Comparative Analysis of Microsatellite and SNP Markers for Genetic Management of Red Deer.

The analysis of population genetic structure and individual multilocus heterozygosity are crucial for wildlife management and conservation. Microsatellite markers have traditionally been used to assess these genetic parameters. However, single-nucleotide polymorphisms (SNPs) are becoming increasingly popular. Our goal here was to determine to what extent SNPs can provide better insights than microsatellites into the overall genetic status and population genetic processes in the species. To this end, we genotyped 210 red deer (Cervus elaphus) in the Spanish wild population with both 11 microsatellites and 31,712 SNPs. We compared parameters related to population genetic structure and individual multilocus heterozygosity obtained with both types of markers. Our results showed correlations between parameters measured using both microsatellites and SNPs, particularly those related to the level of genetic diversity and genetic differentiation. However, we found notably lower precision of microsatellites in measuring the distribution of genetic diversity among individuals. We conclude that microsatellites can be used to monitor the overall genetic status and detect broad patterns in red deer populations. Nevertheless, the greater precision of SNPs in inferring genetic structure and multilocus heterozygosity leads us to encourage scientists and wildlife managers to prioritize their use whenever possible.

Evaluation of candidate reference genes for quantitative real-time PCR normalization in blood from red deer developing antlers.

Sexual selection favors male traits that increase their ability to monopolize the breeding access to several females. Deer antlers are cranial appendages that regenerate annually in males. Throughout life, the phenology of antler growth advances and antler mass increases until the stag reaches, between 8 and 10 years old, maximum body mass and highest reproductive success. The molecular mechanisms of antler development are of great interest in both evolutionary and regenerative medicine studies. To minimize errors in the assessment of gene expression levels by qRT-PCR, we analyzed the stability of a panel of eight candidate reference genes and concluded that qRT-PCR normalization to three stable genes is strongly convenient in experiments performed in red deer antler blood. To validate our proposal, we compared the expression level of three genes linked to red deer antler growth (ANXA2, APOD and TPM1) in fifteen male red deer classified as young (up to 4 years old) and adults (4-6 years old). Our data confirms that B2M, ACTB and RPLP0 are valuable reference genes for future gene expression studies in red deer antler blood, which would provide increased insight into the effects of intrinsic factors that determine antler development in red deer.

Astigmatism correction with toric implantable collamer lens in low and high astigmatism groups.

PURPOSE: To analyze the target induced astigmatism (TIA), surgically induced astigmatism (SIA), difference vector (DV), and correction index (CI) in the correction of astigmatism with phakic lenses, and its influence on visual acuity, and to analyze the safety and efficacy indexes of the correction of high and low power astigmatism with toric phakic lenses. DESIGN: Retrospective comparative study. METHODS: The medical records of patients that were operated on at the research center during the period were analyzed. Results were divided into Low Astigmatism Group - LAG (33 eyes) and High Astigmatism Group - HAG (93 eyes) according to the implanted toric ICL lens power. Preoperative refraction and resultant postoperative refraction were analyzed by vector analysis. Visual acuity pre and postop, with and without optical correction, were compared. RESULTS: A total of 126 eyes were studied. The average preop refraction was -5.02 D sphere with -2.61 D cylinder. The average ICL lens power implanted was -8.31 D sphere +2.77 D cylinder. Refractive remaining was -0.01 ± 0.11 D sphere -0.15 ± 0.28 D cylinder. The arithmetic average angle of error in the astigmatism correction was 1.08°. The resultant cylinder was -0.03 ± 0.12 D and -0.19 ± 0.30 D in the low and high astigmatism groups, respectively, with a mean UDVA -0.01 ± 0.10 and 0.01 ± 0.16 and CDVA -0.03 ± 0.08 and -0.01 ± 0.17 for each group. The safety and efficacy indexes for the low astigmatism group were 1.09 ± 0.16 and 1.05 ± 0.17, respectively, with 1.11 ± 0.17 and 1.06 ± 0.16 for the high astigmatism group. CONCLUSIONS: The correction of astigmatism by the implantation of toric phakic lenses of the posterior chamber is safe and effective, independently of the amount of cylinder corrected.

Aberrometric, Keratometric, and Visual Outcomes After Trans-Epithelial Topography-Guided Phototherapeutic Keratectomy for the Treatment of Irregular Corneas.

PURPOSE: To assess the safety, aberrometric and keratometric changes, and stability of trans-epithelial topography-guided phototherapeutic keratectomy (TE-TG-PTK) with mitomycin C (MMC) using the ALLEGRO Topolyzer platform for the treatment of irregular astigmatism. METHODS: This is a retrospective case series including 57 eyes that underwent TE-TG-PTK + MMC using the ALLEGRO Topolyzer platform for the treatment of irregular astigmatism. CDVA, manifest refraction (MR), keratometry readings, and aberrometry readings were analyzed at 1, 3, 6, and 12 months. RESULTS: Causes of corneal irregularity included non-infectious leucoma (n=23), infectious leucoma (n=7), adenoviral keratitis (n=20), corneal haze (n=2), post-penetrant keratoplasty (PKP) (n=1), and others (n=4). Overall, 76% of the eyes (n=40) gained lines of vision; patients gained 1, and 2 or more lines of vision in 76%, and 38% of cases, respectively. Only 1 patient (2%) lost 5 lines of vision. Mean preoperative CDVA (LogMAR) was 0.37 ±0.31 and improved to 0.14 ±0.18 (p<0.001) at final follow-up (12 months). CDVA remained unchanged in 10 eyes (21%). No significant changes were observed in mean keratometry (Kmean) and keratometric astigmatism readings. Regarding aberrometry, only changes in coma proved to be significant 6 months after surgery (P<0.01). No intraoperative/postoperative complications were reported. CONCLUSION: At final follow-up, significant improvements were observed in CDVA and coma. TE-TG-PTK + MMC proved to be an effective and safe procedure for the treatment of corneal irregular astigmatism due to several causes.

Rainfall decrease and red deer rutting behaviour: Weaker and delayed rutting activity though higher opportunity for sexual selection.

In the last decades, climate change has caused an increase in mean temperatures and a reduction in average rainfall in southern Europe, which is expected to reduce resource availability for herbivores. Resource availability can influence animals' physical condition and population growth. However, much less is known on its effects on reproductive performance and sexual selection. In this study, we assessed the impact of three environmental factors related to climate change (rainfall, temperature and vegetation index) on Iberian red deer Cervus elaphus hispanicus reproductive timing and sexual behaviour, and their effects on the opportunity for sexual selection in the population. We measured rutting phenology as rut peak date, the intensity of male rutting activity as roaring rate, and the opportunity for sexual selection from the distribution of females among harem holding males in Doñana Biological Reserve (Southwest Spain), from data of daily observations collected during the rut over a period of 25 years. For this study period, we found a trend for less raining and hence poorer environmental conditions, which associated with delayed rutting season and decreased rutting intensity, but that appeared to favour a higher degree of polygyny and opportunity for sexual selection, all these relationships being modulated by population density and sex ratio. This study highlights how climate change (mainly rainfall reduction in this area) can alter the conditions for mating and the opportunity for sexual selection in a large terrestrial mammal.

An open platform system based on SNP type genetic markers for discrimination between Alectoris rufa and Alectoris chukar.

The red-legged partridge (Alectoris rufa) is one of the most emblematic game species in Southern Europe. For the conservation of its natural populations against hybridization with chukar partridges (Alectoris chukar) a public and agreed control system able to detect genetic introgression between the two species should be established. As the already available method has not been implemented yet, this paper presents an improvement of the genetic analysis technique by using an open platform system to optimize the diagnostic procedure. Here we present the results obtained from the design of an Open Array™ platform with the available SNPs with proved diagnosis capacity between the two species of interest. By this procedure we genotyped 380 partridge samples, both from farms and field populations, which resulted in an overall percentage of genotyping performed with success of 99.64%. The Open Array genotyping plates showed high performance, specificity and an easy reproducibility compared to conventional techniques of genotyping.

Social environment modulates investment in sex trait versus lifespan: red deer produce bigger antlers when facing more rivalry.

Theory predicts that the plastic expression of sex-traits should be modulated not only by their production costs but also by the benefits derived from the presence of rivals and mates, yet there is a paucity of evidence for an adaptive response of sex-trait expression to social environment. We studied antler size, a costly and plastic sex trait, and tooth wear, a trait related to food intake and longevity, in over 4,000 male Iberian red deer (Cervus elaphus hispanicus) from 56 wild populations characterized by two contrasting management practices that affect male age structure and adult sex-ratio. As a consequence, these populations exhibit high and low levels of male-male competition for mating opportunities. We hypothesized that males under conditions of low intra-sexual competition would develop smaller antlers, after controlling for body size and age, than males under conditions of high intra-sexual competition, thus reducing energy demands (i.e. reducing intake and food comminution), and as a consequence, leading to less tooth wear and a concomitant longer potential lifespan. Our results supported these predictions. To reject possible uncontrolled factors that may have occurred in the wild populations, we carried out an experimental design on red deer in captivity, placing males in separate plots with females or with rival males during the period of antler growth. Males living with rivals grew larger antlers than males living in a female environment, which corroborates the results found in the wild populations. As far as we know, these results show, for the first time, the modulation of a sexual trait and its costs on longevity conditional upon the level of intra-sexual competition.

Scanning Electron Microscope Analysis of Stromal Surface Regularity in DSAEK Using Manual or Automated Microkeratomes or DMEK.

PURPOSE: To analyze differences in the stromal bed according to the method used to obtain the disc for endothelial transplant with 1) an automated microkeratome for Descemet stripping automated endothelial keratoplasty (DSAEK), 2) homogeneous dissection of Descemet membrane (Descemet membrane endothelial keratoplasty [DMEK]), or 3) a manual microkeratome used for DSAEK. METHODS: We analyzed the stromal bed of 25 consecutive corneas used for endothelial transplantation, divided into 3 groups: 1) 11 samples cut with an automated microkeratome, 2) 5 samples used for DMEK, 3) 9 samples cut with a manual microkeratome. A scanning electron microscope was used to obtain an image of the center of the stromal bed. The irregularity index of the surface complementary to the stromal surface of the disc obtained for grafting was calculated with the Canny algorithm (0: completely smooth; 1: completely irregular). RESULTS: At all thresholds studied (20, 30, 50, and 70), the mean irregularity index for group C (35.2, 24.4, 13.7, and 8.8, respectively) was higher than that of group A (26.2, 14.8, 6.7, and 4.0, respectively), which in turn was higher than that of group B (7.0, 4.2, 2.4, and 1.8, respectively). Differences were statistically significant among all groups and for all thresholds. CONCLUSIONS: Irregularity of the stromal bed after any dissection can be quantified using the Canny method. The use of an automated microkeratome for DSAEK reduces irregularity and helps obtain a surface more similar to that obtained with DMEK than with a manual microkeratome.

Sex reversal syndrome in the horse: four new cases of feminization in individuals carrying a 64,XY SRY negative chromosomal complement.

Horses are characterized as having a greater rate of chromosomal abnormalities than other species, which are mainly related to the sex chromosome pair and produce a series of different anomalies known as disorders in sexual development (DSD). In the present study, three Pura Raza Española (PRE) and one Menorquín (MEN) horses were studied and an incompatibility in their genetic and phenotypic sex were detected. Animals were karyotyped by conventional and molecular cytogenetic analyses and characterized using genomic techniques. Although all individuals, were totally unrelated, these animals had the same abnormality (64,XY SRY negative DSD) despite having an anatomically normal external mare phenotype. Therefore, this syndrome could remain undiagnosed in a large percentage of cases because the physiological and morphological symptoms are rare. In the present study, a slight gonadal dysgenesis was observed only in older individuals. Interestingly this chromosomal abnormality has been previously reported less than twenty times, and never in the PRE or MEN horses. With the present research, it is demonstrated that the use of genetic and cytogenetic diagnostic tools in veterinary practice could be an important complementary test to determine the origin of unexplained reproductive failures among horses.