestradiol, possibly by the use of aromatase inhibitors. Photograph by M. P. Kowalewski. [ABSTRACT FROM AUTHOR]

Summary: Indigenous cattle breeds in northern Eurasia have adapted to harsh climate conditions. The local breeds are important genetic resources with cultural and historical heritages, and therefore, their preservation and genetic characterization are important. In this study, we profiled the whole‐blood transcriptome of two native breeds (Northern Finncattle and Yakutian cattle) and one commercial breed (Holstein) using high‐throughput RNA sequencing. More than 15 000 genes were identified, of which two, 89 and 162 genes were significantly upregulated exclusively in Northern Finncattle, Yakutian cattle and Holstein cattle respectively. The functional classification of these significantly differentially expressed genes identified several biological processes and pathways related to signalling mechanisms, cell differentiation and host–pathogen interactions that, in general, point towards immunity and disease resistance mechanisms. The gene expression pattern observed in Northern Finncattle was more similar to that of Yakutian cattle, despite sharing similar living conditions with the Holstein cattle included in our study. In conclusion, our study identified unique biological processes in these breeds that may have helped them to adapt and survive in northern and sub‐arctic environments. [ABSTRACT FROM AUTHOR]

Contents Carnivora is one of the most species-rich order of Mammalia. Some species, e.g. domestic cats, dogs and ferrets, are among the most popular pets; others, such as minks and farmed foxes, have economic value for the fur breeding industry. Still others, such as tigers, bears and other top predators, have great impact on the health of natural ecosystems. Most if not all Carnivora species have great cultural and aesthetic importance for man. There are enormous differences between mammalian species in reproductive physiology, and it is not surprising that reproductive technologies can be used with high efficiency with some animal groups, e.g. most farm animals and laboratory rodents, but are very laborious when used with Carnivora species, which often possess unique reproductive traits. The efficiency of assisted reproductive technology (ART) applied to semi-domestic, non-domestic and especially to endangered species of Carnivora remains extremely low in most cases, and often the first positive result reported is the only instance when ART has been successful with that species. Although there are approximately 270 species in the Carnivora order, to the best of our knowledge, successful published attempts to apply ART have been reported for only four families: Mustelidae, Felidae, Canidae and Ursidae. The main achievements in ART, embryo technology in particular, for these families of Carnivora, together with challenges and problems, are reviewed in the relevant sections. [ABSTRACT FROM AUTHOR]

Contents Despite efforts undertaken to conserve the endangered European mink, its reproduction is still poorly studied. The aim was to study its reproductive cyclicity, faecal progesterone concentration and ovarian changes during early pregnancy, with the emphasis on the pre-implantation period and implantation. During the 2004 breeding season, oestrous cycle was monitored in 39 females as well as ovarian changes during early pregnancy in 22 females. During the 2007 breeding season, faecal progesterone concentration measured by radioimmunoassay was monitored during pregnancy in 10 females throughout their pregnancy. The breeding season 2004 started on March 18 and ended on May 10, with the peak recorded in April. The duration of first oestrus was 1–12 days. If not mated, the vast majority of females entered second oestrus after 12–55 days. In general, relatively low faecal progesterone values were detected in European mink; an average of 42.69 ± 4.70 ng/g faeces in oestrous females with a maximum of 176.44 ± 23.01 ng/g faeces on pregnancy day 12.anova indicated a significant effect of the pregnancy stage. Post hoc comparisons with Fisher least significant difference (LSD) test revealed that faecal progesterone concentrations on days 8 and 12 post coitum (p.c.), but not at the end of pregnancy (day 40), were higher when compared with the initial oestrous level. Implantation in this species occurs on day 12 p.c. and was indicated by prominent uterine swellings and failure to flush the uterine horns beyond this day. Advanced luteogenesis was observed with prominent corpora lutea found in ovaries around the time of implantation. To conclude, European mink is a seasonally polyoestrous species; the early pregnancy of European mink resembles that of European polecat, i.e. in both species, implantation occurs on day 12 p.c. without any implantation delay. [ABSTRACT FROM AUTHOR]

Contents The European mink ( Mustela lutreola) is a small mammal, which belongs to the Mustelidae family ( Carnivora). Earlier, the range of distribution of this species encompassed much of the European continent. During the 20th century, the numbers of European mink declined and the range of its distribution became reduced to three fragmented populations; today this species faces extinction. The urgent necessity for effective conservation efforts to protect the European mink is accepted by the governmental organizations as well as scientific communities of most European countries. In this paper, the reasons for the disappearance of European mink are reviewed and results of past conservation efforts based on captive breeding and reintroduction programmes are critically evaluated in the broad context of modern concepts of conservation genetics and reproductive biology. The data recently obtained on the reproduction and pre-implantation development of European mink and the prospects of incorporation of modern reproductive technologies into the conservation programme of this species are discussed. [ABSTRACT FROM AUTHOR]

Contents The domestic ferret is a seasonally polyoestrous species. Females reach puberty at the age of 8–12 months. Females exhibit a constant oestrus between late March and early August if they are not bred. Increasing tumescence in the pink-coloured vulva is a sign of pro-oestrus. Oestrus can persist for up to 5 months, but once ovulation is induced, either pregnancy or pseudopregnancy ensues. Follicular development and atresia overlap in such a manner that there is a recent cohort of follicles available for ovulation whenever copulation might occur. Copulation may last from 15 min to 3 h, the average being 1 h. Ovulation is induced by pressure on the cervix associated with copulation. After sufficient LH release, the pre-ovulatory follicles mature and an average of 12 oocytes (5–13) per female are ovulated 30–40 h after copulation into the ovarian bursa. The ferret oocytes are most capable of being fertilized up to 12 h after ovulation, i.e. 42–52 h after copulation. Ferret oocytes ovulate at the metaphase of the second meiotic division (MII) embedded in three layers of corona radiata cells. Embryos enter the uterus over a period of several days starting on day 5 after mating. Between days 12 and 13 after mating, the embryos have become implanted in the endometrium. Implantation in the ferret is central with rapid invasion of the uterine epithelium by the trophoblast over a broad area that eventually becomes a zonary band of endotheliochorial placenta. Gestation length is 41 days (39–42 days). The domestic ferret gives birth to an average of eight kits (1–18 kits), which weigh 6–12 g at birth. [ABSTRACT FROM AUTHOR]

Interest in the use of assisted reproductive technology in reindeer husbandry has gradually increased during the last decades. This article reviews Western and Russian literature on reindeer semen collection, semen cryopreservation and artificial insemination. In addition, literature on the synchronisation of the stage of reindeer oestrous cycle among females, recovery of in vivo embryos, embryo transfer, the production of in vitro -produced embryos and pregnancy diagnosis is reviewed. • Several of the reindeer subspecies are regarded as vulnerable or near threatened. • Genome resource banking would be possible for reviving endangered deer species. • Five reindeer offspring have been produced by AI of frozen-thawed spermatozoa. • One reindeer offspring has been produced by transfer of fresh in vivo -produced embryos. • The first reindeer IVP blastocysts have been produced using the bovine IVP protocol. [ABSTRACT FROM AUTHOR]

The effect of addition of glycine betaine to a lactose-EDTA freezing medium on the post-thaw motility of stallion semen was determined. The first three semen-rich fractions of nine stallions were collected with an open-end Krakow artificial vagina on consecutive weekdays. Semen was frozen using the Hannover method with freezing media containing glycine betaine in various concentrations from 0 to 5 %. After thawing, sperm motility was analysed both by a light microscope and by a Hamilton-Thorn Motility Analyser. Total and progressive post-thaw motilities of semen containing 0.25–3 % glycine betaine did not differ significantly from the total and progressive post-thaw motilities of semen frozen without glycine betaine. The total and progressive post-thaw motilities of semen containing 4 or 5 % glycine betaine were significantly lower (P < 0.001) than those of semen without glycine betaine. In conclusion, glycine betaine did not show any beneficial effect on the post-thaw motility of stallion semen when semen was frozen using the Hannover method. [ABSTRACT FROM AUTHOR]

126 hasal ccli carcinomas from 7 patients with nevoid basal cell carcinoma syndrome were examined histologically and - using a scoring system - compared with 54 solitary basal cell carcinomas. The solid subtype formed 78.9% of all tumors in the head and neck region and 48.9% of those located elsewhere. A broader spectrum of histological subtypes was noticed in tumors from patients with the syndrome. As multiple, keratinizing odontogenic and epidermal cysts are often seen in the nevoid basal cell carcinoma syndrome it was interesting to see that multiple keratinizing cysts within skin tumors were recorded more often in this group. [ABSTRACT FROM AUTHOR]

In a first experiment, 11 Finnhorse mares were examined every six hours during late oestrus by rectal palpation and ultrasonography to determine the time of ovulation. The mares were inseminated over one to three subsequent cycles, 6-12 (n=5), 12-18 (n=5), 18-24 (n=5) and 24-30 (n=5) hours after ovulation. Pregnancies were terminated by prostaglandin injection 21 days after insemination. All mares inseminated within 18 hours of ovulation conceived but no mare inseminated 24 hours or more after ovulation conceived. In a second experiment, 14 mares were examined every day at about the same time and inseminated after ovulation had been detected, within 0-27 hours of ovulation. Pregnancies were terminated by prostaglandin injection 25 days after insemination and the animals inseminated again. Forty-six per cent of the mares (13 out of 28) were pregnant 16 days after insemination but there was early embryonic death in five mares, i. e. only 29% of the mares became and remained pregnant. The equine ovum seems to be capable of being fertilized for at least 18 hours after ovulation. However, the high incidence of early embryonic death in the second experiment suggests that the viability of the embryo may have decreased. [ABSTRACT FROM AUTHOR]

Follicular growth and hormone determinations were used to predict ovulation in the mare. Thirty Finnhorse mares were used for the investigation and 38 oestrous cycles were studied. The mares were examined by rectal palpation and ultrasonography every 6 hours during late oestrus. Daily blood samples were obtained for progesterone and oestrone sulphate determination. The preovulatory follicle grew 3 mm a day up to 2 days before ovulation. The size then remained constant, before diminishing by 2-3 mm during the last 12 hours. The maximal diameter of the follicle was 43 ± 4 mm. In 89 % of the follicles the round shape became more irregular before ovulation. During the last 24 hours before ovulation 37 of 38 follicles were regarded as mature on rectal palpation. The oestrone sulphate level was highest 24-48 hours before ovulation, the first decrease being observed most commonly around ovulation (± 1 day). The size of the follicle was the most reliable criterion in the prediction of ovulation. Zusammenfassung Vorhersage der Ovulation bei der Stute aufgrund des Follikelwachstums, der östronsulfat- sowie der Progesteronkonzentration des Serums Zur Vorhersage der Ovulation wurden bei der Stute das Follikelwachstum sowie Hormonbestimmungen herangezogen. Für die Untersuchungen wurden 30 Stuten verwendet. Insgesamt wurden 38 Brunstzyklen untersucht. Die Stuten wurden während der späten Phase des östrus alle 6 Stunden mittels rektaler Palpation und Ultraschall untersucht. Für die Progesteron- und östronsulfatbestim-mungen wurde täglich Blutproben entnommen. Der präovulatorische Follikel wuchs bis 2 Tage vor der Ovulation 3 mm pro Tag. Die Größe blieb dann konstant, bevor sich der Follikel in den letzten 12 Stunden um 2-3 mm verkleinerte. Der maximale Follikeldurchmesser betrug 43 ± 4 mm. Bei 89% der Follikel wurde die runde Form vor der Ovulation unregelmäßig. Bei rektaler Palpation wurden während der letzten 24 Stunden vor der Ovulation 37 von 38 Follikeln als reif beurteilt. Der östronsulfatspiegel war 24-48 Stunden vor der Ovulation am höchsten; die erste Abnahme wurde gewöhnlich um die Ovulation herum (± 1 Tag) beobachtet. Die Follikelgröße war für die Vorhersage der Ovulation das verläßlichste Kriterium. [ABSTRACT FROM AUTHOR]

• Reindeer exhibit a short intense rutting season. • Males have an annual cycle of sexual maturation and involution, similar to other cervids. • A deeper understanding of the male reproductive physiology is crucial for developing better strategies in reindeer husbandry. The present review provides a comprehensive assessment on the basic aspects of the reproduction of the Rangifer male with a special focus on pubertal development, and the seasonal dynamics of gametogenesis, androgenesis, antler cycle and other physiological and behavioural characteristics of the rutting male. A greater understanding of the reproductive physiology of the males is needed to improve existing management strategies of semi-domestic stocks in the changing climate and to develop species-specific assisted reproductive technologies for improving the in situ and ex situ conservation programs to preserve endangered subspecies as well as to facilitate the transfer of genetic resources between fragmented wild or semi-domestic populations. [ABSTRACT FROM AUTHOR]

The drastic change in global climate has led to in-depth studies of the geneticresources of native cattle adapted to challenging environments. Native cattle breeds may harbor unique genetic mechanisms that have enabled them adapt to their given environmental conditions. Adipose tissues are key factors in the regulation of metabolism and energy balance and are crucial for the molecular switches needed to adapt to rapid environmental and nutritional changes. The transcriptome landscape of four adipose tissues was used in this study to investigate the differential gene expression profiles in three local breeds, Yakutian cattle (Sakha Republic), Northern Finncattle (Finland), Mirandesa cattle (Portugal) and commercial Holstein cattle. A total of 26 animals (12 cows, 14 bulls) yielded 81 samples of perirenal adipose tissue (n = 26), metacarpal adipose tissue (n = 26), tailhead adipose tissue (n = 26) and prescapular adipose tissue (n = 3). More than 17,000 genes were expressed in our dataset. Principal component analysis of the normalized expression profiles revealed a differential expression profile of the metacarpal adipose tissue. We found that the genes upregulated in the metacarpal adipose tissue of Yakutian cattle, such as NR4A3, TEKT3, and FGGY, were associated with energy metabolism and response to cold temperatures. In Mirandesa cattle, the upregulated genes in perirenal adipose tissue were related to immune response and inflammation (AVPR2, CCN1, and IL6), while in Northern Finncattle, the upregulated genes appeared to be involved in various physiological processes, including energy metabolism (IGFBP2). According to the sex-based comparisons, the most interesting result was the upregulation of the TPRG1 gene in three tissues of Yakutian cattle females, suggesting that adaptation is related to feed efficiency. The highest number of differentially expressed genes was found between Yakutian cattle and Holstein, several of which were associated with immunity in Yakutian cattle, indicating potential differences in disease resistance and immunity between the two breeds. This study highlights the vast difference in gene expression profiles in adipose tissues among breeds from different climatic environments, most likely highlighting selective pressure and the potential significance of the uniquely important regulatory functions of metacarpal adipose tissue., (© 2024. The Author(s).)

Domestic reindeer (Rangifer tarandus) play a vital role in the culture and livelihoods of indigenous people across northern Eurasia. These animals are well adapted to harsh environmental conditions, such as extreme cold, limited feed availability and long migration distances. Therefore, understanding the genomics of reindeer is crucial for improving their management, conservation and utilisation. In this study, we have generated a new genome assembly for the Fennoscandian domestic reindeer with high contiguity, making it the most complete reference genome for reindeer to date. The new genome assembly was utilised to explore genetic diversity, population structure and selective sweeps in Eurasian Rangifer tarandus populations which was based on the largest population genomic dataset for reindeer, encompassing 58 individuals from diverse populations. Phylogenetic analyses revealed distinct genetic clusters, with the Finnish wild forest reindeer (Rangifer tarandus fennicus) standing out as a unique subspecies. Divergence time estimates suggested a separation of ~ 52 thousand years ago (Kya) between the northern European Rangifer tarandus fennicus and Rangifer tarandus tarandus. Our study identified four main genetic clusters: Fennoscandian, the eastern/northern Russian and Alaskan group, the Finnish forest reindeer, and the Svalbard reindeer. Furthermore, two independent reindeer domestication processes were inferred, suggesting separate origins for the domestic Fennoscandian and eastern/northern Russian reindeer. Notably, shared genes under selection, including retroviral genes, point towards molecular domestication processes that aided adaptation of this species to diverse environments., (© 2023. The Author(s).)

Mycoplasma bovis (M. bovis) causes several costly diseases in cattle and has a negative effect on cattle welfare. There is no effective commercial vaccine, and antimicrobial resistance is common. Maintaining a closed herd is the best method to minimize the risk of introduction of M. bovis. Assisted reproduction is crucial in a closed herd to make genetic improvements. M. bovis has been found in commercial semen, and contaminated semen has been the source of disease in naïve dairy herds. The objective of this study was to evaluate M. bovis transmission in bovine in vitro embryo production (IVP) using several possible exposure routes. We used a wild-type M. bovis strain isolated from semen at a final concentration of 10 6  CFU/mL to infect cumulus-oocyte complexes, spermatozoa, and 5-day-old embryos. We also used naturally contaminated semen in fertilization. Blastocysts were collected on day 7-8 and zona pellucida (ZP)-intact embryos were either washed 12 times, including trypsin washes as recommended by the International Embryo Technology Society (IETS), or left unwashed. Washed and unwashed embryos, follicular fluids, maturation medium, cumulus cells, fertilization medium, and G1 and G2 culture media, as well as all wash media were analyzed using enrichment culture followed by real-time PCR detection of M. bovis. Altogether, 76 pools containing 363 unwashed embryos and 52 pools containing 261 IETS washed embryos were analyzed after oocytes, spermatozoa, or 5-day-old embryos were infected with M. bovis or naturally contaminated semen was used in fertilization. We could not detect M. bovis in any of the embryo pools. M. bovis was not found in any of 12 wash media from different exposure experiments. M. bovis did not affect the blastocyst rate, except when using experimentally infected semen. Contrary to an earlier study, which used a cell co-culture system, we could not demonstrate M. bovis in embryo wash media or tight adherence of M. bovis to ZP-intact embryos. Naturally infected semen did not transmit M. bovis to embryos. We conclude that by using our IVP system, the risk of M. bovis transmission via IVP embryos to recipient cows is very low., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Context: An accurate staging of sexual cycle is essential for the optimum timing of medical interventions., Aims: Here, an updated insight into clinical, endocrinological and vagino-cytological parameters, and their correlation with histomorphology of ovarian and uterine tissue samples is presented., Methods: Samples from 39 dogs were collected at various stages of the oestrous cycle: pro-oestrus (n =8), oestrus (n =12), dioestrus (n =9) (luteal phase) and anoestrus (n =10), according to clinical observations. Final allocation of samples was done after histomorphological evaluation of all tissues. Peripheral oestradiol-17ß (E2) and progesterone (P4) concentrations were measured, P4 by both chemiluminescent immunoassay (CLIA) and radioimmunoassay (RIA)., Key Results: Differences were observed between determination of the stage of the oestrous cycle, either by clinical, endocrinological or histomorphological evaluation. Individuals considered to be in clinical and endocrinological oestrus, had entered the luteal phase according to histomorphology. P4 concentrations measured by two different assays differed, underlying the importance to understand that absolute P4 concentrations may deviate depending on the used assay. Comparison of E2 and P4 concentrations is suggested to be useful when defining the transition from early follicular phase to the time of ovulation., Conclusions and Implications: Based on parallel histomorphological observations, combined with clinical and endocrinological findings on the same individuals, the present study emphasises that an accurate classification of the stage of the cycle in female dogs based solely on clinical and endocrinological assessments can be difficult. The histomorphological findings presented herein provide new insights into the transitional phases between the different stages of the oestrous cycle in the dog.

Para-aminopropiophenone (PAPP) is a potent methaemoglobin (MetHb) forming agent used for the lethal control of exotic carnivores and mustelids. To assess the sensitivity of Australian wildlife to PAPP we developed an in vivo assay that did not use death as an endpoint. Sub-lethal dose-response data were modelled to predict PAPP doses required to achieve an endpoint set at 80% MetHb (MetHb 80 ). The comparative sensitivity of non-target mammals referenced to this endpoint was found to be highly variable, with southern brown bandicoots ( Isoodon obesulus ) the most sensitive species (MetHb 80 = 6.3 mg kg -1 ) and bush rats ( Rattus fuscipes ) the most tolerant (MetHb 80 = 1035 mg kg -1 ). Published LD 50 estimates were highly correlated with PAPP doses modelled to achieve the MetHb 80 endpoint (r 2 = 0.99, p < 0.001). Most dose-response data for native mammals were collected in the field or in semi-natural enclosures, permitting PAPP and placebo dosed animals to be fitted with tracking transmitters and transponders and released at their point of capture. A protracted morbidity and mortality was observed only in Australian ravens ( Corvus coronoides ). The combination of sub-lethal dose-response assay and survival data collected in the field provided more relevant information about the actual hazard of pest control agents to non-target wildlife species than laboratory-based lethal-dose bioassays. We discuss the need to replace lethal-dose data with biologically meaningful insights able to define a continuum of toxicological hazards that better serve the needs of conservation and veterinary scientists and wildlife managers.

Reindeer (Rangifer tarandus) are semi-domesticated animals adapted to the challenging conditions of northern Eurasia. Adipose tissues play a crucial role in northern animals by altering gene expression in their tissues to regulate energy homoeostasis and thermogenic activity. Here, we perform transcriptome profiling by RNA sequencing of adipose tissues from three different anatomical depots: metacarpal (bone marrow), perirenal, and prescapular fat in Finnish and Even reindeer (in Sakha) during spring and winter. A total of 16,212 genes are expressed in our data. Gene expression profiles in metacarpal tissue are distinct from perirenal and prescapular adipose tissues. Notably, metacarpal adipose tissue appears to have a significant role in the regulation of the energy metabolism of reindeer in spring when their nutritional condition is poor after winter. During spring, genes associated with the immune system are upregulated in the perirenal and prescapular adipose tissue. Blood and tissue parameters reflecting general physiological and metabolic status show less seasonal variation in Even reindeer than in Finnish reindeer. This study identifies candidate genes potentially involved in immune response, fat deposition, and energy metabolism and provides new information on the mechanisms by which reindeer adapt to harsh arctic conditions., (© 2021. The Author(s).)

Uterine secretory proteins protect the uterus and conceptuses against infection, facilitate implantation, control cellular damage resulting from implantation, and supply pre-implantation embryos with nutrients. Unlike in humans, the early conceptus of the European polecat ( Mustela putorius ; ferret) grows and develops free in the uterus until implanting at about 12 days after mating. We found that the proteins appearing in polecat uteri changed dramatically with time leading to implantation. Several of these proteins have also been found in pregnant uteri of other eutherian mammals. However, we found a combination of two increasingly abundant proteins that have not been recorded before in pre-placentation uteri. First, the broad-spectrum proteinase inhibitor α 2 -macroglobulin rose to dominate the protein profile by the time of implantation. Its functions may be to limit damage caused by the release of proteinases during implantation or infection, and to control other processes around sites of implantation. Second, lipocalin-1 (also known as tear lipocalin) also increased substantially in concentration. This protein has not previously been recorded as a uterine secretion in pregnancy in any species. If polecat lipocalin-1 has similar biological properties to that of humans, then it may have a combined function in antimicrobial protection and transporting or scavenging lipids. The changes in the uterine secretory protein repertoire of European polecats is therefore unusual, and may be representative of pre-placentation supportive uterine secretions in mustelids (otters, weasels, badgers, mink, wolverines) in general.

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., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Chondrogenic differentiation of human embryonic (hESCs) or induced pluripotent stem cells (hiPSCs) has been achieved in embryoid bodies (EBs) by adding selected growth factors to the medium. Also chondrocyte-secreted factors have been considered to promote the chondrogenic differentiation. Hence, we studied whether co-culture with primary chondrocytes can induce hESCs or hiPSCs to differentiate into chondrocyte lineage. Co-culture of hESCs or hiPSCs was established in a transwell insert system in feeder-free culture conditions, while hESCs or hiPSCs grown alone in the wells were used as controls. After 3-week co-culture with weekly replenished chondrocytes, the chondrogenically committed cells (hCCCs) were evaluated by morphology, immunocytochemistry, quantitative real-time RT-PCR, and analysis of chondrogenic, osteogenic and adipogenic differentiation markers. The expressions of chondrocyte- and pluripotency-associated genes were frequently measured during the monolayer expansion of hCCCs from passage 1 to 10. Human CCCs displayed morphology similar to chondrocytes, and expressed chondrocyte-associated genes, which were declined following passaging, similarly to passaged chondrocytes. They also formed a chondrogenic cell pellet, and differentiated into chondrocytic cells, which secreted abundant extracellular matrix. Human CCCs also proliferated rapidly. However, they did not show osteogenic or adipogenic differentiation capacity. Our results show that co-culture of hESCs or hiPSCs with primary chondrocytes could induce specific chondrogenic differentiation., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

We have investigated whether 5% oxygen tension (O(2)) is beneficial for neocartilage formation when chondrocytes are cultured in transwell-COL inserts. Six million bovine primary chondrocytes were cultured in an insert with DMEM supplemented with 10% fetal bovine serum and antibiotics, with or without glucosamine sulphate (GS) in a 5% or 20% O(2) environment for 2, 4, or 6 weeks. The samples were collected for the histological staining of proteoglycans (PGs) and type II collagen, quantitative reverse transcription with the polymerase chain reaction (RT-PCR) analyses of the mRNA expression of aggrecan and procollagen α(1)(II), procollagen α(2)(I) and hyaluronan synthase 2, quantitation of PGs, and agarose gel electrophoresis. Neocartilage produced at 20% O(2) appeared larger than that at 5% O(2). Histological staining showed that more PGs and type II collagen and better native cartilage structure were produced at 20% than at 5% O(2). The thickness of neocartilage increased during the culture period. Quantitative RT-PCR showed that the procollagen α(1)(II) mRNA expression level was significantly higher at 20% than at 5% O(2). However, no significant difference in gene expression and PG content was found between control and GS-treated cultures at either 20% or 5% O(2). Thus, in contrast to monolayer cultures, engineered cartilage from scaffold-free cultured chondrocytes at 20% O(2) produced better extracellular matrix (ECM) than that at 5% O(2). PGs were mainly large. Exogenous GS was not beneficial for the ECM in scaffold-free chondrocyte cultures.

The European mink is an endangered Mustelidae species and thus requires effective conservation measures, although little is known about reproduction in this species. In particular, preimplantation development has not been studied and, therefore, embryonic development and the growth of embryos was documented in the present study for European mink using light and fluorescent microscopy. Embryos develop in the oviducts and then migrate into the uterus on Day 6 post coitum (p.c.) at the morula stage. Embryos expanded as blastocysts from Day 7 until implantation on Day 12 p.c. Based on these findings, the use of embryo transfer for a conservation programme for the European mink was evaluated. Embryos were flushed from European mink resource females and transferred into the uterine horns of recipient hybrid females (honoriks and nohoriks). These hybrids were obtained by mating European polecat males with European mink females and vice versa. A total of 40 embryos was transferred and 20 live kits were born. The rates of pre- and postnatal survival were 50% and 70%, respectively. Both male and female offspring were lighter at birth in the embryo transfer group compared with naturally born controls, but there was no difference at 3 months of age.

Stoats (Mustela erminea), an exotic pest in New Zealand, threaten the conservation of several ground-nesting bird species and broad-scale methods for their control are sought. Females are seasonally monestrous, show a 9-month period of obligatory diapause and usually do not breed more than once in their lives. A bait-delivered agent that terminates diapause and results in a non-viable embryo may have a significant impact on their reproductive success. Prolactin (PRL) is hypothesised to be the only gonadotrophin required for renewal of luteal activity and blastocyst implantation in some mustelids. We investigated the effects of bait-delivered dopamine (DA) antagonists (which stimulate the release of PRL) using a mink model (Mustela vison), a species that maintains a short period of diapause. A bait dose of 0.8 mg kg(-1) of pimozide was more effective in elevating PRL levels than equivalent doses of fluphenazine, sulpiride (P < 0.01) or haloperidol (P < 0.05). Bait doses of 1.6 mg kg(-1) pimozide given at Days 0, 3, 9 and 11 after mating caused a significant reduction in the length of pregnancy compared with a positive control and placebo (46 days v. 51 days), indicating early termination of diapause (P < 0.01). Pimozide doses caused higher elevations in PRL concentration relative to the oral placebo by Day 12, but mean PRL levels fell below all other groups by Day 18. A borderline significant increase in progesterone (P4) secretion compared with the oral placebo was detected at Day 18. These results suggest that bait-delivered pimozide can elevate PRL outside of the normal breeding season and doses of 1.6 mg kg(-1) are effective in terminating embryonic diapause in mink. The implications and limitations of these data are discussed with reference to the use of bait-delivered DA antagonists as a possible means to affect the reproductive success of wild stoats.

Objective: The purpose was to compare the therapeutic effect, the required overall treatment duration, and patients' subjective perception of treatment of candidosis with miconazole chewing gum 20 mg/piece (3.6 mg released) and 2% Brentan gel 2.5 g (50 mg released), both administered 4 times daily., Study Design: Randomly selected patients (chewing gum: 96; gel: 28; placebo 16) were seen at baseline and at 4 and 6 weeks. They were evaluated clinically and with culture and cytology assays., Results: There was no significant difference between the 2 drugs after 6 weeks, but miconazole released in small doses from chewing gum was at least as effective as a larger gel dose. Both active drugs demonstrated efficiency and efficacy (P=1.9%). The hypothesis of equal proportions was rejected (P=.0001). Patients preferred the chewing gum. There was a statistically significant difference in patient preference for the chewing gum treatment., Conclusion: The chewing gum medication is clinically promising.