Prion protein gene (PRNP) variation in German and Danish cervids.

The structure of cellular prion proteins encoded by the prion protein gene (PRNP) impacts susceptibility to transmissible spongiform encephalopathies, including chronic wasting disease (CWD) in deer. The recent emergence of CWD in Northern European reindeer (Rangifer tarandus), moose (Alces alces alces) and red deer (Cervus elaphus), in parallel with the outbreak in North America, gives reason to investigate PRNP variation in European deer, to implement risk assessments and adjust CWD management for deer populations under threat. We here report PRNP-sequence data from 911 samples of German red, roe (Capreolus capreolus), sika (Cervus nippon) and fallow deer (Dama dama) as well as additional data from 26 Danish red deer close to the German border and four zoo species not native to Germany. No PRNP sequence variation was observed in roe and fallow deer, as previously described for populations across Europe. In contrast, a broad PRNP variation was detected in red deer, with non-synonymous polymorphisms at codons 98, 226 and 247 as well as synonymous mutations at codons 21, 78, 136 and 185. Moreover, a novel 24 bp deletion within the octapeptide repeat was detected. In summary, 14 genotypes were seen in red deer with significant differences in their geographical distribution and frequencies, including geographical clustering of certain genotypes, suggesting "PRNP-linages" in this species. Based on data from North American CWD and the genotyping results of the European CWD cases, we would predict that large proportions of wild cervids in Europe might be susceptible to CWD once introduced to naive populations.

Integrated omics analysis reveals the alteration of gut microbiota and fecal metabolites in Cervus elaphus kansuensis.

The gut microbiota is the largest and most complex microecosystem in animals. It is influenced by the host's dietary habits and living environment, and its composition and diversity play irreplaceable roles in animal nutrient metabolism, immunity, and adaptation to the environment. Although the gut microbiota of red deer has been studied, the composition and function of the gut microbiota in Gansu red deer (Cervus elaphus kansuensis), an endemic subspecies of red deer in China, has not been reported. In this study, the composition and diversity of the gut microbiome and fecal metabolomics of C. elaphus kansuensis were identified and compared for the first time by using 16S rDNA sequencing, metagenomic sequencing, and LC-MS/MS. There were significant differences in gut microbiota structure and diversity between wild and farmed C. elaphus kansuensis. The 16S rDNA sequencing results showed that the genus UCRD-005 was dominant in both captive red deer (CRD) and wild red deer (WRD). Metagenomic sequencing showed similar results to those of 16S rDNA sequencing for gut microbiota in CRD and WRD at the phylum and genus levels. 16S rDNA and metagenomics sequencing data suggested that Bacteroides and Bacillus might serve as marker genera for CRD and WRD, respectively. Fecal metabolomics results showed that 520 metabolites with significant differences were detected between CRD and WRD and most differential metabolites were involved in lipid metabolism. The results suggested that large differences in gut microbiota composition and fecal metabolites between CRD and WRD, indicating that different dietary habits and living environments over time have led to the development of stable gut microbiome characteristics for CRD and WRD to meet their respective survival and reproduction needs. KEY POINTS: • Environment and food affected the gut microbiota and fecal metabolites in red deer • Genera Bacteroides and Bacillus may play important roles in CRD and WRD, respectively • Flavonoids and ascorbic acid in fecal metabolites may influence health of red deer.

Detection of small ruminant Lentivirus proviral DNA in red deer from Poland.

Small ruminant lentiviruses (SRLVs) are widespread and infect goats and sheep. Several reports also suggest that SRLVs can infect wild ruminants. The presence of specific antibodies against SRLVs has been identified in wild ruminants from Poland, but no studies have been conducted to detect proviral DNA of SRLVs in these animals. Therefore, the purpose of this study was to examine samples from Polish wild ruminants to determine whether these animals can serve as reservoirs of SRLVs under natural conditions. A total of 314 samples were tested from red deer (n = 255), roe deer (n = 52) and fallow deer (n = 7) using nested real-time PCR. DNA from positive real-time PCR samples was subsequently used to amplify a CA fragment (625 bp) of the gag gene, a 1.2 kb fragment of the pol gene and an LTR-gag fragment. Three samples (0.95%) were positive according to nested real-time PCR using primers and probe specific for CAEV (SRLV group B). All the samples were negative for the primers and probe specific for MVV (SRLV A group). Only SRLV LTR-gag sequences were obtained from two red deer. Phylogenetic analysis revealed that these sequences were more closely related to CAEV than to MVV. Our results revealed that deer can carry SRLV proviral sequences and therefore may play a role in the epidemiology of SRLVs. To our knowledge, this is the first study describing SRLV sequences from red deer.

A Cost-Benefit Analysis of Alternative Management Strategies for Red Deer in Denmark.

In this paper, we conduct a cost-benefit analysis (CBA) of five alternative management strategies for red deer in Denmark: free harvest, trophy hunting, maximum harvest and two cases for natural demographic population compositions. To capture the outcome under each strategy we use a biological sex- and age-structured population model. The net benefit function includes meat values, recreational values, browsing damage costs and traffic damage costs and these values and costs are assumed to differ for the various sex and age classes of red deer. We show that the maximum harvest strategy leads to a reasonably high positive total net benefit, while the free harvest strategy yields a small positive net benefit. On the other hand, the trophy hunting strategy generates a high negative net benefit, while small negative net benefits are obtained under the two strategies for natural demographic population compositions.

Theileria orientalis Ikeda infection detected in red deer but not dogs or horses in New Zealand.

AIMS: To determine whether evidence for infection with Theileria orientalis (Ikeda) could be identified in samples of commercial red deer (Cervus elaphus), horses, and working farm dogs in New Zealand. METHODS: Blood samples were collected during October and November 2019 from a convenience sample of red deer (n = 57) at slaughter. Equine blood samples (n = 50) were convenience-sampled from those submitted to a veterinary pathology laboratory for routine testing in January 2020. Blood samples, collected for a previous study from a convenience sample of Huntaway dogs (n = 115) from rural regions throughout the North and South Islands of New Zealand between August 2018 and December 2020, were also tested. DNA was extracted and quantitative PCR was used to detect the T. orientalis Ikeda major piroplasm surface protein (MPSP) gene. A standard curve of five serial 10-fold dilutions of a plasmid carrying a fragment of the T. orientalis MPSP gene was used to quantify the number of T. orientalis organisms in the samples. MPSP amplicons obtained by end-point PCR on positive samples were isolated and subjected to DNA sequencing. The resulting sequences were compared to previously published T. orientalis sequences. RESULTS: There were 6/57 (10%) samples positive for T. orientalis Ikeda from the deer and no samples positive for T. orientalis Ikeda from the working dogs or horses. The mean infection intensity for the six PCR-positive deer was 5.1 (min 2.2, max 12.4) T. orientalis Ikeda organisms/µL. CONCLUSIONS AND CLINICAL RELEVANCE: Red deer can potentially sustain low infection intensities of T. orientalis Ikeda and could act as reservoirs of infected ticks. Further studies are needed to determine whether naïve ticks feeding on infected red deer can themselves become infected. ABBREVIATIONS: Cq: Quantification cycle; LOQ: Limits of quantification; MPSP: Major piroplasm surface protein; qPCR: Quantitative polymerase chain reaction.

Comparison of genome-wide DNA methylation patterns between antler precartilage and cartilage.

Deer antlers are the only mammalian organs that can fully regenerate after being lost and provide a valuable model for cartilage development. As one of the best-studied epigenetic mechanisms, DNA methylation is known to engage in organ and tissue development. This study aimed to investigate the role of DNA methylation in antler chondrogenesis by comparing whole-genome DNA methylation between precartilage and cartilage. Quantitative reverse transcription PCR (RT-qPCR) showed significant differences in the expression levels of DNA methyltransferase genes (DNMT1, DNMT3A, and DNMT3B) between precartilage and cartilage. Subsequently, we obtained DNA methylation profiles of antler precartilage and cartilage tissues by whole-genome bisulfite sequencing. Although sequencing data indicated that overall methylation levels at CpG and non-CpG sites were similar between precartilage and cartilage, 140,784 differentially methylated regions (DMRs, P < 0.05) and 3,941 DMR-related genes were identified. Gene ontology (GO) analysis of DMR-related genes demonstrated some significantly enriched GO terms (P < 0.05) related to chondrogenesis, including insulin receptor binding, collage trimer, integrin binding, and extracellular matrix structural constituent. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of DMR-related genes uncovered that the PI3K/AKT, cortisol synthesis and secretion, glycosaminoglycan biosynthesis-keratan sulfate, Hippo, and NF-κB signaling pathways might play a pivotal role in the transition of precartilage to cartilage. Moreover, we found that 25 DMR-related genes, including CD44, IGF1, ITGAV, ITGB1, RUNX1, COL2A1, COMP, and TAGLN, were most likely involved in antler chondrogenesis. In conclusion, this study revealed the genome-wide DNA methylation patterns of antler precartilage and cartilage, which may contribute to understanding the epigenetic regulation of antler chondrogenesis.

LiDAR reveals a preference for intermediate visibility by a forest-dwelling ungulate species.

Visibility (viewshed) plays a significant and diverse role in animals' behaviour and fitness. Understanding how visibility influences animal behaviour requires the measurement of habitat visibility at spatial scales commensurate to individual animal choices. However, measuring habitat visibility at a fine spatial scale over a landscape is a challenge, particularly in highly heterogeneous landscapes (e.g. forests). As a result, our ability to model the influence of fine-scale visibility on animal behaviour has been impeded or limited. In this study, we demonstrate the application of the concept of three-dimensional (3D) cumulative viewshed in the study of animal spatial behaviour at a landscape level. Specifically, we employed a newly described approach that combines terrestrial and airborne light detection and ranging (LiDAR) to measure fine-scale habitat visibility (3D cumulative viewshed) on a continuous scale in forested landscapes. We applied the LiDAR-derived visibility to investigate how visibility in forests affects the summer habitat selection and the movement of 20 GPS-collared female red deer Cervus elaphus in a temperate forest in Germany. We used integrated step selection analysis to determine whether red deer show any preference for fine-scale habitat visibility and whether visibility is related to the rate of movement of red deer. We found that red deer selected intermediate habitat visibility. Their preferred level of visibility during the day was substantially lower than that of night and twilight, whereas the preference was not significantly different between night and twilight. In addition, red deer moved faster in high-visibility areas, possibly mainly to avoid predation and anthropogenic risk. Furthermore, red deer moved most rapidly between locations in the twilight. For the first time, the preference for intermediate habitat visibility and the adaption of movement rate to fine-scale visibility by a forest-dwelling ungulate species at a landscape scale was revealed. The LiDAR technique used in this study offers fine-scale habitat visibility at the landscape level in forest ecosystems, which would be of broader interest in the fields of animal ecology and behaviour.

Distribution of large lungworms (Nematoda: Dictyocaulidae) in free-roaming populations of red deer Cervus elaphus (L.) with the description of Dictyocaulus skrjabini n. sp.

Lungworms of the genus Dictyocaulus are causative agents of parasitic bronchitis in domestic and wild ungulates. This study investigates the distribution, morphology and genetic diversity of D. cervi and a new lungworm species, Dictyocaulus skrjabini n. sp. infecting red deer Cervus elaphus, fallow deer Dama dama and moose Alces alces in Poland and Sweden. The study was conducted on 167 red deer from Poland and on the DNA of lungworms derived from 7 fallow deer, 4 red deer and 2 moose collected in Sweden. The prevalence of D. cervi and D. skrjabini n. sp. in dissected red deer in Poland was 31.1% and 7.2%, respectively. Moreover, D. skrjabini n. sp. was confirmed molecularly in 7 isolates of fallow deer lungworms and 1 isolate of red deer lungworms from Sweden. Dictyocaulus skrjabini n. sp. was established based on combination of their distinct molecular and morphological features; these included the length of cephalic vesicle, buccal capsule (BC), buccal capsule wall (BCW), distance from anterior extremity to the nerve ring, the width of head, oesophagus, cephalic vesicle, BC and BCW, as well as the dimensions of reproductive organs of male and female. Additionally, molecular analyses revealed 0.9% nucleotide sequence divergence for 1,605 bp SSU rDNA, and 16.5­17.3% nucleotide sequence divergence for 642 bp mitochondrial cytB between D. skrjabini n. sp. and D. cervi, respectively, and 18.7­19% between D. skrjabini n. sp. and D. eckerti, which translates into 18.2­18.7% amino acid sequence divergence between D. skrjabini n. sp. and both lungworms.

Classification of brain tumours from MRI images using deep learning-enabled hybrid optimization algorithm.

Brain tumours are produced by the uncontrolled, and unusual tissue growth of brain. Because of the wide range of brain tumour locations, potential shapes, and image intensities, segmentation of the brain tumour by magnetic resonance imaging (MRI) is challenging. In this research, the deep learning (DL)-enabled brain tumour detection is developed by hybrid optimization method. The pre-processing stage used adaptive Wiener filter for minimizing the noise from input image. After that, the abnormal section of the image is segmented using U-Net. Afterwards, the data augmentation is accomplished to recover the random erasing, brightness, and translation characters. The statistical, shape, and texture features are extracted in feature extraction process. In first-level classification, the abnormal section of the image is sensed as brain tumour or not. Here, the Red Deer Tasmanian Devil Optimization (RDTDO) trained DenseNet is hired for brain tumour detection process. If tumour is identified, then second-level classification provides the brain tumour classification, where deep residual network (DRN)-enabled RDTDO is employed. Furthermore, the system performance is assessed by accuracy, true positive rate (TPR), true negative rate (TNR), positive predictive value (PPV), and negative predictive value (NPV) with the maximum values of 0.947, 0.926, 0.950, 0.937, and 0.926 are attained.

Molecular characterization of Dictyocaulus nematodes in wild red deer Cervus elaphus in two areas of the Italian Alps.

Nematodes of the genus Dictyocaulus are the causative agents of parasitic bronchitis and pneumonia in several domestic and wild ungulates. Various species have been described in wild cervids, as the case of Dictyocaulus cervi in red deer, recently described as a separate species from Dictyocaulus eckerti. In Italy, information on dictyocaulosis in wildlife is limited and often outdated. In this work, 250 red deer were examined for the presence of Dictyocaulus spp. in two areas of the Italian Alps (n = 104 from Valle d'Aosta, n = 146 from Stelvio National Park), and the retrieved lungworms were molecularly characterized. Lungworms were identified in 23 and 32 animals from Valle d'Aosta and Stelvio National Park, respectively. The nematodes, morphologically identified as D. cervi, were characterized molecularly (18S rDNA, ITS2, and coxI). Consistently, almost all specimens were found to be phylogenetically related to D. cervi. Three individuals, detected from both study sites and assigned to an undescribed Dictyocaulus sp., clustered with Dictyocaulus specimens isolated from red deer and fallow deer in previous studies. Within each of D. cervi and the undescribed Dictyocaulus sp., the newly isolated nematodes phylogenetically clustered based on their geographical origin. This study revealed the presence of D. cervi in Italian red deer, and an undetermined Dictyocaulus sp. that should be more deeply investigated. The results suggest that further analyses should be focused on population genetics of cervids and their lungworms to assess how they evolved, or co-evolved, throughout time and space and to assess the potential of transmission towards farmed animals.

A survey of handling and transportation of UK farmed deer.

Studies on the transport of deer (Cervidae), in the UK, were published > 15 years ago. A more recent study of deer transport is required to allow for assessments and improvements to the transport of farmed deer. Sixteen deer farmers participated in a survey describing their management practices related to transport. Their responses showed that most vehicles used to transport deer were designed for other livestock. Participating farmers estimated journey times to slaughter as 1-8 h, with an arithmetic mean of 4.8 (± 2.38) h. Specific concerns raised by the respondents, relating to the transport of deer, included a need for deer-specific vehicles, stop-off areas for long journeys, market locations and haulier experience. Furthermore, data were collected from two abattoirs between July 2019 and June 2020 comprising journey times, slaughter times, bruising, location of origin, vehicle type and the number of animals. In total, 4,922 deer were transported across 133 journeys (from farm to abattoir) from 61 farms. Median and range for journey length were 3.2 (0.4-9.8) h and 154.2 (7.1-462.2) km, whereas group size and time spent in the lairage were 24 (1-121) and 17.8 (10.2-68.9) h, respectively. Group size was found to be significantly associated with both the presence of bruising in a group and the amount of bruising per deer. This study provides a much-needed update on the transport of farmed deer in the UK and highlights key areas for future research including the welfare impact of transport in larger groups and for longer durations.

Innovative Spectrum Handoff Process Using a Machine Learning-Based Metaheuristic Algorithm.

A cognitive radio network (CRN) is an intelligent network that can detect unoccupied spectrum space without interfering with the primary user (PU). Spectrum scarcity arises due to the stable channel allocation, which the CRN handles. Spectrum handoff management is a critical problem that must be addressed in the CRN to ensure indefinite connection and profitable use of unallocated spectrum space for secondary users (SUs). Spectrum handoff (SHO) has some disadvantages, i.e., communication delay and power consumption. To overcome these drawbacks, a reduction in handoff should be a priority. This study proposes the use of dynamic spectrum access (DSA) to check for available channels for SU during handoff using a metaheuristic algorithm depending on machine learning. The simulation results show that the proposed "support vector machine-based red deer algorithm" (SVM-RDA) is resilient and has low complexity. The suggested algorithm's experimental setup offers several handoffs, unsuccessful handoffs, handoff delay, throughput, signal-to-noise ratio (SNR), SU bandwidth, and total spectrum bandwidth. This study provides an improved system performance during SHO. The inferred technique anticipates handoff delay and minimizes the handoff numbers. The results show that the recommended method is better at making predictions with fewer handoffs compared to the other three.

How Is Arachidonic Acid Metabolism in the Uterus Connected with the Immune Status of Red Deer Females (Cervus elaphus L.) in Different Reproductive Stages?

Reproductive and condition parameters' dependency on immune status in seasonally reproducing ruminants such as red deer have not been outlined to date. We determined T and B blood lymphocytes; the concentration of IgG, cAMP, haptoglobulin, and 6-keto-PGF1α in blood plasma; and the mRNA and protein expression of PG endoperoxide synthase 2, 5-lipoxygenase, PGE2 synthase (PGES), PGF2α synthase (PGFS), PGI2 synthase (PGIS), leukotriene (LT)A4 hydrolase, and LTC4 synthase (LTC4S) in the uterine endo- and myometrium, on the 4th (N = 7) and 13th (N = 8) days of the estrous cycle, in anestrus (N = 6) and pregnancy (N = 8) in hinds. An increase in CD4+ T regulatory lymphocyte percentage during the estrous cycle and anestrus compared with pregnancy was recorded; the opposite effect was observed for CD21+ B cells (p < 0.05). cAMP and haptoglobin concentration were elevated during the cycle, as was IgG on the fourth day of the cycle, whereas 6-keto-PGF1α concentration was the highest in pregnancy, and the nearest in anestrus similarly were LTC4S, PGES, PGFS, and PGIS protein expression in the endometrium (p < 0.05). We showed an interaction between the immune system activation and AA-metabolite production in the uterus throughout different reproductive stages. IgG, cAMP, haptoglobin, and 6-keto-PGF1α concentrations are valuable candidates for markers of reproductive status in hinds. The results help expand our knowledge of the mechanisms underlying seasonal reproduction in ruminants.

A study of the incidence of bovine tuberculosis in the wild red deer herd of Exmoor.

Bovine tuberculosis (bTB) is an infectious disease which thrives at the wildlife-livestock interface. Exmoor has the largest herd of wild red deer (Cervus elaphus) in England, and also a large number of dairy and beef farms. The population, health and well-being of the herd are managed by a combination of hunting with hounds and by stalking. This study used a serological assay to determine the incidence of bTB in the population of 106 wild red deer of Exmoor, the relationship between regional deer densities and the presence of bTB in deer, and domestic cattle. The overall number of bTB positive deer was 28.3% (30/106). Stalked deer had a slightly higher incidence of bTB (19 out of 55, 34.55%) than hunted deer (11 out of 51, 21.57%). There was no clear pattern of distribution except for one region which showed an incidence of 42.22% compared with 16.4% in the remainder of the moor. There was little difference in the incidence of bTB between male and female animals. The age of animals in the study ranged from < 1 year to over 10 years. There was no clear difference in the incidence across the age range (< 1 year- > 10 years) with the exception of a particularly high incidence in those animals aged 1 year or less. There was a significant correlation between the presence of deer with bTB and the number of farms reporting bTB positive cattle, but not between the regional population of red deer and bTB in deer or cattle.

Temporal stability of fecal cortisol metabolites in mountain-dwelling ungulates.

Fecal cortisol metabolites (FCMs) are widely used to track stress responses in wildlife and captive species. Rules of thumb suggest that samples should be collected as soon as possible after defecation, to avoid decay of FCMs. To date, however, only a few studies investigated the stability of defecated FCMs over time, and most of them were conducted in controlled laboratory conditions. Here, we investigated the stability of FCMs over seven consecutive days, in two mountain-dwelling ungulates, under natural environmental conditions using a semi-experimental approach. Fecal samples from Northern chamois Rupicapra rupicapra (n = 24) and red deer Cervus elaphus (n = 22) were collected in summer of 2020 within the Stelvio National Park, Italy, and placed in an open area above 2000 m a.s.l. For the next 7 days, we collected a portion of each sample, and all sub-samples were analyzed with an 11-oxoetiocholanolone enzyme immunoassay. Exposure, temperature, and precipitation were fitted as covariates in non-linear generalized mixed models to assess FCM variation over time, and competing models were selected using AICc. For chamois, the best model included only time as a predictor, while for red deer, it included time, precipitation, and exposure. For both species, FCM values decreased rapidly from the first days after deposition until the fourth day. For red deer, in northern-exposed samples, FCM values decreased slower than in south-exposed ones; furthermore, FCM values increased with increasing precipitation. Our results offer a solid methodological basis to wildlife researchers and practitioners interested in the investigation of the ecological factors affecting stress variation in wildlife and support the recommendation to collect samples as fresh as possible, to avoid misleading inference. Further studies are necessary to evaluate the stability of FCMs when other enzyme immunoassays are used.

The importance of direct and indirect trophic interactions in determining the presence of a locally rare day-flying moth.

Ecosystem engineers affect other organisms by creating, maintaining or modifying habitats, potentially supporting species of conservation concern. However, it is important to consider these interactions alongside non-engineering trophic pathways. We investigated the relative importance of trophic and non-trophic effects of an ecosystem engineer, red deer, on a locally rare moth, the transparent burnet (Zygaena purpuralis). This species requires specific microhabitat conditions, including the foodplant, thyme, and bare soil for egg-laying. The relative importance of grazing (i.e., trophic effect of modifying microhabitat) and trampling (i.e., non-trophic effect of exposing bare soil) by red deer on transparent burnet abundance is unknown. We tested for these effects using a novel method of placing pheromone-baited funnel traps in the field. Imago abundance throughout the flight season was related to plant composition, diversity and structure at various scales around each trap. Indirect effects of red deer activity were accounted for by testing red deer pellet and trail presence against imago abundance. Imago abundance was positively associated with thyme and plant diversity, whilst negatively associated with velvet grass and heather species cover. The presence of red deer pellets and trails were positively associated with imago abundance. The use of these sites by red deer aids the transparent burnet population via appropriate levels of grazing and the provision of a key habitat condition, bare soil, in the form of deer trails. This study shows that understanding how both trophic and non-trophic interactions affect the abundance of a species provides valuable insights regarding conservation objectives.

Finding a suitable coat: The ecology of the invasive deer ked (Lipoptena cervi (Linnaeus, 1758); Diptera: Hippoboscidae), an ectoparasite of large mammals in the Czech Republic.

Some parasitic diseases of wild animals are transmissible to humans and cause health problems. A widespread ectoparasite of vertebrates is the deer ked, Lipoptena cervi ((L.); Diptera: Hippoboscidae). We studied the interaction of this parasite with habitat-related, host-related, and temporal factors in the Czech Republic. We analysed observations of its abundance in hunted wild animals in the context of their environment. Our results confirmed that cervids are a preferred host species. Most of the other relationships between ked abundance and the studied factors were probably due to the olfactory preferences of keds. Males and older individuals of the game host species were more attractive to the parasites for this reason. The higher abundance of keds at the beginning of the hunting season can be explained by their phenology. The deer ked can significantly affect the management of game species. We conclude that its preference for older male cervids is important for the future conditions of game animals. The population densities of deer keds in areas with high parasitization rates could thus be influenced by active game management.

Epidemic abortions due to Neospora caninum infection in farmed red deer (Cervus elaphus).

This study describes for the first time an abortion outbreak caused by Neospora caninum in farmed red deer. During a 5-year period, farmed hinds, naturally mated, were regularly ultrasound monitored to detect reproductive losses over their gestation. During the 4 years previous to the outbreak, abortion rates ranged from 4.7 to 8.6% (average 6.5%), and serology for indirect diagnosis of neosporosis and toxoplasmosis was performed. At the fifth year, the abortion rate increased to 25.3%. During this outbreak, three aborted foetuses and their placentas were recovered and submitted to laboratory for etiological diagnosis. Blood samples were collected from the 81 hinds at the end of the gestational period and the seropositivity rate for N. caninum, Toxoplasma gondii, Brucella abortus, bovine viral diarrhoea virus and bovine alphaherpesvirus type 1 was 66.7%, 67.9%, 0.0%, 8.6% and 0.0%, respectively. Neospora caninum-seropositive hinds (OR = 5.7, P = 0.0271) and hinds with high antibody titres to N. caninum (OR = 7.4, P = 0.0130) were more likely to abort than seronegative hinds. In addition, N. caninum seropositivity rate in the aborted hinds was higher (OR = 5.4, P = 0.033) than the non-aborted hinds. No association was found between T. gondii nor BVDV-seropositivity and abortions. Typical protozoal histopathologic findings (necrotizing non suppurative encephalitis, meningitis, myocarditis, hepatitis, among others) were observed in all foetuses. Neospora caninum was immunolabelled by immunohistochemistry in several tissues from two foetuses, and infection was also confirmed in the three foetuses by serology and/or DNA detection. No other abortifacient agent was detected in the foetuses. Their dams showed high N. caninum antibody titres (≥ 6400). Serologic evidence and epidemiological data recorded suggested a point-source of N. caninum infection before the occurrence of the outbreak, probably related with contaminated feedstuff with oocysts. Moreover, the intensive production system with a high stocking rate could be also considered a factor which might have increased the risk of horizontal N. caninum infection in this herd.

Melatonin affects red deer spermatozoa motility and physiology in capacitating and non-capacitating conditions.

Melatonin affects sperm physiology, possibly through membrane receptors. Effects were tested at low concentrations (1 pM, 100 pM, 10 nM and 1 µM) in red deer epididymal spermatozoa as a model for high-seasonality species. Samples were incubated with melatonin as uncapacitated or capacitating conditions (heparin) and evaluated for motility and physiology (flow cytometry). Most effects occurred at low concentrations (nM-pM), mainly protecting from apoptosis and maintaining acrosomal integrity, suggesting a role for membrane receptors rather than a direct antioxidant effect. Intracellular calcium was not affected, differing from other studies and perhaps because of the epididymal origin. This study supports the relevance of melatonin on sperm physiology and could contribute to the application of reproductive technologies in wild ruminants.

The Quality and Fertilizing Potential of Red Deer (Cervus elaphus L.) Epididymal Spermatozoa Stored in a Liquid State.

The aim of this study was to assess the quality and fertilizing potential of red deer epididymal spermatozoa stored in a liquid state for up to 11 days (D11). In Experiment 1, sperm quality was determined. In Experiment 2, the efficiency of in vitro fertilization (IVF) and artificial insemination (AI) of stored sperm were evaluated. An analysis of sperm quality on D5 of storage revealed a decrease (p < 0.05) in motility and morphology, and a higher proportion of apoptotic spermatozoa. On D1, D7 and D10, the total motility of sperm for IVF and AI was determined to be 82.6%, 71.0% and 64.8%, respectively. The results of IVF and AI demonstrated that the fertilizing potential of spermatozoa differs between days of storage. The percentage of blastocysts was higher when oocytes were fertilized on D1 (17.4 %) compared to D7 (8.5%) and D10 sperm (10.5%). Differences were noted in the pregnancy rates of inseminated hinds. The insemination with D1, D7 and D10 sperm led to live births (33% from D7 and D10). The results indicate that the quality of red deer epididymal spermatozoa remains satisfactory during ten days of storage in a liquid state, and that these spermatozoa maintain their fertility potential.