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.

Forage quality of consecutive years interact to affect body condition, reproductive rate and rut phenology in Iberian red deer.

Body condition for reproduction in capital breeders such as the red deer (Cervus elaphus) is mostly determined by their stored energy reserves. Thus, environmental conditions and resource availability may affect reproductive performance and breeding success. In warm Mediterranean regions, current climate change is driving to a hotter and drier scenario that is expected to affect the biology and dynamics of many populations. We examined the impact of these local climate variations on red deer body condition and the relationship with female reproductive phenology and breeding success. We used satellite information of landscape vegetation along with a 22-year data series of direct field behavioural observations during the rutting season in Doñana National Park (SW Spain). We analyzed faecal nitrogen content (FN) from faeces collected during the rut. We found that poor vegetation availability in drier years was related to worse body condition of deer (measured by FN) and a delay in the rutting season, which associated with lower reproductive rates (measured by the proportion of females with calves observed the next year). We also evidenced an interesting interaction between environmental conditions in consecutive years on the timing of breeding season, with timing of breeding being more delayed when previous year resource availability was high and many females bred, and the consecutive one was poor, so females hardly recovered condition and the rut occurred later. These findings highlight the carry-over effect of reproduction in capital breeders and the potential impact of climate-change conditions on red deer breeding.

Genetic Diversity of Wild Boar and Deer.

Genetic diversity provides the long-term capacity of species, communities, and the biosphere to persist under change [...].

Host Genetic Diversity and Infectious Diseases. Focus on Wild Boar, Red Deer and Tuberculosis.

Host genetic diversity tends to limit disease spread in nature and buffers populations against epidemics. Genetic diversity in wildlife is expected to receive increasing attention in contexts related to disease transmission and human health. Ungulates such as wild boar (Sus scrofa) and red deer (Cervus elaphus) are important zoonotic hosts that can be precursors to disease emergence and spread in humans. Tuberculosis is a zoonotic disease with relevant consequences and can present high prevalence in wild boar and red deer populations. Here, we review studies on the genetic diversity of ungulates and determine to what extent these studies consider its importance on the spread of disease. This assessment also focused on wild boar, red deer, and tuberculosis. We found a disconnection between studies treating genetic diversity and those dealing with infectious diseases. Contrarily, genetic diversity studies in ungulates are mainly concerned with conservation. Despite the existing disconnection between studies on genetic diversity and studies on disease emergence and spread, the knowledge gathered in each discipline can be applied to the other. The bidirectional applications are illustrated in wild boar and red deer populations from Spain, where TB is an important threat for wildlife, livestock, and humans.

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.

The dark-ventral-patch of male red deer, a sexual signal that conveys the degree of involvement in rutting behavior.

BACKGROUND: In polygynous mammals, signalling may play a decisive role in mating behavior, mediating the intensity of male fights and female mate choice. During the rutting season, male red deer may show a visible dark patch in their ventral fur. Recently, this patch has been suggested to act as a flexible sexual signal, due to its relationships with other variables such as age, body size, antler development, volatile compounds, or the competitive environment. The analysis of fur pigmentation at the ventral patch suggests that this might also visually indicate the male intrinsic predisposition to take part in mating competition. RESULTS: To assess the possible role of this trait as a communicative signal related to mate competition, we used red deer behavioral observations during the rut in Doñana National Park (Spain) to examine the link between the degree of expression of the dark ventral patch and the rutting activity (assessed from both intra-and-inter-sexual behaviors). Consistent with our predictions, we found in a field study that males with large dark patches showed a higher frequency of rutting behaviors (mainly roaring and flehmen), more interactions with females, and attained larger harem sizes. CONCLUSIONS: The dark ventral patch was a better predictor of male behavior than antler tines or territory holding, thus standing as a short-term indicator of male willingness to invest in mating competition.

The dark ventral patch: A bimodal flexible trait related to male competition in red deer.

Sexual signals play a central role in male-male competition in polygynous species. In red deer (Cervus elaphus), male's ventral area become dark during the rutting season due to urine spraying behaviour and retains many chemical compounds potentially revealing individual features. Here we investigate the variation in size of this trait, exploring its relationship with age and male competitive features such as antlers or body size, as well as populational level of intrasexual competition for mates. We found that the size of the dark ventral patch followed a clearly bimodal distribution, i.e. males mostly expressed the full-size trait or just developed a very small one. For these two groups of males according to trait expression, the relationships of trait size with age and antler size differed. Populational level of intrasexual competition appeared to affect the relationship between antler size and the probability of a fully developed ventral patch. These results indicate that the trait encodes information on body size, antler size, age and populational level of mate competition, thus suggesting a role in signalling male's competitive features and willingness to allocate reproductive effort within a particular season.

The exceptional finding of Locus 2 at Dehesilla Cave and the Middle Neolithic ritual funerary practices of the Iberian Peninsula.

There is a significant number of funerary contexts for the Early Neolithic in the Iberian Peninsula, and the body of information is much larger for the Late Neolithic. In contrast, the archaeological information available for the period in between (ca. 4800-4400/4200 cal BC) is scarce. This period, generally called Middle Neolithic, is the least well-known of the peninsular Neolithic sequence, and at present there is no specific synthesis on this topic at the peninsular scale. In 2017, an exceptional funerary context was discovered at Dehesilla Cave (Sierra de Cádiz, Southern Iberian Peninsula), providing radiocarbon dates which place it at the beginning of this little-known Middle Neolithic period, specifically between ca. 4800-4550 cal BC. Locus 2 is a deposition constituted by two adult human skulls and the skeleton of a very young sheep/goat, associated with stone structures and a hearth, and a number of pots, stone and bone tools and charred plant remains. The objectives of this paper are, firstly, to present the new archaeological context documented at Dehesilla Cave, supported by a wide range of data provided by interdisciplinary methods. The dataset is diverse in nature: stratigraphic, osteological, isotopic, zoological, artifactual, botanical and radiocarbon results are presented together. Secondly, to place this finding within the general context of the contemporaneous sites known in the Iberian Peninsula through a systematic review of the available evidence. This enables not only the formulation of explanations of the singular new context, but also to infer the possible ritual funerary behaviours and practices in the 5th millennium cal BC in the Iberian Peninsula.

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.

Efecto de un programa lúdico de actividad física general de corta duración y moderada intensidad sobre las cifras de presión arterial y otros factores de riesgo cardiovascular en hipertensos mayores de 50 años / Effects of a recreational general physical activity program with short term and moderate intensity of blood pressure and other cardiovascular risk factors in hypertensive patients over 50 years old

Objetivo: Evaluar los efectos de un programa recreativo de actividad física general, de intensidad moderada y corta duración, sobre las cifras de hipertensión arterial y otros factores de riesgo cardiovascular (FRCV) en hipertensos mayores de 50 años. Diseño: Estudio cuasi-experimental no aleatorizado con diseño pre-post. Emplazamiento: Íllora (Granada). Participantes: Sesenta sujetos sedentarios de 50-75 años pertenecientes al programa de hipertensos del Centro de Salud. Intervención: Programa lúdico de actividad física general donde predomina la capacidad aeróbica, 3días/semana, durante 4 semanas, y una intensidad del 45-55% de la FC Reserva. Mediciones principales: PAS, PAD, FC, IMC, colesterol total, HDL, LDL, TG y glucosa. Resultados y conclusiones: Disminuciones estadísticamente significativas (p < 0,05) en el IMC (-0,51%; IC95%: 30,26-31,93 unid), la FC (-5,57 lat/min; IC95%: 68,76-71,73 lat/min), la PAS (-14,82 mmHg; IC95%: 131,57-137,52 mmHg), la PAD (-5,33 mmHg; IC95%: 78,94-83,68 mmHg), la glucosa (-7,63 mg/dl; IC95%: 125,06-153,73 mg/dl) y el riesgo REGICOR (-20,46%; IC95%: 5,45-6,90%). Aumentos estadísticamente significativos en el HDL (+2,82mg/dl; IC95%: 46,78-52,11 mg/dl) y los TG (+8,27 mg/dl; IC95% 133,89-152,60 mg/dl). Según el sexo, los hombres presentaron la mayor variación en la FC y la PAD, y las mujeres en la PAS (p < 0,05). Sujetos con valores iniciales de PAS≥160mmHg experimentaron mayores descensos de FC, PAS, PAD, glucosa y TG (-(-10,67lat/min, -31mmHg, -8,27mmHg, -10,86mg/dl y 34,66mg/dl, respectivamente) que aquellos con PAS inicial < 160 mmHg, donde aumentó el HDL y disminuyó el LDL. Tras este programa se obtuvieron mejoras en la presión arterial y otros FRCV en sujetos hipertensos mayores de 50 años (AU)

Objectives: To evaluate the effects of a recreational general physical activity program with moderate intensity and short duration on blood pressure and other cardiovascular risk factors (BMI, cholesterol, Rest Heart Rate, HDL, LDL, Triglycerides) in hypertensive patients older than 50 years. Design: Non-randomised pre-post design, quasi-experimental study. Location: Íllora, Granada, Spain. Participants: A total of 60 subjects aged 50-75 years taking part in the Health Hypertensive Program in the Medical Centre were selected. Intervention: A recreational general physical activity program, mainly aerobic capacity, of 4weeks duration, 3days/week, and an intensity of 45-55% HR Reserve. Main measurements: SBP, DBP, HR, BMI, total cholesterol, HDL, LDL, TG, and Glucose. Results and conclusions: Statistically significant decreases (P < .05) were observed in BMI (-0.51%; 95%CI: 30.26 to 31.93 units), HR (-5.57 beats/min; 95% CI: 68.76 to 71.73 beats/min), SBP (-14.82 mmHg; 95%CI: 131.57 to 137.52 mmHg), DBP (-5.33 mmHg; 95%CI: 78.94 to 83.68 mmHg), Glucose (-7.63 mg/dL; 95%CI: 125.06 to 153.73 mg/dL) and REGICOR risk (-20.46%; 95%CI: 5.45 to 6.90%). Statistically significant increases were observed in HDL (+2.82 mg/dl; 95%CI: 46.78 to 52.11 mmHg), and TG (+8.27 mg/dl; 95%CI: 133.89 to 152.60 mg/dL). Men had a wider variation in HR and DBP, and women in SBP (P<.05). Subjects with baseline SBP≥160 mmHg experienced greater declines in HR, SBP, DBP, Glucose and TG (-10.67 beats/min, -31 mmHg, -8.27 mmHg, -10.86mg/dL, and 34.66mg/dL, respectively) than those with an initial SBP<160 mmHg, where there was an increase in HDL and a decrease in LDL. After this program, improvements in BP and other cardiovascular risk factors were obtained in hypertensive subjects over 50 years (AU)

[Effects of a recreational general physical activity program with short term and moderate intensity of blood pressure and other cardiovascular risk factors in hypertensive patients over 50 years old]. / Efecto de un programa lúdico de actividad física general de corta duración y moderada intensidad sobre las cifras de presión arterial y otros factores de riesgo cardiovascular en hipertensos mayores de 50 años.

OBJECTIVES: To evaluate the effects of a recreational general physical activity program with moderate intensity and short duration on blood pressure and other cardiovascular risk factors (BMI, cholesterol, Rest Heart Rate, HDL, LDL, Triglycerides) in hypertensive patients older than 50years. DESIGN: Non-randomised pre-post design, quasi-experimental study. LOCATION: Íllora, Granada, Spain. PARTICIPANTS: A total of 60 subjects aged 50-75years taking part in the Health Hypertensive Program in the Medical Centre were selected. INTERVENTION: A recreational general physical activity program, mainly aerobic capacity, of 4weeks duration, 3days/week, and an intensity of 45-55% HR Reserve. MAIN MEASUREMENTS: SBP, DBP, HR, BMI, total cholesterol, HDL, LDL, TG, and Glucose. RESULTS AND CONCLUSIONS: Statistically significant decreases (P<.05) were observed in BMI (-0.51%; 95%CI: 30.26 to 31.93units), HR (-5.57beats/min; 95%CI: 68.76 to 71.73beats/min), SBP (-14.82mmHg; 95%CI: 131.57 to 137.52mmHg), DBP (-5.33mmHg; 95%CI: 78.94 to 83.68mmHg), Glucose (-7.63mg/dL; 95%CI: 125.06 to 153.73mg/dL) and REGICOR risk (-20.46%; 95%CI: 5.45 to 6.90%). Statistically significant increases were observed in HDL (+2.82mg/dl; 95%CI: 46.78 to 52.11mmHg), and TG (+8.27mg/dl; 95%CI: 133.89 to 152.60mg/dL). Men had a wider variation in HR and DBP, and women in SBP (P<.05). Subjects with baseline SBP≥160mmHg experienced greater declines in HR, SBP, DBP, Glucose and TG (-10.67beats/min, -31mmHg, -8.27mmHg, -10.86mg/dL, and 34.66mg/dL, respectively) than those with an initial SBP<160mmHg, where there was an increase in HDL and a decrease in LDL. After this program, improvements in BP and other cardiovascular risk factors were obtained in hypertensive subjects over 50years.

Iberian red deer: paraphyletic nature at mtDNA but nuclear markers support its genetic identity.

Red deer populations in the Iberian glacial refugium were the main source for postglacial recolonization and subspecific radiation in north-western Europe. However, the phylogenetic history of Iberian red deer (Cervus elaphus hispanicus) and its relationships with northern European populations remain uncertain. Here, we study DNA sequences at the mitochondrial control region along with STR markers for over 680 specimens from all the main red deer populations in Spain and other west European areas. Our results from mitochondrial and genomic DNA show contrasting patterns, likely related to the nature of these types of DNA markers and their specific processes of change over time. The results, taken together, bring support to two distinct, cryptic maternal lineages for Iberian red deer that predated the last glacial maximum and that have maintained geographically well differentiated until present. Haplotype relationships show that only one of them contributed to the northern postglacial recolonization. However, allele frequencies of nuclear markers evidenced one main differentiation between Iberian and northern European subspecies although also supported the structure of both matrilines within Iberia. Thus, our findings reveal a paraphyletic nature for Iberian red deer but also its genetic identity and differentiation with respect to northern subspecies. Finally, we suggest that maintaining the singularity of Iberian red deer requires preventing not only restocking practices with red deer specimens belonging to other European populations but also translocations between both Iberian lineages.

Genetic Structure and Effective Population Sizes in European Red Deer (Cervus elaphus) at a Continental Scale: Insights from Microsatellite DNA.

We analyzed more than 600 red deer (Cervus elaphus) from large parts of its European distribution range at 13 microsatellite loci, presenting the first continent-wide study of this species using nuclear markers. Populations were clearly differentiated (overall F ST = 0.166, Jost's D est = 0.385), and the BAPS clustering algorithm yielded mainly geographically limited and adjacent genetic units. When forced into only 3 genetic clusters our data set produced a very similar geographic pattern as previously found in mtDNA phylogeographic studies: a western group from Iberia to central and parts of Eastern Europe, an eastern group from the Balkans to Eastern Europe, and a third group including the threatened relict populations from Sardinia and Mesola in Italy. This result was also confirmed by a multivariate approach to analyzing our data set, a discriminant analysis of principal components. Calculations of genetic diversity and effective population sizes (linkage disequilibrium approach) yielded the lowest results for Italian (Sardinia, Mesola; N e between 2 and 8) and Scandinavian red deer, in line with known bottlenecks in these populations. Our study is the first to present comparative nuclear genetic data in red deer across Europe and may serve as a baseline for future analyses of genetic diversity and structuring in this widespread ungulate.

Better Fitness in Captive Cuvier's Gazelle despite Inbreeding Increase: Evidence of Purging?

Captive breeding of endangered species often aims at preserving genetic diversity and to avoid the harmful effects of inbreeding. However, deleterious alleles causing inbreeding depression can be purged when inbreeding persists over several generations. Despite its great importance both for evolutionary biology and for captive breeding programmes, few studies have addressed whether and to which extent purging may occur. Here we undertake a longitudinal study with the largest captive population of Cuvier's gazelle managed under a European Endangered Species Programme since 1975. Previous results in this population have shown that highly inbred mothers tend to produce more daughters, and this fact was used in 2006 to reach a more appropriate sex-ratio in this polygynous species by changing the pairing strategy (i.e., pairing some inbred females instead of keeping them as surplus individuals in the population). Here, by using studbook data we explore whether purging has occurred in the population by investigating whether after the change in pairing strategy a) inbreeding and homozygosity increased at the population level, b) fitness (survival) increased, and c) the relationship between inbreeding and juvenile survival, was positive. Consistent with the existence of purging, we found an increase in inbreeding coefficients, homozygosity and juvenile survival. In addition, we showed that in the course of the breeding programme the relationship between inbreeding and juvenile survival was not uniform but rather changed over time: it was negative in the early years, flat in the middle years and positive after the change in pairing strategy. We highlight that by allowing inbred individuals to mate in captive stocks we may favour sex-ratio bias towards females, a desirable managing strategy to reduce the surplus of males that force most zoos to use ethical culling and euthanizing management tools. We discuss these possibilities but also acknowledge that many other effects should be considered before implementing inbreeding and purging as elements in management decisions.

Males and females contribute unequally to offspring genetic diversity in the polygynandrous mating system of wild boar.

The maintenance of genetic diversity across generations depends on both the number of reproducing males and females. Variance in reproductive success, multiple paternity and litter size can all affect the relative contributions of male and female parents to genetic variation of progeny. The mating system of the wild boar (Sus scrofa) has been described as polygynous, although evidence of multiple paternity in litters has been found. Using 14 microsatellite markers, we evaluated the contribution of males and females to genetic variation in the next generation in independent wild boar populations from the Iberian Peninsula and Hungary. Genetic contributions of males and females were obtained by distinguishing the paternal and maternal genetic component inherited by the progeny. We found that the paternally inherited genetic component of progeny was more diverse than the maternally inherited component. Simulations showed that this finding might be due to a sampling bias. However, after controlling for the bias by fitting both the genetic diversity in the adult population and the number of reproductive individuals in the models, paternally inherited genotypes remained more diverse than those inherited maternally. Our results suggest new insights into how promiscuous mating systems can help maintain genetic variation.

Microsatellite markers for identification and parentage analysis in the European wild boar (Sus scrofa).

BACKGROUND: The wild boar (Sus scrofa) is among the most widespread mammal species throughout the old world. Presently, studies concerning microsatellites in domestic pigs and wild boars have been carried out in order to investigate domestication, social behavior and general diversity patterns among either populations or breeds. The purpose of the current study is to develop a robust set of microsatellites markers for parentage analyses and individual identification. FINDINGS: A set of 14 previously reported microsatellites markers have been optimized and tested in three populations from Hungary, Portugal and Spain, in a total of 167 samples. The results indicate high probabilities of exclusion (0.99999), low probability of identity (2.0E(-13) - 2.5E(-9)) and a parentage assignment of 100%. CONCLUSIONS: Our results demonstrate that this set of markers is a useful and efficient tool for the individual identification and parentage assignment in wild boars.

Low heterozygosity at microsatellite markers in Iberian red deer with small antlers.

Deer antlers are costly structures subjected to directional sexual selection that may be sensitive to heterozygosity. However, a relationship between heterozygosity and antler development has only been found for select protein-coding loci and MHC genes in one deer species (the white-tailed deer Odocoileus virginianus). Here, we study the relationship between multilocus heterozygosity at 11 microsatellite markers and antler size (AS) in a sample of 367 Iberian red deer males (Cervus elaphus hispanicus) from two study areas with different ecological and genetic conditions. We found that males with very small antlers (10% of the sampled individuals with the lowest values of AS) had lower levels of heterozygosity than those with bigger antlers (significant effect in an analysis of variance, P = 0.011). This relationship was noticeable mainly in situations of low genetic diversity, where the differences in heterozygosity between groups of males were greater. Finally, we conducted analyses to address the hypotheses proposed by the heterozygosity-fitness correlation, and we found the local effect as the most likely hypothesis. Our findings reveal an expected but not previously detected association between low heterozygosity and reduced AS, with implications for red deer evolution and management.