Free-living Populations Research Articles

Distribution and Evolutionary History of Sialic Acid Catabolism in the Phylum Actinobacteria.

ABSTRACTSialic acids are present in humans and other metazoans, playing essential roles in physiological and pathological processes. Commensal and pathogenic bacteria have evolved the capacity to utilize sialic acids as nutrient and energy sources. However, in some actinobacteria, sialic acid catabolism (SAC) is associated with free-living populations. To unravel the distribution and evolutionary history of SAC in the phylum Actinobacteria, we analyzed the presence and diversity of the putative SAC gene cluster (nan) in 7,180 high-quality, nonredundant actinobacterial genomes that covered 1,969 species. The results showed that ∼13% of actinobacterial species had the potential to utilize sialic acids, with 45 species capable of anhydro-SAC, all except two of them through the canonical pathway. These species belonged to 20 orders and 81 genera, with ∼36% of them from four genera, Actinomyces, Bifidobacterium, Corynebacterium, and Streptomyces. Moreover, ∼40% of the nan-positive species are free living. Phylogenetic analysis of the key nan genes, nanA, nanK, and nanE, revealed a strong signal of horizontal gene transfer (HGT), accompanied with vertical inheritance and gene loss. This evolutionary pattern led to high diversity and differential distribution of nan among actinobacterial taxa and might cause the cluster to spread to some free-living species while losing in some host-associated species. The evolution of SAC in actinobacteria probably represents the evolution of certain kinds of noncore bacterial functions for environmental adaptation and lifestyle switch, in which HGT plays a dominant role.IMPORTANCE Sialic acids play essential roles in the physiology of humans and other metazoan animals, and microbial sialic acid catabolism (SAC) is one of the processes critical for pathogenesis. To date, microbial SAC is studied mainly in commensals and pathogens, while its distribution in free-living microbes and evolutionary pathway remain largely unexplored. Here, by examining all actinobacterial genomes available, we demonstrate that putative SAC is present in a small proportion of actinobacterial species, of which, however, ∼40% are free-living species. We also reveal remarkable difference in the distribution of SAC among actinobacterial taxa and high diversity of the putative SAC gene clusters. HGT plays a significant role in the evolution of SAC, accompanied with vertical inheritance and gene loss. Our results provide a comprehensive and systematic picture of the distribution and evolutionary history of SAC in actinobacteria, expanding the current knowledge on bacterial adaptation and diversification.

Targeted Metabolomic Approach to Assess the Reproducibility of Plasma Metabolites over a Four Month Period in a Free-Living Population.

Metabolomics is increasingly applied to investigate diet–disease associations in nutrition research. However, studies of metabolite reproducibility are limited, which could hamper their use within epidemiologic studies. The objective of this study was to evaluate the metabolite reproducibility during 4 months in a free-living population. In the A-DIET Confirm study, fasting plasma and dietary data were collected once a month for 4 months. Metabolites were measured using liquid chromatography tandem mass spectrometry, and their reproducibility was estimated using the intraclass correlation coefficient (ICC). Regularized canonical correlation analysis (rCCA) was employed to examine the diet–metabolite associations. In total, 138 metabolites were measured, and median ICC values of 0.49 and 0.65 were found for amino acids and biogenic amines, respectively. Acylcarnitines, lysophosphatidylcholines, phosphatidylcholines, and sphingomyelins had median ICC values of 0.69, 0.66, 0.63, and 0.63, respectively. The median ICC for all metabolites was 0.62, and 54% of metabolites had ICC values ≥0.60. Additionally, the rCCA heat map revealed positive correlations between dairy/meat intake and specific lipids. In conclusion, more than half of the metabolites demonstrated good to excellent reproducibility. A single measurement per subject could appropriately reflect the metabolites’ long-term concentration levels and may also be sufficient for assessing disease risk in epidemiologic studies. The study data are deposited in MetaboLights (MTBLS3428 (www.ebi.ac.uk/metabolights)).

Impairments in glycemic control during Eastbound transatlantic travel in healthy adults.

Study ObjectivesRepeated bouts of circadian misalignment impair glucose tolerance. However, whether circadian misalignment associated with travel and jet lag impair glucose homeostasis in a free-living population is not known. The goal of the present study was to examine glycemic control during one week of Eastbound transatlantic travel in healthy men and women.MethodsSeven healthy participants (5 women; age: 35.6 ± 2.5 years, BMI: 23.9 ± 2.4 m/kg2) traveled from Colorado, USA (GMT-7) to Europe (GMT and GMT+1) and wore a continuous glucose monitor (Freestyle Libre Pro) for 8–14 days before, during, and after travel. Indices of glycemic control were summarized over 24-hour periods and by day and night.ResultsMean glucose, peak glucose, and time spent in hyperglycemia increased linearly throughout the travel period relative to baseline levels. Mean glucose concentrations rose 1.03 mg/dL (95% CI: 0.34, 1.74) and duration of hyperglycemia increased by 17 min (95% CI: 5.5, 28.6) each 24-hour period. Increases in 24-hour glucose were primarily driven by increases in daytime parameters with rising mean glucose (0.72 mg/dL per day, [95% CI: −0.1, 1.5]) and duration of hyperglycemia (13.2 min per day [95% CI: 4.3, 22.1]). Mean glucose, but not peak glucose or time spent in hyperglycemia, increased each night (0.7 mg/dL per night [95% CI: 0.2, 1.2]).ConclusionsEastbound transatlantic travel induced a progressive worsening of glucose metrics during 24-hour, day, and night periods. Future research on managing glycemic control during jet lag in people with metabolic disorders is warranted.Clinical Trial RegistrationNone

Parasitological monitoring of European bison in the West Pomeranian voivodeship on the basis of coproscopical examination

The aim of the study was to determine the level of parasitic infections in West Pomeranian European bison on the basis of coproscopic examinations. A total of 172 faecal samples of European bison from the Mirosławiec, Drawsko and Jałowcówka herds were examined by the flotation and sedimentation methods as well as the Baermann technique. Faecal examinations revealed the presence of gastrointestinal nematode eggs belonging to the family Trichostrongylidae and the genera Nematodirus, Aonchotheca, Trichuris and Strongyloides, eggs of tapeworms of the genus Moniezia, trematodes Fasciola hepatica and Paramphistomum cervi as well as larvae of the lungworm Dictyocaulus viviparus and seven species of coccidia of the genus Eimeria (E. bovis, E. pellita, E. zuernii, E. auburnensis, E. cylindrica, E. ellipsoidalis, and E. brasiliensis). The composition of parasite species in the population of European bison in the West Pomeranian voivodeship was similar to that observed in other free-living European bison populations in Poland.

The Effect of Cigarette Smoking on Penile Vasculature and Cardiodynamics among Young Saudi Population

Erectile and cardiovascular dysfunctions in cigarette smokers are important signs resulting probably due to pelvic autonomic neuropathy with damage to the parasympathetic nervi erigentes. Evidence related to neuropathic etiology comes from studies that show structural changes in autonomic nerve fibers supplying the corpora cavernosa. The objective of this study was to evaluate the effect of cigarette smoking on penile mid shaft circumference and length, penile pulse amplitude, both systolic and diastolic blood pressures, and heart rate in response to erotic stimulation in young Saudi men living in Western region of Saudi Arabia (Makkah). In this free-living population study data was collected from100 men each (mild, moderate and heavy cigarette smokers) ages between 20 and 35 years with a mean age of 26.42 ± 10.69 years along with 100 age matched healthy non smokers who served as controls. Heavy cigarette smoker group showed a significant decrease (p<0.0005) in mid shaft penile circumference and length and penile pulse amplitude where as both systolic and diastolic blood pressures and heart rate exhibited a significant increase (P<0.025, P<0.0005 and P<0.005 respectively. However this difference was found to be non significant in mild smoker group when compared with non smoker controls. We hypothesized that heavy cigarette smoking seems to be associated with sub-fertility in these males with significant alteration in the penile vasculature and cardiodynamics in a dose-dependent manner.

Diel Protein Regulation of Marine Picoplanktonic Communities Assessed by Metaproteomics.

The diel cycle is of enormous biological importance in that it imposes temporal structure on ecosystem productivity. In the world’s oceans, microorganisms form complex communities that carry out about half of photosynthesis and the bulk of life-sustaining nutrient cycling. How the functioning of microbial communities is impacted by day and night periods in surface seawater remains to be elucidated. In this study, we compared the day and night metaproteomes of the free-living and the particle-attached bacterial fractions from picoplanktonic communities sampled from the northwest Mediterranean Sea surface. Our results showed similar taxonomic distribution of free-living and particle-attached bacterial populations, with Alphaproteobacteria, Gammaproteobacteria and Cyanobacteria being the most active members. Comparison of the day and night metaproteomes revealed that free-living and particle-attached bacteria were more active during the day and the night, respectively. Interestingly, protein diel variations were observed in the photoautotroph Synechococcales and in (photo)-heterotrophic bacteria such as Flavobacteriales, Pelagibacterales and Rhodobacterales. Moreover, our data demonstrated that diel cycle impacts light-dependent processes such as photosynthesis and UV-stress response in Synechococcales and Rhodobacterales, respectively, while the protein regulation from the ubiquitous Pelagibacterales remained stable over time. This study unravels, for the first time, the diel variation in the protein expression of major free-living and particle-attached microbial players at the sea surface, totaling an analysis of eight metaproteomes.

Unprecedented Density and Persistence of Feral Honey Bees in Urban Environments of a Large SE-European City (Belgrade, Serbia).

Simple SummaryThe western honey bee, a pollinator species that is essential for modern agriculture and food production sustainability, is under various anthropogenic pressures. In the last few decades, these have led to serious worldwide problems concerning the health and stability of honey bee colonies. The importance of wild or feral honey bees has only recently been recognized, as these populations are crucial for research on the processes that enable their survival under such pressures. Here, we present a case of an unmanaged free-living population of honey bees that, unlike in other known studies, live in a highly populated urban environment. This extraordinarily dense feral honey bee population, which is not directly associated with managed apiaries, provided us with a large dataset of various life history parameters that will considerably fill in the knowledge gaps on unmanaged colonies. Furthermore, we want to underline the importance of citizen science in the data collection process and suggest it as a suitable approach to study feral honey bees in urban landscapes. We believe that highly populated urban landscapes can support and reinforce feral honey bees and that similar citizen science projects should be set up in other urban areas or other countries.It is assumed that wild honey bees have become largely extinct across Europe since the 1980s, following the introduction of exotic ectoparasitic mite (Varroa) and the associated spillover of various pathogens. However, several recent studies reported on unmanaged colonies that survived the Varroa mite infestation. Herewith, we present another case of unmanaged, free-living population of honey bees in SE Europe, a rare case of feral bees inhabiting a large and highly populated urban area: Belgrade, the capital of Serbia. We compiled a massive data-set derived from opportunistic citizen science (>1300 records) during the 2011–2017 period and investigated whether these honey bee colonies and the high incidence of swarms could be a result of a stable, self-sustaining feral population (i.e., not of regular inflow of swarms escaping from local managed apiaries), and discussed various explanations for its existence. We also present the possibilities and challenges associated with the detection and effective monitoring of feral/wild honey bees in urban settings, and the role of citizen science in such endeavors. Our results will underpin ongoing initiatives to better understand and support naturally selected resistance mechanisms against the Varroa mite, which should contribute to alleviating current threats and risks to global apiculture and food production security.

Association with Temperature Variability and Physical Activity, Sedentary Behavior, and Sleep in a Free-Living Population.

The present study examines the temperature variability in physical activity (PA), sedentary behavior (SB), and sleep in a free-living population. A representative sample of 1235 adults (ages 21–70) from Iowa, U.S.A., wore a SenseWear Mini Armband (SWA) for a randomly assigned day. Koppen’s weather climate classification was used to precisely classify the temperature: cold (−13 to 32 °F), cool (32 to 50 °F), mild (50 to 64 °F), warm (64 to 73 °F), and hot (73 to 95 °F). The main effect of three-way ANOVA (age × gender × temperature) had differences for SB and sleep, with older adults having higher levels than younger adults (p < 0.05). However, moderate to vigorous PA (MVPA) did not vary systematically by age or gender, and contrary to expectations, the main effect of the weather was not significant for MVPA (p > 0.05). Participants spent more time participating in PA at cold than at hot temperatures. The results clarify the impact of temperature on shaping PA and SB patterns in adults. The variable impacts and differential patterns by age suggest that weather should be considered when interpreting differences in PA patterns in research or surveillance applications.

Health assessments uncover novel viral sequences in five species of Galapagos tortoises.

Emerging infectious diseases (EIDs) have been reported as causes of morbidity and mortality in free-living animal populations, including turtles and tortoises, and they have even resulted in species extinctions, with human activities contributing to the spread of many of these diseases. In the Galapagos, giant tortoises are endangered due to habitat change, invasive species, and other human impacts; however, the impact of EIDs on Galapagos tortoise conservation remains understudied. To fill this gap, we conducted health assessments of five tortoise species from the islands of Santa Cruz, Isabela and Española. We performed health evaluations of 454 animals and PCR testing for pathogens known to be relevant in other tortoise species. We identified two novel sequences of adenoviruses and four of herpesviruses. Based on alignments of the DNA polymerase gene and maximum likelihood phylogenetic analyses, we found both novel adenoviruses to be most closely related to red footed tortoise adenovirus 2, by nucleotide sequence and red footed tortoise adenovirus 1, based on amino acid sequence. Three of the herpesvirus sequences translated into the same deduced amino acid sequence; therefore, they may be considered the same viral species, closely related to terrapene herpesvirus 2. The fourth herpesvirus sequence was highly divergent from any sequence previously detected and is related to an eagle owl herpesvirus based on nucleotide sequence and to loggerhead oro-cutaneous herpesvirus based on amino acids. These novel viruses may be pathogenic for giant tortoises under specific conditions (e.g., stress). Continued screening is crucial to determine if these viruses play a role in tortoise fitness, morbidity and survival. This information allows us to provide recommendations to the Galapagos National Park Directorate and other institutions to improve the management of these unique species both in Galapagos and globally, and for tortoise reintroduction plans throughout the archipelago.

Associations between MHC class II variation and phenotypic traits in a free-living sheep population.

Pathogen-mediated selection (PMS) is thought to maintain the high level of allelic diversity observed in the major histocompatibility complex (MHC) class II genes. A comprehensive way to demonstrate contemporary selection is to examine associations between MHC variation and individual fitness. As individual fitness is hard to measure, many studies examine associations between MHC variation and phenotypic traits, including direct or indirect measures of adaptive immunity thought to contribute to fitness. Here, we tested associations between MHC class II variation and five phenotypic traits measured in free-living sheep captured in August: weight, strongyle faecal egg count, and plasma IgA, IgE and IgG immunoglobulin titres against the gastrointestinal nematode parasite Teladorsagia circumcincta. We found no association between MHC class II variation and weight or strongyle faecal egg count. We did, however, find associations between MHC class II variation and immunoglobulin levels which varied with isotype, age and sex. Our results suggest associations between MHC and phenotypic traits are more likely to be found for traits more closely associated with pathogen defence than integrative traits such as bodyweight and highlight the association between MHC variation and antibodies in wild populations.

Multispecies Populations of Methanotrophic Methyloprofundus and Cultivation of a Likely Dominant Species from the Iheya North Deep-Sea Hydrothermal Field.

ABSTRACTThe Methyloprofundus clade is represented by uncultivated methanotrophic bacterial endosymbionts of deep-sea bathymodiolin mussels, but only a single free-living species has been cultivated to date. This study reveals the existence of free-living Methyloprofundus variants in the Iheya North deep-sea hydrothermal field in the mid-Okinawa Trough. A clade-targeted amplicon analysis of the particulate methane monooxygenase gene (pmoA) detected 647 amplicon sequence variants (ASVs) of the Methyloprofundus clade in microbial communities newly formed in in situ colonization systems. Such systems were deployed at colonies of bathymodiolin mussels and a galatheoid crab in diffuse-flow areas. These ASVs were classified into 161 species-like groups. The proportion of the species-like groups representing endosymbionts of mussels was unexpectedly low. A methanotrophic bacterium designated INp10, a likely dominant species in the Methyloprofundus population in this field, was enriched in a biofilm formed in a methane-fed cultivation system operated at 10°C. Genomic characterization with the gene transcription data set of INp10 from the biofilm suggested traits advantageous to niche competition in environments, such as mobility, chemotaxis, biofilm formation, offensive and defensive systems, and hypoxia tolerance. The notable metabolic traits that INp10 shares with some Methyloprofundus members are the use of lanthanide-dependent XoxF as the sole methanol dehydrogenase due to the absence of the canonical MxaFI, the glycolytic pathway using fructose-6-phosphate aldolase instead of fructose-1,6-bisphosphate aldolase, and the potential to perform partial denitrification from nitrate under oxygen-limited conditions. These findings help us better understand the ecological strategies of this possibly widespread marine-specific methanotrophic clade.IMPORTANCE The Iheya North deep-sea hydrothermal field in the mid-Okinawa Trough is characterized by abundant methane derived from organic-rich sediments and diverse chemosynthetic animal species, including those harboring methanotrophic bacterial symbionts, such as bathymodiolin mussels Bathymodiolus japonicus and “Bathymodiolus” platifrons and a galatheoid crab, Shinkaia crosnieri. Symbiotic methanotrophs have attracted significant attention, and yet free-living methanotrophs in this environment have not been studied in detail. We focused on the free-living Methyloprofundus spp. that thrive in this hydrothermal field and identified an unexpectedly large number of species-like groups in this clade. Moreover, we enriched and characterized a methanotroph whose genome sequence indicated that it corresponds to a new species in the genus Methyloprofundus. This species might be a dominant member of the indigenous Methyloprofundus population. New information on free-living Methyloprofundus populations suggests that the hydrothermal field is a promising locale at which to investigate the adaptive capacity and associated genetic diversity of Methyloprofundus spp.

Interpopulation differences in male reproductive effort drive the population dynamics of a host exposed to an emerging fungal pathogen.

Compensatory recruitment is a key demographic mechanism that has allowed the coexistence of populations of susceptible amphibians with Batrachochytrium dendrobatidis (Bd), a fungus causing one of the most devastating emerging infectious disease ever recorded among vertebrates. However, the underlying processes (e.g. density-dependent increase in survival at early life stages, change in reproductive traits) as well as the level of interpopulation variation in this response are poorly known. We explore potential mechanisms of compensatory recruitment in response to Bd infection by taking advantage of an amphibian system where male reproductive traits are easy to quantify in free-living populations. The Southern Darwin's frog Rhinoderma darwinii is a vocal sac-brooding species that exhibits a high susceptibility to lethal Bd infection. Using a 7-year capture-recapture study at four populations with contrasting Bd infection status (one high prevalence, one low prevalence and two Bd-free populations), we evaluated whether Bd-positive populations exhibited a higher adult recruitment and a higher male reproductive effort than Bd-negative populations. We also estimated population growth rates to explore whether recruitment compensated for the negative impacts of Bd on the survival of adults. In addition, we evaluated a potential demographic signal of compensatory recruitment (i.e. positive relationship between the proportion of juveniles and Bd prevalence) in response to Bd infection using raw count data from 13 R. darwinii populations. The high Bd prevalence population exhibited the highest male reproductive effort and the highest recruitment among the four monitored populations. This led to a growing population during the study period despite high mortality of adult hosts. In contrast, males from the population with low Bd prevalence had a low reproductive effort and this population, which had the lowest adult recruitment, was declining during the study period despite adults having a higher survival in comparison to the high Bd prevalence population. We also found a demographic signal of compensatory recruitment in response to Bd infection in our broader analysis of 13 R. darwinii populations. Our study underlines the importance of interpopulation variation in life-history strategies on the fate of host populations after infectious disease emergence. Our results also suggest that an increase in reproductive effort can be one of the processes underlying compensatory recruitment in populations of Bd-susceptible amphibians.

Body temperature is a repeatable trait in a free-ranging passerine bird.

Body temperature (Tb) affects animal function through its influence on rates of biochemical and biophysical reactions, the molecular structures of proteins and tissues, and, ultimately, organismal performance. Despite its importance in driving physiological processes, there are few data on how much variation in Tb exists within populations of organisms, and whether this variation consistently differs among individuals over time (i.e. repeatability of a trait). Here, using thermal radio-frequency identification implants, we quantified the repeatability of Tb, both in the context of a fixed average environment (∼21°C) and across ambient temperatures (6-31°C), in a free-living population of tree swallows (Tachycineta bicolor, n=16). By experimentally trimming the ventral plumage of a subset of female swallows (n=8), we also asked whether the repeatability of Tb is influenced by the capacity to dissipate body heat. We found that both female and male tree swallow Tb was repeatable at 21°C (R=0.89-92), but female Tb was less repeatable than male Tb across ambient temperature (Rfemale=0.10, Rmale=0.58), which may be due to differences in parental investment. Trimmed birds had on average lower Tb than control birds (by ∼0.5°C), but the repeatability of female Tb did not differ as a function of heat dissipation capacity. This suggests that trimmed individuals adjusted their Tb to account for the effects of heat loss on Tb. Our study provides a first critical step toward understanding whether Tb is responsive to natural selection, and for predicting how animal populations will respond to climatic warming.

Action Research on Advocating Use of Fortified Foods in the Free-Living Population of Urban Vadodara using Diffusion of Innovation Model

Worldwide, unaffordable healthy diets, dependability over staple food items, and lack of knowledge have been the major cause of malnutrition. Government programs, promoting dietary diversity and supplementation have not given promising results in improving the micronutrient status of the population. Government of India, along with Food Safety and Standards Authority of India have chosen five vehicles, namely, rice, wheat flour, salt, milk, and oil for the fortification process to combat the micronutrient deficiency. Fewer studies aimed at creating advocacy about fortified foods among the free living population and stake holders. The present study undertook the advocacy for fortified foods amongst the free living population using the Diffusion of Innovation Model (DIM) for the purchase of fortified foods. An E- Intervention using graphics, audios and videos were given to the participants on WhatsApp for 1 month. The results in the study have shown significant difference in the awareness, perception and purchase of fortified foods for all the five staples post intervention, which provides enough evidence that the proposed strategy for creating awareness and promoting purchase of fortified foods amongst the selected participants was highly effective. Use of different E-communication channels can be used by researchers at large for creating the awareness about the safe consumption of fortified foods. Besides this, conducive environment is needed for fortification program to be a success, which will ensure the proper supply and demand to make the food fortification program viable.

The association between female reproductive performance and leukocyte telomere length in wild Soay sheep.

Telomere length (TL), typically measured across a sample of blood cells, has emerged as an exciting potential marker of physiological state and of the costs of investment in growth and reproduction within evolutionary ecology. While there is mounting evidence from studies of wild vertebrates that short TL predicts raised subsequent mortality risk, the relationship between reproductive investment and TL is less clear cut, and previous studies report both negative and positive associations. In this study, we examined the relationship between TL and different aspects of maternal reproductive performance in a free-living population of Soay sheep. We find evidence for shorter TL in females that bred, and thus paid any costs of gestation, compared to females that did not breed. However, we found no evidence for any association between TL and litter size. Furthermore, females that invested in gestation and lactation actually had longer TL than females who only invested in gestation because their offspring died shortly after birth. We used multivariate models to decompose these associations into among- and within-individual effects, and discovered that within-individual effects were driving both the negative association between TL and gestation, and the positive association between TL and lactation. This suggests that telomere dynamics may reflect recent physiologically costly investment or variation in physiological condition, depending on the aspect of reproduction being investigated. Our results highlight the physiological complexity of vertebrate reproduction, and the need to better understand how and why different aspects of physiological variation underpinning life histories impact blood cell TL.

Population Studies of Deer (Odocoileus virginianus, Mazama americana) in Southern Yucatán, Mexico

Aims: Estimate the population density of deer in the municipality of Tzucacab, Yucatán in the periods of 2003-2004, 2007-2008 and 2008-2009, determine the use of the habitat by these populations and the sustainability of the deer harvest from the estimated population densities.&#x0D; Study Design: A descriptive and vertical free-living deer population study was carried out in southern Yucatan, Mexico over a three-year period.&#x0D; Methodology: The map of the municipality of Tuzcacab was zoned in quadrants of 36 km2, completing a total of 36 quadrants; Unrestricted random sampling was applied to select seven quadrants in the period from 2003 to 2004 and 18 in each annual period between 2007 and 2009. Population samplings were carried out by applying three population estimation methods: direct sighting in a linear transect of 5 km in length, count of tracks in transect except period 2003-2004 and faecal pellets group count in plots. The evaluation of the use of habitat was carried out using the Bonferroni intervals, from the data of faecal pellets count. The evaluation of the deer harvest was carried out using the sustainable harvest model.&#x0D; Results: The population densities were different in each method, the density by the excreta count was 4.63 ± 2.49 deer / km2 in 2003-2004, 0.294 ± 0.198 deer / km2 in 2007-2008, and in the year 2008-2009 was 0.419 ± 0.0000085 deer / km2. Habitat use in 2007-08 and 2008-2009 was higher in the tropical forest, lower in agriculture and similar to that expected in secondary succession forest (acahual). The values of sustainable harvest, taking as a value the density per count of excreta in the plot because it showed the highest statistical precision, in the period 2003-04 it is sustainable, but in the period from 2007 to 2009 it is not sustainable.&#x0D; Conclusion: The population densities of deer (O. virginianus and M. americana) in Tuzcacab by means of the excreta count method, have decreased significantly. The habitat use preference is the tropical forest. The deer harvest in the period from 2007 to 2009 is not sustainable.

Thiamine deficiency in Gambian women of reproductive age.

Thiamine deficiency disorders are associated with a variety of clinical symptoms affecting the nervous and cardiovascular systems. There is growing recognition that thiamine deficiency can occur in populations well beyond the classical region of South Asia, and at‐risk populations include those who receive a large proportion of their energy from polished white rice (or other low‐thiamine staple foods) and with low dietary diversity. Reports of thiamine deficiency in West Africa over the last century have suggested that this has historically been an issue in this population, but in more recent decades, these reports have been limited to prison populations. To understand if thiamine deficiency might be an unrecognized problem in the communities of this region, erythrocyte samples collected during the wet and dry seasons from 226 women of reproductive age (mean age = 28 years old) were assessed for thiamine status by measuring the erythrocyte transketolase activity coefficient (ETKac). Overall, 35.8% of the sample was at high risk of thiamine deficiency (ETKac ≥ 1.25). Risk of thiamine deficiency was significantly higher in the wet (47.9%) compared with the dry season (22.9%) (P < 0.001). To our knowledge, this is the first report of biochemical thiamine deficiency in a free‐living population in West Africa in the 21st century and suggests that further investigation is warranted.

A lesson from the wild: The natural state of eosinophils is Ly6Ghi.

With a long history of promoting pathological inflammation, eosinophils are now emerging as important regulatory cells. Yet, findings from controlled laboratory experiments so far lack translation to animals, including humans, in their natural environment. In order to appreciate the breadth of eosinophil phenotype under non‐laboratory, uncontrolled conditions, we exploit a free‐living population of the model organism Mus musculus domesticus. Eosinophils were present at significantly higher proportions in the spleen and bone marrow of wild mice compared with laboratory mice. Strikingly, the majority of eosinophils of wild mice exhibited a unique Ly6Ghi phenotype seldom described in laboratory literature. Ly6G expression correlated with activation status in spleen and bone marrow, but not peritoneal exudate cells, and is therefore likely not an activation marker per se. Intermediate Ly6G expression was transiently induced in a small proportion of eosinophils from C57BL/6 laboratory mice during acute infection with the whipworm Trichuris muris, but not during low‐dose chronic infection, which better represents parasite exposure in the wild. We conclude that the natural state of the eosinophil is not adequately reflected in the standard laboratory mouse, which compromises our attempts to dissect their functional relevance. Our findings emphasize the importance of studying the immune system in its natural context – alongside more mechanistic laboratory experiments – in order to capture the entirety of immune phenotypes and functions.

Decline in body size and female fraction in the grass snake (Natrix natrix, Linnaeus 1758) population after 40 years (Southern Poland)

Depletion of free-living populations is often associated with changes in fitness-related traits, e.g., body size. Ongoing decrease in body size has been reported in most vertebrates, but reptiles remain understudied. Moreover, sexual size dimorphism, commonly observed in reptiles, indicates that environmental pressures on body size may appear sex-specific. This can also result in shifts in sex ratio, an aspect even less studied. We investigated body size and sex ratio in population of grass snake (Natrix natrix) surveyed over 40 years ago in comparison with the current state. We found that both sexes express similar magnitude in body size decline. The current sex ratio does not deviate from 1:1, while in the past, females outnumbered males. The observed changes are likely an outcome of several non-mutually exclusive factors. In the studied area, an increase in road traffic and human presence and a drop in prey availability have been documented. Both factors may exert higher pressure on larger individuals, particularly females, due to their high costs of reproduction. It is recorded here that increase in ambient temperatures and summer duration may additionally enhance the mortality risk and resource requirements. Shifts in body size and sex ratio can catalyze further declines in abundance and reproductive potential of the population.

Developing a non-invasive method of detecting elephant endotheliotropic herpesvirus infections using faecal samples.

Elephant endotheliotropic herpesvirus (EEHV)-associated haemorrhagic disease (EEHV-HD) is a leading cause of death in Asian elephant calves across the world. Cases of EEHV-HD have been detected in free-living calves through post-mortem examination (PME) indicating the presence of the virus in the wild. In the absence of a non-invasive sampling method, little research into free-living populations has been possible. This study aimed to provide evidence that faeces can be used as a non-invasive sampling method for the detection of EEHV excretion using quantitative polymerase chain reaction. Serial saliva swabs and faecal samples were taken from five captive Asian elephants in Thailand over 12weeks. To ensure the presence of detectable elephant DNA within the sample, qPCR was run for amplification of the Asian elephant tumour necrosis factor (TNF-α) gene, EEHV1 and EEHV4. Of 28 sample pairs, seven saliva samples were positive for EEHV, of which two had paired positive faecal samples. This study presents the first evidence that EEHV is excreted in faeces at detectable levels. This method may in future be used for improved understanding of the epidemiology of EEHV in free-living elephant populations, as well as detection of EEHV excretion in captive herds.