bioRxiv Science⌕ Search

Biology subjects

Portugal, S.

Publications and source records attributed to Portugal, S..

16 recordsLinked to original sources

Sociality, diurnal temperature range and isothermality: Significant determinants of mass-independent resting metabolic rate in subterranean African mole-rats (Superfamily Bathyergidae)

Animals exhibit a diverse range of sociality from the strictly solitary to the highly social. Different forms of sociality have evolved in response to ecological constraints and selective habitat pressures, which are governed by the energetic and fitness costs to an individual. Uniquely among mammals, the clade of African mole-rats (Bathyergidae and Heterocephalidae) covers three distinct life-history forms of sociality: solitary, social and eusocial species. This variety in social structure makes them a model clade to study how metabolic traits vary between different forms of sociality. Resting metabolic rates (RMR) of seven African mole-rat species, ranging from solitary to eusocial, were measured using open-flow respirometry. Results were combined with published data, enabling the inclusion and statistical analysis of 16 species in total. We identified distinct allometric scaling of RMR, with eusocial species exhibiting a considerably greater rise in RMR with increases in body mass. This is likely attributable to reproductive and behavioural divisions of labour, and mass-dependent colony roles in eusocial species. Phylogenetically-informed analyses further identified that sociality, in addition to select bioclimatic traits - diurnal temperature range ({degrees}C) and isothermality (%) - significantly explain variation in the mass-independent RMR of African mole-rats. These findings elucidate, for the first time, that sociality can be a determinant of RMR, and calls for further study to identify the wider significance of sociality on mammalian metabolism, as well as exploring the allometric scaling of metabolic rate with respect to mammalian sociality.

physiology↗

Continental-scale genomic surveillance of Plasmodium falciparum malaria with rapid nanopore sequencing

In sub-Saharan Africa, continental-scale genomic surveillance of Plasmodium falciparum malaria is needed to track the spread of antimalarial drug resistance and diagnostic test evasion, as well as to monitor parasite evolutionary responses to vaccine rollout. Yet implementation of malaria genomic surveillance at a continental-scale is hindered by resource constraints, the vastness of the continent, and the lack of sequencing protocols suitable for most local laboratories. To address this, we developed an approach to enable a decentralized scale-up of P. falciparum genomic surveillance and established it in six African countries in one year, locally sequencing 1,065 samples. The approach includes a rapid ([~] 5 hours) and cost-efficient (<$25 USD/sample) nanopore sequencing protocol that provides surveillance of drug resistance-associated genes, hrp2/3 deletions, the vaccine target csp, and the polymorphic gene ama1. We coupled this to a bioinformatics dashboard that runs offline on a laptop and displays mapping and variant calling results in real-time. We demonstrate robust sequencing coverage across parasitemia levels and laboratories, accurate identification of antimalarial resistance markers and hrp2/3 deletions; and, with a novel variant caller, sensitive detection of mutations carried by minor clones. Our approach will accelerate genomic surveillance of P. falciparum malaria across sub-Saharan Africa at a time of urgent need.

genomics↗

Uncovering the genetic diversity of the malaria parasite antigen MSP2 across Sub-Saharan Africa

Genetic diversity in Plasmodium falciparum poses a significant challenge to malaria control and elimination. This is particularly important for developing fully efficacious vaccines, which should include valuable blood stage antigens. Several antigen candidates are highly diverse and require further understanding. We surveyed the genetic diversity of the highly polymorphic merozoite surface protein 2 (MSP2) in 2761 P. falciparum isolates collected across Sub-Saharan Africa. Using PCR-based genotyping and long-read sequencing, we identified extensive diversity among msp2 size variants and sequences. Some size variants were more prevalent than others across different geographical regions, transmission intensities, and time points. These variants comprised multiple unique sequences, of which several were geographically and temporally widespread. Our study reveals greater msp2 sequence diversity than previously known, while also identifying interesting similarities in sequence and gene length across Sub-Saharan Africa. These findings support the further exploration of common msp2 variants in relation to parasite virulence and vaccine development.

microbiology↗

Seasonal transcriptional profiling of malaria parasites using a new single-cell atlas of a Malian Plasmodium falciparum isolate

Asymptomatic persistence is crucial for the survival of malaria parasites in seasonal transmission settings, where transmission halts during dry months, while mosquitos are scarce and new infections rare. How Plasmodium falciparum avoids causing host symptoms or being cleared is not fully understood. Parasites that persist through the dry season circulate longer within the [~]48h replication cycle compared to parasites isolated from clinical cases, where only early stages circulate. The mechanisms promoting increased circulation and avirulent infections bridging two wet seasons remain unclear. We generated a single-cell RNA-seq reference atlas with P. falciparum erythrocytic stages of a recently adapted isolate, and with it defined the developmental stage of over 25000 parasites from six asymptomatic children at the dry-wet season transition, and nine children with clinical malaria in the wet season. Our data confirms that older parasites circulate in the dry season, and reveals transcriptional modulation of exported proteins that affect cytoadhesion of infected erythrocytes, which we validated by immuno-fluorescence microscopy.

molecular biology↗

A Metabolite-Based Resistance Mechanism Against Malaria

Whether jaundice, a common presentation of Plasmodium (P.) falciparum malaria (1-3) arising from the accumulation of circulating bilirubin, represents an adaptive or maladaptive response to Plasmodium spp. infection is not understood (1-3). We found that asymptomatic P. falciparum infection was associated with a >10-fold higher ratio of unconjugated bilirubin over parasite burden, compared to symptomatic malaria. Genetic suppression of bilirubin synthesis by biliverdin reductase A (BVRA) (4) increased parasite virulence and malaria mortality in mice. Accumulation of unconjugated bilirubin in plasma, via genetic inhibition of hepatic conjugation by UDP glucuronosyltransferase family 1 member A1 (UGT1A1) (5), was protective against malaria in mice. Unconjugated bilirubin inhibited P. falciparum proliferation in red blood cells (RBC) via a mechanism that suppressed mitochondrial pyrimidine synthesis. Moreover, unconjugated bilirubin inhibited hemozoin (Hz) crystallization and compromised the parasites food vacuole. In conclusion, jaundice represents a metabolic response to Plasmodium spp. infection that limits malaria severity.

immunology↗

Plasmodium falciparum expresses fewer var genes at lower levels during asymptomatic dry season infections than clinical malaria cases

In seasonal transmission areas, clinical malaria occurs during the wet season when mosquitoes are present, while in the dry season, malaria transmission is interrupted and clinical cases are rare. In Mali, Plasmodium falciparum can persist in low parasitaemic asymptomatic individuals through the six-month dry season and shows reduced cytoadhesion of infected erythrocytes, evidenced by the circulation of further developed parasite stages compared to clinical malaria cases. How prolonged circulation of infected erythrocytes is achieved remains unknown. Here, we explored var gene expression in subclinical infections and clinical malaria cases of Malian children, collected during the dry and wet seasons. We sequenced expressed var DBL-tags, used bioinformatic tools to predict their domain composition, binding phenotype and upstream sequence type; and determined their relationship to seasonality and clinical presentation. We found that parasites of asymptomatic infections expressed fewer var genes, with a larger proportion of var transcripts attributed to one or a few vars. In contrast, clinical cases exhibited expression of many var genes at lower proportions. We found that parasites of asymptomatic carriers expressed a mixture of CD36- and EPCR-binding PfEMP1, which changed over time. We confirmed that vars encoding CD36-binding PfEMP1 dominated in non-severe malaria cases, and found no significant difference in expressed var types between dry and wet seasons. Asymptomatic carriers were older, had higher titers of anti-P. falciparum antibodies, and broader reactivity to PfEMP1, suggesting that host immunity was the main determinant limiting var transcript variation in asymptomatic carriers. However, by RNAseq and qRT-PCR we also observed significantly higher total var transcript levels in malaria cases compared to asymptomatic carriers, suggesting that in addition to the parasites switching and the hosts immune selection of expressed var genes, parasites able to sustain long-term infections may be poised for reduced PfEMP1 expression.

microbiology↗

Iron transport pathways in the human malaria parasite Plasmodium falciparum revealed by RNA-sequencing

Host iron deficiency is protective against severe malaria as the human malaria parasite Plasmodium falciparum depends on bioavailable iron from its host to proliferate. The essential pathways of iron acquisition, storage, export, and detoxification in the parasite differ from those in humans, as orthologs of the mammalian transferrin receptor, ferritin, or ferroportin, and a functional heme oxygenase are absent in P. falciparum. Thus, the proteins involved in these processes may be excellent targets for therapeutic development, yet remain largely unknown. Here, we show that parasites cultured in erythrocytes from an iron-deficient donor displayed significantly reduced growth rates compared to those grown in red blood cells from healthy controls. Sequencing of parasite RNA revealed diminished expression of genes involved in overall metabolism, hemoglobin digestion, and metabolite transport under low-iron versus control conditions. Supplementation with hepcidin, a specific ferroportin inhibitor, resulted in increased labile iron levels in erythrocytes, enhanced parasite replication, and transcriptional upregulation of genes responsible for merozoite motility and host cell invasion. Through endogenous GFP tagging of differentially expressed putative transporter genes followed by confocal live-cell imaging, proliferation assays with knockout and knockdown lines, and protein structure predictions, we identified six proteins that are likely required for ferrous iron transport in P. falciparum. Of these, we localized PfVIT and PfZIPCO to cytoplasmic vesicles, PfMRS3 to the mitochondrion, and the novel putative iron transporter PfE140 to the plasma membrane for the first time in P. falciparum. PfNRAMP/PfDMT1 and PfCRT were previously reported to efflux Fe2+ from the digestive vacuole. Our data support a new model for parasite iron homeostasis, in which PfE140 is involved in iron uptake across the plasma membrane, PfMRS3 ensures non-redundant Fe2+ supply to the mitochondrion as the main site of iron utilization, PfVIT transports excess iron into cytoplasmic vesicles, and PfZIPCO exports Fe2+ from these organelles in case of iron scarcity. These results provide new insights into the parasites response to differential iron availability in its environment and into the mechanisms of iron transport in P. falciparum as promising candidate targets for future antimalarial drugs.

cell biology↗

Distributed Compact Plasma Reactor Sterilization for Planetary Protection in Space Missions

This paper presents a proof-of-concept study establishing effectiveness of the Active Plasma Sterilizer (APS) for sterilization in planetary protection. The APS uses Compact Portable Plasma Reactors (CPPRs) to produce surface dielectric barrier discharge, a type of cold plasma, using ambient air to generate and distribute reactive species like ozone used for decontamination. Sterilization tests were performed with pathogenic bacteria (Escherichia coli and Bacillus subtilis) on materials (Aluminum, Polycarbonate, Kevlar and Orthofabric) relevant to space missions. Results show that the APS can achieve 4 to 5 log reductions of pathogenic bacteria on four selected materials, simultaneously at 11 points within 30 minutes, using power of 13.2 {+/-} 2.22 W. Spatial sterilization data shows the APS can uniformly sterilize several areas of a contaminated surface within 30 minutes. Ozone penetration through Kevlar and Orthofabric layers was achieved using the CPPR with no external agent assisting penetration. Preliminary material compatibility tests with SEM analysis of the APS exposed materials showed no significant material damage. Thus, this study shows the potential of the APS as a light-weight sustainable sterilization technology for planetary protection with advantages of uniform spatial decontamination, low processing temperatures, low exposure times, material compatibility and the ability to disinfect porous surfaces.

microbiology↗

FLOCK STASIS DRIVES FLYING SPEED IN PIGEONS, WHILE ARTIFICIAL MASS ADDITIONS DO NOT

Animals are characterised, in part, by their use of voluntary movement, which is used to explore and exploit resources from their surrounding environment. Movement can therefore benefit animals, but will cost them their energetic reserves. Thus, adaptations for faster movements with negligible increases in energy expenditure will likely evolve via natural selection. Individual and social-level mechanisms have been shown to optimise this speed/energetic trade-off. Nevertheless, studies of social-level traits typically ignore individual variation, which is a cornerstone principle in evolutionary ecology. Furthermore, how individual phenotype interacts with the phenotypic composition of the group to govern the cost of transport may have been entirely overlooked. We investigate speed and the energetic consequences of individual-level phenotypic differences using body mass (both natural and artificially manipulated with additional weights) of homing pigeons (Columba livia) (N =16 birds; N = 193 useable flight trajectories). We then turn to social level phenomena, and manipulate the composition of pigeon groups by body mass (N= 12 birds in four treatments; N = 192 useable flight trajectories) and leadership rank (N = 30 birds in three groups, N = 286 useable flight trajectories) following earlier leadership identification flights (N = 33 birds, N = 306 useable flight trajectories). "Natural" body mass was predictive of flying speed in solo flights, but not in groups of greater mass by composition; "artificial" mass loading had no impact on speed in solo fliers, and was not tested in groups. Groups of leader phenotypes, showed faster speeds, and greater cohesion than follower phenotype groups, both in terms of flock spread, but also in consistency of positioning within the flock ("flock stasis") across the flight. Flock stasis was further analysed across all other group flights. Its positive impact on speed was found to be consistent across all experimental treatments. Therefore, predicting flock stasis may be critical to understanding optimal phenotypic compositions of birds, and thus the social evolution of birds which fly together. We provide evidence that greater stasis may be driven by phenotypic compositions (i.e. groups of leaders, and homogeneous mass groups) and also discuss the implications of stasis for different flocking structures (e.g. V-formations) and human crowd control.

animal behavior and cognition↗

Homing pigeon (Columba livia) dominance hierarchies are stable over time and resistant to perturbation

Dominance hierarchies are a social dynamic common in many species, which help balance the costs and benefits of social living. Which individuals in a group occupy high ranking positions is influenced by a multitude of different intrinsic and/or extrinsic factors. While homing pigeons (Columba livia) have been a model system for their navigational abilities and flight dynamics, less is known about their dominance hierarchies and general social behaviour when not in flight. Here we document the dominance hierarchies in two closed populations of homing pigeons, and investigate the physiological factors associated with dominance rank, including body size, metabolic rate, personality, and iridescent plumage % reflectance. Measurements of body size and resting metabolic rate all positively correlated with dominance rank in accordance with the performance model of energetic management. Iridescent plumage % reflectance was negatively correlated with rank, suggesting a potential cost of dominance. Attempts to alter hierarchy structure through manipulations of body mass and feather iridescence were ineffective which hierarchy dynamics remaining stable throughout the perturbations and across measurement sessions.

animal behavior and cognition↗

Constant rate of Plasmodium falciparum sexual commitment promotes variable gametocyte proportions as infection progresses over time

Plasmodium falciparum infections persist through long dry seasons at low parasitaemia without causing malaria symptoms and thus remain untreated. In asymptomatic children, increased circulation of infected erythrocytes without adhering to the vascular endothelium is observed during the dry months, compared to febrile malaria in the wet season. However, alterations of parasite sexual commitment and gametocytogenesis have not been investigated. Here, we compared the expression of genes related to sexual commitment and gametocytogenesis, the proportion and density of P. falciparum gametocytes, and the blood concentration of phospholipids in dry season asymptomatic individuals versus symptomatic subjects in the wet season. Additionally, we adapted a within-host mathematical model considering asexual and sexually-committed parasites and gametocytes to understand the dynamics of gametocyte number and proportion as infections progress. Compared to clinical malaria cases, transcripts of late-stage gametocytes were predominantly upregulated in the dry season, associating with increased proportions of mature gametocytes; while transcription of genes related to parasite sexual commitment was unaltered throughout the year. Our data suggest that gametocyte density and proportion diverge as infections progress from recent transmission to chronic carriage, without alterations in the sexual commitment rate over time.

microbiology↗

Pre-winter fattening and fat loss during wing moult: the annual cycle of fat deposition in captive barnacle geese (branta leucopsis)

Many different physiological changes have been observed in wild waterfowl during the flightless stage of wing moult, including a loss of body mass. Previously we established that captive barnacle geese (Branta leucopsis) underwent this characteristic decrease in body mass during their wing moult, even though they had unlimited and unrestricted access to food. In the present study we aimed to determine if this body mass loss during moult comprised mainly a reduction in fat stores, and to ascertain if the captive geese undergo pre-migratory and pre-winter fattening over a similar temporal scale to their wild conspecifics. The non-destructive technique of deuterium oxide isotope dilution was employed to provide repeated measurements of estimated fat deposition from a captive flock of fourteen barnacle geese. Birds were injected with deuterium oxide at 7 distinct intervals for one annual cycle. During the flightless period of the moult, body fat decreased by approximately 40% from the pre-moult value. During late-September and early October, body fat reached its highest point in the annual cycle, both as an absolute value and as a percentage of total body mass. We propose that while the energetic cost of wing moult is not the ultimate cause of fat loss in moulting barnacle geese, the approximate 212 g of fat catabolised during moult would provide sufficient energy to cover the cost of the replacement of the flight feathers, estimated to be 6384 kJ, over an approximate 42 day period. We conclude that the previously recorded increase in metabolism during moult in the geese, led to the use of endogenous fat reserves because the birds reduced rather than increased their food intake rates owing to the increased risk of predation when flightless. We also conclude that captive barnacle geese do undergo pre-winter and pre-migratory fattening, providing further evidence of the innate nature of these fat deposition cycles.Competing Interest StatementThe authors have declared no competing interest.View Full Text

zoology↗

The determinants of aggression in male Siamese fighting fish, Betta splendens

Siamese fighting fish, Betta splendens, have been extensively studied due to their aggression and stereotypical displays. Many studies have focused on their characteristic opercular flaring, while the less aggressive and less energetically costly lateral display have been comparatively understudied. Many factors have been shown to influence aggression in Bettas, notably body length and the personality trait of boldness, however, the role that colour plays in determining an individuals aggressiveness is much less clear. The role of colour has only been briefly studied, and based on human interpretations of colour, i.e. limited to what the receivers eyes and sensory systems actually can process and discriminate, with results suggesting blue males are more aggressive than red males. Using male-male interactions, measuring opercular flaring and lateral display we found that colour and personality do play a role in determining the degree of aggressiveness in Betta splendens. Blue-finned males were more aggressive, performing longer lateral displays more frequently. Blue fins are a phenotype observed in wild type males and is likely selected for to allow visual cues to travel through the murky water they inhabit. Body mass was positively correlated with lateral display frequency, and opercular flare frequency and duration. Finally, neophobic individuals - individuals that were less willing to approach a novel object - were more aggressive, performing significantly more lateral displays. This indicates that personality may impact fighting strategy, with males either choosing to end conflicts quickly with more aggressive displays or to outlast their opponent with less energetically costly displays.

animal behavior and cognition↗

Overall dynamic body acceleration as an indicator of dominance in homing pigeons

The benefits of dominance are well known and numerous, including first access to resources such as food, mates and nesting sites. Less well studied are the potential costs associated with being dominant. Here, the movement of two flocks of domestic homing pigeons (Columba livia) - measured via accelerometry loggers - was recorded over a period of two weeks. Movement was then used to calculate each individuals daily overall dynamic body acceleration (ODBA, G), which can be used as a proxy for energy expenditure. The dominance hierarchy of the two flocks was determined via group-level antagonistic interactions, and demonstrated a significantly linear structure. The most dominant bird within each flock was found to move significantly more than conspecifics - on average, c.39% greater than the individual with the next highest degree of movement - indicating a possible cost to possessing the top rank within a hierarchy. Despite the dominance hierarchy being linear, mean daily total ODBA did not reflect a linear nature, with no pattern observed between rank and ODBA, once the top ranked individuals had been accounted for. This suggest that energy expenditure may be more reflective of a despotic hierarchy. These results show the potential for the future use of accelerometery as a tool to study the fusion of energetics and behaviour. Subject Categorybehaviour Subject Areasbehaviour, physiology

animal behavior and cognition↗

Mechanics Of Flight Feathers: Effects Of Captivity?

ABSTRACTFeathers act as aerodynamic cantilevers, and to withstand the prolonged cyclical loading that occurs during flight, feathers must be stiff, lightweight and strong. We experimentally tested the differences in feather structure, primarily stiffness and size, between (a) wild and captive Barnacle Geese Branta leucopsis, and (b) primary feathers dropped during the annual flight feather moult, and those feathers freshly regrown during the moult process. We found that, despite having undergone a 5,000km round-trip migration, flight feathers dropped during moult in the wild geese were stiffer than those measured in the captive geese, both for those dropped during moult and those re-grown. We propose that this may be related to diet or stress in the captive geese.Competing Interest StatementThe authors have declared no competing interest.View Full Text

zoology↗

Plasmodium falciparum-specific IgM B cells dominate in children, expand with malaria and produce parasite inhibitory IgM

IgG antibodies are known to play a central role in naturally acquired immunity to blood-stage malaria in humans, but little is known about the IgM response to blood-stage malaria, the mechanisms by which IgM may protect, or the underlying biology of Plasmodium falciparum (Pf)-specific IgM B cells. In a Mali cohort spanning infants to adults we conducted a longitudinal analysis of B cells specific for the Pf blood-stage antigens AMA1 and MSP1, as well as the comparator antigen influenza hemagglutinin (HA). At the uninfected baseline, before the malaria season, Pf-specific memory B cells (MBCs) in children are disproportionally IgM+ and only gradually shift to IgG+ with age, in contrast to HA-specific MBCs that are predominantly IgG+ from infancy to adulthood. In response to acute febrile malaria, Pf-specific IgM B cells increase in frequency and upregulate activation and co-stimulatory markers. B cell receptor (BCR) analysis showed that Pf-specific IgM B cells are somatically hypermutated at levels comparable to HA-specific IgG B cells. Finally, IgM antibodies from the plasma of malaria-exposed individuals were comparable to IgG in inhibiting Pf blood-stage growth in vitro, and significantly better at enhancing phagocytosis of Pf merozoites, suggesting that IgM may protect through both direct neutralization and opsonization. Thus, somatically hypermutated Pf-specific IgM MBCs dominate in early life, are activated and expand during acute malaria and are associated with plasma IgM that inhibits parasite growth in vitro.

immunology↗