Ontogenetic skull variation in a shovel-headed amphisbaenian species.

Leposternon microcephalum is a species belonging to the Amphisbaenia, a group of burrowing reptiles. Amphisbaenia present various morphological and physiological adaptations that allow them to penetrate the ground and live underground, through a system of galleries and permanent chambers that they build themselves. Among the morphological adaptations in this group, those of the skull stand out as it serves as the main excavation tool. Four basic skull shapes are recognized: rounded, keeled, shovel-shaped, and spade-shaped. The skull of L. microcephalum belongs to this last type, which is considered the most specialized. The species inhabits soils that are highly compacted and difficult to penetrate. Among the species of Leposternon present in South America, L. microcephalum has the widest distribution, being found in all Brazilian biomes and neighboring countries such as Bolivia, Argentina, Paraguay, and Uruguay. The analysis of the skull of this species was carried out using three-dimensional geometric morphometrics (3D-GMM), a technique that allows comparative analysis, through robust statistical methods, of shape and its variations, using Cartesian coordinate data from a configuration of homologous landmarks. The technique allows the size and shape components of a structure to be analyzed separately. From an ontogenetic point of view, this methodology had also been used to investigate variations in Cynisca leucura, a member of the Amphisbaenidae with a rounded head. Our hypothesis is that the patterns of morphological differentiation in the skull, mainly in the intermediate and occipital regions, are similar in different Amphisbaenia species. Therefore, the objective of this study was to analyze cranial morphological variations in an ontogenetic series of L. microcephalum using 3D-GMM. Computed Tomographic scans of 13 specimens were analyzed: juveniles (N = 8) and adults (N = 5), based on 20 landmarks that characterize the skull. Principal components and regression analyses between shape (dependent variable) and size (independent variable) showed a clear difference between the cranial morphological pattern of juvenile individuals and that of adults. For instance, young specimens tend to have a dorsoventrally tall neurocranium, with the tip of the snout more anteriorly oriented and its dorsal border subtly curved. Dorsally, the parietal region is thicker and smoothly dome-shaped in juveniles. As in C. leucura, the variation was strongly correlated with the size change from juvenile to adult, indicating a dominant role for ontogenetic allometry in determining skull shape.

Common carp as an ecological indicator of environmental pollution in reservoirs of southern Spain: inferring the environmental risks of anthropogenic activities.

Extraction and mineral processing, as well as the waste generated by old abandoned mining sites, are the main sources of contamination of water bodies and lands by potentially toxic elements (PTEs). The common carp (Cyprinus carpio Linnaeus 1758) has been reported to be a good ecological indicator of environmental pollution in water bodies. Hence, we evaluated the concentration of eleven PTEs (As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb, and Zn) in different tissues of common carp in two reservoirs of the province of Jaén, southern Spain: El Tranco de Beas (S1) and La Fernandina (S2). We also assessed the concentration of PTEs in water and sediment samples. We used inductively coupled plasma mass spectrometry for all the collected samples. We found high concentrations of As and Fe in water in the S2 reservoir, above the maximum limits allowed by the sanitary criteria in Spain; however, the analysis of sediments indicated low ecological risk in S1 and moderate ecological risk for As in S2. The concentration of PTEs in common carp was higher in the S2 reservoir, exceeding the permissible limits in the case of As, Cd, Pb, and Zn. As and Cd showed higher concentrations in the kidney; Cu, Fe, and Zn showed higher concentrations in the liver; and Pb and Mn presented higher concentrations in the gill and gill bone. There was a good correlation between the concentrations found in water/sediment samples and those in common carp, corroborating its usefulness as a good ecological indicator, allowing the detection of environmental pollution and inferring previous or current anthropogenic activities such as mining.

Epidemiology of Trichinella infection in wild boar from Spain and its impact on human health during the period 2006-2019.

Trichinellosis is a notifiable zoonotic disease caused by parasitic nematode larvae belonging to the genus Trichinella. Domestic pig and wild boar are important hosts within the natural cycle of T. spiralis, the last one being an animal whose populations have experienced an important growth. Therefore, this paper studies the prevalence of Trichinella infection in wild boar in Spain, as well as its relation with hunting and its impact on public health during the period 2006-2019. For this purpose, different sources of information were consulted and analyzed depending on the autonomous communities of Spain and years. During the fourteen years of study, the number of wild boars hunted and the number of cases of Trichinella infection in them increased (from 172 cases in 2006 to 421 in 2019), although prevalence values remained low as the number of animals analyzed also increased. On the other hand, trichinellosis in humans tended to decrease (from a peak of 107 cases in 2007 to 11 cases in 2019). Nevertheless, the numbers of both wild boars and humans infected with Trichinella in Spain are among the highest in Europe, and this emphasizes the importance of food safety, sanitary controls of game meat and citizen awareness campaigns, which prevent the spread of Trichinella through the human population.

Modeling the age-related shift in the mineral content of hard tissues in two Mediterranean deer species.

OBJECTIVE: In this paper we aim to provide baseline data and model the changes of Ca, P and Mg throughout life in the mandibular bone, enamel and dentin of red (Cervus elaphus) and fallow deer (Dama dama) in Mediterranean ecosystems. DESIGN: Through a cross-sectional study of cervids from 1.5 to 20 yrs old, hunted between 1990 and 1997, we apply generalized additive models (GAMs) with data from scanning-electron-microscope with energy-dispersive X-ray (FESEM-EDX) and inductively coupled plasma-mass spectrometry (ICP-MS) analyses. RESULTS: The mineral content varied in a similar range to that reported for other ruminants. However, we detected lower Ca content values, while more similar results were obtained for P and Mg contents, which led to relatively lower Ca/P ratios and higher Ca/Mg in our deer at that time. A significantly lesser pattern of decreasing mineral content with aging was detected in the fallow deer males, similarities were found between the sexes, and significantly less resistance to demineralization was observed in dentin compared to bone. We discuss how the basic macromineral elements involved in the biomineralization process vary with age throughout life depending on deer species, sex and hard tissues. CONCLUSION: Allowing for possible inferences of differential changes in the mineralization state at the main stages in life history, our methodological approach opens up new possibilities in zooarchaeological, paleontological, and wildlife research.

Individual stress responses to Sarcoptes scabiei infestation in Iberian ibex, Capra pyrenaica.

In this study we have monitored the stress of Iberian ibex at individual level within the course of an experimental infection with Sarcoptes scabiei mites. For this purpose we have measured faecal 11-ketoetiocholanolone (11-k) using high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS). We used linear mixed models to explore the effects of host sex and age, clinic (mange status) and time (number of days post-infection) on the concentration of faecal 11-k. The most parsimonious model included clinic, time and host age, which explained 76.6% of the variance of the response variable. Moreover, the concentration of faecal 11-k varied greatly between individuals. Our results evidence the stressor nature of the disease and highlight the negative effects on hosts due to cortisol release and activity.

HPLC-QTOF method for quantifying 11-ketoetiocholanolone, a cortisol metabolite, in ruminants' feces: Optimization and validation.

Studies of animal ecology can benefit from a quantified understanding of eco-physiological processes and, in particular, of the physiological responses in free-ranging animals to potential stressors. The determination of fecal cortisol metabolites as a noninvasive method for monitoring stress has proved to be a powerful tool. High-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) has emerged as the most accurate method for avoiding problems related to the nonspecificity of immunoassays. In this study, we optimize and validate a reliable method using HPLC-MS/MS for quantifying 11-ketoetiocholanolone (11-k), a representative fecal cortisol metabolite in ruminants. An appropriate extraction and purification procedure was developed taking into account the complex nature of feces. The final extract obtained was then analyzed with HPLC-MS/MS using a quadrupole-time-of-fly (QTOF) tandem mass spectrometer with an electrospray ionization interface operating in positive mode, which allowed an unequivocal determination of the metabolite due to its accurate mass capabilities. After rigorous optimization of both sample extraction and the HPLC-QTOF parameters, making use of feces from free-ranging Iberian ibex, ideal conditions were established. Matrix-matched standards were used to calibrate the method. The limit of detection and quantification was 13- and 40- ng/g, respectively. The validation of the method was performed with recoveries in the range of 85-110%, a figure much higher than the 60% obtained with the previous extraction methods used in our laboratory, and with relative standard deviations (RSDs) no higher than 15% for the complete analytical procedure, including extraction and analysis. The time required for the fecal 11-k analysis was greatly reduced in comparison with the previous work carried out in our laboratory. This is the first time that QTOF mass detection coupled with HPLC has been validated for 11-k quantification in feces from free-ranging ruminants such as Iberian ibex. Given the high selectivity and sensitivity attained, our method could become a useful tool for noninvasive stress quantification in ruminants.

Growth in fossil and extant deer and implications for body size and life history evolution.

BACKGROUND: Body size variation within clades of mammals is widespread, but the developmental and life-history mechanisms by which this variation is achieved are poorly understood, especially in extinct forms. An illustrative case study is that of the dwarfed morphotypes of Candiacervus from the Pleistocene of Crete versus the giant deer Megaloceros giganteus, both in a clade together with Dama dama among extant species. Histological analyses of long bones and teeth in a phylogenetic context have been shown to provide reliable estimates of growth and life history patterns in extant and extinct mammals. RESULTS: Similarity of bone tissue types across the eight species examined indicates a comparable mode of growth in deer, with long bones mainly possessing primary plexiform fibrolamellar bone. Low absolute growth rates characterize dwarf Candiacervus sp. II and C. ropalophorus compared to Megaloceros giganteus displaying high rates, whereas Dama dama is characterized by intermediate to low growth rates. The lowest recorded rates are those of the Miocene small stem cervid Procervulus praelucidus. Skeletal maturity estimates indicate late attainment in sampled Candiacervus and Procervulus praelucidus. Tooth cementum analysis of first molars of two senile Megaloceros giganteus specimens revealed ages of 16 and 19 years whereas two old dwarf Candiacervus specimens gave ages of 12 and 18 years. CONCLUSIONS: There is a rich histological record of growth across deer species recorded in long bones and teeth, which can be used to understand ontogenetic patterns within species and phylogenetic ones across species. Growth rates sensu Sander & Tückmantel plotted against the anteroposterior bone diameter as a proxy for body mass indicate three groups: one with high growth rates including Megaloceros, Cervus, Alces, and Dama; an intermediate group with Capreolus and Muntiacus; and a group showing low growth rates, including dwarf Candiacervus and Procervulus. Dwarf Candiacervus, in an allometric context, show an extended lifespan compared to other deer of similar body size such as Mazama which has a maximum longevity of 12 years in the wild. Comparison with other clades of mammals reveals that changes in size and life history in evolution have occurred in parallel, with various modes of skeletal tissue modification.

Age and sex-related geometrical variation of ramus mandibulae of Sus scrofa's (Mammalia: Artiodactyla) jaws / Variaciones geométricas del ramus mandibulae en mandíbulas de Sus scrofa (Mammalia: Artiodactyla) según edad y sexo

Morphological variation of the ramus mandibulae of wild Sus scrofa was described and correlated to sex and age differences. Young forms (n=27, 15 males, 12 females) and adults (n=23, 13 males, 10 females) were analyzed through Geometric Morphometrics (2-D) to assess differences between groups. We found significant differences between age- and sex-related shape variations at ramus mandibulae level. Young wild boars present bulky angular process with backward oriented coronoid process, while adults show slender jaw vertical ramus, and orthogonally oriented coronoid process. The ramus mandibulae in female wild boars is more robust and bulky than in males. The morphological changes between young and adult forms are related to alterations in the osteo-muscular jaw complex along wild boars ontogeny. The jaw morphological variations observed in adult wild boars could also be related to dietary differences between sexes.

Se describen las variaciones morfológicas a nivel de la rama mandibular del jabali (Sus scrofa) en función del sexo y la edad. Se analizó material procedente de jabalíes juveniles (n=27, 15 machos y 12 hembras) y adultos (n=23, 13 machos y 10 hembras) a través de la morfometría geométrica (2-D) para valorar diferencias entre grupos. Se encontraron diferencias a nivel de la forma del ángulo mandibular entre edades y sexos, de manera que los individuos juveniles presentaron un ángulo más abultado y un proceso coronoides orientado hacia atrás, mientras que los adultos muestran una rama mandibular más estrecha y vertical, así como, un proceso coronoides orientado ortogonalmente. En las hembras la parte posterior de la mandíbula es más abultada y compacta que en los machos. Los cambios morfológicos entre juveniles y adultos están relacionados con los procesos ontogenéticos que afectan al complejo osteomuscular durante el crecimiento. Por otra parte, las variaciones morfológicas observadas en los adultos podrían tener relación con diferencias en la dieta entre ambos sexos.

Mandibular osteomyelitis in red deer (Cervus elaphus hispanicus) and fallow deer (Dama dama): occurrence and associated factors in free-living populations in southern Spain.

The prevalence of mandibular osteomyelitis, which results in a condition called lumpy jaw, and factors associated with its occurrence were investigated in syntopic free-living populations of red deer (Cervus elaphus hispanicus) and fallow deer (Dama dama) in Spain. The study material consisted of 3,586 mandibles from 2,548 red deer and 1,038 fallow deer shot during sport hunting, herd management culls, and programs for population control between 1988 and 1997 (period 1) and 2002 and 2009 (period 2) in eastern Sierra Morena, southern Spain. Disease prevalence ranged from 0.36% to 10.91% among age groups. Older animals were significantly more likely to be affected than younger ones. Red deer stags had higher prevalence than other groups. There was a significantly higher prevalence in period 1, probably associated with differences in climatic and population conditions. High population densities of female red deer contributed significantly to occurrence of disease. Intensive herd management and poor environmental conditions were considered risk factors that increased susceptibility to disease. The study of this affliction could be useful for monitoring general herd welfare and ecologic changes in Mediterranean ecosystems.